nanoBioMed2016 Conference Book by Phantoms Foundation

nov. 22-24, barcelona (spain) www.nanobiomedconf.com

abstracts book organisers

On behalf of the Organizing Committee, we take great pleasure in welcoming you to Barcelona (Spain) for the NanoBio&Med2016 International Conference.

Foreword Organising

Committee

Sponsors

Exhibitors

Speakers

Posters

This event, after successful editions organized within ImagineNano in Bilbao 2011 & 2013, and in Barcelona in 2014 & 2015, is going to present the most recent international developments in the field of Nanobiotechnology and Nanomedicine and will provide a platform for multidisciplinary communication, new cooperations and projects to participants from both science and industry. Emerging and future trends of the converging fields of Nanotechnology, Biotechnology and Medicine will be discussed among industry, academia, governmental and non-governmental institutions. NanoBio&Med2016 will be the perfect place to get a complete overview into the state of the art in those fields and also to learn about the research carried out and the latest results. The discussion in recent advances, difficulties and breakthroughs will be at his higher level. As in previous editions, an industrial forum will be organized to promote constructive dialogue between business and public leaders and put specific emphasis on the technologies and applications in the nanoBioMed sector. We are indebted to the following Companies, Scientific Institutions and Government Agencies for their financial support: Institute for Bioengineering of Catalonia (IBEC), Bicosome, PlasmaChem GmbH, FEI, NanoSciences Grand Sud-Ouest (Câ&#x20AC;&#x2122;Nano GSO) and ICEX Spain Trade and Investment. We would also like to thank NANOVEX Biotechnologies S.L. and ReDIB for their participation. In addition, thanks must be given to the staff of all the organising institutions whose hard work has helped planning this conference.

NanoBio&Med2016

november 22-24, 2016 - Barcelona (Spain)

Antonio CORREIA President of the Phantoms Foundation (Spain) Dietmar PUM Deputy Head of the Biophysics Institute – BOKU (Austria) Josep SAMITIER Director of the Institute for Bioengineering of Catalonia – IBEC (Spain)

NanoBio&Med2016

november 22-24, 2016 - Barcelona (Spain)

Speakers list: Alphabetical order Authors Juan Pablo Agusil Antonoff (IMB-CNM (CSIC), Spain) Suspended planar-array chips for molecular multiplexing at the microscale Jouni Ahopelto (VTT Technical Research Centre of Finland Ltd, Finland) Graphene biosensor with fusion protein receptor modules Lorenzo Albertazzi (Institute for Bioengineering of Catalonia (IBEC), Spain) Nanoscopy for Nanomedicine: looking at nanomaterials one molecule at a time Lisa Almonte (Universidad de Murcia, CIOyN, Spain) Ideal Atomic Force Microscopy imaging of heterogeneous biological samples in liquids: Topography and Chemical information Lide Arana (Instituto Biofisika (UPV/EHU, CSIC), Spain) Solid lipid nanoparticles for delivery of calendula officinalis extract Gianluca Arauz Garofalo (Universitat de Barcelona, Spain) Prospects of Microwave Spectrometry for vascular stents monitoring: Towards a noninvasive and non-ionizing follow-up alternative Ana I. Ballestar Balbás (GPNT, Spain) Highest Quality Graphene for bio-sensing Rafael Bernad (BICOSOME S.L., Spain) Antioxidant stabilization through Bicosomes Mattia Bramini (Istituto Italiano di Tecnologia/Center for Synaptic Neuroscience and Technologies & Graphene Labs, Italy) Towards graphene for biomedical applications: evaluation of neuronal biocompatibility Inmaculada Campos Sanchez (Universitat Autónoma de Barcelona, Spain) Voltammetric sensor based on Molecularly Imprinted Polymers for 4-Ethylphenol detection Virginia Cebrián (Mecwins SA, Spain) Ultrasensitive Scanning Laser Analyser for Biomedical Applications Alexandre Chicharo (International Iberian Nanotechnology Laboratory / IST-UL, Portugal) Lab-on-a-chip for purification and automated counting of Circulating Tumor Cells from peripheral blood of metastatic cancer patients Gianni Ciofani (Polytechnic Univ. of Torino & IIT, Italy) Smart materials for nanomedicine Gavin Coleman (Centre for BioNano Interactions, UCD Dublin, Ireland) Study of Shape-dependent Nanoparticle Interactions Frederique Cunin (Institut Charles Gerhardt Montpellier-CNRS, France) Porous silicon: a biodegradable semiconductor for nanomedecine Bruno De Geest (Ghent University, Belgium) Biomimetic nanoparticles for lymphe node focused immune activation Maria de la Fuente (Fundación Ramón Domínguez / Health Research Institute of Santiago de Compostela, Spain) Nanotheranostics to interfere the process of metastasis Lucia Gemma Delogu (University of Sassari, Italy) Immune characterization of graphene oxide and amino functionalized graphene using new high-throughput analysis. Tal Dvir (Tel Aviv University, Israel) Engineering strategies for building hearts

Session

Page

Oral

Plenary Session

Invited Plenary Session

Keynote Plenary Session

Oral Plenary Session

Oral Parallel Session I

Oral Plenary Session

Oral Parallel Session I

Oral Plenary Session

Oral Parallel Session I

Keynote Plenary Session

Oral Parallel Session I

Keynote Plenary Session

Oral Parallel Session II

Oral Plenary Session

Keynote Plenary Session

9 10 48 50 51 52 54 56 58 60 61 12 63 13 15 65 67 16

NanoBio&Med2016

november 22-24, 2016 - Barcelona (Spain)

Authors Yuval Ebenstein (Tel Aviv University, Israel) Genome mapping in nanochannel arrays Georg Fantner (EPFL, Switzerland) Up Close and Personal: The Process of Mycobacterial Cell Division Neus Feliu (Karolinska Institutet, Sweden) Interaction of Nanoparticles with Cells: Nanomaterials For Stem Cell Tracking Applications Giancarlo Franzese (Universitat de Barcelona / Institut de Nanociencia i Nanotecnologia (IN2UB), Spain) Contribution of Water to Protein Stability and Strategies for Protein Design Juan Gallo (International Iberian Nanotechnology Laboratory, Portugal) Magnetic Solid Lipid Nanocomposites as ultra-high MRI contrast enhancers and magnetic hyperthermia induced drug delivery vehicles Paul Galvin (Tyndall National Institute, University College Cork, Ireland) Digital Health Technology: From Precision Engineering to Precision Medicine Jose Antonio Garrido (ICREA-ICN2, Spain) Graphene flexible electronics for neuroprosthetics Giada Graziana Genchi (Istituto Italiano di Tecnologia, Italy) Arrays of titanium dioxide nanotubes with different diameter and surface chemistry influence C2C12 skeletal myoblast adhesion and differentiation Véronique Gigoux (LPCNO/INSA - INSERM ERL1226, France) Targeted nanotherapy of endocrine tumors by magnetic intra-lysosomal hyperthermia (MILH) Arnaud Gissot (Bordeaux University, France) Nucleic acid based supramolecular systems: a route to biomedical applications Judith Guasch (Materials Science Institute of Barcelona (ICMAB-CSIC)/CIBER-BBN, Spain) Nanostructured functionalized surfaces for adoptive cell therapy Christoph Guger (g.tec medical engineering GmbH, Austria) Current and future applications of brain-computer interfaces Fernando Herranz (Centro Nacional de Investigaciones Cardiovasculares (CNIC), Spain) Radioisotope Core-doped Magnetic Nanoprobes; a chelator-free approach for PET/MR molecular imaging with extremely small nanoparticles Pascal Jonkheijm (University of Twente, The Netherlands) Dynamic cell-material interactions Antonios Kanaras (University of Southampton, United Kingdom) Synergistic actions of colloidal nanoparticles: sensing and drug delivery Didier Letourneur (Inserm U1148/CNRS, France) Design and in vivo Validations of Nanosystems in Cardiovascular. Tissue Engineering and Molecular Imaging Ke Liu (École Polytechnique Fédérale de Lausanne, (EPFL), Switzerland) Democratizing solid-state nanopores for high throughput bio-sensing Mika Lindén (Ulm University, Germany) Intracellular drug release using mesoporous silica nanoparticles as drug vectors Andreas Manz (KIST Europe, Germany) Biomimetic channels and channel networks for microfluidics Arben Merkoçi (ICREA/ICN2, Spain) Paper-based nanobiosensors in diagnostics: from health, safety and security to environment monitoring Daniel Navajas (Universitat de Barcelona / IBEC, Spain) Nanomechanics of Soft Biological Tissues Probed with Atomic Force Microscopy Salima Nedjari (Institute for Bioengineering of Catalonia (IBEC), Spain) Geometry of electrospun nanofibers determine the initial interaction and subsequent osteogenic differentiation of mesenchymal stem cell Wolfgang Parak (Philipps University of Marburg, Germany) Interaction of colloidal hybrid nano- and microparticles with cells

NanoBio&Med2016

november 22-24, 2016 - Barcelona (Spain)

Session

Page

Keynote

Plenary Session

Keynote Plenary Session

Oral Parallel Session II

Oral Plenary Session

Oral Parallel Session II

Keynote Plenary Session

Oral Plenary Session

Keynote Plenary Session

Oral Plenary Session

Invited Plenary Session

Oral Plenary Session

Keynote Plenary Session

Oral Plenary Session

Keynote Plenary Session

Oral Plenary Session

Keynote Plenary Session

Oral Parallel Session II

Keynote Plenary Session

18 69 70 71 19 20 73 21 23 75 25 77 26 79 28 80 30 31 32 33 81 34

Authors Francesca Peiro (Institute of Nanoscience and Nanotechnology (In2UB), University of Barcelone, Spain) A journey through the secret life of nanoparticles: from 2D high resolution images to 3D oxidation state mapping Ilaria Pezzini (Scuola Superiore Sant´Anna, BioRobotic Institute, Italy) Cerium oxide nanoparticles, a promising powerhouse in bioenergetic imbalance Danny Porath (The Hebrew University of Jerusalem, Israel) Charge Transport in single DNA-Based Molecules Maurizio Prato (CIC biomaGUNE, Spain) Novel Functional Carbon Bio-Interfaces Valerio Pruneri (ICFO, Spain) Sensing of particles, micro-organisms and biomarkers using the camera of a mobile phone Imma Ratera (Nanomol Group/Institut de Ciencia de Materials de Barcelona (CSIC), Spain) Organic Radical Nanoparticles for Bioimaging Applications Meital Reches (The Hebrew Univ of Jerusalem, Israel) Interactions between peptides and inorganic matter: From basic science to applications Ana Rita Rodrigues (University of Minho, Portugal) Magneto-sensitive liposomes containing manganese ferrite nanoparticles as nanocarriers for new promising antitumor thienopyridin-amine derivates Mafalda Rodrigues (Universitat de Barcelona, Spain) Nanostructured materials for intracellular sensing and delivery Silvina Samy (INL - International Iberian Nanotechnology Laboratory, Portugal) Microfluidic isolation of cancer cells from blood: towards point-of-care liquid biopsy Samuel Sánchez (IBEC, Spain) Chemical Nanomachines as active drug nanovehicles Ronit Satchi-Fainaro (Tel Aviv University, Israel) Identifying molecular signatures of tumor dormancy as a basis for the rational design of precision nanomedicines Avi Schroeder (Technion, Israel) Personalized Cancer Nanomedicine Sylwia Sekula-Neuner (Karlsruhe Institute of Technology / Institute of Nanotechnology, Germany) Interdigitated Bioink Micropatterns Generated by Multiplexed Polymer Pen Lithography Michael Tadros (Botín Foundation, Spain) The innovation imperative: turning research results into socioeconomic impact Nagamalai Vasimalai (INL-International Iberian Nanotechnology Laboratory, Portugal) Facile one-pot synthesis of highly luminescent carbon dots from food and its application of in-vitro cancer growth inhibition Ester Vazquez Fernandez-Pacheco (University of Castilla La Mancha, Spain) Graphene for Bioapplications: Preparation, Cytotoxicity and Integration in 3D-scaffolds

Session

Page

Oral

Plenary Session

Oral Parallel Session II

Keynote Plenary Session

Oral Plenary Session

Keynote Plenary Session

Oral Parallel Session I

Oral Parallel Session II

Oral Parallel Session I

Keynote Plenary Session

Oral Plenary Session

Invited Plenary Session

Oral Parallel Session II

Keynote Plenary Session

85 35 37 38 87 39 88 90 92 40 41 42 93 43 95 44

NanoBio&Med2016

november 22-24, 2016 - Barcelona (Spain)

Graphene biosensor with fusion protein receptor modules J. Ahopelto, Ahopelto M. Soikkeli, A. Paananen, M. Kainlauri, H. Arola, J. J. Joensuu, S. Arpiainen, T. Nevanen VTT Technical Research Centre of Finland Ltd, Espoo, Finland

Health care and well-being are becoming more and more important as the fraction of the elderly of the population is constantly increasing. This calls for easy-to-use appliances to monitor the physical condition preferably at home on daily or weekly basis with minimum requirements in sample preparation and with connectivity via internet or mobile phone. Most of the biological molecules carry charge providing a route to label-free and fast sensing. We have developed a sensor concept based on graphene field effect transistor and fusion protein receptor modules [1], shown schematically in Figure 1. The sensor can be sensitised to various analytes, allowing use of pre-fabricated sensor chips and/or changing the receptor modules in-situ for reprogramming of the device. Relatively standard microfabrication processes are used in the fabrication and the sensors can also be manufactured on flexible substrates. The response is fast and detection of concentrations down to femtomolar range has been demonstrated. In this talk we will discuss the potential and challenges of this label-free biosensor approach.

Figures

Figure 1. GFET biosensor shown schematically. The receptor modules form a dense monolayer on the graphene channel. The receptors selectively bind the desired analyte from the sample and the charge of the analyte molecules is detected by the highly chargesensitive graphene FET.

References [1] Miika Soikkeli, Katri Kurppa, Markku Kainlauri, Sanna Arpiainen, Arja Paananen, David Gunnarsson, Jussi J. Joensuu, PĂ¤ivi Laaksonen, Mika Prunnila, Markus B. Linder, Jouni Ahopelto, Graphene biosensor programming with genetically engineered fusion protein monolayers, ACS Appl. Mater. Interfaces 8 (2016) pp 8257â&#x20AC;&#x201C;8264.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nanoscopy for Nanomedicine: looking at nanomaterials one molecule at a time Lorenzo Albertazzi, Albertazzi Nanoscopy for Nanomedicine Group Institute for Bioengineering of Catalonia (IBEC) Barcelona, Spain

The development of nanocarriers for intracellular delivery of therapeutic moieties is a great challenge for synthetic chemistry and nanotechnology. A crucial factor limiting the design of effective materials is the lack of understanding about material-cell interactions that hampers the rational design of nanosized carriers. Therefore obtaining detailed information about the behavior of nanomaterials in the biological environment is a key issue in nanobiotechnology. Here we discuss the use of super resolution microscopy to image materials in vitro and in mammalian cells. This novel technique, allowing to obtain a resolution down to 20nm, had a dramatic impact in the field of cell biology, however its use in the field of chemistry and nanotechnology is poorly explored. Super resolution microscopy offers nanometric resolution and multicolor ability, therefore it is an ideal tool to study nano-sized supramolecular assemblies of multiple components in vitro and in cells. We employed Stochastic Optical

Reconstruction Microscopy (STORM) to image biomaterials in vitro, with special emphasis on supramolecular polymers and nanoparticles, unveiling novel information on materials structure and dynamics. Moreover we propose a methodology to image nano-sized materials in cells, tracking them during their membrane targeting, cell uptake and intracellular targeting. We show how 2color STORM can be used to perform nanometric-accurate colocalization unveiling at the molecular level materials-cell interactions. These methodologies are used to answer open questions in the nanobiotechnology field related to the interactions of nanosystems with the cellular environment. This allows looking at nanomaterials in action with new eyes and use the information obtained for the â&#x20AC;&#x153;STORMguidedâ&#x20AC;? design of novel nanomaterials for drug delivery and other targeted therapies.

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Multicomponent Supramolecular Polymers as a Modular Platform for Intracellular Delivery ACS Nano, 2016, 10 (2), 1845â&#x20AC;&#x201C;1852 [2] Super Resolution Imaging of Nanoparticles Cellular Uptake and Trafficking ACS Appl. Mater. Interfaces, 2016, 8 (10), pp 6391â&#x20AC;&#x201C;6399 [3] Probing exchange pathways in onedimensional aggregates using super resolution microscopy Science, 2014, 344(6183): 491-5

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Smart Materials in Nanomedicine Gianni Ciofani1,2 Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Torino, Italy 2 Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, Pontedera (Pisa), Italy gianni.ciofani@iit.it 1

Nanotechnology has enabled unprecedented control of the interactions between materials and biological entities, from the microscale down to the molecular level. For example, nanosurfaces and nanostructures have been used to mimic or interact with biological microenvironments, to support specific biological functions such as cell adhesion, mobility and differentiation, as well as tissue healing. Recently, a new paradigm has been proposed for nanomedicine to exploit the intrinsic properties of nanomaterials as active devices rather than as passive structural units or carriers for medications. In this view, the nanomaterial itself is the active device that responds to external stimuli by modifying its intrinsic chemical and/or physical characteristics, so as to provide useful bio-stimulation and/or biosignaling. Our research approach falls into this latter, “active” category: we develop “smart” nanomaterials that change their structural/functional properties in response to specific external stimuli (electric or magnetic fields, electromagnetic radiation, ultrasound, etc.). Specifically, we develop multifunctional nanostructured materials that are pharmacologically active and that can be actuated by virtue of their magnetic, dielectric, optically active, or piezoelectric properties.

as active components for substrates and scaffolds for tissue engineering and regenerative medicine, with a special attention on stimulation of neuronal cells [4]. Moreover, a stimuli-responsive hybrid lipid/magnetic nanovector, for advanced theranostic applications, will be introduced [5]. This presentation will summarize our main results to date, and highlights the most promising examples that could have a practical translation into the clinical or technological realms. Acknowledgments: Acknowledgments This research is partially supported by the Italian Ministry of Health Grant Number RF-2011-02350464, and by the Compagnia di San Paolo Starting Grant Number 55_AI16GC01

References [1] Genchi G.G., [...], Ciofani G. Nanotechnology, 27(23): 232001 (2016) [2] Marino A., [...], Ciofani G. ACS Nano 8(11): 11869-11882 (2014) [3] Rocca A., [...], Ciofani G. Nanomedicine 11(7): 1725-1734 (2015) [4] Genchi G.G., [...], Ciofani G. Advanced Healthcare Materials, 5(14): 1808-1820 (2016) [5] Grillone A., [...], Ciofani G. Advanced Healthcare Materials 4(11): 1681-1690 (2015)

More in details, in this talk I will approach applications of smart nanomaterials such as piezoelectric barium titanate nanoparticles [1,2] and anti-oxidant cerium oxide nanoparticles [3],

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Porous silicon: a biodegradable semiconductor for nanomedecine Frederique Cunin Institut Charles Gerhardt Montpellier-CNRS, France

The choice and interest of nanotechnologies is now well established for applications in nanomedecine and for the development of targeted and personalized therapies. Besides the interest in the possibility to miniaturize the therapeutic or diagnostic tools, nanomaterials exhibit chemical, physical and textural properties interesting for the encapsulation, the targeting and the controlled release of active molecules, as well as for cell labeling and medical imaging. Despite the rapid growth and development of the field of functional nanostructured materials, the current main reason to the limited clinical progress of nanotherapeutics is the potential toxicity of the materials or their degradation byproducts, and their unknown fate once administered. Porous silicon (pSi) nanostructures are fully biodegradable, and nontoxic in vivo [1]. The primary biodegradation product of pSi is orthosilicic acid, which is the bioavailable form of silicon. PSi is easily prepared by electrochemical etching of crystalline silicon, it displays tunable textural properties (surface area, pore diameter, etc). Chemical

modification provides a means to adjust the degradation rate of pSi, as well as to load drugs or molecules of interest. PSi has an interesting osteoconductive potential and is especially favorable for osteoblast adhesion, growth and mineralization [2]. Due to their electronic semiconductor-derived properties, pSi nanostructures exhibit intrinsic photoluminescence amenable for imaging. Moreover, they can be excited by near infrared (NIR) two photon excitation light [1,3] offering possibilities for phototherapies, and for light triggered and targeted treatment, based on tissue-penetrable NIR light response. The development of photoactive pSi nanostructures functionalized with organic ligands for applications in imaging, drug delivery and photo-activated therapies, as well as for bone tissue engineering will be presented [4-9].

Figures

Figure 1. Functionalized luminescent pSi nanoparticles in cells

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] J.-H. Park, L. Gu, G. von Maltzahn, E. Ruoslahti, S.N. Bhatia, M. J. Sailor, Nature Mat., 2009, 8(4), 331. [2] P-Y. Collart-Dutilleul, E. Secret, I. Panayotov, D. Deville de Perrière, R. Martin-Palma, V. TorresCosta, M. Martin, C. Gergely, J-O. Durand, F. Cunin, F. Cuisinier, ACS Applied Materials & Interfaces, 2014, 6 (3), 1719. [3] E. Secret; M. Maynadier; A. Gallud; A. Chaix; E. Bouffard; M. Gary-Bobo; N. Marcotte; O. Mongin; K. El Cheikh; V. Hugues; M. Auffan; C. Frochot; A. Morère; P. Maillard; M. BlanchardDesce; M.J. Sailor; M. Garcia; J.O. Durand; F. Cunin Adv. Mater. 2014, 26(45), 7643. [4] 4. A. Chaix, K. El Cheikh, E. Bouffard, M. Maynadier, D. Aggad, V. Stojanovic, N. Knezevic, M. Garcia, P. Maillard, A. Morère, M. Gary-Bobo, L. Raehm, S. Richeter, J. O. Durand and F Cunin, J. Mater. Chem. B, 2016, 4, 3639.

[5] N.K. Knezevic, V. Stojanovic, A. Chaix, E. Bouffard, K. El Cheikh, A. Morère, M. Maynadier, G. Lemercier, M. Garcia, M. GaryBobo, J.O. Durand, F. Cunin, J. Mater. Chem. B, 4, 2016, 1337. [6] V. Stojanovic, F. Cunin, J. O. Durand, M. Garcia and M. Gary-Bobo, J. Mater. Chem. B, accepted. [7] E. Secret, M. Maynadier, A. Gallud, M. GaryBobo, A. Chaix, E. Belamie, P. Maillard, M. J. Sailor, M. Garcia, J.O. Durand, F. Cunin, Chem. Commun., 2013, 49(39), 4202. [8] E. Secret, K. Smith, V. Dubljevic, E. Moore, P. Macardle, B. Delalat, M.L. Rogers, T. G. Johns, J.O. Durand, F. Cunin, N. H. Voelcker, Advanced Healthcare Materials, 2013, 2(5), 718. [9] P-Y. Collart-Dutilleul, I. Panayotov, E. Secret, F. Cunin, C. Gergely, F. Cuisinier, M. Martin, Initial stem cell adhesion on porous silicon surface: molecular architecture of actin cytoskeleton and filopodial growth Nanoscale Research Letters, 2014, 9, 564.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Biomimetic nanoparticles for lymphe node focused immune activation Bruno G. De Geest Department of Pharmaceutics, Ghent University, Ghent, Belgium

Agonists of Toll-like receptors are potent activators of the innate immune system and hold promise as vaccine adjuvant and for anticancer immunotherapy. Unfortunately, in soluble form, they readily enter systemic circulation and cause systemic inflammatory toxicity. Here we demonstrate that by covalent ligation of a small molecule imidazoquinolinebased TLR7/8 agonist to 50 nm sized degradable polymeric nanoparticles, the potency of the agonist to activate TLR7/8 in in vitro cultured dendritic cells is largely retained. Importantly, imidazoquinoline-ligated nanoparticles focused the in vivo immune-activation on the draining lymph nodes whilst dramatically reducing systemic inflammation. Mechanistic studies revealed a prevalent passive diffusion of the nanoparticles to the draining lymph node.

Clinical relevance of this therapeutic strategy is demonstrated as vaccine adjuvant and for tumour immunotherapy. Immunization studies in mice show that, relative to soluble TLR7/8 agonist, imidazoquinoline-ligated nanoparticles induce superior antibody and T cell responses against an admixed antigen. Currently, we are evaluating the potential of imidazoquinolineligated nanoparticles for co-delivery of tumourassociated peptide antigens. On the other hand, we also demonstrated that imidazoquinoline-ligated nanoparticles are capable to reduce tumour growth upon peritumoral injection by activation of DCs in the tumour-draining lymph node. In summary, our approach opens avenues to enhance the therapeutic benefit of small molecule TLR agonists for a variety of applications in anti-cancer immunotherapy.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Engineering strategies for building hearts Tal Dvir Head, Laboratory for Tissue Engineering and Regenerative Medicine. Department of Molecular Microbiology & Biotechnology, Department of Materials Science and Engineering, The Center for Nanoscience and Nanotechnology Tel Aviv University. Tel Aviv, 69978, Israel

The heart is a non-regenerating organ. Consequently, the loss of cardiac cells and formation of scar tissue after extensive myocardial infarction frequently leads to congestive heart failure. Given the scarcity of cardiac donors, a potential approach to treat the infarcted heart is to repopulate the â&#x20AC;&#x2DC;dead zoneâ&#x20AC;&#x2122; with cells capable of spontaneous contraction. Cellular therapy evolved to introduce cells into diseased areas and regain function. However, two main drawbacks of this approach are the lack of control of cell accumulation site after injection, and cell death before forming cell-cell or cell-matrix interactions. These shortfalls motivated the development of the tissue engineering concept, where 3-dimensional (3D) biomaterials serve as extracellular matrix-like scaffolds to the cells, enabling the cells to assemble into effective tissue substitutes, that may restore tissue or organ function. After transplantation the scaffolds either degrade or metabolize, eventually leaving a vital tissue instead of the defected tissue. In this talk I will discuss the recent advancements in the field of cardiac tissue engineering. I will describe cutting-edge technologies for engineering functional cardiac tissues, focusing on the design of new biomaterials mimicking the natural microenvironment of the heart, or releasing biofactors to promote stem cell recruitment and cardioprotection. In addition, I will discuss the development of patient-specific materials and

3D-printing of personalized vascularized cardiac patches and whole hearts. Finally, I will show a new direction in tissue engineering, where, micro and nanoelectronics are integrated within engineered tissues to form cyborg tissues. In this new concept the built-in electronic network is used to on-line record cellular electrical activity and when needed to provide electrical stimulation for synchronizing cell contraction. Furthermore, electroactive polymers containing biological factors can be deposited on designated electrodes to release drugs in the cellular microenvironment on demand, affecting the engineered tissue or the host.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Genome mapping in nanochannel arrays Yuval Ebenstein Department of Chemical Physics, Tel-Aviv University, Tel-Aviv, Israel

Silicon nano channel arrays with channel cross section on the order of 50-100nm are now routinely fabricated and have recently been commercialized. These nanostructures allow stretching individual DNA molecules inside the channels by applying an electric field. By attaching fluorescent molecules to genetic and epigenetic marks on the DNA molecule, the stretched DNA is visualized as a pattern of fluorescent spots along the DNA molecules. We show how physical extension of long DNA molecules on surfaces and in nanofluidic channels reveals this information in the form of a linear, optical â&#x20AC;&#x153;barcodeâ&#x20AC;?, like beads threaded on a string, where each bead represents a distinct type of observable. Recent results from our lab demonstrate our ability to detect genetic structural variations (SVs), DNA repeats, DNA epigenetic modifications and various forms of DNA damage on individual genomic DNA molecules.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Up Close and Personal: The Process of Mycobacterial Cell Division Georg E. Fantner, Fantner Pascal D. Odermatt, Mélanie T.M. Hannebelle, Haig-Alexander Eskandarian, Joëlle Ven, John McKinney Interfaculty Institute for Bioengineering –Polytechnique Fédéral de Lausanne, Lausanne, Switzerland georg.fantner@epfl.ch Bacterial cell growth and division occurs as a sequence of events for which many processes remain only partially understood. Simple questions such as "do single cells grow linearly or exponentially", or "do cells grow symmetrically or asymmetrically" are still a subject of intense debate. What governs the position and timing of cell division is still largely unclear for many bacteria. In most bacterial models, FtsZ-ring formation is believed to be the first event leading to division. Nucleoid segregation, septum formation, cytokinesis, and physical cell separation are all successive events in the process of division [2]. To date, many of these processes have been characterized using static methods. Establishing the sequence and possible dependence of these processes however requires time resolved live-cell imaging [3, 4], at high resolution. Here we present a concise time-sequence of events describing division of Mycobacterium smegmatis, a nonpathogenic cousin of Mycobacterium tuberculosis. We used a combination of multiday time-lapse atomic force microscopy (AFM), time-lapse fluorescence microscopy with real time measurements of the cell separation at timescales down to 10s of milliseconds. Combining the nanoscale 3D information from AFM and the biochemical specificity from fluorescence microscopy we have characterized cell division from the early stage pre-selection of division sites, to assembly and subsequent disassembly of the FtsZ ring, localization of Wag31 and cytokinesis all the way to the rapid

cell separation. Contrary to what is believed to be true in many bacterial cell types, cell separation in Mycobacterium smegmatis isn’t a gradual event, but occurs abruptly within 10s of milliseconds, resembling more a mechanical fracture than a cellular remodeling process. Using mechanical stimulation, we demonstrate that the build-up of mechanical stress governs the time and place of cell separation. By applying additional mechanical stress, we were able to initiate cell separation at times in the cell cycle well before it would occur in the nonstimulated case. These observations suggest a new model for the late stages of cell division in Mycobacterium smegmatis, where cell mechanical properties and local stress concentration govern the timing and place of cell separation.

References [1] Adams DW, Errington J. Nature reviews Microbiology (2009);7:642-53. [2] Errington J, Daniel RA, Scheffers DJ. MMBR (2003);67:52-65, [3] Santi I, Dhar N, Bousbaine D, Wakamoto Y, McKinney JD. Nature communications (2013);4:2470. [4] Santi I, McKinney JD. mBio (2015);6:e0199914. [5] Zhou X, Halladin DK, Rojas ER, Koslover EF, Lee TK, Huang KC, et al. Science (2015);348:574-8.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Digital Health Technology: From Precision Engineering to Precision Medicine Paul Galvin Head of ICT for Health Strategic Programmes and Head of Life Sciences Interface Group, Tyndall National Institute, University College Cork, Cork, Ireland paul.galvin@tyndall.ie Breakthrough research in key enabling technologies including photonics, micro-nanoelectronics, nanotechnology, biotechnology, advanced manufacturing and advanced materials is driving innovative solutions to healthcare. This digitisation of healthcare requires an “ABC ecosystem”, in which Academic, Business and Clinical stakeholders align and collaborate and leverage the natural synergies between academic research groups, global leaders and SMEs in medtech and pharma industries, and the relevant clinical expert. This approach ensures that research delivers maximum impact in terms of clinical utility and commercial opportunity. ESTHER is an industry driven initiative initiated by the European Commission and the Medtech industry within the framework of H2020, bringing together key European stakeholders involved in Emerging and Strategic Technologies for Healthcare. ESTHER’s key objective is to raise the competitiveness of European medical technology industries by developing and manufacturing innovative healthcare technologies, in a changing environment. ESTHER should support the transition of the EU medical technology industry from predominantly passive devices manufactured by precision engineering, into smart medical technologies with additional

functionalities enabled by leveraging a multi KET (Key Enabling Technologies) development and manufacturing ecosystem. This includes the development, manufacture and translation of multi-KET enabled medical technologies and the development of the medical technologies ecosystem. Tyndall National Institute in Cork Ireland is engaged in selected research and development activities, involving leading Medtech and Pharma companies, and several innovative SMEs through EU, national and direct contract research funded programmes, to advance nextgeneration healthcare technologies. Research capabilities at Tyndall allow concepts to be taken through the complete development cycle from defining the problems statements with clinical and medtech / pharma industry partners, through to design, device fabrication, prototype system integration, and right up to product development and validation, using state-of-the-art research infrastructure and working through collaborators such as industrial designer and contract manufacturers. Tyndall’s ABC health partnership model, is a strategic innovation hub engaging with smart medtech stakeholders to ensure impact based on clinical utility and commercial opportunity for research outputs.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Graphene flexible electronics for neuroprosthetics Jose A. Garrido Catalan Institute of Nanoscience and Nanotechnology (ICN2), Bellaterra, Spain ICREA, Instituciรณ Catalana de Recerca i Estudis Avanรงats, 08070 Barcelona, Spain

Graphene and graphene-based materials possess a rather exclusive set of physicochemical properties holding great potential for biomedical applications, in particular neural prostheses. In this presentation, I will provide an overview on fundamentals and applications of several graphene-based technologies and devices aiming at developing an efficient bidirectional communication with electrogenic cells and nerve tissue. To this end, I will discuss several device technologies based on graphene that are used to investigate the electrical activity in cell cultures and in acute experiments (nerve tissue slices); finally, we will disclose recent in-vivo experiments in which flexible graphene devices are used to record brain activity. The main goal of this talk is to highlight the great potential of graphene technologies in neuroprosthetics and, at the same time, to identify the main challenges ahead.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Targeted nanotherapy of endocrine tumors by magnetic intra-lysosomal hyperthermia (MILH) Veronique Gigoux Team INSERM ERL1226: Receptology and Targeted Therapy of Cancers Laboratory LPCNO, CNRS UMR5215-INSA, University of Toulouse, Toulouse, France veronique.gigoux@inserm.fr Cancer is a leading cause of death with millions of new people diagnosed with cancer every year. One major difficulty in anti-cancer therapy is the multidrug resistance which appears during treatments. Recently, studies have shown that cancer cells resistant to traditional therapies are sensitive to agents that induce lysosome membrane permeabilization (LMP) causing lysosomal cell death (LCD) which displays necrotic or apoptotic features. To date, LCD has been obtained using lysosomotropic agents which could not selectively target lysosomes of tumoral cells. Thus, the development of an efficient LCD-targeting therapy constitutes a promising challenge as a new targeted anti-tumoral therapeutic approach. Magnetic intra-lysosomal hyperthermia (MILH) represents an effective way to trigger LCD specifically in cancer cells. Indeed, magnetic nanoparticles (MNPs) offer the potential to be driven into the lysosomes of cancer cells by grafting them with ligands or antibodies recognizing receptors overexpressed in tumors and able to internalize after activation. Moreover, targeted MNPs eradicate cancer cells by MILH upon application of a high frequency alternating magnetic ďŹ eld (AMF). As a proof-of-concept, we showed that minute amounts of iron oxide MNPs targeting gastrin receptor (CCK2R) are internalized by tumoral cells through CCK2R-dependent physiological process, accumulated into their lysosomes and killed tumoral cells through LCD upon AMF application (275 kHz, 40 mT) [1,2]. Moreover, no perceptible temperature rise in the cell medium

occurred during AMF application {Sanchez, 2014 #60;Domenech, 2013 #51;Creixell, 2011 #56}. We thus termed this approach: magnetic intralysosomal hyperthermia (MILH) which differs from magnetic fluid hyperthermia whereby tumor eradication is achieved with large doses of MNPs which cause temperature elevation of the whole tumor. We investigated the mechanism of cell death and demonstrated that MILH induced reactive oxygene species production, lysosomal damage leading to the leakage of lysosomal enzymes into the cytoplasmic compartment and to cell death. Moreover, ROS production and lysosome membrane permeabilization were detected only 30 minutes after AMF application, demonstrating that they occur at an early stage in the cascade of events leading eventually to cell death. We are currently seeking to clarify the mechanisms involved in cell death in MILH condition, especially the signaling pathway activated downstream of lysosomal damage. In conclusion, our results strongly support the potential of lysosomal damage induced by MILH as a therapeutic strategy to eradicate specifically tumoral cells by using lysosometargeting MNP. These data are very promising in light of the recent concept that induction of LMP has wildly appeared as an efficient way to eliminate apoptosis-resistant cancer cells and that some lysosome-targeting drugs can also resensitize multi-drug resistant cancer cells to classical chemotherapy.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Sanchez C et al. (2014) Targeting a G-ProteinCoupled Receptor Overexpressed in Endocrine Tumors by Magnetic Nanoparticles To Induce Cell Death. ACS Nano 8(2):1350-63. [2] Connord V et al. (2015) Real-time Analysis of Magnetic Hyperthermia Experiments on Living Cells under Confocal Microscope. Small 11(20):2437-45.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nucleic acid based supramolecular systems: a route to biomedical applications Arnaud Gissot, Gissot Brune Vialet and Philippe Barthelemy UniversitĂŠ de Bordeaux, UnitĂŠ ARNA Inserm U1212, UMR 5320 CNRS, Bordeaux, France arnaud.gissot@u-bordeaux.fr Amphiphilic molecules spontaneously selfassemble in water to form supramolecular assemblies at the nanometer scale. These aggregates are finding more and more applications in the context of biotechnologies and nanomedicine. Our laboratory is primarily concerned with the synthesis and applications of nucleoside- and oligonucleotide-based amphiphiles. Besides being inherently

biocompatible, the presence of the nucleic acidbased polar head in their structure brings about new interesting properties to these hybrid molecules. The presentation will mainly focus on Lipid OligoNucleotides (LONs) and their unique self assembling and recognition properties as well as their use as nanocargoes for biomedical applications.

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Patwa, A.; Gissot, A.; Oumzil, K.; Barthélémy, P. “Lipid Oligonucleotide Bioconjugates: Applications in Medicinal Chemistry”. In DNA in Supramolecular Chemistry and Nanotechnology; E. Stulz and G. Clever, 2015; pp 276–293. [2] Gissot, A., Oumzil, K., Patwa, A., Barthélémy, P. " A hybrid lipid oligonucleotide: a versatile tool for supramolecular chemistry” New. J. Chem., 2014, 38, 5129-5134. [3] Patwa, A., Gissot, A., Bestel, I., Barthélémy, P. "Hybrid lipid oligonucleotide conjugates: synthesis, self-assemblies and biomedical applications” Chem. Soc. Rev., 2011, 40, 58445854. [4] Gissot, A., Camplo, M., Grinstaff, M. W., Barthélémy, P. "Nucleoside, nucleotide and oligonucleotide based amphiphiles: a successful marriage of nucleic acids with lipids” Org. Biomol. Chem., 2008, 6, 13241333.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Current and future applications for brain-computer interfaces Christoph Guger g.tec medical engineering GmbH, Austria

Research groups all over the world have been working enthusiastically on Brain-Computer Interfaces (BCIs), which provide a direct connection from the human brain to a computer. BCIs translate brain activity into control signals for numerous applications, including tools to help severely disabled users communicate and improve their quality of life. BCIs have been used to restore movement, assess cognitive functioning, and provide communication and environmental control. Dr. Christoph Guger, CEO of g.tec medical engineering GmbH and developer of real-time brain-computer interfaces, will shortly explain the three major BCI approaches - motor imagery, P300 and steady state visual evoked potentials (SSVEP) - for spelling, assessment, rehabilitation and robot control. He will also explain new

directions like active and dry electrodes, invasive ECoG systems and advanced VR control. The audience will get informed about all the required hardware and software, and BCI operation. g.tec is also an active member in a number of national and international research projects and is active in scientific publishing. One of these projects is called Neurographene (http://www.neurographene.eu/), which focuses on the fabrication of a graphene-based multielectrode array (MEA) prototype for local stimulation and recording of brain activity. It is our hypothesis that characteristics of graphene such as flexibility, high biocompatibility and conductivity can be exploited into engineering an optimal brain interface.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Dynamic cell-material interactions Pascal Jonkheijm1,2 1 Bioinspired molecular engineering laboratory, MIRA Institute for Biomedical Technology and Technical Medicine 2 Molecular nanofabrication group, MESA+ Institute for Nanotechnology p.jonkheijm@utwente.nl www.jonkheijm.org Supramolecular chemistry provide nowadays an excellent prospect to construct reversible biological interfaces that can be employed for supramolecular cell manipulation experiments [1]. Making use of supramolecular chemistry is rewarding to develop functional materials and devices. Knowing the limitations involved in ordering proteins at different length scales will surely hasten developing future applications, supramolecular bionanotechnology being the most prominent. The construction of synthetic supramolecular assemblies of proteins provides an excellent tool to fabricate organized bioactive components at surfaces. I will present new synthetic procedures for site-specific noncovalent anchoring of proteins to surfaces and polymers [25]. Special attention is paid to orientational and conformational aspects at the surface and will be demonstrated. Using concepts of multivalency the interactions between proteins and surfaces can be modulated by design. Many of the protein complexes were patterned on surfaces using microcontact printing or nanolithography and visualized using fluorescence microscopy [6]. Furthermore, supramolecular linkers that are sensitive to remote electrochemical stimuli will be presented, using cucurbituril (CB) [2] and cyclodextrin (CD)-modified surfaces [4]. Electrochemical switching was studied using surface embedded electrodes [7]. Cell release was studied in detail in the case of cell-adhesive peptides and growth factors [7,8]. Lastly, supramolecular linkers were compared to reversible covalent linkers, using imine chemistry, providing insight in the cell receptor signaling pathway [8]. With the development of reversible bioactive platforms on surfaces serving as a

reversible dynamic interfaces to cells, improved scaffolds for tissue regeneration will become in hand. First steps into this directions will be introduced as well.

Figures

Figure 1. Supramolecular platform for cell adhesion employing host-guest chemistry suitable for electroresponsive cell manipulation

References [1] J. Brinkmann, E. Cavatorta, S. Sankaran, B. Schmidt, J. van Weerd and P. Jonkheijm, Chem. Soc. Rev. 43, 4449 (2014). [2] A. Gonzalez-Campo, M. Brasch, D. Uhlenheuer, A. Gรณmez-Casado, L. Yang, L. Brunsveld, J. Huskens and P. Jonkheijm, Langmuir 28, 16364 (2012). [3] A. Gonzรกlez-Campo, B. Eker, H.J.G.E. Gardeniers, J. Huskens and P. Jonkheijm, Small 8, 3531 (2012).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

[4] L. Yang, A. Gómez-Casado, J.F. Young, H.D. Nguyen, J. Cabanas-Danés, J. Huskens, L. Brunsveld and P. Jonkheijm, J. Am. Chem. Soc. 134, 19199 (2012). [5] D. Wasserberg, N. Nicosia, E.E. Tromp, V. Subramaniam, J. Huskens and P. Jonkheijm, J. Am. Chem. Soc. 135, 3104 (2013). [6] J. Voskuhl, J. Brinkmann and P. Jonkheijm, Curr. Opin. Chem. Biol. 18, 1 (2014). [7] Q. An, J. Brinkmann, J. Huskens, S. Krabbenborg, J. de Boer and P. Jonkheijm, Angew. Chem. Int. Ed. 51, 12233 (2012). [8] J. Cabanas-Danés, J. Huskens and P. Jonkheijm, J. Mater. Chem. B 2, 2381 (2014).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Design and in vivo Validations of Nanosystems in Cardiovascular Tissue Engineering and Molecular Imaging Didier Letourneur Cardiovascular Bio-engineering - INSERM U1148 - LVTS X. Bichat Hospital, University Paris 7 - Denis Diderot, 75018 Paris Institut GalilĂŠe, University Paris 13 - Paris Nord, 93430 Villetaneuse, France didier.letourneur@inserm.fr This presentation intends to present nanosystems for regenerative medicine and drug delivery systems as well as targeted contrast agents for molecular imaging with an emphasis on cardiovascular pathologies. We will present examples of innovative nano medical imaging tools. In the context of atherothrombotic diseases, there is a need for new approaches for early diagnosis and improved therapies. This is the focus of NanoAthero, an European large scale project, started in February 2013. The aim is to demonstrate that nanotechnologies can be developed and clinically proven to be effective in tackling cardiovascular diseases. The NanoAthero consortium is a unique opportunity to extend the frontiers of knowledge on atherothrombosis management. NanoAthero aims to demonstrate the preliminary clinical feasibility of the use of nanosystems for targeted imaging and treatment of advanced atherothrombotic disease in humans. NanoAthero combines in-depth knowledge of nanocarrier bioengineering and production with state-of-the-art expertise in imaging and treatment of cardiovascular patients providing a full framework of 16 partners within one collaborative European consortium (16 partners from 10 countries - see www.nanoathero.eu).

rats to achieve rapid and strong closure and healing of deep wounds in skin and liver. Nanoparticles can also be used to fix polymer membranes to tissues even in the presence of blood flow, such as occurring after liver resection, yielding permanent hemostasis within a minute. Furthermore, medical devices and tissue engineering constructs could be fixed to organs such as a beating heart.

Figures

We will also present how to use nanomaterials for regenerative medicine. Indeed, adhesion by aqueous nanoparticle solutions can be used in vivo in

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Suzuki, M., Bachelet-Violette, L., Rouzet, F., Beilvert, A., Autret, G., Maire, M., Menager, C., Louedec, L., Choqueux, C., Saboural, P., Haddad, O., Chauvierre, C., Chaubet, F., Michel, J. B., Serfaty, J. M., and Letourneur, D. (2015) Ultrasmall superparamagnetic iron oxide nanoparticles coat-ed with fucoidan for molecular MRI of intraluminal thrombus. Nanomedicine, 10(1):73-87 [2] Varna, M., Juenet, M., Bayles, R., Mazighi, M., Chauvierre, C., Letourneur D. (2015) Nanomedicine as a strategy to fight thrombotic diseases. Future Science OA 10.4155/FSO.15.46, 1-14 [3] Silva, A.K., Juenet, M., Meddahi-Pellé, A., and Letourneur, D. (2015) Polysaccharide-based strate-gies for heart tissue engineering. Carbohydr Polym 116, 267-77. [4] Bonnard, T., Serfaty, J. M., Journe, C., Ho Tin Noe, B., Arnaud, D., Louedec, L., Derkaoui, S. M., Letourneur, D., Chauvierre, C., and Le Visage, C. (2014) Leukocyte mimetic polysaccharide micropar-ticles tracked in vivo on activated endothelium and in abdominal aortic aneurysm. Acta Biomat 10, 3535-3545 [5] Letourneur, D., Trohopoulos, P. (2014) Atherosclerotic disease and management challenges with nanomedicine: EU FP7 NMP funded “NanoAthero” and “CosmoPHOSnano” large-scale projects. Eur J Nanomed 6(2), 115-119 [6] Bonnard, T., Yang, G., Petiet, A., Ollivier, V., Haddad, O., Arnaud, D., Louedec, L., BacheletViolette, L., Derkaoui, S. M., Letourneur, D., Chauvierre, C., and Le Visage, C. (2014) Abdominal aortic aneurysms targeted by functionalized polysaccharide microparticles: a new tool for SPECT imaging. Theranostics 4, 592-603 [7] Silva, A. K., Letourneur, D., and Chauvierre, C. (2014) Polysaccharide nanosystems for future pro-gress in cardiovascular pathologies. Theranostics 4, 579-591

[8] Meddahi-Pelle, A., Legrand, A., Marcellan, A.,

Louedec, L., Letourneur, D., and Leibler, L. (2014) Organ repair, hemostasis, and in vivo bonding of medical devices by aqueous solutions of nanoparti-cles. Angew Chem Int Ed Engl 53, 6369-6373 + Cover

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Intracellular drug release using mesoporous silica nanoparticles as drug vectors Mika LindĂŠn Inorganic Chemistry II, Ulm University, Albert-Einstein-Allee 11, D-89081 Ulm, Germany mlinden@uni-ulm.de Keywords: biomaterial, biomedical, sol-gel, Si Mesoporous silica nanoparticles have attracted immense interest as drug carriers over the last years, and successful targeted drug delivery has been demonstrated in vitro and occasionally in vivo in small animal models. The flexibility in terms of particle size, shape, and surface chemistry makes it possible to fine-tune the physical and chemical properties to correspond to the requirements of a specific administration route. The presentation will critically discuss biodistribution issues found when moving from in vitro to in vivo studies, and relate these to the physicochemical properties of the nanoparticles controlling the extent of protein adsorption to the particles. Special focus will be put on the optimization of passive targeting of these particles to tumors. In the second part of the presentation approaches towards local administration of the particles for local therapies or tissue engineering applications will be discussed, with focus on organic scaffolds serving as hosts for the mesoporous silica nanoparticles. Efficient control of stem cell differentiation based on local, intracellular drug delivery will be demonstrated.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Biomimetic channels and channel networks for microfluidics Andreas Manz KIST Europe, Campus E71, 66123 Saarbrücken, Germany

The idea to shrink a laboratory down to chip size, or at least to integrate some critical lab tasks onto micro devices has been around since over 20 years now. In the meantime, the field has grown significantly, many of my students are now professors in this field, the early patents are all expired, small companies use microfluidics for their products, and the main application focus has shifted from analytical chemistry to cell biology and tissue engineering [1].

References [1] P Neužil, S Giselbrecht, K Länge, TJ Huang, A Manz, “Revisiting lab-on-a-chip technology for drug discovery”, Nature Reviews Drug Discovery 11 (8), 620-632 (2012). [2] W.Wu, R.M.Guijt, Y.E.Silina, M.Koch, A.Manz, “Plant leaves as templates for soft lithography”, RSC Advances 6, 22469-22475 (2016).

Microfluidic chips are usually defined by photolithography masks which are generated from straight lines and CAD programs. The manufacturing process usually gets more complex if multiple depths, i.e. multiple masks have to be used, and variations in depth profile are difficult to achieve. I will present a simple way of obtaining channel structures which feature gradually increasing or decreasing channel depths, and which also can feature irregularities in its surface. At first sight, this may seem inappropriate, may look “ugly” and not engineering-like. However, in biological surroundings, we can see such structures, and they are fully functional. Plant leaves are used as templates for channel patterns, including its fine structure and including its macroscopic network pattern. Structures are formed in PDMS and covered by glass slides for microscopic observation [2]. Structures are used for investigating cell behaviour.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Paper-based nanobiosensors in diagnostics: from health, safety and security to environment monitoring Arben Merkoçi Catalan Institute of Nanoscience and Nanotechnology (ICN2), CSIC and The Barcelona Institute of Science and Technology, Barcelona, Spain. ICREA - Institucio Catalana de Recerca i Estudis Avançats, Barcelona, Spain arben.merkoci@icn2.cat Biosensors field is progressing rapidly and the demand for cost efficient platforms is the key factor for their success. Physical, chemical and mechanical properties of cellulose in both micro and nanofiber-based networks combined with their abundance in nature or easy to prepare and control procedures are making these materials of great interest while looking for cost-efficient and green alternatives for device production technologies. Both paper and nanopaper-based biosensors are emerging as a new class of devices with the objective to fulfil the “World Health Organization” requisites to be ASSURED: affordable, sensitive, specific, user-friendly, rapid and robust, equipment free and deliverable to end-users. How to design simple paper-based biosensor architectures? How to tune their analytical performance upon demand? How one can ‘marriage’ nanomaterials such as metallic nanoparticles, quantum dots and even graphene with paper and what is the benefit? How we can make these devices more robust, sensitive and with multiplexing capabilities? Can we bring these low cost and efficient devices to places with low resources, extreme conditions or even at our homes? Which are the perspectives to link these simple platforms and detection technologies with mobile phone communication? I will try to give responses to these questions through various interesting applications related to protein, DNA and even contaminants detection

all of extreme importance for diagnostics, environment control, safety and security.

References [1] C. Parolo, A. Merkoçi, “Paper based nanobiosensors for diagnostics”, Chem. Soc. Rev., 42, 2013, 450-457 ; [2] D. Quesada-González, A. Merkoçi. «Nanoparticle-based lateral flow biosensors», Biosensors and Bioelectronics, 73, 2015, 47–63 [3] E. Morales-Narváez, H.Golmohammadi, T. Naghdi, H. Yousefi, U. Kostiv, D. Horak, N. Pourreza, A.Merkoçi. “Nanopaper as an optical sensing platform “, ACS Nano, 9, 2015, 7296-7305 ; [4] E. Morales-Narvaez, T. Naghdi, E. Zor, A. Merkoci, “Photoluminescent Lateral-Flow Immunoassay Revealed by Graphene Oxide: Highly Sensitive Paper-Based Pathogen Detection”, Anal. Chem. 2015, 87, 8573−8577. [5] L. Baptista-Pires, C. C. Mayorga-Martínez, M.M. Sanchez, H. Monton, A. Merkoçi, “Water Activated Graphene Oxide Transfer Using Wax Printed Membranes for Fast Patterning of a Touch Sensitive Device”, ACS Nano 2016, 10, 853−860 [6] A.M. López_Marzo, A. Merkoçi, “Paper-based sensors and assays: a success of the engineering design and the convergence of knowledge areas”, Lab Chip, 2016, 16, 3150– 3176

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nanomechanics of Soft Biological Tissues Probed with Atomic Force Microscopy Daniel Navajas Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain School of Medicine, University of Barcelona, Barcelona, Spain CIBER of Respiratory Diseases. Madrid, Spain dnavajas@ub.edu Cell biomechanics mediates critical cell functions including proliferation, differentiation, gene expression, contraction, and migration. Moreover, cells sense and actively respond to the mechanical features of their microenvironment. Therefore, a precise characterization of the mechanical properties of cells and extracellular matrix (ECM) is needed to further our understanding of the cellmicroenvironment interplay. We use atomic force microscopy (AFM) to study nanomechanical properties of cells and EMC of lung and heart tissues [1-4]. The Young’s modulus (E) is computed by fitting the tipsample contact model to force-indentation curves recorded on sample. The complex shear modulus (G*) is measured by placing the tip at an operating indentation of ∼500 nm and superimposing small amplitude (∼75 nm) multifrequency oscillations composed of sine waves (0.1-11.45 Hz). G* is computed in the frequency domain from the complex ratio between oscillatory force and indentation signals. Both cells and ECM exhibit a viscoelastic behavior with a complex shear modulus that increases with frequency as a two power law composed of a weak power law (exponent 0.05), accounting for a viscoelastic solid regime dominant at physiological frequencies, and a second power law with an exponent of 3/4, accounting for a viscoelastic liquid regime at high frequencies. Our AFM measurements define intrinsic mechanical properties of the ECM at the length scale in which cells sense and

probe their microenvironment. Regional changes in mechanical properties of the ECM could provide differential mechanical cues to regulate the spatial distribution, differentiation and function of lung and heart cells. (Funded in part by the Spanish Ministry of Economy and Competitiveness grant FIS-PI1400280)

References [1] Alcaraz J, Buscemi L, Grabulosa M, Trepat X, Fabry B, Farré R, Navajas D. Microrheology of human lung epithelial cells measured by atomic force microscopy. Biophys J 2003; 84: 2071-9. [2] Luque T, Melo E, Garreta E, Cortiella J, Nichols J, Farré R, Navajas D. Local micromechanical properties of decellularized lung scaffolds measured with atomic force microscopy. Acta Biomater 2013; 9:6852-9. [3] Andreu I, Luque T, Sancho A, Pelacho B, Iglesias-García O, Melo E, Farré R, Prósper F, Elizalde MR, Navajas D. Heterogeneous micromechanical properties of the extracellular matrix in healthy and infarcted hearts. Acta Biomater 2014; 10:3235-42. [4] Ignasi

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Interaction of colloidal hybrid nano- and microparticles with cells Wolfgang J. Parak Fachbereich Physik, Philipps Universität Marburg, Marburg, Germany, and CIC Biomagune, San Sebastian, Spain wolfgang.parak@physik.uni-marburg.de What happens to inorganic nanoparticles (NPs), such as plasmonic gold or silver, superparamagnetic iron oxide, or fluorescent quantum dot NPs, after they have been administrated to an animal or a human being? The review discusses the integrity, biodistribution, and fate of NPs after in vivo administration. First the hybrid nature of the NPs is described, by conceptually dividing them into the inorganic NP core, an engineered surface coating around the core which comprises the ligand shell and optionally also bioconjugation, and into the corona of adsorbed biological molecules. It is shown that in vivo all of these three compounds may degrade individually and that each of them can drastically modify the life-cycle and biodistribution of the whole hetero-structure. The NPs thus may be disintegrated into different parts, of which biodistribution and fate would need to be analyzed individually. Multiple labelling and quantification strategies for such purpose will be discussed. All reviewed data indicate that in vivo NPs no longer should be considered as homogeneous entity, but should be seen as inorganic/organic/biological nanohybrids with complex and intricately linked degradation pathways.

References [1] M. Chanana, P. Rivera Gil, M. A. CorreaDuarte, L. M. Liz-Marzán. W. J. Parak, "Physicochemical Properties of ProteinCoated Gold Nanoparticles in Biological Fluids and Cells before and after Proteolytic Digestion", Angewandte Chemie International Edition 52, 4179–4183 (2013). [2] W. G. Kreyling, A. M. Abdelmonem, Z. Ali, F. Alves, M. Geiser, N. Haberl, R. Hartmann, S. Hirn, K. Kantner, D. Jimenez de Aberasturi, G. Khadem-Saba, J.-M. Montenegro, J. Rejman, T. Rojo, I. Ruiz de Larramendi, R. Ufartes, A. Wenk, W. J. Parak, "In vivo integrity of polymer-coated gold nanoparticles", Nature Nanotechnology 10, 619–623 (2015).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Charge Transport in single DNA-Based Molecules Danny Porath Institute of Chemistry and Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, 91904 Israel danny.porath@mail.huji.ac.il DNA is primarily and with no doubt the most important biological molecule. But its doublestrand recognition, as well as the ability to control its sequence and manipulate its structure open a multitude of ways to make it useful for molecular electronics. Step by step we improve the synthesized constructs and the measurement methods of single DNA-based molecules. In this lecture I will report on our progress in producing and measuring DNAbased building blocks towards the construction of DNA-based programmable circuits.

Figures

References [1] "Direct measurement of electrical transport through DNA molecules", Danny Porath, Alexey Bezryadin, Simon de Vries and Cees Dekker, Nature 403, 635 (2000). Cited 1312 times [2] "Charge Transport in DNA-based Devices", Danny Porath, Rosa Di Felice and Gianaurelio Cuniberti, Topics in Current Chemistry Vol. 237, pp. 183-228 Ed. Gary Shuster. Springer Verlag, 2004. Cited 193 times [3] “Direct Measurement of Electrical Transport Through Single DNA Molecules of Complex Sequence”, Hezy Cohen, Claude Nogues, Ron Naaman and Danny Porath, PNAS 102, 11589 (2005). Cited 201 times [4] “Long Monomolecular G4-DNA Nanowires”, Alexander Kotlyar, Nataly Borovok, Tatiana Molotsky, Hezy Cohen, Errez Shapir and Danny Porath, Advanced Materials 17, 1901 (2005). Cited 70 times [5] “Electrical characterization of self-assembled single- and double-stranded DNA monolayers using conductive AFM”, Hezy Cohen et al., Faraday Discussions 131, 367 (2006). Cited 42 times [6] “High-Resolution STM Imaging of Novel Poly(G)Poly(C)DNA Molecules”, Errez Shapir, Hezy Cohen, Natalia Borovok, Alexander B. Kotlyar and Danny Porath, J. Phys. Chem. B 110, 4430 (2006). Cited 24 times [7] "Polarizability of G4-DNA Observed by Electrostatic Force Microscopy Measurements", Hezy Cohen et al., Nano Letters 7(4), 981 (2007). Cited 56 times

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

[8] “Electronic structure of single DNA molecules resolved by transverse scanning tunneling spectroscopy”, Errez Shapir et al., Nature Materials 7, 68 (2008). Cited 93 times [9] “A DNA sequence scanned”, Danny Porath, Nature Nanotechnology 4, 476 (2009). [10] “The Electronic Structure of G4-DNA by Scanning Tunneling Spectroscopy”, Errez Shapir, et.al., J. Phys. Chem. C 114, 22079 (2010). [11] “Energy gap reduction in DNA by complexation with metal ions”, Errez Shapir, G. Brancolini, Tatiana Molotsky, Alexander B. Kotlyar, Rosa Di Felice, and Danny Porath, Advanced Maerials 23, 4290 (2011). [12] "Quasi 3D imaging of DNA-gold nanoparticle tetrahedral structures", Avigail Stern, Dvir Rotem, Inna Popov and Danny Porath, J. Phys. Cond. Mat. 24, 164203 (2012). [13] "Comparative electrostatic force microscopy of tetra- and intra-molecular G4-DNA", Gideon I. Livshits, Jamal Ghabboun, Natalia Borovok, Alexander B. Kotlyar, Danny Porath, Advanced materials 26, 4981 (2014). [14] "Long-range charge transport in single G4-DNA molecules", Gideon I. Livshits et. al., Nature Nanotechnology 9, 1040 (2014). [15] "Synthesis and Properties of Novel Silver containing DNA molecules", Gennady Eidelshtein et. al., Advanced Materials 28, 4839 (2016).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Novel Functional Carbon Bio-Interfaces Maurizio Prato Department of Chemical and Pharmaceutical Sciences, University of Trieste, Italy CIC BiomaGUNE, Parque Tecnológico de San Sebastián, San Sebastián (Guipúzcoa), Spain prato@units.it, mprato@cicbiomagune.es Connecting nanostructured materials to biological compartments is a crucial step in prosthetic applications, where the interfacing surfaces should provide minimal undesired perturbation to the target tissue. Ultimately, the (nano)material of choice has to be biocompatible and promote cellular growth and adhesion with minimal cytotoxicity or disregulation of, for example, cellular activity and proliferation. In this context, carbon nanomaterials, including nanotubes and graphene, are particularly well suited for the design and construction of functional interfaces. This is mainly due to the extraordinary properties of these novel materials, which combine mechanical strength, thermal and electrical conductivity.

In particular, we have recently shown that carbon nanotubes and graphene can act as active substrates for neuronal growth, a field that has given so far very exciting results. Nanotubes and graphene are compatible with neurons, but, especially, they play a very interesting role in interneuronal communication. Improved synaptic communication is just one example. During this talk, we will show the latest and most exciting results obtained in our laboratories in these fast developing fields.

Our group has been involved in the organic functionalization of various types of nanocarbons, including carbon nanotubes, fullerenes and, more recently, graphene. The organic functionalization offers the great advantage of producing soluble and easy-tohandle materials. As a consequence, since biocompatibility is expected to improve upon functionalization, many modified carbon nanomaterials may be useful in the field of nanomedicine.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Sensing of particles, micro-organisms and biomarkers using the camera of a mobile phone V. Pruneri1,2, M. Jofre1, R. A. Terborg1 , J.- M. Pérez1, T. Coll1, P. Martínez1, W. Amaya1 and J. Pello1 1 ICFO-Institut de Ciencies Fotoniques, Castelldefels, The Barcelona Institute of Science and Technology, Spain 2 ICREA-Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain valerio.pruneri@icfo.eu By exploiting full field of view CMOS or CCD image sensor array (ISA) at the highest resolution, we have designed and developed low cost and portable devices for measuring light scattering, reading microarrays, as well as detecting and counting microorganisms, cells and particles [1,2]. In addition to CMOS-ISA, the devices are made of other off-the-shelf components, including a light emitting diode (LED) source, and are based on proprietary geometries that allow the processing of large sample areas or volumes. The devices are also portable and affordable, which allows for the extension of traditionally lab-based solutions towards point-of-care applications. Such applications are numerous, from bedside diagnostics to quality control of pharmaceutical powders and testing of drinking water sources in areas lacking accessibility to laboratory infrastructures.

Figures

References [1] [1] J. M. Pérez et al., Analyst 140, 7734-7741 (2015) [2] [2] R. Terborg et al., Science Advances 2, e1600077 (2016)

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Interactions between peptides and inorganic matter: From basic science to applications Meital Reches The Hebrew University of Jerusalem, Israel

Several natural processes are mediated by the interactions between organic and inorganic materials. The immune response towards an implant inserted into the body is mediated by proteins. Composite materials are formed by the interactions of organic materials (usually proteins) and minerals. Biofouling, the process in which organisms attached to surfaces, is also mediated by organic molecules. Understanding the nature of interactions between organic and inorganic materials will bring to the development of improved implants, new composites and antifouling materials. This lecture will present single-molecule force spectroscopy measurements of the interactions between individual biomolecules (either amino acid residues or short peptides) and inorganic surfaces in aqueous solution. Using this method, we were able to measure low adhesion forces and could clearly determine the strength of interactions between individual amino acid residues and inorganic substrates. Our results with peptides also shade light on the factors that control the interactions at the organicinorganic interface. Based on our knowledge from single molecule experiments, we designed a short peptide (tripeptide) that can spontaneously form a coating that resists biofilm formation. Our results clearly demonstrate the formation of a coating on various surfaces (glass, titanium, silicon oxide, metals and polymers). In addition,

we showed that this coating prevents the first step of antifouling, which involves the adsorption of bioorganic molecules to the substrate. Moreover, the coating significantly reduces the attachment of various organisms such as bacteria and fungi to surfaces.

References [1] Das, P. & Reches, M. (2016) Nanoscale Revealing the Role of Catechol Moieties in the Interactions between Peptides and Inorganic Surfaces. DOI: 10.1039/C6NR04550B. [2] Das, P. & Reches, M. (2016) Nanoscale Single-Stranded DNA Detection by SolventInduced Assemblies of a Metallo-PeptideBased Complex. 8, 9527-9536. [3] Maity, S., Zanuy, D., Razvag, Y., Aleman, C. & Reches, M. (2015) Phys. Chem. Chem. Phys. Elucidating the mechanism of interaction between peptides and inorganic surfaces. 17, 15305-15315. [4] Maity, S., Nir, S., Zadaa, T. & Reches, M. (2014) Self-Assembly of a Tripeptide into a Functional Coating that Resists Fouling. Chem. Comm. 50, 11154-11157. [5] Razvag, Y., Gutkin, V.& Reches, M. (2013) Probing the Interaction of Individual Amino Acids with Inorganic Surfaces Using Atomic Force Spectroscopy. Langmuir 29, 10102â&#x20AC;&#x201C; 10109.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Chemical Nanomachines as active drug nanovehicles Samuel Sánchez1,2,3 1 Smart Nano-Bio-Devices Group, Institute for Bioengineering of Catalonia, Barcelona, Spain 2 Max Planck Institute for Intelligent Systems Institution, Stuttgart, Germany 3 Institució Catalana de Recerca i Estudis Avancats (ICREA), Barcelona, Spain sanchez@is.mpg.de; ssanchez@ibecbarcelona.eu; Engineering tiny machines that convert chemistry into motion is of huge impact in multidisciplinary chemistry as indicated by the current Nobel Prize in Chemistry to “Nanomachines”. Self-propelled nanomachines are opening many avenues in fields such as robotics, biosensing, nanomedicine, microfluidics, and environmental field [1, 2]. One example could be in nanomedicine, where the active and direct transport of drugs to specific locations enabled by hybrid micronano-bots, which are powered either by enzymatic catalytic reactions [3-5], by motile cells such as sperms or bacteria or by multiple 3D bioprinted cells. [6] Here, I will present our recent developments in the field of bio- and nano-engineering systems that can be autonomously activated and perform complex tasks. We fabricate nano-bots from mesoporous silica nanoparticles, microcapsules, microtubes up to 3D Bioprinted structures. Our types of hybrid Micro-bio-bots combine the best from the two worlds, biology and (nano)materials providing remote control with biocompatible fuels with the hope to make them suitable for bio-related applications in the near future.

References [1] S. Sanchez, Ll. Soler and J. Katuri. Angew.Chem.Int.Edit. 2015, 54,1414-1444 [2] D. Vilela, J. Parmar, YF Zheng, Y. Zhao and S. Sanchez Nano Lett., 2016, 16 (4), 2860–2866 [3] X. Ma, A.C. Hortelao, T. Patiño and S. Sanchez. ACS Nano 2016. DOI: 10.1021/acsnano.6b04108 [4] X. Ma, A. Jannasch, U-R Albrecht, K. Hahn, A. Miguel López, E. Schäffer and S. Sanchez. Nano Lett. 2015, 15, 7043–7050. [5] X. Ma, X. Wang, K. Hahn, S. Sanchez. ACS Nano. 2016, 10, 3597–3605 [6] T. Patino, R. Mestre and S. Sanchez, Lab Chip, 2016. DOI: 10.1039/C6LC90088G

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Identifying molecular signatures of tumor dormancy as a basis for the rational design of precision nanomedicines Ronit SatchiSatchi-Fainaro Head, Cancer angiogenesis and Nanomedicine Laboratory; Chair, Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 69978, Israel. ronitsf@post.tau.ac.il Tumor progression is dependent on a number of sequential steps, including initial tumorvascular interactions and recruitment of blood vessels, as well as established interactions of tumor cells with their surrounding microenvironment and its different immune, endothelial and connective cellular and extracellular components. Failure of a microscopic tumor, either primary, recurrent or metastatic, to complete one or more of these early stages may lead to delayed clinical manifestation of the cancer. Micrometastasis, dormant tumors, and minimal residual disease, contribute to the occurrence of relapse, and constitute fundamental clinical manifestations of tumor dormancy that are responsible for the majority of cancer deaths. However, although the tumor dormancy phenomenon has critical implications for early detection and treatment of cancer, it is one of the most neglected areas in cancer research and its biological mechanisms are mostly unknown. To that end, we created several models of patient-derived cancer models mimicking pairs of dormant versus fast-growing, primary versus metastatic and drug-sensitive versus drugresistant cancers using cutting-edge techniques of patient-derived xenografts, 3D printing and genetically-modified mouse models. We investigated the molecular changes in tumorhost interactions that govern the escape from drormancy and contribute to tumor

progression. Those led to the discovery of novel targets and provided important tools for the design of novel cancer nano-sized theranostics (thera therapeutics and diagnostics nostics) thera nostics [1]. Our libraries of precision nanomedicines are synthesized as highly controlled micellar, nanogels, coiled or globular particulated supramolecular structures consisting of linear, hyperbranched and dendritic polymers based on polyglutamic acid, polyethyleneglycol, poly(N-(2hydroxypropyl)methacrylamide), polyglycerol, and hybrid systems [2-6]. We hypothesize that the acquired knowledge from this multidisciplinary research strategy will revolutionize the way we diagnose and treat cancer.

References [1] Tiram, G., et al. ACS Nano 10, 2028-2045 (2016). [2] Baabur-Cohen, H., et al. J Control Release, in press (2016). [3] Ofek, P., et al., Nanomedicine : nanotechnology, biology, and medicine 12, 2201-2214 (2016). [4] Polyak, D., et al., J Control Release (2016). [5] Scomparin, A., et al., J Control Release 208, 106-120 (2015). [6] Shatsberg, Z., et al. J Control Release 239, 159-168 (2016).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Personalized Cancer Nanomedicine Avi Schroeder Assistant Professor of Chemical Engineering, Technion â&#x20AC;&#x201C; Israel Institute of Technology, Israel avids@technion.ac.il The field of medicine is taking its first steps towards patient-specific care. Our research is aimed at tailoring treatments to address each personâ&#x20AC;&#x2122;s individualized needs and unique disease presentation. Specifically, we are developing nanoparticles that target disease sites, where they perform a programmed therapeutic task. These systems utilize molecular-machines and cellular recognition to improve efficacy and reduce side effects. Two examples will be described: the first involves a nanoscale theranostic system for predicting the therapeutic potency of drugs against metastatic cancer. The system provides patient-specific drug activity data with singlecell resolution. The system makes use of barcoded nanoparticles to predict the therapeutic effect different drugs will have on the tumor microenvironment.

The second system makes use of enzymes, loaded into a biodegradable chip, to perform a programed therapeutic task â&#x20AC;&#x201C; surgery with molecular precision. Collagenase is an enzyme that cleaves collagen, but not other tissues. This enzyme was loaded into the biodegradable chip and placed in the periodontal pocket. Once the collagenase releases from the chip, collagen fibers that connect between the teeth and the underlying bone are relaxed, thereby enabling enhanced orthodontic corrective motion and reducing pain. This new field is termed BioSurgery. The clinical implications of these approaches will be discussed.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

The innovation imperative: turning research results into socioeconomic impact Michael R. Tadros Science & Technology Transfer FundaciĂłn BotĂn CastellĂł 18C, 28001 Madrid (Spain)

Dr. Michael Tadros will share insight on the innovation imperative and underlines the role and importance of knowledge based economies. Dr. Tadros will share experiences and practices turning science into products geared towards socioeconomic impact.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Graphene for Bioapplications: Preparation, Cytotoxicity and Integration in 3D-scaffolds Ester Vázquez Departamento de Química Orgánica, Facultad de Ciencias y Tecnologías Químicas-IRICA, Universidad de Castilla-La Mancha, Ciudad Real, (Spain) ester.vazquez@uclm.es Graphene has emerged as a new material, with outstanding mechanical and electronic properties that will permit a broad range of applications, from microelectronics to composite or even medicine. Although there has been a huge effort directed in the area of nanomedicine, biomedical applications of graphene derivatives have, so far, mainly focused on graphene oxide and reduce graphene oxide. The main reason for this fact is the difficult to obtain pristine graphene flakes, directly in water or in culture media, due to the intrinsic hydrophobicity of this material. Our group have recently described an interesting approach for the preparation of stable dispersions of graphene in water, without

detergents or any other additives, driven by an easy and eco-friendly ball milling approach. These aqueous suspensions can be rapidly frozen and, subsequently, lyophilized giving rise to a very soft and low-density black powder. Powders of graphene can be safely stored or shipped and they can be readily dispersed in culture media within the presence or absence of serum and antibiotics. During this talk, we will discuss (i) optimized ways to generate graphene dispersions in culture media; (ii) studies of interaction of soprepared solutions with cells. (ii) the use of graphene in polymeric 3D structures for drug delivery purposes and for 3D cell culture media.

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Suspended planar-array chips for molecular multiplexing at the microscale L. Pérez-García1*, N. Torras2, J. P. Agusil2,3, P. Vázquez4, M. Duch2, A. M. Hernández-Pinto4, J. Samitier3, E. J. de la Rosa4, J. Esteve2, T. Suárez4, J. A. Plaza2 1

Departament de Farmacologia, Toxicologia i Química Terapèutica, Institut de Nanociència i Nanotecnologia (IN2UB), Universitat de Barcelona, Barcelona, Spain 2 Instituto de Microelectrónica de Barcelona, IMB-CNM (CSIC), Barcelona, Spain 3 Nanobioengineering Group, Institute for Bioengineering of Catalonia (IBEC) and Department d’Electrònica, Universitat de Barcelona, Spain 4 Centro de Investigaciones Biológicas, CIB (CSIC), Madrid, Spain.

*mlperez@ub.edu Bio-functionalized microparticles are hybrid materials capable of sensing and tagging living cells [1]. As our approach, we report how novel suspended planar-array chips allow molecular multiplexing using a single suspended chip to analyze extraordinarily small volumes. The suspended chips are fabricated by combining silicon-based technology and polymer-pen lithography, obtaining increased molecular pattern flexibility, and improving miniaturization and parallel production. The chip miniaturization is so dramatic that it permits the intracellular analysis of living cells. Here, we will describe the technology to produce and functionalize suspended chips, and as an example, how to use them for sensing pH in HeLa cells [2].

Acknowledgements: This work was supported by the EU ERDF (FEDER) funds and the Spanish Government grant TEC2014-51940-C2.

References [1] O. Penon, S. Novo, S. Durán, E. Ibañez, C. Nogués, J. Samitier, M. Duch, J. A. Plaza, L. Pérez-García, Bioconjugate Chem., 23 (2012) 2393-2402. [2] N. Torras, J. P. Agusil, P. Vázquez, M. Duch, A. M. Hernández-Pinto, J. Samitier, E. J. de la Rosa, J. Esteve, T. Suárez, L. Pérez-García, J. A. Plaza, Adv. Mater. 28 (2016) 1449–1454.

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Ideal Atomic Force Microscopy imaging of heterogeneous biological samples in liquids: Topography and Chemical information L. Almonte and J. Colchero* Centro de Investigación en Óptica y Nanofísica (CIOyN), Departamento Física, Facultad de Química (Campus Espinardo), Universidad de Murcia, E-30100 Murcia (Spain).

*colchero@um.es Scanning Probe Microscopy techniques have revolutionized biological Nanoscience allowing to work in liquid environment and to study biological samples “at work”. A variety of imaging modes have been optimized or even explicitly developed to improve Atomic Force Microscopy (AFM) operation in biological applications. Nevertheless, we believe that the issue of best AFM imaging mode and its corresponding operation parameters is still a field of debate. Indeed, we believe that AFM operation should be considered in many cases more “art than science”. In the present work Dynamic AFM (DAFM) has been used to study in detail tip-sample interaction and to compare different acquisition modes (see figure below), in particular constant amplitude DAFM (AMDAFM), constant frequency DAFM (FM-DAFM) and constant normal force DAFM (NF-DAFM). We combine imaging and locally resolved AFM-spectroscopy to study the conservative as well as the dissipative components of tip-sample interaction. From their analysis we conclude that the dissipation (=reduction of oscillation amplitude) does not depend on surface chemistry, while the conservative interaction (force and phase/frequency) has a very strong dependence on surface chemistry. Therefore, on heterogeneous samples AM-DAFM is the only imaging mode that allows for correct topography imaging, while at the same time

giving correct information about the chemical composition of the sample. Finally, we show that imaging in liquids is possible in a non-contact regime where essentially no forces are applied, rendering DAFM a truly non-invasive technique. In this thermal noise limited regime the interaction with the sample is of the order of the energy fluctuations kT≈0.025meV induced by thermodynamics. Samples stable at room temperature should therefore not be damaged by the tipsample interaction induced during AFM imaging in the non-contact regime proposed here. The methodology for dynamic AFM operation described will greatly improve AFM experiments, rendering AFM operation in liquids much more quantitative, more stable, less invasive and more easily interpretable; and thus more “science than art”, as compared to many cases where the contrary is true.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. Topography images and line profiles of DOTAP-DPPC (4:1) supported planar bilayer adsorbed on mica in water. Data has been acquired using AM-DAFM (A), FM-DAFM (B) and FN-DAFM (C) modes. The lateral size of all images is about 2x1.6 Âľm2. The total color scale is exactly the same for all images and corresponds to height differences of 8 nm. As clearly observed from the different color in the images, and the different height values in the profiles the DPPC domains have a similar height when measured in the different modes. The height of the DOTAP domains, however, changes drastically. Only AM-DAFM gives correct results; in addition it is is the least invasive imaging mode.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Solid lipid nanoparticles for delivery of calendula officinalis extract Lide Arana1, Clarisa Salado2, Sandra Vega3, Oier Aizpurua-Olaizola4, Igor de la Arada1,Tatiana Suarez3, Aresatz Usobiaga4, José Luis R. Arrondo1, Alicia Alonso1, Félix M. Goñi1,Itziar Alkorta1 1

Instituto Biofisika (UPV/EHU, CSIC), and Departamento de Bioquímica y Biología Molecular, (UPV/EHU), Spain 2 Innoprot, Parque Tecnológico de Bizkaia, Elexalde Derio, Spain 3 Bioftalmik Applied Research, Bizkaia Science and Technology Park Derio, Spain 4 Analytical Chemistry Department, University of the Basque Country (UPV/EHU), Leioa, Spain.

lide.arana@ehu.eus Solid Lidid Nanoparticles (SLN) composed of long-chain saturated fatty acids (arachidic acid, stearic acid and palmitic acid), Epikuron 200 (purified phosphatidylcholine) and bile salts (cholate, taurocholate and taurodeoxycholate) have been obtained by microemulsion method. All nanoparticle suspensions have been characterized by dynamic light scattering, differential scanning calorimetry, infra-red spectroscopy and transmission electron microscopy in order to select the most promising nanoparticle suspension for Calendula officinalis extract delivery. This plant presents anti-inflammatory, emollient and wound-repairing activity mainly because of two hydrophobic components (faradiol and carotenoids). Loading capacity of SLN was determined for these components and the citotoxicity and wound-healing efficiency of calendula-loaded SLN were tested in the conjunctival epithelium cell line WKD. Our results suggest that this SLN formulation is a safe and solvent-free Calendula extract delivery system which could provide a controlled therapeutic alternative for reducing diseaserelated symptoms and improving epithelium repair in ocular surface.

[3] Gastaldi, L., Battaglia, L., Peira, E., Chirio, D., Muntoni, E., Solazzi, I., Gallarate, M. and Dosio, F. Eur J Pharm Biopharm. 87(3) (2014), 433-44. [4] Arana, L., Salado, C., Vega, S., AizpuruaOlaizola, O., de la Arada, I., Suarez, T., Usobiaga, A. Arrondo, J.L., Alonso, A., Goñi, F.M., Alkorta, I. Colloids Surf B Biointerfaces, 135 (2015) 18-26.

Figures

References [1] Mehnert, W. and Mader, K. Adv Drug Deliv Rev. 47(2001) 165-96. [2] Ugazio E, Cavalli R, Gasco MR. Int J Pharm. 241(2) (2002) 341-4

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Prospects of Microwave Spectrometry for vascular stents monitoring: Towards a noninvasive and non-ionizing follow-up alternative Gianluca ArauzArauz-Garofalo1, Antoni García-Santiago1,2, Flavio David Gerez-Britos1, Ferran Macià1, Javier Tejada1,2, Susana Amorós-García-de-Valdecasas3, Juan Manuel O’Callaghan3, Carolina GálvezMontón4, Carolina Soler-Botija4, Oriol Rodriguez-Leor5,6 and Antoni Bayes-Genis4,5,6 1

Grup de Magnetisme, Departament de Física de la Matèria Condensada, Universitat de Barcelona, Barcelona, Spain 2 Institut de Nanociència i Nanotecnologia IN2UB, Universitat de Barcelona, Barcelona, Spain 3 Department of Signal Theory and Communications, Universitat Politècnica de Catalunya, Barcelona, Spain 4 ICREC Research Program, Fundació Institut d'Investigació en Ciències de la Salut Germans Trias i Pujol, Barcelona, Spain 5 Servei de Cardiologia. Hospital Universitari Germans Trias i Pujol, Barcelona, Spain 6 Departament de Medicina, Universitat Autònoma de Barcelona, Barcelona, Spain

Gian.Arauz@gmail.com Stent failure is an open issue in interventional cardiology. Some conditions are well known to be directly associated with stent failure, including in-stent restenosis, stent fracture, and in-stent neoatherosclerosis. Identification of these high-risk conditions currently requires invasive, complex procedures, such as coronary angiography, intravascular ultrasound, or optical coherence tomography. These invasive techniques are expensive, and they require hospital admission and trained physicians. Our study explores the value of microwave spectrometry (MWS), a new non-invasive and non-ionizing stent monitoring approach for the assessment of stent structural integrity [1,2] and changes in stent environment [3]. A future implementation of this non-invasive and nonionizing methodology will not imply patient sedation, hospitalization or specialized medical personnel. The achievement of this landmark would improve the secondary prevention of stented patients as well as the long-term outcome of stenting. Further studies in experimental models, like coronary stent monitoring in large animals, are required to confirm our encouraging results before undertaking clinical translation of MWS.

References [1] Arauz-Garofalo, Gianluca et al., Medical Physics, 41 (2014) 041902. [2] Arauz-Garofalo, Gianluca et al., Conference: 2014 IEEE MTT-S International Microwave Symposium (IMS2014), 41 (2014) 1-4. [3] Arauz-Garofalo, Gianluca et al., Biomedical Physics & Engineering Express, 1 (2015) 035202. [4] Rodriguez-Leor, Oriol et al., Eurointervention, (in press).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Highest Quality Graphene for bio-sensing A. Ballestar1,2, A. GarcĂa-GarcĂa1,3, L. Serrano1,2, J. M. de Teresa2,4, P. Godignon3, M. R. Ibarra2 1

GPNT - Graphene Nanotech, S.L., Miguel Villanueva 3, 26001 LogroĂąo, Spain INA, LMA, Universidad de Zaragoza, Mariano Esquillor, 50018 Zaragoza, Spain 3 CNM-IMB-CSIC, Campus UAB, Bellaterra, 08193 Barcelona, Spain 4 ICMA, Universidad de Zaragoza, 50009 Zaragoza, Spain

ana@gpnt.es It seems that we are entering the era of two dimensional materials, that ever since they have been isolated showed amazing potential for developing new devices based on their extraordinary properties. Due to the combination of its properties (flexibility, transparence, strength, thermal and electric conductivity, biocompatibility, etc.), graphene appears as the best positioned among them, particularly in the bio-sensing area, to lead a new generation of high performance devices. When it comes to industrialization and commercialization, two main issues have to be faced: utilization of reproducible and affordable standardized manufacturing processes and high quality of the final product. For that purpose, we fabricate graphene by selective sublimation of Silicon Carbide (SiC), using an optimized protocol developed in our company. Our typical graphene layers grown on Si face SiC present mobilities in the range of 3000-4000 cm2V-1s-1 at RT and up to almost 40000 at low temperature. Furthermore, pursuing the inclusion of graphene in the field of electronics, we produced a novel graphene product grown on top of pre-processed SiC substrates with a buried conducting layer at a depth of 300 nm, formed by ion implantation. By these means, we can provide a substrate where graphene is fully exposed to the relevant environment and can be easily back gated by biasing the semiconductor buried layer, which sets the basis of a Graphene-Field-Effect-Transistor (GFET). This device can be directly used in highly sensitive and robust biological sensors that will improve healthcare thorough detection of, e.g.

DNA hybridization [1], cancer markers [2] or immunoglobulin G [3] and E [4]. Our quality inspections, in terms of Raman investigations, revealed that we obtained SiC substrates covered with more than 75% of graphene of superior quality. Moreover, measurements performed on Hall bar devices fabricated along the SiC terraces corroborated those results. That is: mobilities of the electrons as high as 13000 cm2V-1s-1 at low temperature and 2300 cm2V-1s-1 at RT, as well as, low magnetic field plateaux on the Quantum Hall Effect have been observed. Even if these performances are pretty good in this first generation material, the pre-processing of the semiconductor substrate is still impacting the graphene performances and we are working to improve the SiC processing to reach the electrical values obtained on graphene grown on virgin SiC substrates. All those premises make our graphene an ideal candidate to overcome the difficulties in bringing its potential to the desired industrial scale.

References [1] Dong, X., et al. (2010). Adv Mater 22(14): 1649. [2] Tehrani, Z., et al. (2014. 2D Materials 1(2): 025004. [3] Mao, S., et al. (2010). Adv Mater 22(32): 3521 [4] Ohno, Y., et al. (2010). J Am Chem Soc 132(51): 18012..

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures Figure 1. Raman spectra measured at the position indicated by the blue crossin the optical image at the inset. The lower inset figure consists on a maping of the FWHM of the 2D peak, in which only the yellow dots indicate positions where single layer graphene is not found, while all the rest corresponds to the presence of high quality monolayer graphene, as it can be inferred from the color scale to the right.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Antioxidant stabilization through Bicosomes Rafael Bernad, Bernad Lucyanna Barbosa-Barros, Gelen Rodríguez, Mercedes Cócera, Rosana Saldaña, Olga López Bicosome S.L. c/ Jordi Girona 18, 08034 Barcelona, Spain

Rafael.Bernad@bicosome.com There is an increasing health concern in aging societies which are in demand of healthier products with antiaging properties. Antioxidants are great candidates for satisfying this demand as they are molecules able to inhibit the oxidation of other molecules which is the main mechanism behind aging and health loss. Antioxidants are commonly used in many products including dermatological. The limitation of antioxidants resides on their poor stability. If not formulated in adequate systems, these molecules are quickly degraded. Bicosome technological platform was firstly developed to provide products able to penetrate gently the skin, remain there, and deliver their content in the targeted layers, overcoming one of the biggest challenges for formulators of skin care products: skin penetration. We have studied whether Bicosome double encapsulation systems are capable of providing protection to antioxidants from degradation. Our results indicate that Bicosomes can prevent antioxidant from premature degradation and therefore can improve the stability of the products containing them.

References [1] A.N. Ellepola, G.J. Panagoda, L.P. Samaranayake, Microbiol. Immunol., 1999, 14:358-363. [2] C. Surber, P. Elsner, M. Farage, Curr. Probl. Dermatol., 2011, 40. [3] C.S. Boon, et al. Crit. Rev. Food Sci. Nutr., 2010, 50, 215-532. [4] E. Fernández, et al., RCS Advances, 2014, 4, 53109-53121. [5] Elias, P.M. J. Control. Release, 1991, 15: 199-208. [6] G. Rodríguez, G. Soria, E. Coll, L. Rubio, L. Barbosa-Barros, C. López-Iglesias, A. M. Planas, J. Estelrich, A. de la Maza, O. López, Biophys. J., 2010, 99,480. [7] G. Rodríguez, L. Barbosa-Barros, L. Rubio, M. Cócera, C. López-Iglesias, A. de la Maza, O. López, Colloids and Surfaces B: Biointerfaces, 2011, 84, 390. [8] J. W. Wiechers, Science and Applications of Skin Delivery Systems, Allured Publishing Corporation, 2008. [9] L. Barbosa-Barros, G. Rodríguez, C. Barba, M. Cócera, L. Rubio, J. Estelrich, C. López-Iglesias, A. de la Maza, O. López, Small, 2012, 8, No 6, 807. [10] L. Rubio, G. Rodríguez, L. Barbosa-Barros, C. Alonso, M. Cócera, A. de la Maza, J.L. Parra, O. López, Colloids and Surfaces B: Biointerfaces, 2012, 92, 322. [11] M. Cócera, et al. Soft Matter, 2011, 7, 8605-8611. [12] M.E. Darwin, et al., J Dermatol. Sci., 2011, 64, 5358. [13] A. Holzer, M. Athar, C. Elmets, J. Invest. Dermatol., 2010, 130(6), 1496-1499. [14] C. Calles et al., J. Invest. Dermatol., 2010, 130: 1424-36. [15] T. Herrling, et al., Spectrochim Acta A Mol Biomol Spectrosc., 2006, 63(4), 840-845.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Towards graphene for biomedical applications: evaluation of neuronal biocompatibility Mattia Bramini, Bramini Fabrizia Cesca, Fabio Benfenati Center for Synaptic Neuroscience and Technologies & Graphene Labs, Istituto Italiano di Tecnologia, Genova, Italy

mattia.bramini@iit.it Graphene has the potential to make a very significant impact on society, with new interesting and beneficial applications for both individuals and industrial organizations [1]. However, this new material may also pose risks to the environment and raise health and safety concerns. On the other side, the ability of nanomaterials to interact with and enter into cells affecting their biochemical functions makes them important tools at the molecular level. The emerging interest toward applying this nanomaterial within the central nervous system prompted us to focus our attention on the toxicity and biocompatibility of graphene nanostructures in contact with primary neural cells. In this scenario, we study potential effects of different types of graphene (in particular pristine graphene and graphene oxide) and 2D materials with different types of biological matter. The main goal is to evaluate graphene biocompatibility and accordingly any unwanted effects that graphene materials could potentially induce to living systems. We are currently pursuing two lines of research inside the Graphene Flagship European project, focusing our attention on the biocompatibility and toxicity of graphene-based materials to primary neurons (Figure 1, [2]) and glial cells. We are characterizing (i) the molecular mechanisms of graphene flakes internalization together with the possible inflammatory responses [2], and (ii) the possibility of using graphene-SiC and CVD supports and graphene-inks as biocompatible scaffolds for biomedical applications, trying to exploit the conductive properties of graphene to modulate and control the activity of neural networks grown in strict contact with such

scaffolds (i.e. for artificial retina approach [3]). In a separate set of experiments, we moved to more complex experimental systems to address the issue of graphene related materials and the physiology of blood brain barrier [4]. To achieve a more comprehensive overview of the interaction of graphene related materials with this barrier we investigated both the biology of blood-brain barrier when in contact with graphene-particles and blood-brain barrier penetration of graphene nanosheets by reconstitution in vitro of the tripartite endothelium/glial/neuronal interface. In conclusion, our results show that although graphene flake exposure does not impact cell viability and network formation, it does nevertheless have important effects on neuronal transmission and network functionality, thus warranting caution when planning to employ this material for neurobiological applications. It is also important to point out that planar graphene (not nanosheet suspension) could, on the other hand, be a promising tool for medical implants, making â&#x20AC;&#x153;shape and sizeâ&#x20AC;? key parameters for biological application of graphene.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References

Figures

[1] Zhang et al., J. Mater. Chem. B, 1, (2013), 2542-2567 [2] Bramini et al., ACS Nano, 10 (7), (2016), 7154-7171 [3] Ghezzi et al., Nat. Photon., 7, (2013), 400-406 [4] Bramini et al., ACS Nano, 8 (5), (2014), 43044312

Figure 1. Effects of graphene oxide nanosheet exposure on excitatory and inhibitory synapsis of primary cortical neuronal cells.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Voltammetric sensor based on Molecularly Imprinted Polymers for 4-Ethylphenol detection Inmaculada Campos, Campos Anna Herrera-ChacĂłn, Ferdia Bates and Manel del Valle Sensors and Biosensors Group, Department of Chemistry, Universitat AutĂ˛noma de Barcelona, Barcelona, Spain

inmaculada.campos@uab.cat 4-Ethylphenol (4-EP) is a volatile phenolic compound, principal defect in wine contaminated by proliferation of Brettanomyces yeast during fermentation. Presence of 4-EP in early stages of the fermentation is difficult to detect using on-site analysis. Molecularly imprinted polymers (MIPs) are plastic materials able to recognize biological and chemical species, which are synthesized using host-guest principles. In these, chemicals that one wants to bind are firstly used as templates during the polymerization to generate molecular motifs, i.e. tailored made cavities allowing for the selective capture of the desired molecules. The present work reports a MIP material able to capture 4-EP and readily to be used in an electrochemical sensor in a near future. Synthesis of MIPs was done by co-precipitation. Template removal was performed with a Soxhlet using MeOH. Control non-imprinted polymers (NIPs) were also synthesized for comparison purpose. A miniature home-made voltammetric cell was used for the electrochemical measurements using the Differential Pulse Voltammetry technique (DPV). Series of thermodynamic equilibrations of the synthesized polymers were performed to obtain the ligand binding characteristics (Scatchard plots) [Fig.1].

with 100 mg of MIP materials (or NIP, for control purpose). Before measuring the MeOH:ACN fraction 10 mL of phosphate buffer were added to ensure the conductivity of the media. In order to ensure reproducible signals, an electrochemical cleaning stage was carried out between measurements. A1/A2 figures show how the MIP and NIP have a similar structure, as observed by SEM microscopy; equivalent size particle distributions are represented in the histograms (Figures B1/B2) not displaying any differences between them. Also, equivalent MIP and NIP behaviour can be observed in the Scatchard Plot used to characterise the polymeric materials (Figures C1/C2). The final sensor assembly consisted on a solgel deposition on a Graphite Epoxy Composite (GEC). Afterwards a calibration curve was obtained showing promising results. The presented work reports a polymeric material able to capture 4-EP and readily to be implemented as recognition element for an electrochemical sensor. All the studies to characterise this material give us valuable information but at this point the preconcentration column shows contradictory probably because the NIPs shows a faster kinetics than the MIPs one.

Preconcentration columns were constructed using disposable plastic syringe bodies, packed

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Bates, F., & del Valle, M. (2015). Voltammetric sensor for theophylline using solâ&#x20AC;&#x201C;gel immobilized molecularly imprinted polymer particles. Microchimica Acta, 182(5-6), 933-942. [2] Matsui, J., Miyoshi, Y., Doblhoff-Dier, O., & Takeuchi, T. (1995). A molecularly imprinted synthetic polymer receptor selective for atrazine. Analytical Chemistry, 67(23), 44044408.

Figures

Figure 1. (A1/A2) Scanning electron microscopy of the obtained MIP (top) and NIP (down). (B1/B2) Histograms of the MIP (top) and NIP (down). (C1/C2) Scatchard plot for MIP (top) and NIP (down).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Ultrasensitive Scanning Laser Analyser for Biomedical Applications Virginia Cebriรกn Mecwins S.A., Plaza de la Encina 10-11, Tres Cantos (Madrid), Spain

vcebrian@mecwins.com Mecwins was founded in 2008 by Dr. Javier Tamayo and Dr. Montserrat Calleja, Bionanomechanics group leaders from the Institute of Microelectronics of Madrid (IMMCSIC) [1]. Since then, we have been developing cutting edge technology for nanomechanical sensing. This technology, based on detecting variations in the resonance frequency of microcantilevers and plasmon emission was the groundwork for the technical improvements that led us to our new ultrasensitive detection device, ScalaBio [2]. ScalaBio combines the previous mechanical detection technique (microcantilevers resonance frequency analysis) with a new optical detection technique that increases current levels of sensitivity in clinical practice a million times [3], while requiring for the test a small biological sample and less time than techniques currently used in hospitals and central laboratories, without increasing current cost per sample. The molecular recognition on the surface of a biofunctionalized cantilever results in a nanomechanical response, that produces cantilever bending of a few nanometers (static mode) or changes in cantilever resonance frequency (dynamic mode) where the added mass due to selective molecular recognition decreases resonance frequency value. The microcantilevers also exhibit enough structural quality to be used as optical resonators. Biomolecular interactions on the cantilever surface can modify its optical spectra. It allows for the first time easy and fast detection of protein biomarkers at

concentrations as low as 10-7 ng/ml, which is a million times lower than the achieved in current clinical practice (0.1 ng/ml) [3]. In the present study, ScalaBio has been used for the successful detection of different protein biomarkers in clinical diagnosis, focusing our preliminary experiments on oncology (PSA, CEA), cardiac (Troponin I) and infectious diseases biomarkers (p24). The adoption of ultrasensitive detection equipment will enable screening for early detection of a wide range of diseases with established diagnostic biomarkers from a droplet of blood.

References [1] Spanish Research Council: http://www.imm-cnm.csic.es/bionano/es [2] http://mecwins.com/ [3] Kosaka et al. Nat Nanotechnol. 9(12),104753 (2014)

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Lab-on-a-chip for purification and automated counting of Circulating Tumor Cells from peripheral blood of metastatic cancer patients A. Chícharo 1,2, M. Martins1, S. Cardoso3, B. Espina1, L. Diéguez1, P.P. Freitas1,2 1

International Iberian Nanotechnology Laboratory, Braga, Portugal 2 Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal 3 INESC-MN, Lisbon, Portugal

alexandre.chicharo@inl.int Novel systems for fast and specific quantification of rare cells have gained extraordinary interest in a wide spread of applications, being a basic tool for biomedical research, as well as for its integration in pointof-care diagnostic devices [1]. Such systems can be relevant for the quantification of circulating tumor cells (CTCs), present in peripheral blood of metastatic cancer patients and suspected to play a crucial role in cancer progression. Even though CTCs can be used as a metastatic biomarker for cancer diagnosis, prognosis, and therapy monitoring; current methodologies for CTC analysis lack sensitivity and specificity. Towards this goal, we present a low-cost, compact high-throughput platform for cell purification from complex body fluids and single rare cell enumeration. Our cytometer biochip is integrated by two separate modules: i) microfluidic size filter for sample processing and purification of rare cells, and ii) miniaturized cytometer for automatic detection of labelled cells [2,3] (Fig.1a,b). A colorectal adenocarcinoma cell line (SW480) was chosen as a CTC model, since it presents a morphology similar to CTCs and expresses EpCAM receptors in the cell membranes. 50 SW480 cells were spiked in i) whole blood from healthy donors, or ii) isolated buffy coat from

healthy donor, to evaluate the CTC isolation and quantification yield. The samples were incubated with 1µm superparamagnetic beads functionalized with anti-EpCAM antibodies (Fig.1c) and processed in the microfluidic cell size filter to purify and enrich the cancer cells. . The number of cells trapped in, and recovered from the filter was counted in order to calculate a 71%the efficiency of the device.

References [1] Costa C, Abal M, López-López R, MuineloRomay L., Sensors (Basel). MDPI, Biosensors for the Detection of Circulating Tumour Cells, 14 (2014) 4856–75. [2] Vila A, Martins VC, Chicharo A, et al., IEEE Transactions on Magnetics, Customized Design of Magnetic Beads for Dynamic Magnetoresistive Cytometry, 50 (2014), 1-4. [3] Loureiro J, Andrade PZ, Cardoso S, da Silva CL, Cabral JM, Freitas PP., Lab on a Chip, Magnetoresistive chip cytometer, 11 (2011) 2255-61.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. a) integrated microfluidic cytometry chip. b) the sample (red) is hydrodynamically focused (blue) over the magnetoresistive sensors. c) a SW480 cancer cell is labelled with 1Âľm magnetic beads.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Study of Shape-dependent Nanoparticle Interactions Gavin Coleman, Coleman Ester Polo, Luca Boselli, Kenneth A. Dawson Centre for BioNano Interactions, School of Chemistry, University College Dublin, Belfield, Dublin 4, Ireland

gavin.coleman@cbni.ucd.ie The interaction of nanoparticles with the biological environment results in the formation of a biomolecular corona, which substantially modifies the nanoparticleâ&#x20AC;&#x2122;s pristine surface properties [1]. This complex multilayer of biomolecules ultimately mediates the interactions of nanoparticles with cells and organisms, but the exact mechanisms that govern these interactions are still not completely understood. Certain sequences of these (bio-) molecules when presented on the surface are implicated in specific cellular responses (for example uptake by a specific receptor pathway), cell signaling or other biological processes [2, 3]. The organization and arrangement of these molecules on the surface depends on nanoparticle features such as shape, size and surface chemistry [4, 5]. The composition and architecture of biomolecules on the surface in turn affect the bio-interactions and thus in vivo destination, often leading to undesirable or even potentially toxic results, such as rapid clearance by the liver [6, 7]. It is therefore of incredible importance and interest to deeply understand this bionanointerface, and the effects it may have on particle destination, from both a developmental and regulatory point of view. Characterizing in molecular detail the information encoded at the surface of these nano-structures is crucial to understand the link between the presentation of certain endogenous molecules and their biological response. However, meaningful measurements of complex interfaces are nontrivial, requiring advancement of current approaches.

This work aims to investigate the interplay between the shape and the biological identity of the nanostructures. Understanding this interplay will provide an avenue of control for desirable nanoparticle-cell interactions, through control of shape and functionalisation. The development of platforms for screening the exposed motifs of biomolecules on the surface nanoparticles in realistic scenarios will lead to the ability to identify and quantify relevant motifs available to engage with specific cellular receptors. To this end, gold nanoparticles of varying shapes are synthesised, functionalised with different ligands and/or proteins of interest, and thoroughly characterised. The biological activity of the particles is then examined in simple and complex media, through development of label-free highthroughput quantitative interaction assays on an Attana Quartz Crystal Microbalance (QCM).

References [1] T. Cedervall, et al., Proceedings of the National Academy of Sciences. 104 (2007) p. 2050 [2] A. Salvati, et al., Nat Nano. 8 (2013) p. 137 [3] P. M. Kelly, et al., Nat Nano. 10 (2015) p. 472 [4] C. D. Walkey, et al., ACS Nano. 8 (2014) p. 2439 [5] N. P. Truong, et al., Expert Opinion on Drug Delivery. 12 (2015) p. 129 [6] M. P. Monopoli, et al., Nat Nano. 7 (2012) p. 779 [7] S. Tenzer, et al., Nat Nano. 8 (2013) p. 772

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nanotheranostics to interfere the process of metastasis Alonso-Nocelo M, Vazquez-RĂos AJ, Reimondez-TroitiĂąo S, Bouzo BL, Nagachinta S, Gutierrez C, Lopez-Lopez R, de la Fuente M. M Nano-oncologicals Lab, Translational Medical Oncology Group, Health Research Institute of Santiago de Compostela (IDIS), SERGAS, Santiago de Compostela, Spain

maria.de.la.fuente.freire@sergas.es / maria.fuente.freire@gmail.com Metastases are the major cause of death in cancer patients with solid tumors mainly because of the ineffectiveness of current therapies once tumor cells spread and metastases are formed. Disseminated cells, i.e. circulating tumor cells and occult micrometastases, the intermediate pathogenic link between the primary tumor and macroscopic metastases in all solid cancers, are present in a relevant number of patients without any clinical or even histopathological sign, meaning that the true prevalence of metastatic disease is therefore unknown [1,2]. The development of a rapid, non-invasive detection tool that can give accurate wholebody readouts of occult micrometastases might therefore have a substantial clinical impact in the management of disseminated disease before the burden of metastases. Additionally, there is a pressing need in oncology for the development of novel therapeutic interventions addressed to the treatment of low-level disseminated malignant disease. Nanotheranostics composed by biodegradable materials, have the potential to increase the sensibility of current diagnosis techniques and selectively deliver new therapeutic entities to disseminated cancer cells. We aim to develop novel nanotheranostics to confront different stages of the metastatic process. In this regards, we have engineered lipidic nanoestructures by using simple and scalable methodologies. Additionally, we have

decorated these nanoestructures with peptides and proteins targeted against specific biomarkers, which we have previously identified in disseminated cancer cells, more in detail in Circulating Tumor Cells (CTCs). We have successfully encapsulated novel therapeutic entities, such as microRNAs, as well as conventional anticancer drugs, and optimized the release conditions to cancer cells. We have also developed in vitro 3D models that mimic the tumor microenvironment, of utility to explore the potential of our nanoestructures, and are also using zebrafish models for that purpose. Current experiments are aimed to associate imaging elements, which will allow tracking of our nanoestructures after intravenous injection, such as magnetic nanoparticles and 18F.

References [1] Bali M et al. Micrometastasis: detection methods and clinical importance. Cancer Biomark: 9 (1-6): 397-419, 2010. [2] Chaffer CL & Weinberg RA. A perspective on cancer cell metastasis. Science 331: 15591564, 201 [3] Alonso-Nocelo M et al. Development and characterization of a three-dimensional coculture model of tumor T cell infiltration. Biofabrication 8 (2): 025002.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. Our nanosystems have a mean size around 100 nm, as measured by Dynamic Light Scattering and Transmission Electron Microscopy (A). Nanoparticles functionalized with monoclonal antibodies against a cell membrane receptor (red) were efficiently and selectively taken-up by metastatic lung cancer cells (B). 3D cell-culture models that include tumor and immune cells were used to explore the selective interaction of the nanosystems with tumor cells under dynamic flow conditions (C). Fluorescent nanosystems (red) accumulate in tumor xenografts (green) in zebrafish embryos (D).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Immune characterization of graphene oxide and amino functionalized graphene using new high-throughput analysis Marco Orecchioni1, Davide Bedognetti2, Wouter Hendrickx 2, Claudia Fuoco3, Filomena Spada3, Francesco Sgarrella1, Gianni Cesareni3, Francesco M Marincola2, Alberto Bianco3, Kostas Kostarelos4 and Lucia Gemma Delogu1 1

University of Sassari, 07100 Sassari, Italy Research Branch, Sidra Medical and Research Center, Doha, Qatar CNRS, Institute de Biologie MolĂŠculaire et Cellulaire 67000 Strasbourg, France 3 University of Roma Tor Vergata, 00133, Roma, Italy 4 Faculty of Medical & Human Sciences, University of Manchester Manchester M13 9PT, UK

lgdelogu@uniss.it There is an enormous interest in exploring the potentialities of novel nanomaterials such as graphene and its derivates in biomedical applications [1]. The understanding of the biomolecular interactions of graphene nanomaterials with human cells is critical for their implementation as diagnostic or therapeutic agents. In this context understand the immune impact mediated by GO or nanomaterials in general is mandatory for any biological or medical application [2]. Here, we propose an integrative analytical pipeline encompassing molecular and cellular characterization of the impact of graphene nanomaterials on immune cells. For the first time in the context of nanotechnology, we employed single cell mass cytometry to deconvolute the effect of GO flakes (between 12 graphene layers) functionalizated by us through the addition of 1,3 dypolar cycloaddiction of amino groups (GONH2) on 15 immune cell populations looking at 30 markers at single cell level. We then used whole transcriptomic analysis for further functional molecular characterization. We showed that functionalization is able to increase the biocompatibility of GO in all the sub-population analyzed (Fig 1). Intriguingly, GONH2 was found to be a potent immune activator of monocytes and dendritic cells. This effect was confirmed

and explained by the transcriptomic analysis. Notably, GO mainly modulated cell metabolism. The functionalized GONH2 instead did not have an impact on the cell metabolism. Rather, they induce, in both monocytes and T cells, the activation of immunomodulatory pathways mainly related with innate and adaptive system and centered on interferon (IFN) signaling (Fig 2). Immune activations resulted in increased expression of the T helper 1 chemokines (i.e., CXCR3 and CCR5 ligands), which are critical for the development of an effective anti-tumor immune response [3]. These idiosyncratic immune modulatory proprieties of GONH2 represents new interesting data to develop new nanoscale platforms in medicine as novel immunemodulator tools.

References [1] Sechi G, Bedognetti D, Sgarrella F, Van Eperen L, Marincola FM, Bianco A, Delogu LG,. Nanomed. (Lond) 9 (2014)1475-86. [2] Goldberg MS. Cell 2 (2015) 201-4. [3] Galon J, Angell HK, Bedognetti D, Marincola FM. Immunity 1 (2016) 11-2

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Interaction of Nanoparticles with Cells: Nanomaterials For Stem Cell Tracking Applications Neus Feliu1,2, Wolfgang Parak2 1

Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Stockholm, Sweden 2 Fachbereich Physik, Philipps Universität Marburg, Marburg, Germany

Neus.feliu@ki.se Nanomaterials offer promising opportunities for a wide range of applications including medicine. Their novel properties make them excellent imaging and diagnostic agents. However, nanoparticles could be potentially harmful for humans and environment. To understand the possible effects of nanoparticles exposed to biological systems, a detail association of such effects to the physicochemical properties of the nanoparticles is needed. Therefore, we will provide an overview of the common denominators to evaluate the nanosafety research, and discuss the potential use of an advanced system based on nanoparticle-based imaging labels to monitor stem cells non-invasively in vivo. A deeper understanding in these areas will help to improve the development of nanomaterials for nanomedicine research, including regenerative medicine.

References [1] N. Feliu, D. Docter, M. Heine, P. del Pino, S. Ashraf, J. Kolosnjaj-Tabi, P. Macchiarini, P. Nielsen, D. Alloyeau, F. Gazeau, R. H. Stauber, W. J. Parak, “In vivo degeneration and the fate of inorganic nanoparticles”, Chemical Society Reviews 45, 2440-2457 (2016). [2] N. Feliu, J. Hühn, M. Zyuzin, S. Ashraf, D. Valdeperez, A. Masood, A. H. Said, A. Escudero, B. Pelaz, E. Gonzalez, M. A. Correa Duarte, S. Roy, I. Chakraborty, M. L. Lim, S. Sjöqvist, P. Jungebluth, W. J. Parak, “Quantitative uptake of colloidal particles by cell cultures”, Science of the Total Environment, 2016 568, 819-828 (2016).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Contribution of Water to Protein Stability and Strategies for Protein Design Giancarlo Franzese SecciĂ˛ de Fisica Estadistica i Interdisciplinariaâ&#x20AC;&#x201D;Departament de Fisica de la Materia Condensada, Facultat de Fisica, & Institut de Nanociencia i Nanotecnologia (IN2UB), Universitat de Barcelona, Barcelona, Spain

gfranzese@ub.edu The mechanisms of cold and pressure denaturation of proteins are matter of debate and are commonly understood as due to watermediated interactions. Here, we study several cases of proteins, with or without a unique native state, with or without hydrophilic residues, by means of a coarse-grain protein model in explicit solvent. We show, using Monte Carlo simulations, that taking into account how water at the protein interface changes its hydrogen bond properties and its density fluctuations [1-5] is enough to predict protein stability regions with elliptic shapes in the temperature-pressure plane, consistent with previous theories. Our results [6] clearly identify the different mechanisms with which water participates to denaturation and allow us to develop an advanced computational design protocol for protein engineering. In particular, we apply our design analysis to understand why proteins that are functional at ambient conditions do not necessarily work at extreme conditions of temperature T and pressure P, and why there are limits of T and P above which no protein has a stable functional state. We show that the hydropathy profile of proteins is a consequence of evolutionary pressure exerted by water and can lead the way for engineering working proteins and drugs at extreme conditions.

References [1] E. G. Strekalova, M. G. Mazza, H. E. Stanley, and G. Franzese, Large decrease of fluctuations for supercooled water in hydrophobic nanoconfinement, Physical Review Letters 106, 145701 (2011). [2] E. G. Strekalova, J. Luo, H. E. Stanley, G. Franzese, S. V. Buldyrev, Confinement of Anomalous Liquids in Nanoporous Matrices, Physical Review Letters 109, 105701 (2012). [3] G. Franzese, V. Bianco, and S. Iskrov, Water at interface with proteins, Food Biophysics, 6, 186 (2011). [4] G. Franzese, and V. Bianco Water at Biological and Inorganic Interfaces Food Biophysics, 8, 153 (2013). [5] V. Bianco and G. Franzese, Critical behavior of a water monolayer under hydrophobic confinement, Scientific Reports (Nature Publishing Group) 4, 4440 (2014). [6] V. Bianco and G. Franzese, Contribution of Water to Pressure and Cold Denaturation of Proteins, Physical Review Letters 115, 108101 (2015).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Magnetic Solid Lipid Nanocomposites as ultrahigh MRI contrast enhancers and magnetic hyperthermia induced drug delivery vehicles Juan Gallo1, Carolina L. Moura1,2, Nágila M. P. S. Ricardo2 and Manuel Bañobre-López1 1

Advanced (magnetic) theranostic nanostructures Group, International Iberian Nanotechnology Laboratory, Braga, Portugal 2 Department of Organic and Inorganic Chemistry, Federal University of Ceará, CX 12200 Fortaleza, Brazil

juan.gallo@inl.int Magnetic hybrid nanocomposites have opened new perspectives in biomedical and environmental applications [1]. Solid lipid nanoparticles (SLNs) have attracted great interest within the scientific community as bioactive encapsulating agents and carriers due to their biocompatibility, low toxicity and ability to influence the delivery profile of pharmacological agents [2,3]. Hybrid organicinorganic SLNs are being explored to synergistically combine the modified bioactive release provided by the lipidic encapsulation and the intrinsic physico-chemical properties from the inorganic counterpart [4]. In this context, we present the preparation of drug loaded magnetic solid lipid nanocomposites (MSLNs) showing good multifunctional performance as ultra-high T2-contrast agents and heat generating sources in magnetic resonance imaging (MRI) and magnetic hyperthermia (MH), respectively. A method based on oil-in-water emulsion was employed to prepare MSLNs from natural carnauba wax containing different concentration of ~10 nm superparamagnetic oleic acid coated magnetite (Fe3O4@OA) nanoparticles. Successful incorporation of the magnetic nanoparticles within the spherical nanocomposites was confirmed by transmission electron microscopy (TEM). Dynamic light scattering (DLS) and ζ-potential measurements showed a relatively narrow size distribution with an average particle size of

~180 nm and a surface charge around -60 mV. In terms of magnetic characterization, hysteresis loops showed MSLNs to behave as superparamagnetic particles. Interestingly, MSLNs showed an anomalous ultra-high transversal relaxivity (r2) with values higher than 900 mM(Fe)-1s-1, what clearly translated into dark contrast effects when sample phantoms were imaged by MRI at 3T. Simultaneously, the MSLNs were tested as vehicles for the delivery of an anticancer drug and its release profile was assessed under the application of MH. Results show that the delivery profile can be modified through MH protocols. In vitro results will also be shown and discussed.

References [1] C. Sanson, O. Diou, J. Thévenot, E. Ibarboure, A. Soum, A. Brûlet, S. Miraux, E. Thiaudière, S. Tan, A. Brisson, V. Dupuis, O. Sandre and S. Lecommandoux. ACS Nano, 5 (2011), 1122. [2] W. Mehnert and K. Mader., Adv. Drug Delivery Rev. 47 (2001), 165. [3] E. Andreozzi, P. Wang, A. Valenzuela, C. Tu, F. Gorin, M. Dhenain and A. Louie, Bioconjugate Chem., 24 (2013), 1455. [4] K. Oumzil, M. A. Ramin, C. Lorenzato, A. Hemadou, J. Laroche, M. J. Jacobin-Valat, S. Mornet, C.-E. Roy, T. Kauss, K. Gaudin, G. Clofent-Sanchez and P. Barthelemy, Bioconjugate Chem., 27 (2016), 569.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Arrays of titanium dioxide nanotubes with different diameter and surface chemistry influence C2C12 skeletal myoblast adhesion and differentiation Giada G. Genchi1,2, Ioannis Liakos3, Virgilio Mattoli1 and Tejal A. Desai2* 1

Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, Pontedera, Italy University of California, San Francisco, Department of Bioengineering and Therapeutic Sciences, California, USA 3 Istituto Italiano di Tecnologia, Smart Materials, Nanophysics Department, Genoa, Italy

giada.genchi@iit.it, *tejal.desai@ucsf.edu TiO2 nanotubes (NTs) have attracted huge interest in several fields of nanotechnology, recently finding also application in the biomedical research [1]. Here, we propose the use of TiO2 NT arrays for C2C12 skeletal myoblast culture, and the array binding with a celladhesive protein to improve implementation in biological environments. Arrays with different nanotube diameters and surface chemistry were obtained by anodization (4, 20, and 40 V), annealing (400°C, 1 h), and silanization with an alkaline 20% APTES solution (4 h, 80°C). Incubations with a 5 mg/ml genipin solution (1 h, 37°C) and with an acidic 50 µg/ml laminin solution followed. Surface morphology and chemistry were then investigated with SEM and XPS. SEM demonstrated open-top nanotubes and different nanotube diameters (10, 50 and 100 nm) were achieved, and a thin conformal layer of laminin was bound to the surfaces (Figure 1a,b). XPS demonstrated anatase phase was achieved in annealed samples, and silane/protein were bound (the Ti2p peak neatly decreased, whereas the Si2p and N1s peaks appeared) in laminin coated samples.

metabolism. Focal adhesion clustering was improved on coated substrates (mostly on 10 nm diameter NTs), whereas no dead cells were found on all substrates. Cell metabolism decreased on laminin coated arrays. For differentiation studies, myoblast fusion into myotubes and myotube width were assessed, demonstrating that the 10 nm diameter nanotubes promoted the formation of wider myotubes in laminin coated arrays as an evidence of better cell differentiation (Figure 1c,d). Our results thus support the use of TiO2 NT arrays with skeletal muscle cells. Upon suitable modification, these arrays may prove excellent smart interfaces for addressing cell response, and eventually for cell stimulation [2].

References [1] Roy, Berger, Schmucki, Angewandte Chemie, 50 (2011) 2904. [2] Genchi, Nuhn, Liakos, Marino, Marras, Athanassiou, Mattoli, Desai, RSC Advances, 6 (2016) 18502.

For proliferation studies, cell adhesion and viability were investigated in terms of focal adhesion clustering, live/dead cell number and

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. Scanning electron microscopy images of 10 nm diameter nanotube arrays, without (a) and with laminin coating (b). Fluorescence microscopy image of TRITC-phalloidin and DAPI stained C2C12 cells after 3-day differentiation on 10 nm diameter nanotube arrays without (c) and with laminin coating (d). Arrows evidence the higher width of differentiated cells on laminin coated substrates.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nanostructured functionalized surfaces for adoptive cell therapy J. Guasch, Guasch C. Belz, J. Diemer, H. Riahinezhad, J. P. Spatz Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Stuttgart, Germany Molecular Nanoscience and Organic Materials (Nanomol), Institute of Materials Science of Barcelona (ICMAB-CSIC) / CIBER-BBN, Spain

jguasch@icmab.es The worldwide incidence of cancer is expected to reach nearly 25 million cases over the next two decades according to the International Agency for Research on Cancer. Encouragingly, the remission of otherwise terminal leukemia patients thanks to novel adoptive T cell therapies, based on genetically modified autologous T cells, has recently been reported [13]. These therapies would greatly benefit from more powerful cell culture systems to efficiently provide the large amounts of autologous T cells that they require. Quasi-hexagonal arrays of passivated glass surfaces decorated with gold nanoparticles (AuNPs) functionalized with the T cell stimulating antibody antiCD3 have been prepared. They have been demonstrated to be more efficient for T cell activation than uniformly coated antiCD3 Au substrates due to their capacity of presenting biomolecules to cells in a relevant orientation and spacing [4]. Building on these findings, we transferred the ordered nanostructures prepared on glass to poly(ethylene glycol) (PEG) hydrogels, which were cross-linked with small fibronectin-derived peptide ligands. We studied the influence of celladhesive backgrounds on the activation of primary human CD4+ T cells when using arrays of antiCD3 immobilized on AuNPs. These AuNPs were prepared with controlled interparticle distances ranging from 20 to 150 nm. Early activation markers, cytokine secretion, proliferation, and differentiation were analyzed demonstrating the influence of both interparticle distances and integrin-mediated adhesion.

Furthermore, we prepared PEG hydrogels decorated with arrays of Au and TiO2 NPs [5], which can provide control on the nanoscale not only of antiCD3 but also of the secondary costimulatory antibodies needed to provide an effective T cell response. In summary, we will discuss the preparation and functionalization of nanostructured hydrogels for activating primary human CD4+ T cells.

References [1] D. L. Porter, et. al. , Sci. Transl. Med., 7 (2015) 303ra139. [2] M. Kalos, B. L. Levine, D. L. Porter, S. Katz, S. A. Grupp, A. Bagg, C. H. June, Sci. Transl. Med., 3 (2011) 1. [3] D. L. Porter, B. L. Levine, M. Kalos, A. Bagg, C. H. June, N. Eng. J. Med., 365 (2011), 725. [4] J. Matic, J. Deeg, A. Scheffold, I. Goldstein, J. P. Spatz, Nano Lett., 13 (2013), 5090. [5] J. Guasch, J. Diemer, H. Riahinezhad, S. Neubauer, H. Kessler, J. P. Spatz, Chem. Mater., 28 (2016), 1806.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. T cell on a nanostructured hydrogel

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Radioisotope Core-doped Magnetic Nanoprobes; a chelator-free approach for PET/MR molecular imaging with extremely small nanoparticles Fernando Herranz, Herranz Juan Pellico, Irene Fernรกndez-Barahona, Riju Bavesh, Jesus Ruiz-Cabello Centro Nacional de Investigaciones Cardiovasculares (CNIC) and Spanish Lung Diseases Network (CIBERES), Melchor Fernรกndez-Almagro 3, 28029 Madrid, Spain

fherranz@cnic.es The use of nanomaterials for combined PET/MR imaging offers the possibility of combining the best of both topics; the size-dependent properties of nanomaterials and the excellent sensitivity of nuclear imaging. The design of dual-modality PET/MRI nanoscale systems present at least two key questions: which radioisotope and nanocarrier to use, and where the radioisotope will be attached to the nanoparticle [1,2]. Here, we describe the synthesis and PET/MRI performance of two different nano-radiochemicals; 68Ga/89Zr coredoped iron oxide nanoparticles with an extremely small size [3]. The particles are synthesised through a fast microwave-driven process that yields very large specific activities and r1 values [4]. Microwave (MW) technology allows fast and reproducible synthesis of multifunctional NPs and their doping with a variety of radioisotopes in a single synthetic step.

References [1] Judenhofer MS, Wehrl Pichler HF, Pichler BJ, et al. Nature Medicine, 14 (2008) 459. [2] Groult H, Ruiz-Cabello J, Herranz F, et al. Bioconjugate Chem. 26 (2015) 153. [3] Pellico J., Ruiz-Cabello J., Herranz F. et. al. Contrast Media & Mol. Imaging 11 (2016) 203. [4] R. Bhavesh, A. A. V. Lechuga-Vieco, J. RuizCabello, F. Herranz, Nanomaterials 5 (2015) 1880.

We will show how reproducible the approach is, full characterisation of physicochemical and imaging properties and, finally, we will demonstrate the utility of this approach by multimodal imaging in mice and rabbits for a number of cardiovascular applications with unprecedented quality for in vivo hot spot MRI (T1 weighted contrast) and PET imaging.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Synergistic actions of colloidal nanoparticles: Sensing and drug delivery Amelie Heuer-Jungemann1, Afaf H. El-Sagheer2, Peter M. Lackie4, Tom Brown2 and Antonios G. Kanaras1,3 1

Physics and Astronomy, Faculty of Physical Sciences and Engineering, University of Southampton, UK 2 Departement of Chemistry, University of Oxford, Oxford, OX13TA, UK 3 Institute for Life Sciences, University of Southampton, Southampton, UK 4 Faculty of Medicine, University of Southampton, Southampton, UK,

Nanoparticles are of importance in biomedical applications due to their intrinsic properties. The ligand coating of the nanoparticles is critical for colloidal robustness and nanoparticle function and the chemical composition/morphology of the inorganic core is important in defining/tuning the optical, electrical and magnetic properties of the nanoparticulate system. As the chemical synthesis of nanoparticles and nanoparticle functionalization techniques advance during the years, the design of nanoparticulate systems become more sophisticated facilitating multitasking nanoparticle roles.

References [1] Heuer-Jungemman, A.; El-Sagheer, A.H.; Lackie, P. M.; Brown, T.; Kanaras, A. G. "Selective killing of cells triggered by their mRNA signature in the presence of smart nanoparticles" Nanoscale 2016, ASAP DOI: 10.1039/c6nr06154k [2] Heuer-Jungemann, A.; Harimech, P.; Brown, T.; Kanaras A. G. “Gold Nanoparticles and Fluorescently-Labelled DNA as a Platform for Biological Sensing” Nanoscale, 2013,5 (20), 9503–9510 DOI:10.1039/c3nr03707j

In this presentation I will discuss recent progress in our group concerning the design of nanoparticles and their incorporation in biological systems to facilitate sensing, drug delivery and accurate manipulation. My talk will focus on a new class of nanoparticles that can accommodate synergistic actions of sensing and drug delivery in cells. These multitasking particles are able to selectively release their therapeutic payload in response to specific mRNA signatures and selectively kill model cancerous cells as opposed to healthy cells.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Democratizing solid-state nanopores for high throughput bio-sensing Ke Liu, Liu Jiandong Feng, Aleksandra Radenovic Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, EPFL, 1015 Lausanne, Switzerland

ke.liu@epfl.ch Solid state nanopore based bio-sensing is emerging as a rapid single molecule detection and manipulation technique [1]. Conceptually, a single nanometer size aperture located on a membrane can detect electrophoretically driven biomolecules translocation in a high throughput manner, revealing localized information of the analyte. However, the formation of single nanopores relies heavily on expensive instrumentation, i.e., Transmission Electron Microscope (TEM) and well trained TEM users, which renders it still confined to laboratory use. Atomically thin nanopore membranes, graphene and molybdenum disulphide (MoS2) [2, 3], have drawn much attention due to their unprecedented single nucleotide resolution, which holds promise as a candidate for so called 3rd generation DNA sequencers. Dielectric breakdown has been demonstrated as a facile method to make individual nanopores on silicon nitride membranes (10-30 nm thick) without the need for TEM. This can dramatically reduce the cost of device fabrication and scale up the production of devices. However, if the same methodology of nanopore fabrication is applied to the membranes made in 2D materials, we observe that the mechanism that governs the pore formation is not dielectric breakdown but electrochemical reaction that occurs once the transmembrane potential exceeds oxidation voltage [2], which can be readily accessible for commercialization.

References [1] Branton,D, et al., The potential and challenges of nanopore sequencing. Nature Biotechnology, 2008, 26:1146-1153. [2] Feng JD, Liu K, et al. Electrochemical reaction in single layer MoS2: nanopores opened atom by atom. Nano lett. 2015, 15:3431-3438. [3] Feng JD, Liu K, et al. Identification of single nucleotides in MoS2 nanopores. Nature nanotech. 2015, 10:1070-1076.

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Geometry of electrospun nanofibers determine the initial interaction and subsequent osteogenic differentiation of mesenchymal stem cell Salima Nedjari, Nedjari George Altankov IBEC, Institute for Bioengineering of Catalonia, Molecular Dynamics at Cell-Biomaterial Interface, Campus de Bellvitge, Feixa Llarga, Pavello Govern, 1119, 08907 L'Hospitalet de Llobregat, Barcelona, Spain

george.altankov@icrea.cat Stem cells have emerged as very promising solutions for regenerative medicine, disease modeling, developmental biological studies and drug screening. In vivo, stem cells behavior is strongly dependent on their 3D microenvironment called niche - the place where they are constantly challenged by various chemical and physical cues within the extra cellular matrix (ECM). While the effect of chemicals factors on stem cells behaviors is well studied, the impact of physical cues such as the topography required further investigation as they appears to be a key factor in the differentiation. Indeed, it has been suggested that specific patterns can either maintain mesenchymal stem cells in undifferentiated state or drive them towards osteogenic, adipogenic or chondrogenic lineages [1]. In tissue engineering, electrospinning has become a very prospective technique as it allows the elaboration of nanofibers that could mimic the morphology of native ECM fibrils. While nanofibers are generally collected randomly, it is possible to elaborate differently micropatterned scaffolds by controlling the electrostatic forces at the vicinity of the collector [2].

forces during the electrospinning process, we succeed to manufacture three types of architectures: aligned (A), honeycomb (H) and random (R) PLCL/FBG fibers. We further evaluated the biological response of adipose derived mesenchymal stem cells (ADMSC) as a function of these scaffold’s architecture. We demonstrated that ADMSC responded by changing their attachment (focal adhesions), overall morphology and differentiation path toward osteogenic lineage (judged by alkaline phosphatase production, calcification and specific genes expression). Significantly favorable differentiation was observed on honeycomb architectured nanofibers.

References [1] Higuchi A. et al, Chemical Review (2013) 113 3297–3328. [2] Nedjari S. et al, Macromolecular Biosciences (2014) 14 1580– 1589.

In this work, novel type of architectured electrospun scaffolds were elaborated based on poly (L-lactide ε-caprolactone) (PLCL), a biocompatible copolymer and fibrinogen (FBG), an ECM protein. By playing with the electrostatic

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. Overall morphology of ADMSC adhering on (A) random, (B) aligned and (C) honeycomb architectured nanofibers viewed for their actin cytoskeleton (green) at 24 hours of incubation

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

A journey through the secret life of nanoparticles: from 2D high resolution images to 3D oxidation state mapping Pau Torruella1, Raúl Arenal2, Zineb Saghi3, Lluís Yedra1, Alberto Eljarrat1, Josep Rebled1,4, Francisco de la Peña3, Marta Estrader5, Germán Salazar-Alvarez6, Alberto López-Ortega7, Josep Nogués8,9, Paul A. Midgley3, Sònia Estradé1, Francesca Peiró1 1

LENS-MIND, Institute of Nanoscience and Nanotechnology, University of Barcelona, Barcelona, Spain Laboratorio de Microscopia Avanzada, Instituto de Nanociencias de Aragón and Fundacion ARAID, Zaragoza, Spain 3 Department of Materials Science and Metallurgy, University of Cambridge, Cambridge, United Kingdom 4 Centres Científics i Tècnics, Universitat de Barcelona, Barcelona, Spain 5 Laboratoire de Physique et Chimie des Nano-Objets (LPCNO), INSA, UPS, CNRS, Toulouse, France 6 Department of Materials and Environmental Chemistry, Stockholm University, Stockholm, Sweden 7 Dipartamento di Chimica "U. Schiff", Università degli Studi di Firenze, FIrenze, Italy 8 ICREA, Barcelona, Spain 9 Institut Català de Nanociencia i Nanotecnologia (ICN2), Bellaterra, Barcelona, Spain

francesca.peiro@ub.edu Electron tomography has been used in the field of materials science for recovering the three dimensional (3D) structure of materials in the nanoscale. To add chemical information to this 3D volume, Energy Filtering TEM (EFTEM) is the widely spread technique. However, the selection of specific energy channels implies that significant information in the spectrum is lost. Instead, electron energy-loss spectroscopy (EELS) spectrum-images (SI) acquired in tomographic series retain all chemical information for 3D chemical reconstruction [1]. In particular, the near edge energy losses (ELNES) contain information relative to the elemental coordination and thus, recovering of the oxidation state of a transition metal is also possible. In this work we will develop the concept of EELS spectrum volume (EELS-SV), a full EELS dataset in 4D, where every voxel of a whole volume contains a complete spectrum of energy losses. Our approach to EELS-SV reconstruction is based on the acquisition of spectrum images (SI) in tomographic mode SI, thus taking a single SI for every tilt angle and on the identification of independent components and the corresponding abundance maps (ICA

analysis). We will prove the feasibility of EELS tomography from data acquired aberration corrected TEM’s but also in uncorrected instruments thanks to the use of multivariate analysis (MVA), and blind source separation (BSS) methods combined with advanced reconstruction algorithms [2]. We have applied this approach to obtain 3D reconstruction of advanced metal oxide nanostructured materials as FexCo(3-x)O4/Co3O4 mesoporous structures and bimagnetic FeOx/FeOy core/shell nanoparticles, for which the oxidation state of Fe can be discriminated in 3D.

References [1] Yedra, L.; Eljarrat, A.; Arenal, R.; Pellicer, E.; Cabo, M.; López-Ortega, A.; Estrader, M.; Sort, J.; Baró, M. D.; Estradé, S.; Peiró, F. Ultramicroscopy 2012, 122, 12–18. [2] Torruella, P.; Arenal, R.; de la Peña, F.; Saghi, Z.; Yedra, L.; Eljarrat, A.; López-Conesa, L.; Estrader, M,: López-Ortega, A.; SalazarAlvarez, G.; Nogués, J.; Ducati, C.; Midgley, P.; Peiró, F.; Estradé, S. Nano Letters 2016, 16, 5068−5073

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Cerium oxide nanoparticles, a promising powerhouse in bioenergetic imbalance Ilaria Pezzini1, Attilio Marino2, Doccini Stefano3, Filippo Maria Santorelli3 and Gianni Ciofani2,4 1

Scuola Superiore Sant'Anna, The BioRobotics Institute, (Pisa), Italy; Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, (Pisa), Italy 3 IRCCS Stella Maris, Molecular Medicine, (Pisa), Italy 4 Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Torino, Italy 2

ilaria.pezzini@sssup.it Thanks to their catalytic properties as reactive oxygen species scavengers, cerium oxide nanoparticles (nanoceria, NC) have become an extremely promising candidate in both biological and medical application, with a possible exploitation in all those conditions where the oxidative stress has been proposed as one of the main pathogenesis actor [1]. In particular, it has been well established that nanoceria exhibit superoxide dismutase (SOD) mimetic activity [2], catalase mimetic activity [3] and hydroxyl radical scavenging activity [4], which are three of the endogenous enzymatic systems helpful to counteract both reactive oxygen species (ROS) and reactive nitrogen species (RNS) accumulation. Concerning neurological application, NC has been very recently proposed for the treatment of several neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, yet also for inflammatory disorders, diabetes and even cancer [5-7]. Moreover, several studies have been already demonstrated the NC protective role in counteracting ROS accumulation both in vitro and in vivo [8-11]. In this work, we have been investigated the NC antioxidant power in primary skin fibroblasts, derived from healthy individuals, both in basal condition and after hydrogen peroxide stimulation, as a proof of principle of the

antioxidant pharmacological potential of these nanoparticles. Viability was assessed after 24 and 72 h at increasing concentrations of the NC. Moreover, we tested the NC effects on different important bioenergetics parameters such as the ROS production, the adenosine triphosphate (ATP) content, and the effects on both the mitochondrial membrane potential and on the mitochondrial dynamics, as indicators of the bioenergetic power of NC. Consistently with data from the literature, NC treatment preserved cell viability at all the tested concentrations, underling the completely biocompatibility of these nanoparticles and, at the same time, an extensive cellular up-take as demonstrated by transmission electron microscopy analysis. Regarding the general effects of NC on mitochondrial functions and network, our results have demonstrated a general beneficial effects not only after a prooxidative insult, yet also in basal conditions so that NC indeed seem to improve the main physiological mitochondrial functions. The NC antioxidant activity combined with an apparent lack of toxicity makes NC an extremely promising therapeutic tool in all the pathological conditions that are characterized by a global bioenergetics dysfunction.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] Hroudová J, Singh N, Fišar Z. Biomed Res Int. (2014);2014:175062. [2] C. Korsvik, S. Patil, S. Seal, W.T. Self, Chem. Commun.(2007); 1056–1058. [3] T. Pirmohamed, J.M. Dowding, S. Singh, B. Wasserman, E. Heckert, A.S. Karakoti, J.E.S. King, S. Seal, W.T. Self, Chem. Commun. (2010); 46, 2736. [4] Y. Xue, Q.F. Luan, D. Yang, X. Yao, K.B. Zhou, J. Phys. Chem. (2011); 115, 4433. [5] P. D. Ray, B. W. Huang, Y. Tsuji, Cell. Signalling. (2012); 24, 981.

[6] J. Cedergren, T. Forslund, T. Sundqvist, T. Skogh, J. Rheumatol. (2007); 34, 2162. [7] B. Kumar, S. Koul, L. Khandrika, R. B. Meacham, H. K. Koul, Cancer Res. (2008); 68, 1777. [8] J. Niu, K. Wang, P.E. Kolattukudy, J. Pharmacol. Exp. Ther. (2011); 338, 61. [9] J. Chen, S. Patil, S. Seal, J.F. McGinnis, Nat. Nanotechnol. (2006); 1, 142. [10] S. Hussain, F. Al-Nsour, A.B Rice, J. Marshburn, B. Yingling, Z. Ji, J.I. Zink, N.J. Walker, S. Garantziotis, ACS Nano. (2012); 6, 5820. [11] M.S. Wason, J. Zhao, Am. J. Transl. Res. (2013); 5, 126.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Organic Radical Nanoparticles for Bioimaging Applications Davide Blasi1, Domna Maria Nikolaidou2, Francesca Terenziani2, Imma Ratera1, Jaume Veciana1 1

Institut de Ciencia de Materials de Barcelona (ICMAB-CSIC) and Ciber-BBN, Bellaterra (Barcelona), Spain 2 Università degli Studi di Parma, Department of Chemistry, Parma, Italy

iratera@icmab.es Polychlorotriphenylmethyl radicals are considered as an inert carbon free-radicals for their elevate chemical and thermal stability both in solution and in solid state, where their half-lives are on the order of decades. Its “openshell” configuration, due to the presence of an unpaired electron mainly localized on the αcarbon, leads to specific optical characteristics like an emission at long wavelength without the need of elongated π-systems, necessary for the optical biological window, a huge Stokes shift, an emission lifetime on the order of tens ns and a doublet electronic configuration both for ground and excited states. Despite such interesting properties, there are several limiting factors for the use of these molecules as photoactive materials in devices or as probes for bioimaging applications: water compatibility, the low value of fluorescent quantum yield (φF) and the low photostability, in particular in solution. Recently, several molecular approaches have been proposed in order to enhance these two fundamental parameters, but so far, it is not known what happens when radical molecules are confined in rigid organic matrix and processed as organic nanoparticles (ONPs). These kinds of systems are very promising for the fabrication of optoelectronic devices but specially as fluorescent probes for bioimaging due to its biocompatibility and stability in water suspensions.

In this work, we report the optical behavior of ONPs made by the optically neutral tris(2,3,4,5,6-chlorophenyl)methane (TTM-αH), that is acting as a matrix, doped with different amount of the optically active tris(2,4,6trichlorophenyl)methyl radical (TTM). We will see how, the change of the concentration of radical molecules doping the ONPs, affects their emission properties. We will compare the emission characteristics of these ONPs with the emission of the radical in THF and 2-methyl THF frozen solution at 77K. In summary, we will demonstrate for the first time how TTM radical, confined in ONPs shows a clear improvement of luminescence and photostability and moreover, it forms excimers presenting a broadband emission from 530 nm up to 850 nm. These excimers, observed for the first time in the case of a carbon free-radical, are promising for bioimaging applications and also for the fabrication of WOLEDs.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Magneto-sensitive liposomes containing manganese ferrite nanoparticles as nanocarriers for new promising antitumor thienopyridin-amine derivatives Ana Rita O. Rodrigues1, Ricardo C. Calhelha2, Juliana M. Rodrigues3, Isabel C.F.R. Ferreira2, Maria João R. P. Queiroz3, Paulo J. G. Coutinho1, Elisabete M. S. Castanheira1 1

Centre of Physics (CFUM), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Mountain Research Centre (CIMO), Polytechnic Institute of Bragança, 5301-855 Bragança, Portugal 3 Centre of Chemistry (CQ-UM), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal

ritarodrigues@fisica.uminho.pt Magneto-sensitive liposomes, resulting from the encapsulation of magnetic nanoparticles into liposomes, combine the remarkable physical properties of these two types of systems, while preserving the magnetic properties of the nanoparticles. These nanosystems can overcome pharmacokinetics problems of the encapsulated drugs and can be guided and localized to the therapeutic sites of interest by external magnetic field gradients. The use of magneto-sensitive liposomes as nanocarriers allows a safer use of powerful anticancer drugs in therapy with lower drug dosage and a more efficient treatment. In this work, manganese ferrite (MnFe2O4) nanoparticles, with superparamagnetic behaviour at room temperature, were obtained by coprecipitation method and their structural and magnetic properties were evaluated [1]. The synthesized nanoparticles were either entrapped in liposomes, originating aqueous magnetoliposomes (AMLs), or covered with a lipid bilayer (solid magnetoliposomes, SMLs), with sizes below 150 nm (Figure 1), suitable for biomedical applications [1,2]. Membrane fusion between both types of magnetoliposomes and giant unilamellar vesicles, used as models of cell membranes, was confirmed by FRET

(Förster Resonance Energy Transfer) assays [1,2]. New promising antitumor drugs, thienopyridine derivatives containing an amine moiety (compounds 1 and 2, Figure 2) [3], were successfully incorporated in aqueous and solid magnetoliposomes, with encapsulation efficiencies between 75% and 89% for both antitumor compounds. Drug-loaded magnetoliposomes, containing compounds 1 or 2, presented very low growth inhibitory concentrations (GI50), between 0.09 and 5.67 µM, when tested in vitro against several human tumor cell lines; MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer), HeLa (cervical carcinoma) and T3M4 (pancreatic cancer cells) [4]. These results are promising for future drug delivery applications using magnetoliposomes in oncology, allowing a dual therapeutic approach (simultaneous chemotherapy and magnetic hyperthermia).

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

References [1] A. R. O. Rodrigues, J. M. F. Ramos, I. T. Gomes, B. G. Almeida, J. P. AraĂşjo, M.-J. R. P. Queiroz, E. M. S. Castanheira, P. J. G. Coutinho, RSC Advances, 6 (2016) 1730217313. [2] A. R. O. Rodrigues, I. T. Gomes, B. G. Almeida, J. P. AraĂşjo, E. M. S. Castanheira, P. J. G. Coutinho, Phys. Chem. Chem. Phys., 17 (2015) 18011-18021. [3] M.-J. R. P. Queiroz, D. Peixoto, R. C. Calhelha, P. Soares, T. Santos, R. T. Lima, J. F. Campos, R. M. V. Abreu, I. C. F. R. Ferreira, M. H. Vasconcelos, Eur. J. Med. Chem., 69 (2013) 855-862. [4] A. R. O. Rodrigues, C. N. C. Costa, J. M. Rodrigues, M.-J. R. P. Queiroz, R. C. Calhelha, I. C. F. R. Ferreira, P. J. G. Coutinho, E. M. S. Castanheira, submitted to publication.

Figures

Figure 1. TEM image of solid magnetoliposomes.

Figure 2. Structure of compounds 1 and 2.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Nanostructured materials for intracellular sensing and delivery Mafalda Rodrigues1,2, Oriol Penon1,2, María E. Alea-Reyes1,2, David Limón1,2, Ezhil Amirthalingam1,2, and Lluïsa Pérez-García1,2,3 1

Departament de Farmacologia, Toxicologia i Química Terapèutica, Universitat de Barcelona, Barcelona, Spain. 2 Institut de Nanociència i Nanotecnología UB (IN2UB), Universitat de Barcelona, 08028 Barcelona, Spain 3 School of Pharmacy, The University of Nottingham, University Park, Nottingham, NG72RD, England, UK

mafaldanunes@ub.edu Nanostructured materials are promising vehicles in drug delivery applications both for sensing and therapy purposes, as they can be used to carry drugs into their site of action and/or target specific tissues, overcoming adverse side effects derived from non-specific distribution or poor water solubility of the drugs.[1] In our group we use supramolecular chemistry to assemble hybrid nanostructured materials and functionalize them with either sensing or therapeutical elements. Examples include the use of: a) silicon microparticles for intracellular pH sensing [2], b) supramolecular hydrogels for the sustained release of anionic drugs [3], c) incorporation of photosensitizers into inorganic or metallic nanoparticles for photodynamic therapy [4], d) multifunctional microparticles for intracellular delivery,[5] moving molecules on surfaces [6]. Acknowledgements: This work was supported by the EU ERDF (FEDER) funds and the Spanish Government grant TEC2014-51940-C2. M.E. A.-R., D.L. and E.A. thank Universitat de Barcelona, CONACYT and Generalitat de Catalunya, respectively, for predoctoral grants.

References [1] Mishra, D.; Hubenak, J. R.; Mathur, A. B.; J. Biomed. Mater. Res. A 101 (2013) 3646– 3660. [2] Torras, N.; Agusil, J. P.; Vázquez, P.; Duch, M.; Hernández-Pinto, A. M.; Samitier, J.; Rosa, E. J. de la; Esteve, J.; Suárez, T.; PérezGarcía, L.; Plaza, J. A.; Adv. Mater. 28 (2016) 1449–1454. [3] Rodrigues, M.; Calpena, A. C.; Amabilino, D. B.; Garduño-Ramírez, M. L.; Pérez-García, L.; J. Mater. Chem. B 2 (2014) 5419–5429. [4] Penon, O.; Marín, M. J.; Amabilino, D. B.; Russell, D. A.; Pérez-García, L.; J. Colloid Interface Sci. 462 (2016) 154–165. [5] Patiño, T.; Soriano, J.; Amirthalingam, E.; Durán, S.; González-Campo, A.; Duch, M.; Ibáñez, E.; Barrios, L.; Plaza, J. A.; PérezGarcía, L.; Nogués, C.; Nanoscale 8 (2016) 8773–8783 [6] Haq, S.; Wit, B.; Sang, H.; Floris, A.; Wang, Y.; Wang, J.; Pérez-García, L.; Kantorovitch, L.; Amabilino, D. B.; Raval, R. A.; Angew. Chemie Int. Ed. 54 (2015) 7101–7105.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Microfluidic isolation of cancer cells from blood: towards point-of-care liquid biopsy Silvina Samy1, Thais Pereira2, Clotilde Costa2, Marta Oliveira1, Lorena Diéguez1 1

INL - International Iberian Nanotechnology Laboratory, Av. Mestre José Veiga s/n, 4715-330 Braga, Portugal IDIS-CHUS – Instituto de Investigación Sanitaria de Santiago – Complejo Hospitalario Universitario de Santiago de Compostela, Travesia da Choupana s/n, 15706 Santiago de Compostela, Spain

silvina.samy@inl.int Metastatic disease is directly responsible for most of cancer-related deaths worldwide. Circulating tumor cells (CTCs) escape the primary tumor and disseminate through the blood stream potentially invading other organs and causing metastasis. Therefore, the study of CTCs might provide valuable information for the clinical management of cancer patients since they provide a real-time snapshot of the current tumor burden [1]. Additionally, it offers unique opportunities for low invasive sampling in cancer patients, constituting the so-called liquid biopsy, aimed for therapy monitoring, accurate prognosis and personalized treatment. However, these cancer cells are extremely rare – presenting ratios as low as 1 to 10 CTCs per a billion blood cells –, making their isolation finding a needle in a haystack [2]. Current isolation technologies rely on the expression of the EpCAM protein on the cell membrane. However, this is not a CTC ubiquitous biomarker, since CTCs are known to undergo Epithelial to Mesenchymal transition. Hence, the study of CTCs has been hampered so far by the limitations of current isolating technologies and their limited availability. Our goal is to provide efficient inexpensive microfluidic tools to isolate and characterize all CTCs from a peripheral blood sample independently of their phenotype. We have developed a size-based rare cell capture device that comprises several isolation areas. Since tumor cells have a bigger size and are more rigid compared to other cells contained in body fluids, they are expected to stay entrapped

within the device while non-cancerous cells are able to flow through. Immunostaining with fluorescently labelled antibodies was performed to identify specific phenotypes. Results showed good isolation yield of cancer cells while maintaining high purity when isolating CTCs from whole blood of metastatic colorectal cancer patients. Additionally, the device revealed an ability to capture as many or more CTCs when compared to the CTCs enumeration obtained by the current gold standard CELLSEARCH® CTC Test.

References [1] Gorges, T.M., Kuske, A., Röck, K., Mauermann, O., Müller, V., Peine, S., Verpoort, K., Novosadova, V., Kubista, M., Riethdorf, S. and Pantel, K. Clinical Chemistry, pii: clinchem.2016.260299. (2016) [Epub ahead of print] [2] Yu, M., Stott, S., Toner, M., Maheswaran, S. and Haber, D.A. The Journal of Cell Biology, 192 (2011), 373–382.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Interdigitated Bioink Micropatterns Generated by Multiplexed Polymer Pen Lithography S. SekulaSekula-Neuner1, R. Kumar1, S. Weigel2, R. Meyer2, C. M. Niemeyer2, A. Bonicelli3, A. C. B. Cato3, M. Hirtz1, H. Fuchs1,4,5 1

Institute of Nanotechnology, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany Institute of Biological Interfaces (IBZ-1), Karlsruhe Institute of Technology (KIT), Eggenstein -Leopoldshafen, Germany 3 Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, German) 4 Institute of Applied Physics and Center for Functional Nanostructures (CFN), Karlsruhe, Germany 5 Institute of Physics, University of Münster, Münster, Germany

Sylwia.Sekula-Neuner@kit.edu Application and integration of more than one ink in an interdigitated microscale pattern is still a challenge for microcontact printing (µCP). With the growing demand for interdigitated patterns of more than one molecule on subcellular to cellular length scales a new integrative approach is needed. We present a technique for the fabrication of bioactive microarrays and complex multi-ink patterns by polymer pen lithography (PPL). By taking advantage of the strength of microcontact printing (µCP) combined with the spatial control and capability of precise repetition of PPL in an innovative way, a new inking and writing strategy is introduced which enables true multiplexing within each repetitive sub-pattern. This patterning strategy aims specifically at application by cell biologists and biochemists addressing a wide range of relevant pattern sizes, easy pattern generation and adjustment, the use of only bio-friendly, nontoxic chemicals, and mild processing conditions during the patterning steps. The retained bioactivity of the fabricated cm2 area filling multimolecule patterns is demonstrated by showing: − covalently immobilized fluorophore- and oligonucleotide arrays for self-assembly of proteins as demonstrated by use of the arrays to monitor cell-protein interactions of MCF7 cells.

− covalently bound allergen arrays that are stable and easily integrated with microfluidic chips, utilized as a platform for mast cell activation studies and cell sorting applications The large area functionalization offered by PPL allows an easy incorporation of such arrays into microfluidic chips. In such a setup, easy handling of cell suspension, incubation process, and read-out by fluorescence microscopy will allow cell screening to be easily adapted for diagnostics and biomedical research.

References [1] R. Kumar, A. Bonicelli, S. Sekula-Neuner, A. C. B. Cato, M. Hirtz, H. Fuchs, Small (2016), in press [2] R. Kumar, S. Weigel, R. Meyer, C. M. Niemeyer, H. Fuchs, M. Hirtz, Chem. Comm. (2016), in press [3] F. Brinkmann, M. Hirtz, A. M. Greiner, M. Weschenfelder, B. Waterkotte, M. Bastmeyer, H. Fuchs, Small 9 (2013) 3266– 3275 [4] F. Brinkmann, M. Hirtz, A. Haller, T. M. Gorges, M. J. Vellekoop, S. Riethdorf, V. Müller, K. Pantel, H. Fuchs, Sci. Rep. 5 (2015) 15342

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

Figure 1. (a) Cartoon of a segmented multi-color PPL stamp with four different inks and a printed multi-color array. (b) Writing strategy exemplified for four colors. (c) Examples of fluorescent multi-color arrays with different azides containing three (right), four (middle) and five (left) components. Scale bars equal 50Âľm in all images and insets

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Facile one-pot synthesis of highly luminescent carbon dots from food and its application of in-vitro cancer growth inhibition N. Vasimalai1 , V. Vilas-Boasa2, J. Gallo1, M. F. Cerqueira3, M. Menéndez4, J. M. Costa4, L. Diéguez1, B. Espiña1, M. T. Fernández-Argüelles1* 1

Life Sciences Department, International Iberian Nanotechnology Laboratory (INL), Braga, Portugal UCIBIO-REQUIMTE, Laboratory of Toxicology, Biological Sciences Department, Faculty of Pharmacy, University of Porto, Porto, Portugal 3 Center of Physics, University of Minho, Braga, Portugal 4 Department of Physical and Analytical Chemistry, University of Oviedo, Oviedo, Spain

vasi.malai@inl.int *maria.fernandez-arguelles@inl.int Synthesis and application of fluorescent carbon dots (C-dots) is emerging as a new alternative to inorganic semiconductor quantum dots due to their high photoluminescence, low photobleaching, high biocompatibility and excellent stability in aqueous media. All these features make them excellent candidates in biosensing, bioimaging, biomedical or catalysis applications. Nowadays multiple synthesis techniques are described to obtain C-dots, as well as different carbon sources as alternative for graphite source [1]. In fact, natural source materials are being used for production of Cdots due to the simple, cost-effective and environmental friendly syntheses. However, when talking about the toxicity of C-dots, in vitro and in vivo results reported in the literature are inconsistent, and it is considered that the toxicity is mainly determined by the synthesis protocol [2]. Therefore, due to the fascinating medicinal properties of certain compounds, in this work four different C-dots have been synthesized from spices including cinnamon, red chilli, turmeric and black pepper, by one-pot green hydrothermal method. They have been characterized with UV-Vis, Fluorescence, FTIR spectroscopies, DLS and TEM.

These C-dots have been evaluated to study their toxicity to in-vitro human normal and cancer cells. Bioimaging studies confirmed that the Cdots inhibited cell viability dose-dependently, showing higher toxicity in cancer cells than in normal cells after 24 h incubation. Particularly, cell viability studies suggested that the growth of human cancer cells is suppressed up to 35%, 50%, 50% and 75%, when using cinnamon, red chilli, turmeric and black pepper respectively, whereas the growth inhibition of normal cells was not significantly affected. Conversely, Cdots synthesized from citric acid did not show any significant toxicity neither in normal nor in cancer cells, which suggests that the anticancer properties observed in the spice-derived C-dots can be attributed to the starting material employed for their fabrication. The interesting anti-cancer activity of the spice-derived C-dots along with the bioimaging applicability and excellent tolerability in normal HK-2 cells, suggests a promising future potential as efficient theranostic agents with minimal side effects in normal cells.

References [1] S. N. Baker et al., Angew. Chem. Int. Ed. 49 (2010) 6726-6744. [2] P. Pierrat et al., Biomaterials 51 (2015) 290302.

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Figures

NanoBio&Med2016 november 22-24, 2016 - Barcelona (Spain)

Presenting Author

Topic

Andrade Carrera

Drug Delivery

Poster Title

Berenice

Mexico

Berardi Bola Sampol Bondia Raga Cerpa Naranjo Cerpa Naranjo Cobaleda Siles E.A. Teixeira

Ginevra

Italy

In vitro ocular study of natural flavanone loaded in PLGAnanoparticles Nanomaterials for Bio and Activation of magnetic nanoparticles with an antimicrobial Medical Applications polypeptide: a new therapeutic tool

Raúl

Spain

Bio-nano measurement and microscopy

Intracellular Rheological Studies with Optical Tweezers

Patricia

Spain

Bio-nano measurement and microscopy

Hybrid nanoscopy of hybrid nanomaterials

Arisbel

Spain

Nanomaterials for Bio and Rheological behaviour of carbon nanotubes suspensions with Medical Applications biomedical applications

Arisbel Beatriz Macarena

Spain

Nanomaterials for Bio and Penetration of Mesenchymal Stem Cells by Carboxylic Acid Medical Applications Functionalized Carbon Nanotubes

Spain

Nanotoxicology

Feng

Yuping

Optimal strategies for the reduction of nanotechnology associated risks: Safety by design

Freixanet Muriel

Spain

Garcia

Alba

Spain

Quinazolinediones-derivatives SIRT6 inhibitors for targeted selective chemotherapy Nanomaterials for Bio and Novel Fe-Mn-Si-Pd dense and porous alloys: insights on Medical Applications mechanical, magnetic and corrosion performance Nanomaterials for Bio and Metastatic Skin Cancers: Identification of Chemotherapeutic Medical Applications Targets for Nanotherapeutics Nanoassemblies / Amphiphobic coatings for antifouling protection in sea Surfaces environment Nanomaterials for Bio and Multicomponent aliphatic polyester-based nanoparticles for Medical Applications antigen delivery and immune modulation against cancer disease NanoMaterials for Multimodal Inorganic Nanoparticles to Overcome Cancer Drug Medicine Resistance Improving an in vitro 3D co-culture as an alternative model of intestinal barrier for the assessment of engineered nanomaterials Nanotoxicology (ENMs) risk

Grillone

Agostina

Italy

Drug Delivery

Maria do Céu Portugal Spain

Fernandes Ana Rita

Portugal

Ferrari

Michele

Italy

Florindo

Helena

Portugal

Drug Delivery

Preparation and Characterization of nanoceria-loaded liposomes Magnetic molecularly imprinted polymer for the detection of methyl parathion in fish

Hassan Herrera Chacon

Amal

Spain

Integrated Systems / Sensors

Anna

Spain

Nanomaterials for Bio and Molecularly imprinted polymers: functional nanoplastics for Medical Applications diagnostics and specific sorption

Jorba

Ignasi

Spain

Bio-nano measurement and microscopy

Kyriazi

Maria-Eleni

Nanomaterials for Bio and Medical Applications Advanced probes for multiplexed intracellular biomarker detection

Kyvik

Adriana

Spain

Nanoassemblies / Surfaces

LaprestaFernández Alejandro Lopez Bouzo Belen

Nanoassemblies /

Switzerland Surfaces Spain

Nanomechanics of the Drosophila embryonic central nervous system

Dynamic Molecular Bio-Interfaces for Controlled Environments Towards Spatial and Temporal Control of Cell Behavior Monitoring a bioengineered surface coating replacement by surface enhanced Raman scattering (SERS) on Ag nanocubes and performance in vesicle trapping

Mira Moran Badenas MorenoSereno

Amalia Maria del Carmen

Spain

Neutral nanostructures for oligonucleotide delivery to cancer cells Development of nanocrriers polyelectrolytes as antimicrobial Nanomaterials for Bio and agents for dual use as drugs and bioimaging probes in human Medical Applications health

Spain

Nanomaterials for Bio and Mammalian Cell Viability on Hydrophobic and Superhydrophobic Medical Applications Fabrics

Mauricio

Spain

Bioelectronics

Nadejde NavarroRequena

Claudia

Romania

Multichannel Optical Waveguide Lightmode Spectroscopy: Sensor and Instrument Nanomaterials for Bio and Influence of stabilization agents on the properties of Medical Applications biocompatible magnetite nanoparticles

Claudia

Spain

Nanomaterials for Bio and Stimulation of Dermal Fibroblasts with Calcium-Phosphate Medical Applications Nanoparticles for the Design of Novel Bioactive Dressings

Drug Delivery

Presenting Author

Topic

Poster Title

Niemi NietoGarai

Inhibiting Notch Activity in Breast Cancer Stem Cells by Glucose Functionalized Nanoparticles Carrying Ȗ-secretase Inhibitors

Rasmus

Finland

Drug Delivery

Jon Ander

Spain

Drug Delivery

Nunez-Bajo Estefania

Spain

Integrated Systems / Sensors

Ozcelik

Spain Czech Republic

Ani

Panacek Ales Peral Guillamon Ariadna

Nanoassemblies / Surfaces

Nanoliposomes Targeting Myeloid Cells to Deliver HIV-1 Reactivation Compounds Paper-based gold-modified electroanalytical devices for glucose and inorganic arsenic sensing Self-Assembled Up-standing Chiral Architectures for the Construction of Functional Surfaces

Nanomaterials for Bio and Silver nanoparticles restore bactericidal activity of inactive Medical Applications antibiotics against multiresistant bacteria

Spain

Nanomaterials for Bio and Medical Applications CeO2NPs Time-evolution stability

Pablo Aitor

Spain

Bio-nano measurement and microscopy

Postigo Pablo Aitor RodríguezPulido Alberto

Spain

Bio-nano measurement and microscopy

Spain

Bio-nano measurement and microscopy

Salvo

Spain

Postigo

Ignacio

Israel Scomparin Anna Serrá i Spain Albert Ramos Silva Marrero Jonas Ismael Spain

Optofluidic chips with nanochannels for dynamic molecular detection using enhanced fluorescence Fluorescence signal enhancement and background suppression in autofluorescence microscopy imaging by using two-dimensional nanostructured glass substrates

Photosensitizing flavoproteins: improved bio-tags for nanoscale imaging with correlative microscopy Nanomaterials for Bio and Control synthesis of metal oxide nanoparticles for medical Medical Applications applications A Comparative Study of Folate Receptor-Targeted Doxorubicin Drug Delivery Delivery Systems: Dosing Regimens and Therapeutic Index Nanomaterials for Bio and In vitro study of gold nanoparticles, nanorods and microparticles, Medical Applications as potential carriers for photodynamic therapy Nanomaterials for Bio and Chitosan-TPP nanoparticles as a DNA carrier to silence gene Medical Applications expression in vivo

Spasojevic Vojislav

Serbia

Nanomaterials for Bio and Magnetic hyperthermia of MgxFe3-xO4 ferrofluid synthesized by Medical Applications hydrothermal procedure

Suñé Pou

Marc

Spain

Drug Delivery

Tejero

Ricardo

Spain

Tilocca

Antonio

Improved Formulation of Cationic Solid Lipid Nanoparticles Displays Cellular Uptake and Biological Activity of Nucleic Acids Nanomaterials for Bio and Importance of calcium ions at bioinspired integrative implant Medical Applications surfaces for tissue regeneration Nanomaterials for Bio and Nanosized bioactive glasses: atomic-scale features from Medical Applications simulations

Vargas

Carolina Bogdan Stefan

Spain

Bioelectronics

Suspended silicon micro-photodiodes for single cell stimulation

Romania

NanoMaterials for Medicine

TiO2 nanostructures as UV blocking components in sunscreens

Drug Delivery

Pentacyclic triterpenes with anti-inflammatory activity formulated in topical nanoemulsion

Vasile Vazquez Prada Vázquez Ríos Vilanova

Karla Ximena Mexico Abi Judit

Spain

Oriol

Spain

Whitehead Debra

Exosome-like nanoparticles for microRNA selective transport to tumoral cells Understanding the Kinetics of Protein-Nanoparticle Corona Formation Nanomaterials for Bio and Titania coating of mesoporous silica nanoparticles- Protective Medical Applications effects on vasodilator function, ex vivo Bio-Inspired nanotechnologies Nanoassemblies / Surfaces

In vitro ocular study of natural flavanones loaded in PLGA-nanoparticles. 1

Berenice Andrade-Carrera , Mireia Mallandrich , Beatriz Clares-Naveros , Helen L. Alvarado , Elena 2 2 1 2 SĂĄnchez-LĂłpez , Ana Calpena-Campmany , MarĂa Luisa GarduĂąo-RamĂrez and MarĂa Luisa GarcĂa . 1

Centro de Investigaciones QuĂmicas, Instituto de InvestigaciĂłn en Ciencias BĂĄsicas y Aplicadas, Universidad AutĂłnoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, 62209, Cuernavaca, Morelos, MĂŠxico 2 Department of Pharmacy, Pharmaceutical Technology and Physical Chemical, Faculty of Pharmacy and Food Sciences, University of Barcelona, Av. Sant Joan s/n, Barcelona, Spain. 3 Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Granada, Granada, Spain. lgarduno@uaem.mx Introduction: Natural products have been regarded as important sources that could produce potential therapeutic agents. Over 50 % of drugs approved were originated from the natural resources. Eysenhardtia. platycarpa is a small trHH GLVWULEXWHG LQ 6RXWKHUQ 0H[LFR ZKHUH LW LV NQRZQ DV ÂłWDUD\Â´ ÂłSDOR GXOFHÂ´ VZHHW ZRRG DQG ÂłSDOR D]XOÂ´ EOXH ZRRG ,W LV ZHOO-known in traditional herbal medicine and used for the treatment of kidney diseases, as well as bladder infections and as a diuretic. It is also 1 used for the treatment from diabetes mellitus. In the present study two flavanones were isolated (2S)5,7-dihidroxy-6-prenylflavanone (1) and (2S)-5-hydroxy-7-methoxy-6-prenylflavanone (2) were evaluated in ocular tolerance assay and loaded in poly(D,L-lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). Materials and methods: From the methanolic extract of Eysenhardtia platycarpa leaves two flavanones 1 13 (1 and 2) were isolated. The flavanones were characterized by 1D and 2D H and C high-resolution 1 solid-state NMR spectroscopy with a Varian INOVA-400 instrument (400 MHz for H and 100 MHz for 13 1 C). The flavanones (1 y 2) were evaluated in ocular tolerance assay HET-CAM and developed as described in the INVITTOX No. 15 protocol, this test is based on the observation of the irritant effects 2 (bleeding, vasoconstriction and coagulation). PLGA NPs containing flavanone 1 and 2 (NPs1 and NPs2) (1.5 mg/mL) were elaborated by the solvent displacement technique described by Dominguez et 3 al. Mean particle size (Z-ave), zeta potential (ZP) and polydispersity index (PI) were determined with a Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). Morphological and structural properties were examined by transmission electron microscopy (TEM) with a Zeiss 902 CEM 902 (Zeiss, Barcelona, Spain). Results and Discussion: In vitro ocular tolerance was using HET-CAM test. In additions of 0.9 % saline solution to the healthy membranes produced no visual response over five minutes period. In contrast, 1 M NaOH produce severe, hemorrhage, which increased over five minutes. Applications of 300 l of the samples (1 and 2) were dissolved 1 mg/ml of Transcutol , in the first 5 min. of application the flavanone 1 was not irritant versus flavanone 2 was produced hemorrhage and showed irritant reaction. Spherical shaped NPs1 and NPs2 with Z-ave 205.2 0.26 and 141.6 0.78 nm, respectively with PI values 0.058 0.053 to NPs1 and 0.086 0.017 to NPs2. ZP values to NPs1 is -8.25 0.346 and NPs2 is -10.6 0.231 mV. Conclusions: Ocular administration for the treatment of pathological eye tissues offers the advantage of delivering the drug. HET-CAM assay results indicate that the flavanone 1 is less toxic and irritating. The NPs1 and NPs2 were spherical shape and the negative Zeta Potential values ensure a good stability with a small Z-Average. Acknowledgements: The authors would like to thank CONACyT, Mexico for the scholarship 253949, the Ministry of Science and Innovation of Spain with the project MAT201459134R and Laboratorio Nacional de Estructuras de MacromolĂŠculas de la Universidad AutĂłnoma del Estado de Morelos, Mexico (LANEM). References [1] V. DomĂnguez-Villegas, V. DomĂnguez-Villegas, M. L. GarcĂa, A. Calpena B. Clares-Naveros, M.L. GarduĂąo-RamĂrez. Natural Products Communications. 8, No. 2 (2013), 177. [2] M. Warren, K. Atkinson, S. Steer, INVITTOX: The ERGATT/FRAME data bank of in vitro techniques in toxicology, Toxicol. Vitro, 4, (1990) 707. [3] V. DomĂnguez-Villegas, B. Clares-Naveros, M.L. GarcĂa-LĂłpez, A.C. Calpena-Campmany, P. Bustos-Zagal, M.L. GarduĂąo-RamĂrez. Colloids and Surfaces B: Biointerface. 116 (2014) 183.

Activation of magnetic nanoparticles with an antimicrobial polypeptide: a new therapeutic tool a

Berardi Ginevra , Grgurina Ingeborg , Barteri Mario

'HSDUWPHQW RI &KHPLVWU\ Âł6DSLHQ]DÂ´ University of Rome, Rome, Italy; 'HSDUWPHQW RI %LRFKHPLFDO 6FLHQFHV Âł$ 5RVVL-)DQHOOLÂ´ Âł6DSLHQ]DÂ´ 8QLYHUVLW\ RI 5RPH 5RPH ,WDO\ a

ginevra.berardi@uniroma1.it Bacterial secondary metabolites often show significant biological activities, like the antimicrobial one, that make them an interesting research field for new drugs in order to overcome the problem of multidrug resistance of some pathogenic organisms. Moreover, the development of new strategies to deliver the drug into the infected tissue using nanotechnology, and in particular nanoparticles, can lead to a more efficient treatment of the pathology. Syringopeptins (SPs) are cyclic lipodepsipeptides produced by certain strains of the phytopathogenic bacterium Pseudomonas syringae. Belonging to this class of metabolites, Syringopeptin 22a (SP22A) has shown to inhibit the growth of gram-positive bacteria, fungi and mycobacteria, particularly Mycobacterium smegmatis, a nonpathogenic surrogate of Mycobacterium tubercolosis. For the first time, we have conjugated this novel antimicrobial agent to polymer coated superparamagnetic nanoparticles (MNPs). The biological activity of functionalized MNPs was tested on a fungal model, Rhodotorula pilimanae. Conjugation of SP22A onto the nanosystem does not interfere with its biological activity; the MNPs, in fact, functionalized with SP22A have shown growth-inhibitory effects of the fungus. Starting from these encouraging preliminary results, we are also investigating the possibility of internalizing functionalized magnetic nanoparticles inside macrophages. These phagocytic cells are the natural environment for mycobacteria in the human body; these organisms can hidden into the macrophages infecting them and surviving into them. The in vitro experiments with THP-1 monocytic cell line differentiated into macrophage demonstrated the possibility to engulf these cells with functionalized magnetic nanoparticles. These interesting results could pave the way to the in loco treatment of microorganisms, inside the cells where they are hidden. This approach can lead to a more effective therapy of difficult and resistant infections. [1] Grgurina I, Bensaci M., Pocsfalvi G., Mannina L., Cruciani O., Fiore A., Fogliano V., Sorensen K.N., Takemoto J.Y., Antimicrobial Agents and Chemotherapy, 12 (2005) 5037; [2] Bensaci M.F., Gurnev P.A., Bezrukov S.M., Takemoto J.Y., Frontiers in Microbiology, 22 (2011) 216; [3] Griffiths G., Nystrom B., Sable S.B., Khuller G.K., Nature Reviews Microbiology, 8 (2010) 827 [4] Clemens D.L., Lee B., Xue M., Thomas C.R., Meng H., Ferris D., Nel A.E., Horwitz M.A., Antimicrobial Agents and Chemotherapy 56 (2012) 2535; [5] Wang Q., Cheng H., Peng H., Zhou H., Li P.Y., Langer R., Advanced Drug Delivery Reviews 91 (2015) 125

Intracellular Rheological Studies with Optical Tweezers RaĂşl Bola, Estela MartĂn-Badosa and Mario Montes-Usategui Optical Trapping Lab Âą Grup de BiofotĂ˛nica, Departament de FĂsica Aplicada, Universitat de Barcelona, C/ MartĂ i FranquĂ¨s 1, 08028 Barcelona, Spain raul.bola@ub.edu Abstract Optical tweezers are tightly focused laser beams capable of exerting pico-Newton forces on micronsized objects and provide a powerful tool for manipulating organelles and vesicles inside living cells without perturbing the cell membrane or harming the cell itself [1]. Apart from micro-manipulating objects, external forces applied on a trapped sample can also be measured for analyzing a wide variety of dynamic processes at the micro scale, from in-cell biophysics and micro-swimmer hydrodynamics, to single-molecule experiments and micro-rheology. Most of force measurement techniques with optical tweezers are calibration-based methods in which the interaction between the trapped object and the optical trap is described as a harmonic potential where the optical trap acts as an elastic spring (F = - Č&#x203A;Ă&#x201A;[ 7KHQ RQFH WKH VSULQJ FRQVWDQW Č&#x203A; LV NQRZQ measuring the relative position between the object and the trap is used to calculate the applied force. Unfortunately, these calibration-based methods become useless when dealing with living cells. On the RQH KDQG WKH VSULQJ FRQVWDQW Č&#x203A; KDV WR EH FDOLEUDWHG DW HDFK H[SHULPHQW VLQFH LW GHSHQGV RQ VHYHUDO parameters such as the refractive index of the trapped object and the surrounding medium, the geometry of the trapped object, the local viscosity or the local temperature. These are quantities that are often unknown when measuring inside the cell, due to the non-homogeneous, viscoelastic and dynamic features of the cell cytoplasm. On the other hand, the action of molecular motors responsible for intracellular transport breaks the thermodynamic equilibrium between the trapped object and the environment, while most calibration methods are based on the fulfilment of the fluctuation-dissipation theorem (FDT), in which thermodynamic equilibrium is assumed. Here, we apply the active-passive calibration method [2] in the interior of living cells, in order to provide significant knowledge of the viscoelastic behaviour of the cytoplasm. Also, we show how the biological activity affects force measurements and we present an alternative method, based on the direct detection of beam momentum changes, which overcomes the aforementioned problems [3,4]. References [1] A. Ashkin, J. M. Dziedzic, J. E. Bjorkholm, and S. Chu, Opt. Lett., 11(5) (1986) 288Âą290. [2] M. Fischer, Andrew C. Richardson, S. Nader S. Reihani, Lene B. Oddershede, and Kirstine BergSĂ¸rensen, Rev. Sci. Instrum., 81(1) (2010) 015103. [3] A. FarrĂŠ and M. Montes-Usategui, Opt. Express, 18 (2010) 11955Âą11968. [4] A. FarrĂŠ, F. MarsĂ , and M. Montes-Usategui, Opt. Express, 20 (2012) 12270Âą1229. Figures

Figure 1. Friction retardation spectrum (a) and calibrated spring constant (b) obtained inside living chinese hamster ovary (CHO) cells. In a), the water friction retardation spectrum is also shown, for comparison with the viscoelastic behaviour of the cell cytoplasm.

Hybrid nanoscopy of hybrid nanomaterials 1

Patricia Bondia, Santiago Casado, Rocío Jurado, José M. Domínguez-Vera, Natividad 2 1 Gálvez, Cristina Flors 1

IMDEA Nanoscience, Ciudad Universitaria de Cantoblanco. Faraday 9, 28049, Madrid, Spain 2

Department of Inorganic Chemistry, University of Granada. Av. Fuentenueva Fuente s/n, 18071, Granada, Spain.

Email: patricia.bondia@imdea.org Recently, research in the field of new nanomaterials has focused its attention on proteins and peptides as nanoscale building blocks that provide a simple route to fabricate structures from zero to three-dimensions (0±3D) [1]. These structures are rich in functional groups allowing specific coupling with different molecules, which is an attractive approach for templating nanoscale elements [2]. In particular, amyloid-like fibers are emerging as an important class of functional materials since they stand out for their rigidity, mechanical strength and controllable fabrication process [3]. Moreover, they have shown excellent properties as templates for the production of 1D inorganic nanostructures [4]. The combination of fibers with inorganic particles confers these hybrid nanomaterials extraordinary properties such as conductivity, magnetism, and complex optical responses [5]. With the aim of improving the design of these hybrid materials, different complementary techniques are necessary to gain a complete picture of their structure and function. Here we applied atomic force microscopy (AFM), which offers high spatial resolution images and topography of the sample surface, in combination with super-resolution fluorescence microscopy to characterize luminescent amyloid-like fibrils. These fibers were made by heat and pH-denatured ȕ-lactoglobulin (ȕLG) functionalized with organic fluorophores (Alexa488) or quantum dots (QD) [3]. Both fluorescence molecules possess specific photophysical properties, namely photoswitching or blinking, enabling to perform super-resolution fluorescence microcopy by single molecule localization techniques [6]. We show that hybrid nanoscopy is specially useful to optimize super-resolution image reconstruction since the topography image is used as D ³JURXQG WUXWK´ [7]. This is particularly important in the case of QDs, which show suboptimal blinking. Importantly, the correlative images provide nanoscale insight into the structure and functionalization of this hybrid material revealing heterogeneous labeling of Alexa488-fibrils which was not detected by standard fluorescence microscopy. Finally, we have achieved twocolor super-resolution fluorescence imaging in correlation with topography to characterize fibrils functionalized with QDs of different color. This is the first time that the latter study has been performed and represents an important step forward in the characterization of multifunctionalized hybrid materials, a key challenge in nanoscience. References: 1. Reches, M. and Gazit, E., Nature nanotechnology, 2006, 1(3): p. 195-200. 2. Lakshmanan, A.; Zhang, S.; Hauser, C. A., Trends in biotechnology, 2012, 30(3): p. 155165. 3. Jurado, R.; Castello, F.; Bondia, P.; Casado, S.; Flors, C.; Cuesta, R.; Domínguez-Vera, J. M.; Orte, A.; Gálvez, N., Nanoscale, 2016, 8(18): p. 9648-9656. 4. Wang, X.; Li, Y.; Zhong, C., Journal of Materials Chemistry B, 2015, 3(25): p. 4953-4958. 5. Knowles, T. P. and Mezzenga, R., Advanced Materials, 2016. 6. Huang, B.; Bates, M.; Zhuang, X., Annual review of biochemistry, 2009, 78: p. 993. 7. Monserrate, A.; Casado, S.; Flors, C., ChemPhysChem, 2014, 15(4): p. 647-650.

Figure 1. Correlative AFM and fluorescence nanoscopy for the characterization Č&#x2022;LG fibers functionalized with QDs. AFM and standard fluorescence images are acquired in situ. The AFM image provides additional information to the super-resolution fluorescence image and is useful to optimize image reconstruction of the latter.

Rheological behaviour of carbon nanotubes suspensions with biomedical applications 1

Arisbel Cerpa Naranjo , BegoĂąa IbaĂąez MartĂnez , Isabel Lado TouriĂąo , Rodrigo Moreno 2 3 4 Botella , Paloma Ballesteros GarcĂa , SebastiĂĄn CerdĂĄn GarcĂa-Esteller 1

Department of Industrial and Aerospatial Engineering. Universidad Europea de Madrid, C/Tajo s/n, 28670 Villaviciosa de OdĂłn, Spain 2 Institute of Ceramics and Glass-CSIC, 28049 Madrid, Spain 3 Laboratory of Organic Synthesis and Molecular Imaging by Magnetic Resonance, Faculty of Sciences, UNED, 28040 Madrid, Spain 4 /,(50 ,QVWLWXWH RI %LRPHGLFDO 5HVHDUFK Âł$OEHUWR 6ROVÂ´ &6,& 0DGULG 6SDLQ arisbel.cerpa@universidadeuropea.es

Abstract The rheological behaviour of different carbon nanotubes suspensions in the dilute regime was evaluated. Suspensions of commercial multi-walled carbon nanotubes without functionalization, multi-walled carbon nanotubes oxidized in our laboratory with nitric acid under reflux for 24 hours and commercial multi-walled carbon nanotubes functionalized with carboxylic groups were studied. The study was performed in two dispersing media: water and fetal bovine serum (FBS). In addition, the results were compared with the results of rheological behaviour of singlewalled carbon nanotubes suspensions obtained in previous works [1, 2] to understand the influence of the structure of carbon nanotubes on the rheological properties when a biological medium is present. Structural characterization was performed with different techniques: XRD, IR, TGA, TEM, SEM and AFM. For the colloidal characterization, the zeta potential of suspensions was determined at pH values between 6 and 8, close to physiological pH. Rheological measurements were carried out with a rotational rheometer using a cone and plate system. The measurements were done for different concentrations of suspensions (1 and 1.5 mg/ml), temperatures from 25 to 37 C, and water and FBS as solvents. The results obtained by Atomic Force Microscope show a high tendency to aggregation between carbon nanotubes and bovine serum albumin (BSA) protein present in the fetal bovine serum. This behaviour was also observed by molecular dynamics simulations. The carbon nanotubes suspensions present a Newtonian rheological behaviour. Depending on the type of solvent and carbon nanotubes the viscosity changes. The viscosity values of multiwalled carbon nanotubes suspensions are greater than those of single-walled carbon nanotubes suspensions when water is used as solvent. The viscosity values of multi-walled carbon nanotubes suspensions are between 2 and 6 mPa.s for the concentrations studied. Nevertheless, the viscosity values of single-walled carbon nanotubes suspensions are less than 3 mPa.s. The change of concentration in the suspension prepared with FBS does not cause significant changes in the viscosity values while in the aqueous suspensions the viscosity increases with increasing concentration. Furthermore, the viscosity decreases linearly with the rise of temperature from 25 C to 37 C although the changes are small.

References [1] Arisbel Cerpa, Mariana KĂśber, Daniel Calle, Viviana Negri, JosĂŠ MarĂa Gavira, Antonio Hernanz, Fernando Briones, SebastiĂĄn CerdĂĄn, and Paloma Ballesteros. Med. Chem. Com 473 (2013) 270. [2] A. Cerpa, O. Quiroga, R. Moreno, I. Lado, M.P. Ros, V. Negri, S. CerdĂĄn, P. Ballesteros. DIAM 2014. International Conference on Diamond and Carbon Materials. Madrid (2014)

Penetration of Mesenchymal Stem Cells by Carboxylic Acid Functionalized Carbon Nanotubes 1

Arisbel Cerpa Naranjo , Carlos Bello Álvarez , Beatriz de Lucas Moreno , Beatriz González 2 Gálvez 1

Department of Industrial and Aerospatial Engineering. Universidad Europea de Madrid, C/Tajo s/n, 28670 Villaviciosa de Odón, Spain 2 Escuela de Doctorado e Investigación. Universidad Europea de Madrid, C/Tajo s/n, 28670 Villaviciosa de Odón, Spain arisbel.cerpa@universidadeuropea.es

Abstract The extraordinary properties of carbon nanotubes make them excellent candidates for the development of applications in the biomedicine field. Nowadays the use of carboxylic acid functionalized carbon nanotubes (f-CNT) are considered a promising tool as contrast agents for magnetic resonance imaging [1,2], as nanovectors (gene therapy, drug delivery) and for stem cell therapy and tissue engineering [3]. The aim of this preliminary study was to determine the interactions between single-walled and multi-walled carbon nanotubes (SWCNTs and MWCNTs), oxidized with nitric acid under reflux for 24 hours, and mesenchymal stem cells (MSCs). The results obtained by Transmission Electron Microscopy show that both kinds of carbon nanotubes are able to penetrate into MSCs. CNTs can cross the plasmatic membrane appearing inside the cells and within vesicles. Other important result is the absent of toxic effect of carbon nanotubes in MSCs. Furthermore, we test wound healing capacity of MSCs adding carbon nanotubes. This addiction of carbon nanotubes do not produce significant changes in cell motility and the cells were able to close the wound as control situations. In conclusion, we produce f-CNTs that not only are able to penetrate inside the cells, but also have not negative effect, a priori, in the cells. These results are encouraging, but need much more investigation of their impact in cells and in the exploitation of their properties for biomedical applications. References [1] Arisbel Cerpa, Mariana Köber, Daniel Calle, Viviana Negri, José María Gavira, Antonio Hernanz, Fernando Briones, Sebastián Cerdán, and Paloma Ballesteros. Med. Chem. Com 473 (2013) 270. [2] Viviana Negri, Arisbel Cerpa, Pilar López-Larrubia, Laura Nieto, Sebastián Cerdán, Paloma Ballesteros. Angewandte Chemie Int. Ed. 49 (2010) 1813. [3] Liu, Dandan, Changqing Yi, Dawei Zhang, Jinchao Zhang, and Mengsu Yang. ACS nano 4, 4 (2010) 2185.

Optimal strategies for the reduction of nanotechnology associated risks: Safety by design 1

1,2,3

M. Cobaleda-Siles , A.P. Guillamon , V.F. Puntes . (1) Institut de Recerca Vall dÂ´Hebron (VHIR). Pg. Vall d'Hebron, 119-129 08035 Barcelona (Spain) (2) InstituciĂł Catalana de Recerca i Estudis AvanĂ§ats(ICREA) Pg. LluĂs Companys, 23, 08010 BCN (3) Institut CatalĂ de Nanociencia i Nanotecnologia (ICN2), Campus de la UAB, Av. Serragalliners, s/n, 08193 BCN. beatriz.cobaleda@vhir.org

The development of Nanotechnology in the last decades has brought a new area of research focusing on the synthesis, characterization and development of new materials. However in recent years the increase of projects, congresses and publications related to nanosafety has gained a significant 1Âą3 space in the community of nanotechnology. The main objective of developing innovative methodologies to evaluate and manage human and environmental health risks of nano-enable products has focused the aim of the researchers concerned about the problem of nanoparticles to cause adverse health effects and how the people are expose to ENMs. Although most of the existing nanotoxicology studies have been conducted on pristine NMs, it is a reality that consumers and workers are exposed to NM in later stages of their life cycle such as other manufacturing states (e.g. coated) or aging NMs (e.g. recycling) rather than pristine NMs. Therefore, understanding the hazard of these NMs is necessary for a meaningful risk assessment. The behavior of NMs in the environment and inside the body has to be understood and based on that, we should be capable, under a synthetic point of view, of designing and predicting their possible negative impact and avoid it. As it is well known, the NMs possess unique physico-chemical properties compared with their 4 corresponding bulk materials. Furthermore, these properties lead to toxicities completely different to their reference materials. These physico-chemical characteristics including, surface area, agglomeration states or the pH value of the ENMs can be controlled for reducing their harmful effects. For instance, controlling pH is especially significant, as it is directly related with the state of aggregation and consequently affects cells uptake and cell viability. In this sense, our work is focused on the synthesis and characterization of different NMs WLWDQLXP GLR[LGH VLOYHU 13VÂŤ PRGLI\LQJ WKHLU VL]H VKDSH RU VXUIDFH FRDWLQJ OLJDQGV 3(* $87 393ÂŤ DQG VWXG\LQJ WKHLU SK\VLFo-chemical properties when they are exposed to different environmental conditions (Fig. 1). The outcomes reflected in this work were performed under the ongoing GUIDEnano project funded by European CommissionÂ´s Framework Programme (FP7/2007 XQGHU JUDQW DJUHHPHQW Ę&#x2039;

Fig. 1. Schematic figure of safe by design. Picture adapted from Strategies for the intracellular delivery of nanoparticles, Chem. Soc. Rev. 40, 233Âą245 (2011). 1. 2. 3. 4.

OberdĂśrster, G. Safety assessment for nanotechnology and nanomedicine: concepts of nanotoxicology. J. Intern. Med. 267, 89Âą105 (2010). Ai, J. et al. Nanotoxicology and nanoparticle safety in biomedical designs. Int. J. Nanomedicine 6, 1117Âą27 (2011). J Nanopart Res. et al. Perspectives on the design of safer nanomaterials and manufacturing processes. September 17, 366 (2015). Beer, C., Foldbjerg, R., Hayashi, Y., Sutherland, D. S. & Autrup, H. Toxicity of silver nanoparticlesÂ˛Nanoparticle or silver ion? Toxicol. Lett. 208, 286Âą292 (2012).

Quinazolinediones-derivatives SIRT6 inhibitors for targeted selective chemotherapy Teixeira, M.C.1*, Fernandes, A.R.1, .RYDĂžHYLĂź $ % 1, Sanchez-Lopez, E.2,3, Espina, M.2,3, Garcia, M. L.2,3, Veiga, F.J.1,4, Calpena, A.C.3,5, Souto, E.B.1,4 1

Department of Pharmaceutical Technology, Faculty of Pharmacy, Coimbra University, Azinhaga de Santa Comba 3000-548, Coimbra, Portugal 2 Department of Physical Chemistry, Faculty of Pharmacy, University of Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain 3 Institute of Nanoscience and nanotechnology (IN2UB). Faculty of Pharmacy, University of Barcelona, 08028 Barcelona, Spain 4 REQUIMTE/LAQV, Group of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal 5 Department of Pharmacy and Pharmaceutical Technology, Biopharmaceutical and Pharmacokinetic Unit, Faculty of Pharmacy, University of Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain #Presenting

author: mceuteixeira1@gmail.com

Abstract The occurrence of the high levels of reactive oxygen species (ROS) in tumors is responsible for the enhanced sensitivity of neoplastic cells to oxidative stress [1]. On the other hand, the increase of oxidative stress in these cells may trigger their capacity to adapt to this environment, resulting in ROSmediated cell death. Chemotherapy may be regulated via ROS-mediated signaling and this has been already documented for novel quinazolinediones. Knowing that NAD+-dependent sirtuin SIRT6 is overexpressed in several types of cancer cells [2-4], efforts have been made towards the identification of quinazolinediones-based SIRT6 inhibitors useful for chemotherapy. SIRT6 deficient cells also show higher sensitivity to chemotherapeutics. Indeed, the oncogenic role of SIRT6 in different cancer types have been already described. Thus, combining these two approaches, synergistic effects are expected. In this work, we have carried out a screening of SIRT6-targeting agents based on quinazolinediones GHULYDWLYHV 6HOHFWLRQ FULWHULD LQFOXGHG WKH DYDLODEOH ELRORJLFDO GDWD DERXW WKH GUXJVÂś FDSDFLW\ WR L increase the cellular glucose uptake, (ii) reduce the production of THF-ÄŽ DQG LLL WR LQFUHase acetylation of histone-3-lysine-9. The half maximal inhibitory concentration (IC50) reported in the literature were used to rank the drugs with respect to their selectivity. A set of compounds, that can be used to sensitize tumor cells against conventional anti-neoplastic drugs, have been identified and ranked accordingly to their effectiveness and selectivity. We report on the systematic meta-analysis to document and correlate them with different types of cancer. References [1] E.B. Souto, S. Doktorovova, A.M. Silva, M. Vazzana, A. Santini, C. Faggio, The role of epigenetic regulator SIRT6 in skin tumorigenesis, 88th SIBS National Congress on Biology and Human Health: Theoretical and Translational Research Aspects, Genoa, Italy, 11-12 December (2015), (2015). [2] M. Cea, A. Cagnetta, S. Adamia, C. Acharya, Y.T. Tai, M. Fulciniti, H. Ohguchi, A. Munshi, P. Acharya, M.K. Bhasin, L. Zhong, R. Carrasco, F. Monacelli, A. Ballestrero, P. Richardson, M. Gobbi, R.M. Lemoli, N. Munshi, T. Hideshima, A. Nencioni, D. Chauhan, K.C. Anderson, Evidence for a role of the histone deacetylase SIRT6 in DNA damage response of multiple myeloma cells, Blood, 127 (2016) 1138-1150. [3] D. Lasiglie, S. Boero, I. Bauer, S. Morando, P. Damonte, M. Cea, F. Monacelli, P. Odetti, A. Ballestrero, A. Uccelli, R. Mostoslavsky, A. Poggi, A. Nencioni, Sirt6 regulates dendritic cell differentiation, maturation, and function, Aging, 8 (2016) 34-49. [4] L.K. Ran, Y. Chen, Z.Z. Zhang, N.N. Tao, J.H. Ren, L. Zhou, H. Tang, X. Chen, K. Chen, W.Y. Li, A.L. Huang, J. Chen, SIRT6 Overexpression Potentiates Apoptosis Evasion in Hepatocellular Carcinoma via BCL2-Associated X Protein-Dependent Apoptotic Pathway, Clinical cancer research : an official journal of the American Association for Cancer Research, 22 (2016) 3372-3382. Acknowledgements The authors would like to acknowledge the financial support received through the projects M-ERANET/0004/2015 and UID/QUI/50006/2013, from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and co-financed by FEDER, under the Partnership Agreement PT2020.

Novel Fe-Mn-Si-Pd dense and porous alloys: insights on mechanical, magnetic and corrosion performance Y.P. Feng , N. Gaztelumendi , J. Fornell , H. Y. Zhang , P. Solsona 0 ' %DUČŠ , S. SuriĂąach , E. 1

IbĂĄĂąez , E. GarcĂa-Lecina , L. Barrios , E. Pellicer , C. NoguĂŠs , J. Sort

Departament de FĂsica, Universitat AutĂ˛noma de Barcelona, E-08193 Bellaterra, Spain

Departament de Biologia Cellular, Fisiologia i Immunologia, Universitat AutĂ˛noma de Barcelona,

E-08193 Bellaterra, Spain 3

Surfaces Division, IK4-CIDETEC, Parque TecnolĂłgico de San SebastiĂĄn, E-20009 Donostia,

Spain 4

InstituciĂł Catalana de Recerca i Estudis AvanĂ§ats (ICREA) and Departament de FĂsica, Universitat

AutĂ˛noma de Barcelona, E-08193 Bellaterra, Spain *Presenting author. E-mail address: Yuping.Feng@uab.cat Abstract In this study, new quaternary Fe-Mn-Si-Pd alloys have been investigated, both in bulk and porous forms. Bulk fully-compact Fe-30Mn6Si1Pd alloy has been fabricated by arc-melting followed by copper mold suction casting. The allo\ PDLQO\ FRQVLVWV RI Ä°-PDUWHQVLWH DQG Č&#x2013;-austenite Fe-rich phases with minor content of Pd-rich precipitates. The presence of these two antiferromagnetic phases assures nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) compatibility. Good mechanical response was observed by nanoindentation: a hardness value around 4.2 GPa and a UHGXFHG <RXQJÂśV modulus value of 93 GPa were obtained; KRZHYHU HYHQ LI WKH <RXQJÂśV PRGXOXV LV ORZHU WKDQ WKH RQH of the 316L stainless steel, which is one of the most common Fe-based reference materials for ELRPHGLFDO DSSOLFDWLRQV D <RXQJÂśV PRGXOXV FORVHU WR WKDW RI KXPDQ ERQH -20 GPa) would help to avoid the stress shielding effect and subsequent loosening of the implant. With this purpose, porous Fe30Mn6Si1Pd alloys were fabricated by a simple press and sinter process from blended elemental powders with 10-40 wt.% of NaCl, as porogen (space holding), to obtain different degrees of porosity. %\ LQWURGXFLQJ SRURVLW\ <RXQJÂśV PRGXOXV values ranging from 55 GPa to 7 GPa were subsequently obtained. The degradation performance was evaluated by static immersion and electrochemical test in +DQNÂśV VROXWLRQ The influence of immersion in simulated body fluid on composition, microstructure, mechanical and magnetic properties of both alloys was assessed, and the correlation between microstructure evolution and physical properties will be discussed.

Metastatic Skin Cancers: Identification of Chemotherapeutic Targets for Nanotherapeutics 1*

1,2

3,4

Fernandes, A.R. , Teixeira, M.C. , Santos, A.C. , Sanchez-Lopez, E. , Espina, M. , Garcia, M. L. , 1,5 3,6 1,5 Veiga, F.J. , Calpena, A.C. , Souto, E.B. 1

Department of Pharmaceutical Technology, Faculty of Pharmacy, Coimbra University, Azinhaga de Santa Comba 3000-548, Coimbra, Portugal 2 Institute for Innovation and Health Research, Group Genetics of Cognitive Dysfunction, Institute for Molecular and Cell Biology, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal 3 Department of Physical Chemistry, Faculty of Pharmacy, University of Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain 4 Institute of Nanoscience and nanotechnology (IN2UB). Faculty of Pharmacy, University of Barcelona, 08028 Barcelona, Spain 5 REQUIMTE/LAQV, Group of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal 6 Department of Pharmacy and Pharmaceutical Technology, Biopharmaceutical and Pharmacokinetic Unit, Faculty of Pharmacy, University of Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain #

Presenting author: anaritavfernandes@hotmail.com

Abstract Among the different types of human skin cancers, melanoma is recognized as the major and most aggressive type, being highly invasive and proliferative. Squamous cell carcinoma (SCC) is also known for its high metastatic risk [1, 2]. The treatment of metastatic skin cancers continues to be challenging, and the identification of several targets for chemotherapy is nowadays a demand [3]. Indeed, experimental data recently available demonstrate that the occurrence of metastasis can be triggered by the classical treatment with neoplastic drugs and/or radiotherapy. To reach and colonize distant cluster of healthy cells, the tumor cells have to follow a step-by-step pathway, namely, (i) cell detachment from the primary tumor, (ii) tumor cell invasion and intravasation to achieve systemic circulation, (iii) extravasation and implantation in the target cells, and (iv) angiogenesis. In this pathway, the most important step is the cells detachment from the primary tumor and their adhesion to the healthy cells of the target. Neoplastic cells have a reduced cell-matrix adhesion, being therefore highly invasive. The current treatment of invasive skin cancers make use of chemotherapy of the primary lesions. However, this approach has limited successful rate because the cells of primary lesions and metastatic cells react different against chemotherapeutics. A case is made about the need for more effective, targetedspecific approaches for metastatic skin cancers. We have revised the most recent pre-clinical data on the treatments under development for metastatic skin cancers. The most relevant approaches include the interference with the un-controlled mechanisms involved in the integrity of cell membrane and cell adhesion (e.g. integrin receptors, selectins, matrix metalloproteinases, actin microfilaments, myosis activity, and cell surface immunoglobulins). References [1] K.A. Burton, K.A. Ashack, A. Khachemoune, Cutaneous Squamous Cell Carcinoma: A Review of High-Risk and Metastatic Disease, American journal of clinical dermatology, (2016). [2] L. Nissinen, M. Farshchian, P. Riihila, V.M. Kahari, New perspectives on role of tumor microenvironment in progression of cutaneous squamous cell carcinoma, Cell and tissue research, (2016). [3] E.B. Souto, S. Doktorovova, S. A.M., G. M.L., C. B., New nanotechnological approaches against malignant proliferation of epidermal keratinocytes for the treatment of multifocal Actinic Keratosis (AK) and Skin Squamous Cell Carcinoma (SCC) XI SPLC-CRS Conference, Granada, Spain, 21-23 January, (2016). Acknowledgements The authors would like to acknowledge the financial support received through the projects M-ERANET/0004/2015 and UID/QUI/50006/2013, from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and co-financed by FEDER, under the Partnership Agreement PT2020. The authors also acknowledge FCT for the individual fellowship SFRH/BD/109261/2015 granted to ACS.

Amphiphobic coatings for antifouling protection in sea environment Michele Ferrari, Francesca Cirisano, Alessandro Benedetti, Libero Liggieri, Francesca Ravera, Eva Santini CNR- ,&0$7( ,VWLWXWR GL &KLPLFD GHOOD 0DWHULD &RQGHQVDWD H GL 7HFQRORJLH SHU OÂś(Qergia, via De Marini, 6, 1649 Genova-Italy michele.ferrari@ge.icmate.cnr.it Abstract Amphiphobic materials could be an innovative solution where technological and ecological aspects allow to be merged, taking into account the limitations imposed by international laws in terms of environmental protection. Protection from fouling and corrosion are key issues for drag reduction and fuel consumption, material degradation, desalination plants and so on. In marine field the use of highly hydro and olephobic (amphiphobic) materials is relatively young and not very explored. In this work amphiphobic/superamphiphobic (SA) coatings has been characterized and tested for marine applications both in laboratory and field conditions, since investigations in real seawater are crucial to evaluate the behaviour of SA surfaces because of a complexity not reproducible in laboratory. Because of the real conditions where the surface can operate, preliminary tests of wearing, thermal stress and durability have been also performed in order to study amphiphobic systems for different applications related to the marine environment.

Acknowledgements: The Authors acknowledge the RITMARE (La Ricerca Italiana per il Mare) Flagship Project for the financial support. This work has been done under the umbrella of COST Action MP1106 and CM1101

References [1] M Ferrari, A.Benedetti Advances In Colloid And Interface Science, Vol. 222, P. 291-304, (2015) [2] F.Cirisano, A.Benedetti, L.Liggieri, F.Ravera, E.Santini, M.Ferrari Colloids and Surfaces A: Physicochemical and Engineering Aspects Volume 505, 20 September 2016, Pages 158Âą164 (2016) .

Multicomponent aliphatic polyester-based nanoparticles for antigen delivery and immune modulation against cancer disease Carina Peres

1, 2, 3*

, Vanessa Sainz , Ana I Matos , Ana S Viana , Liane Moura , LuĂs GraĂ§a , VĂŠronique 3 1 PrĂŠat , Helena F Florindo

Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 2 Lisbon, Portugal, Instituto de Medicina Molecular (IMM), Faculty of Medicines, Universidade de Lisboa, 3 Lisbon, Portugal, Louvain Drug Research Institute (LDRI), FacultĂŠ de Pharmacie, UniversitĂŠ Catholique 4 de Louvain, Brussels, Belgium, Centro de QuĂmica e BioquĂmica, Faculty of Sciences, Universidade de Lisboa, Lisbon, Portugal hflorindo@ff.ulisboa.pt Abstract Dendritic-cell (DC) targeting has been used as a promising strategy for the development of vaccines and immune modulation [1]. Several studies proved that aliphatic polyester-based biodegradable nanoparticles (NPs) are potential vaccine delivery systems for cancer and infectious diseases and suitable platforms for immune modulation [2, 3]. We aim at producing biodegradable NPs to target antigen-presenting cells (APCs) and evaluate their potential to be used as vaccine delivery systems. Different formulations of PLGA/PLA NPs with tumor antigens, immune adjuvants, and gene regulators, such as small interfering RNA (siRNA), were developed by the state of the art double emulsion solvent evaporation technique. Mannose-grafted hybrid lipid/polymeric NPs and hyaluronic acid-coated NPs have also been prepared to specifically target antigen-presenting cells (APCs), as DCs, and tumor cells, respectively. NP size, surface charge (ZP) and morphology were analyzed by Dynamic Light Scattering, Laser Doppler Electrophoresis and Atomic Force Microscopy (AFM), respectively. Antigen entrapment efficiency (EE) and loading capacity (LC) were quantified by HPLC, while siRNA EE and LC were determined by PicoGreenÂŽ reagent. Finally, cell viability was determined by Alamar BlueÂŽ assay. NPs demonstrated to be a suitable platform to deliver multiple tumor-associated antigens, including melanoma and breast cancer peptide antigens, immune adjuvant molecules, such as the Tolllike receptor ligands (TLRls) and also gene regulators, as siRNA. Generally, NPs presented a mean diameter below 200 QP ZLWK ORZ SRO\GLVSHUVLW\ LQGH[ 3G, YDOXHV Â&#x201D; DQG DQWLJHQ and siRNA entrapment efficiency (EE) values of 75 % and 95 % respectively. NP formulations did not affect the viability of targeted cells after 72 h of incubation, even at high NP concentrations. In vitro cellular uptake studies evidenced that both polymeric and hybrid NPs were efficiently taken up by DCs, in a time and concentration-dependent manner. In melanoma and breast murine models, the produced NPs vaccine delivery systems demonstrated to significantly decrease tumor growth rate, especially when the delivery of tumor associated antigens and TLRls by NPs was combined with the natural killer T cell (NKT) ligand galactosylceramide or a siRNA to silence immune-suppressive cytokines. Our results confirm the successful development of biodegradable polymeric nanocarriers with great potential to be use for vaccine delivery and immunotherapy.

References [1] Conniot J et al, Front in Chemistry 2 (2014) 1-27 [2] Silva JM et al, J Control Release, 198 (2015) 91-103. [3] Silva JM et al Nanomedicine (Lond.) 9 (2014) 2639-2656. * equal contributors

Multimodal Inorganic Nanoparticles to Overcome Cancer Drug Resistance Muriel Freixanet 9DOO GÂś+HEURQ ,QVWLWXW GH 5HFHUFD 9+,5 3J 9DOO GÂś+HEURQ -129, Barcelona, Spain muriel.freixanet@vhir.org Abstract Cancer is one of the leading causes of morbidity and mortality worldwide and it is expected to become the major cause of death in the coming decades. Despite remarkable progress made in prevention, diagnosis and treatment, efficient cancer therapies remain elusive. The major causes that lead to treatment failure are: late stage diagnosis, drug penetration to tumors, the lack of adequate approaches to treat metastases and tumor resistance. Although notable advances have been produced in the field of cancer treatment, drug resistance and relapse are still frequent. Multi-drug resistance (MDR) is defined as a state of resistance against structurally and/or functionally unrelated drugs. Mechanisms of drug resistance can be differentiated in intrinsic or acquired, if these features were present before chemotherapy or developed as a response of it, respectively. Initially it was related exclusively to efflux transporters, drug kinetics and drug targets; however, more recent studies revealed that tumor heterogeneity is a major driver, which comprises genetic variation, microenvironment and cell plasticity. Current anticancer therapy strategies mainly include chemotherapy, radiotherapy and surgery. So far these therapeutic strategies alone or in combination fail to eradicate cancer MDR cells, instead favour the expansion of the MDR cell clones, which ultimately leads to relapse and the new tumors become more malignant. Thus, the development of effective treatments may address tumor heterogeneity to better design new therapeutic approaches to overcome drug resistance. Nanotechnology offers a novel set of smart drug delivery systems with unique physicochemical characteristics that optimize drug delivery improving treatment outcome and reducing toxic side effects. These nanocarriers are able to selectively target the tumor, and combine two or more therapeutic agents to overcome drug resistance and effectively eradicate the pool of MDR-cells. Consequently: i.- To fight against resistance: multimodality. It is becoming widely known that none of the existing single-modality treatments can cure fatal diseases such as cancer. Consequently, a combination of these treatments have been investigated for their synergistic effects that may dramatically improve outcomes and reduce the side effects of each single modality treatment. This is because therapeutic effects of the different treatments are designed to add up, while their side effects do not, they are distributed. In this context, multimodality appears to be a very promising strategy for both, to create therapies where resistance cannot be developed against, and to beat already resistant tumoral cells. ii.- To develop multimodality: nanoparticles. Inorganic NPs are suitable platforms to: i) combine more than one drug and targeting ligands unifying and synchronizing their pharmacokinetic profiles and ii) absorb different wavelength photons for imaging, radio and thermal therapy. Inorganic NPs which EHKDYH DV ÂłDUWLILFLDO DWRPVÂ´ GXH WR WKHLU KLJK GHQVLW\ RI HOHFWURQLF VWDWHV - which controls many physical and chemical properties - can be extensively and easily adjusted by tuning their composition, size, shape and surface state. NPs can be multi-functionalized with different organic and biological molecules carrying different chemical and biological functionalities. Perhaps more important, however, is the possibility to combine a series of advances in a single unit which enables the creation of multimodal nanosized therapeutical platforms. References [1] Bae, Y. H. & Park, K. Targeted drug delivery to tumors: myths, reality and possibility. J. Control. Release 153, 198Âą205 (2011). [2] Parhi, P., Mohanty, C. & Sahoo, S. K. Nanotechnology-based combinational drug delivery: an emerging approach for cancer therapy. Drug Discov. Today 17, 1044Âą52 (2012). [3] Iyer, A. K., Singh, A., Ganta, S. & Amiji, M. M. Role of integrated cancer nanomedicine in overcoming drug resistance. Adv. Drug Deliv. Rev. 65, 1784Âą802 (2013). [4] Hanahan, D. & Weinberg, R. A. Hallmarks of cancer: the next generation. Cell 144, 646Âą74 (2011).

Improving an in vitro 3D co-culture as an alternative model of intestinal barrier for the assessment of engineered nanomaterials (ENMs) risk Alba García Rodriguez, Laura Rubio, Laura Vila, Alba Hernández, Ricard Marcos Grup de Mutagènesi, Departament de Genètica i de Microbiologia, Facultat de Biociències, Universitat Autònoma de Barcelona, Bellaterra, Spain; CIBER Epidemiología y Salud Pública, ISCIII, Spain. alba.garcia.rodriguez@uab.cat Abstract The use of nano-based consumer products is growing in an exponential way. In particular, engineered nanomaterials (ENMs) are present in the alimentary sector and in many other industries. For instance, silver, silica and titanium dioxide nanoparticles (Ag-, SiO2-, TiO2-NPs) are used in food packaging as permitted food additives. In this way, the small intestinal epithelium became the primary area for absorption and metabolism of the putative ingested NPs. This epithelium is the main barrier preventing molecules from intestinal lumen to reach the blood compartment [1, 2]. Since NPs exposure by the gastrointestinal tract seems to be unavoidable, it is necessary to assess the health impact of human exposure to NPs and to stablish a common regulatory testing. Nowadays to study the mechanism of translocation of nanoparticles over the gastrointestinal wall, several in vitro models for the intestinal epithelium have been developed. Undoubtedly, the human colon adenocarcinoma cell line Caco-2 has been the most widely used in the study of intestinal permeability to foreign compound over the last decade [3]. In the present study, the Caco-2 cell model has been modified and improved in order to mimic the real conditions, trying to set up a realistic intestinal environment and getting a clear understanding of how ENMs interact with the intestinal mucosa. Thus, an alternative in vitro permeability model system based on a triple culture, mixing enterocyte-like cells (Caco-2), mucus-secreting goblet cells (HT29) and M cells with a transcytosis feature, altogether in the same transwell [4]. To validate the model a wide battery of techniques such as the measurement of the transepithelial resistance value and the detection of the Lucifer Yellow permeability was used to characterize the 3D co-culture integrity. On the other hand, innovative techniques to characterize the membrane like the use of confocal fluorescent microscopy or by detecting specific molecular markers have also been tuned up (see figure). Our results lead us to conclude that the 3D intestinal membrane works properly and further support the potential use of this in vitro 3D co-culture system in translocation, kinetic, cytotoxic and genotoxic studies,

mainly

the

nanotoxicology field.

References [1] Chaudhry et al. 2008. Applications and implications of nanotechnologies for the food sector. Food Addit Contam Part A, 25: 241-258. [2] Bouwmeester et al. 2011 Characterization of translocation of silver nanoparticles and effects on whole-genome gene expression using an in vitro intestinal epithelium coculture model. ACS Nano, 5: 4091-4103. [3] Schimpel et al. 2014 Development of an advanced intestinal in vitro triple culture permeability model to study transport of nanoparticles. Mol. Pharm. 11: 808-818

Preparation and characterization of nanoceria-loaded liposomes a,b

Agostina Francesca Grillone , Tianshu Li , Shinji Takeoka , Gianni Ciofani agostina.grillone@iit.it

a,d

Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, Viale Rinaldo Piaggio 34, 56025 Pontedera (Pisa), Italy b Scuola Superiore Sant'Anna, The BioRobotics Institute, Viale Rinaldo Piaggio 34, 56025 Pontedera (Pisa), Italy c Waseda university, 7RN\R :RPHQÂśV 0HGLFDO 8QLYHUVLW\ Center for advanced Biomedical Sciences, 8-1 Kawadacho, 162-8666 Tokyo, Japan d Politecnico di Torino, Department of Mechanical and Aerospace Engineering, Corso Duca degli Abruzzi 24, 10129 Torino, Italy Abstract Introduction: Cerium oxide nanoparticles (nanoceria) represent an interesting class of nanomaterials because presenting both pro- and anti-oxidant features [1]. More specifically, it has been shown that nanoceria act as a reactive oxygen species scavenger by protecting cells against oxidative stress, yet that they also own cytotoxic and anti-invasive properties toward cancer cells, by triggering ROS accumulation at low pH values [2]. Aim of this work is the encapsulation of nanoceria in liposomes, in order to increase nanoparticles colloidal stability, and to selectively target them those scenario where oxidative stress plays a key role, and therefore where nanoceria might represent a potential pharmacological care. Experimental methods: Nanoceria-loaded liposomes were prepared through reverse phase evaporation method, by using as lipid formulation 1,2-dipalmitoyl-sn-glycero-3phosphocholine (DPPC), cholesterol, 1,5-dihexadecyl-succcinic anhydrate (DHSG), and 1,2-distearoylsn-glycero-3-phosphoethanolamine-N-monomethoxy poly(ethylene glycol) (PEG-DSPE) at a molar ratio of 5:5:1:0.03. 5 mg of lipids and 50 mg of nanoceria with size of about 11 nm were dissolved in ethanol. After removal of the solvent by rotary evaporation, the resulting dried lipid-nanoceria mixed film was dissolved in water at room temperature for three hours. Liposome suspension was then extruded with membrane filters and purified through gel chromatography. Obtained hybrid nanovectors were extensively characterized in terms of size distribution and Z-potential by dynamic light scattering, and morphology through transmission electron microscopy (TEM). Finally, loading efficiency was assessed with inductively coupled plasma (ICP) mass spectroscopy analysis. Results: Nanoceria-loaded liposomes showed a size of about 200 nm in diameter, and Z-potential of about 30 mV. TEM imaging highlights the formation of well-defined spherical liposomes and confirms the presence of cerium oxide nanoparticles inside and on the surface of the nanovectors. Loading efficacy was around 15%. Conclusion: Our results, even if preliminary, demonstrate the possibility to exploit liposomes as carriers for cerium oxide nanoparticles. Future mandatory investigations will aim at obtaining higher loading efficiency and at the assessment of the unaltered catalytic activity of the entrapped nanoceria in vitro. Moreover, the possibility to functionalize the surface of the lipid structures with appropriate ligands for a targeted delivery will be also considered.

20 nm

100 nm

Figure 1: TEM imaging of cerium oxide nanoparticles (left) and of nanoceria-loaded liposomes (right) References [1] Ciofani G, Genchi GG, Mazzolai B, Mattoli V. Biochimica et Biophysica Acta. 2014; 1840(1): 495506. [2] Celardo I, Pedersen JZ, Traversa E, Ghibelli L. Nanoscale. 2011; 3(4): 1411-1420.

Magnetic molecularly imprinted polymer for the detection of methyl parathion in fish ac

Amal H.A. Hassan , Laura Martinez Fajardo , Silio Lima Moura , Luciano Sappia , Fatma H.M. Ali , b c Walaa A. Moselhy , Maria Isabel Pividori a

Food Hygiene and Control department, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, 62511, Egypt. b Forensic Medicine and Toxicology department, Faculty of Veterinary Medicine, Beni-Suef University, , Beni-Suef, 62511, Egypt. c Group of sensors and Biosensor, Chemistry department, Faculty of Science, UAB, 08193,Bellaterra (Barcelona),Spain. emyhamdy41@gmail.com

Abstract Fish is one of the most popular foods constituting an exceptionally important component of the human diet. It contains high level of protein, minerals, vitamins and omega 3. Despite of the dietary and economical benefits of fish and due to environmental pollutions, fish are exposed to contamination with different types of chemical hazards. Methyl parathion is a widely used and highly toxic organophosphorus pesticide [1].It tends to accumulate in water, soil and fish due to its low solubility and bioaccumulation properties causing serious health and environmental problems [2]. Ingestion of contaminated fish with methyl parathion is considered one of the most important routes for human toxicity, so fish samples should be accurately and continuously examined. Continuous monitoring of methyl parathion residue in fish muscles required a highly sophisticated, labor and time consuming instruments. Since the early reports on magnetic separation technology [3], magnetic particles (MPs) have been used as a powerful and versatile preconcentration tool in a variety of analytical and biotechnology applications. This technology has been widely incorporated for researchers worldwide in classical methods, in molecular tools (PCR, immunoassays) and in emerging technologies including biosensors [4] and microfluidic devices with application ranging from biomarker detection of infectious diseases [5] foodborne pathogens [6], among many others. The use of MPs greatly improves the performance of the biological reaction by increasing the surface area, improving the washing steps and, importantly, minimizing the matrix effect. MPs also allow reduction of reaction times and reagent volumes. In addition, MPs can be easily magneto-actuated using permanent magnets [7]. Beside the amazing properties of MPs, the main drawback of the biologically-modified MPs is their high cost and low stability. Molecularly Imprinted Polymers (MIPs) are synthetic biomimetic materials mimicking biological receptors. [8].they are highly cross-linked macromolecular structures towards the template which is then extracted after polymerization, originating cavities (binding sites) complementary to the template molecule, acting as plastic antibodies.[9].Although MIPs have in general lower affinity and selectivity compared to the biological counterparts, they show important technological features: i) they can be easily and affordably synthesized on a animal-free large scale procedures, and ii) they show high chemical and mechanical stability, allowing to work in harsh conditions. In order to merge the outstanding properties of MIPs and magnetic particles, a novel hybrid molecularly-imprinted polymer with magnetic properties (magnetic-MIP) is presented with affinity towards methyl parathion for the pre concentration of this pesticide from complex samples. During the synthesis of the magnetic core-shell particles, methyl parathion was used as a template. The magnetic-MIP was synthesized through a coreshell strategy using methacrylic acid as a functional monomer, EGDMA as cross-linking monomer and AIBN as radical initiator (as shown in Fig 1). The selection of the functional monomer was investigated by theoretical calculation of the formation energies of complexes using the density functional theory (DFT) with the program package Gaussian 09. The solvent effect was taken into account by means of a dielectric continuum. The characterization of this material by microscopy techniques including SEM and TEM is presented. Moreover, the binding capacity towards methyl parathion is demonstrated and compared with the corresponding magnetic-NIP (non-imprinted polymers). This material showed the synergic advantages of MIPs and MPs, including low cost of production, stability and magnetic actuation as promising features for their integration in magneto-actuated approaches for the examination of seafood in resource-constrained settings. References: [1] Silva D., Cortez C.M., Cunha-Bastos J., Louro S.R.W., Toxicology Letters, 147(2004) 53. [2] Lagaly, G. Appl. Clay Sci., 18 (2001) 205. [3] Rembaum, A., Yen, R.C.K, Kempner, D.H., Ugelstad, J., J Immunol Methods, 52(1982) 341. [4] Zacco, E., Pividori, M.I., Alegret, S., Galve, R., Marco, M.P., Anal Chem.78 (2006) 1789. [5] Carinelli, S., Martí, M., Alegret, S., Pividori, M. I., New Biotechnol.32 (2015) 521. [6] Brandão, D., Liébana, S., Pividori, M. I., New Biotechnol. 32 (2015a) 511. [7] Zacco, E., Pividori, M.I., Alegret, S., Galve, R., Marco, M.P., Anal Chem.78 (2006) 1789-1798. [8] Vlatakis G, Andersson LI, Müller R, Mosbach K, Nature, 361 (1993) 645. [9] Haupt K, Nature Mate, 9 (2010) 612.

. NH3H2O

MPS

NH3.H2O+TEOS

Fe3O4

o 30 min 80 c

Fe3Cl2.4H2O + FeCl3.6H2O

Figures

ETOHo 12h 25 c

Fe3O4@SiO2

Anhydrous toluene o 12h 25 c

Fe3O4@SiO2-MPS

Rebinding EGDMA,AIIBN

Methyl parathion o

24h 60 c Eluting

MMIP without MP

MP- MMIP

Figure1.Synthesis of MP-magnetic molecular imprinted polymers

Methacrylic acid

Molecularly imprinted polymers: functional nanoplastics for diagnostics and specific sorption Anna Herrera-Chacon, Andreu González-Calabuig, Inmaculada Campos and Manel del Valle Sensors and Biosensors Group, Department of Chemistry, Universitat Autònoma de Barcelona, Edifici Cn, 08193 Bellaterra, Barcelona, Spain anna.herrera@uab.cat Abstract Molecularly imprinted polymers (MIP), alternatively known as nanoplastics, are polymers furnished with supramolecular chemistry principles that have proven their potential as synthetic receptors. These materials are able to recognize biological and chemical species, and they do this applying the same principles as their biological counterparts: the host-guest principles. Due to this fact, these materials have drawn the attention of the scientific community who has already proposed their use in fields like medical diagnostics, solid phase extraction, sensors, specific sorption, or controlled release, among others. For their conception, computational modeling may be used, as this powerful tool can reduce the amount of experimental work choosing among the different options available for the synthesis of the desired material. This computational approach is mainly composed of two steps: the screening of the possible functional monomers and, once the adequate monomer is selected, the determination of the template:monomer stoichiometric ratio. The synthesis starts with the mixing of the monomer and the template in an appropriate solvent, for their prearrangement. Afterwards it is added the crosslinker and the radical initiator into the mixture. The system is purged with nitrogen and then it is shelled and heated in a water bath. After the polymerization, the template is removed from the mixture by a Soxhlet reflux. The non-imprinted polymer (NIP) is synthesized for control purposes in the same conditions than MIP but without the addition of the template. Once the synthesis is completed, these materials need to be characterized, especially concerning its expected functionality. For the characterization, visualization strategies like scanning electron microscopy can be necessary. The scanning electron microscopy give us valuable information that we transform on statistical data to develop a histogram and study the particle size, the distribution and its standard deviation to show its homodispersion. The chemical interaction, in its thermodynamic aspects, can be studied with a binding study and a Scatchard analysis. A Scatchard plot is a saturation binding experiment; you vary the concentration of template and measure binding after an equilibration time. The goal is to determine the Kd (affinity constant) and Bmax (specific binding capacity). Finally, the use as sorbent material can be evaluated using the material in a solid phase extraction process. Packing a column with the MIP, a preconcentration experiment can be performed. A volume of sample that contains the analyte is passed through the column, and afterwards, the captured analyte is eluted and measured by electrochemical techniques; the obtained ratio permits to obtain the imprinting factor. Overall the work represents a multidisciplinary approach that combines different chemistry areas to achieve an optimal polymer with expected functionalities. References [1] Bates, F., & del Valle, M. (2015). Voltammetric sensor for theophylline using sol±gel immobilized molecularly imprinted polymer particles. Microchimica Acta, 182(5-6), 933-942. [2] Matsui, J., Miyoshi, Y., Doblhoff-Dier, O., & Takeuchi, T. (1995). A molecularly imprinted synthetic polymer receptor selective for atrazine. Analytical Chemistry, 67(23), 4404-4408. [3] Bates, F., CelaǦPérez, M. C., Karim, K., Piletsky, S., & LópezǦVilariño, J. M. (2016). Virtual Screening of Receptor Sites for Molecularly Imprinted Polymers. Macromolecular bioscience.

Fig.1: Schematic procedure of molecularly imprinting technique

Nanomechanics of the Drosophila embryonic central nervous system 1,2

Ignasi Jorba , Katerina Karkali , Anand P. Singh , Timothy E. Saunders , Daniel Navajas 3 Enrique Martin-Blanco

1,2,5

Unitat de BiofĂsica i Bioenginyeria, Universitat de Barcelona, Barcelona, Spain 2 Institute for Bioengineering of Catalonia, Barcelona, Spain 3 Instituto de Biologia Molecular de Barcelona (IBMB-CSIC), Barcelona, Spain 4 Mechanobiology Institute, National University of Singapore, Singapore 5 CIBER de Enfermedades Respiratorias, Madrid, Spain dnavajas@ub.edu

Cell functions can be controlled through purely mechanical means and the impact of mechanical forces is decisive in how tissues coordinate the unfolding of their developmental program to build functional and structurally optimized organs. Understanding not only how cells generate forces, but also how cells in turn respond to forces is of fundamental importance in biology. One neglected aspect of current initiatives exploring the brain is the study of nervous system biomechanics. To tackle this subject, we are studying the development of the Drosophila embryonic Central Nervous System (CNS), a model that allows imaging in vivo and where significant genetic tools are available. Within our general aim of establishing a 4D mechanical map of the developing brain, here we characterized the stiffness of the CNS of Drosophila embryos measured by atomic force microscopy (AFM). Embryos at stage 16 (n=5) were placed on top of positively charged glass slides to immobilize them on a rigid substrate. The embryos were immersed in PBS solution and dissected to expose the CNS allowing AFM measurements. Force-indentation curves were obtained with a custom-built AFM mounted on an inverted optical microscope. A 20 Âľm diameter polystyrene bead was glued (2-part epoxy) to the end RI D WLSOHVV FDQWLOHYHU QRPLQDO VSULQJ FRQVWDQW N 1 P )RU HDFK HPEU\R WKH <RXQJÂśV PRGXOXV (E) was measured at 3 positions along the antero-posterior axis in the midline and in lateral regions (left and right separated 20 Âľm from the midline). Five force-indentation curves (ramp amplitude of 20 Âľm and a frequency of 1 Hz) were recorded at each measurement point. E was computed by fitting force-indentation curves for a maximum indentation of 4 Âľm with the Hertz contact model for a sphere indenting an infinite elastic half-space. E of the midline was 237 Âą 63 Pa (mean Âą standard deviation, n=15) and 109 Âą 44 Pa and 120 Âą 36 Pa for each lateral region, respectively. Midline stiffness was significantly higher than those of lateral domains (p < 10-6). No statistical differences were found between the lateral domains (p > 0.4). The Drosophila embryonic CNS is thus extremely soft in correspondence with other measurements in nervous tissues from different organisms or individual neurons. Further, our results indicate that the most central regions of the embryonic CNS, where most axons bundle, are more rigid than the lateral domains, where most of the somata are found. This suggests a role for the fasciculated axons acting as a suspension cable network providing structural support to the developing nervous system.

Advanced probes for multiplexed intracellular biomarker detection a

Maria-Eleni Kyriazi, Afaf H. El-Sagheer, a

Physics and Astronomy,

d,e

Peter Lackie, Tom Brown, Otto Muskens and Antonios a,b G. Kanaras *

Institute for Life Science, University of Southampton, Southampton, SO17 1BJ, UK c Clinical and Experimental Sciences, University of Southampton, Southampton General Hospital, SO16 6YD, UK d Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford, OX1 3TA, UK e Chemistry Branch, Department of Science and Mathematics, Faculty of Petroleum and Mining Engineering, Suez University, Suez 43721, Egypt Corresponding author: a.kanaras@soton.ac.uk

The ability to detect and monitor biomolecules such as mRNA in live cells has been shown to be of utmost importance. For example, changes in mRNA expression levels in individual cells have been associated with the development of diseases such as cancer.[1] Common detection strategies are often based on real-time polymerase chain reaction, microarray analysis or in situ hybridisation. However, the shortcoming of these strategies is the incapability of detecting Âą in the live cell, at real time- cell to cell variations in mRNA expression. Fluorescent nucleic acid probes (such as hairpin based molecular beacons) have been used as an alternative to measure mRNA levels.[2] However, their susceptibility towards nuclease degradation and the fact that they do not penetrate the cell membrane with ease makes nucleic acid delivery into cells a great challenge. Promising developments have been made by associating nucleic acids with gold nanoparticles creating a socalled spherical nucleic acid (SNA).[3, 4] These systems have been shown to successfully penetrate the cell membrane without the aid of transfection agents and resist nuclease degradation.[5] Oligonucleotide-modified gold nanoparticles hybridised to fluorophore complements, which are termed nanoflares, have been successfully used to visualise mRNA in living cells and exhibit high signalling and have low background fluorescence.[3] However, visualising mRNA targets has been limited by utilising one fluorescent output and therefore only one mRNA target can be monitored within the same endocellular microenvironment. This poses a limitation when trying to detect changes in the expression of more than one mRNA target. In this work we present a multiplexed probe consisting of a covalently linked SNA dimer for the simultaneous detection of multiple mRNA targets within the same cellular local microenvironment, focusing on the detection of epithelial cell-based lung cancer. Scheme 1 illustrates the basic concept RI WKLV SUREH Âľ6HQVLQJÂś QXFOHLF DFLGV GHVLJQHG WR FDSWXUH WZR GLIIHUHQW P51$ WDUJHWV DUH DWWDFKHG WR WZR FRYDOHQWO\ OLQNHG JROG QDQRSDUWLFOHV $X13V 6KRUW IOXRURSKRUH EHDULQJ ÂľIODUHÂś VWUDQGV DUH DWWDFKHG WR HDFK ÂľVHQVHÂś VWUDQG 'XH WR WKH FORVH SUR[LPLW\ RI WKH G\H WR WKe AuNP the fluorescence is TXHQFKHG Âľ2)) VWDWHÂś :KHQ D WDUJHW P51$ ELQGV WR WKH VHQVH VHTXHQFH GXH WR FRPSOHPHQWDULW\ the concomitant displacement of the flare can be detected as a corresponding increase in IOXRUHVFHQFH Âľ21 VWDWHÂś YLD FRQIRFDO PLFURVcopy. The use of this probe allows for the simultaneous detection of multiple mRNA targets which is crucial for the early identification of metastatic cancerous

cells as expression levels of specific mRNAs change considerably during tumour progression. We demonstrate that our multiplexed nano-probes are easily taken up by cells whilst resisting nuclease degradation. They exhibit excellent target specificity, can be used for the simultaneous live cell sensing of different mRNAs and can thus be used as a tool to reliably distinguish metastatic cancer cells from healthy ones. In future, we envisage further developing these nano-SUREHV LQWR ÂľVPDUWÂś GUXJ FDUULHUV E\ FRPELQLQJ the detection of metastatic cancer cells with the simultaneous release of a therapeutic drug. Ultimately, these nano-probes may present the next step towards rapid detection of cancer metastasis leading to a new era in personalised medicine. References 1. 2. 3. 4. 5.

Calin, G.A. and C.M. Croce, MicroRNA signatures in human cancers. Nature Reviews Cancer, 2006. 6(11): p. 857-866. Santangelo, P., N. Nitin, and G. Bao, Nanostructured probes for RNA detection in living cells. Annals of Biomedical Engineering, 2006. 34(1): p. 39-50. Seferos, D.S., et al., Nano-flares: Probes for transfection and mRNA detection in living cells. Journal of the American Chemical Society, 2007. 129(50): p. 15477-+. Giljohann, D.A., et al., Oligonucleotide loading determines cellular uptake of DNA-modified gold nanoparticles. Nano Letters, 2007. 7(12): p. 3818-3821. Prigodich, A.E., et al., Nano-flares for mRNA Regulation and Detection. Acs Nano, 2009. 3(8): p. 2147-2152.

Schemes

Scheme 1: Nano-probe illustration

Dynamic Molecular Bio-Interfaces for Controlled Environments Towards Spatial and Temporal Control of Cell Behavior A. R. Kyvik, C. Luque, J. Veciana, I.Ratera Institut de CiĂ¨ncia de Materials de Barcelona (ICMAB-CSIC)/CIBER-BBN Campus de la UAB, 08193 Bellaterra, Spain akyvik@icmab.es Abstract Dynamic molecular interfaces that allow the control of biological interactions and cell behaviour in culture using an external stimulus are relevant for applications in a broad spectrum of scientific areas, such as biology, material sciences and medicine. A dynamic substrate consisting of a self-assembled monolayer (SAM) containing the redox active hydroquinone group has been developed and characterized electrochemically and with XPS. Also, contact angle measurements have been performed. The use of hydroquinone groups enables the spatial and temporal control of the Diels-Alder reaction on surface, which takes place between a cyclopentadiene (cp) moiety and a benzoquinone, the oxidized form of the hydroquinone. Therefore, this presents a platform that at a given time, when a potential is applied, allows for the functionalization of the surface with cp-coupled molecules. In this study, thiols incorporating ethylene glycol units terminated in hydroquinone and hydroxyl were synthesized, and SAMs were formed on gold covered substrates. These where characterized using cyclic voltammetry and the oxidation and reduction peak were clearly visualized. The Diels-Alder reaction with a cp-moiety was also followed with cyclic voltammetry: as expected, the reduction and oxidation peaks of the hydroquinone disappear once the reaction has taken place. Contact angle measurements indicate a change when the SAM is oxidized and XPS results demonstrate that the molecules are anchored onto the gold surface and that the hydroquinone groups are oxidized and react with the cp-moiety, which they are exposed to. Further studies will include the use of micro-contact printing to acquire spatial control and the functionalization of the substrates using cp-fluorescein, cp-RGD and cp-dendrimers (mono and multimodal) for cell culture studies. Figure

BioM

Cyclopentadiene-Biomolecule

Monitoring a bioengineered surface coating replacement by surface enhanced Raman scattering (SERS) on Ag nanocubes and performance in vesicle trapping a,b

Alejandro Lapresta-FernĂĄndez , Paulo J. Silva , Zekiye Pelin GĂźven and Francesco Stellacci

Institute of Materials Science & Engineering, Ă&#x2030;cole Polytechnique FĂŠdĂŠrale de Lausanne, (EPFL) 1015 Lausanne, Switzerland b ECsens, Department of Analytical Chemistry, University of Granada, Granada 18071, Spain alejandro.lapresta@epfl.ch

Abstract Here, we explore the feasibility of using surface enhanced Raman scattering (SERS) to monitor a ligand 1 exchange process on Silver Nanocubes (AgNCs). In this line, we monitored the replacement of Poly(vinyl pyrrolidone), (PVP) by different types of thiolated ligands such as sodium-11mercaptoundecane-sulfonate (MUS) and 1-octanethiol (OT). MUS has an alkane back bone with a hydrophilic anionic sulfonate group, whereas OT is purely hydrophobic and both are known to play a 2 key role in controlling cellular penetration. The PVP is used as capping agent in the shape-assisted synthesis of AgNCs as well as to stabilize their colloidal suspension. During the ligand exchange, different SERS spectra were acquired from 1 min to 96 hours, monitoring a decrease in PVP signal as well as the increase in the Raman signatures regarding MUS and OT over time. We confirmed, by SERS, the successful replacement of PVP in different ligand conditions to develop different monolayers such as (i) all MUS, (ii) different MUS:OT (1:1, 2:1, 5:1 and 9:1) ratios and (iii) all OT. Zeta potential measurements were carried out to verify the variation of surface charge in time on the AgNCs at physiological pH (7.3-7.5), monitoring also their changes in optical properties by UV-vis measurements. Preliminary results provide new insight in the nonspecific AgNCs-lipid interactions by combining the engineered AgNCs with unilamellar lipid vesicles to serve as a model system. The monolayer based on 5:1 (MUS:OT) ratio showed the best characteristics in term of surface charge and stability in high ionic strength media (PBS 1X). On the other hand, homoligand monolayers made only of MUS required a longer reaction time to reach an adequate molecular packing to stabilize the AgNCs, as it issuggested from the Absorbance spectra, thus showing a similar behavior to the 5:1-AgNCs in the presence of vesicles. Therefore, the surface of AgNCs can be engineered via the PVP ligand exchange method, arming the AgNCs with the capacity to trap vesicles in the whole engineered silver surface. b 5:1

Absorbance (a.u)

300

500

Series2 Reference 10m 1h 4h 6h Series1 30h

700 900 Č&#x153; (nm)

100 nm

Figure 1. (a) Raman C-C stretching region for different reaction times in the mix-ligand MUS:OT replacement 5:1 ratio as well as several Raman and SERS used as reference.(b) UV-vis measurements in PBS (1X) and (c) cryo-TEM images showing the vesicle trapping by the AgNCs with a 5:1 monolayer. A. Lapresta-FernĂĄndez acknowledges the AndalucĂa Talent hub Post-doc fellowship supported by the Andalusian Knowledge Agency, COFU1' E\ WKH (8 WK )UDPHZRUN 3URJUDP 0DULH 6NĂĄRGRZVND-Curie DFWLRQV *UDQW $JUHHPHQW QĆ&#x2022; DQG 7KH 0LQLVWU\ RI (FRQRP\ ,QQRYDWLRQ 6FLHQFH DQG Employment of the Junta de AndalucĂa. References [1] C. H. Moran, M. Rycenga, Q. Zhang and Y. Xia, J. Phys. Chem. C, 115,(2011), 21852-21857. [2] R. C. Van Lehn, P. U. Atukorale, R. P. Carney, Y. S. Yang, F. Stellacci, D. J. Irvine and A. Alexander-Katz, Nano Lett., 13, (2013), 4060-4067.

Neutral nanostructures for oligonucleotide delivery to cancer cells 1,4

Bouzo BL , Vazquez-Ríos AJ , Avinjo A , Grijalvo S , Gutiérrez-Lovera C , Sánchez L , López-López 1 4 2 1 R , Alonso MJ , Eritja R , de la Fuente M 1

NanoOncologicals Laboratory, Translational Medical Oncology Group, Health Research Institute of 2 Santiago de Compostela (IDIS), Santiago de Compostela, Spain. Nucleic Acid Chemistry 3 Group, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Barcelona, Spain. Genetic 4 Department, Veterinary Faculty, University of Santiago de Compostela, Lugo, Spain.! Nanobiofar Group, Department of Pharmacy and Pharmaceutical Technology, CIMUS, University of Santiago de Compostela, Santiago de Compostela, Spain.! lopezbouzobelen@gmail.com Abstract Synthetic gene carriers are typically composed of cationic polymers or lipids, which efficiently complex anionic nucleic acids [1]. However, cationic nanocarriers typically interact with serum proteins, lipoproteins, extracellular matrix components, and the complement system, leading to aggregation, early release of the associated nucleic acid, and/or rapid clearance by the reticuloendothelial system. In addition, toxicity showed by this type of formulations has limited their potential in therapeutics [2]. We propose here the design and characterization of a neutral nanostructure composed by natural occurring lipids, non-toxic and able to associate hydrophobically modified oligonucleotides, which is also efficient in delivering them to cancer cells. Our nanostructures can be prepared by a very mild technique, ethanol injection, hence preserving the integrity of these labile biomolecules. They were exhaustively characterized in terms of size, polydispersity and zeta potential, as well as morphologically examined by transmission electron microscopy (TEM) (Figure A). Association efficiency of modified-oligonucleotides to the nanostructures was determined by HPLC. Stability assays were also performed at shelf conditions and in relevant biological media (culture media and plasma). Their interaction with cancer cells was assessed after fluorescent labelling with a hydrophobic fluorophore or with a fluorescent oligonucleotide (Figure B). Transfection experiments were additionally carried out. Furthermore, the toxicity of this formulation, in relation with a formulation that incorporated the cationic transfection agent DOTAP, was explored in zebrafish embryo (Figure C). Results show that following our approach we could obtain monodisperse lipidic nanostructures, prepared in a simple fashion, and avoiding the use of organic solvents other than ethanol. The nanoestructures show good stability under the experimental conditions, and have capacity to interact and enter cancer cells. After association of hydrophobic nucleic acids, we could observe, as they were efficiently deliver to cancer cells in culture and modified the expression of the targeted molecules in vitro. Interestingly, and despite their ability to interact with cells and cross biological membranes, our results show that the nanosystems were non-toxic. After incubation of the nanoestructures with zebrafish embryos, we can similarly conclude that they do not show acute toxicity. Finally, uptake studies in zebrafish embryos indicate that nanoemulsions are apparently able to cross the chorion and reach embryo cells in few hours, confirming their ability to cross biological membranes. In conclusion, the developed nanoestructures shows potential for the intracellular delivery of compounds without infringing toxicity, and therefore have a potential in anticancer gene therapy. References [1] Ozpolat B, et al., Journal of internal medicine, Nanomedicine based approaches for the delivery of siRNA in cancer (2010) 267(1), 44-53. [2] Lv H, et al., Journal of Controlled Release, Toxicity of cationic lipids and cationic polymers in gene delivery (2006) 114(1), 100-109. Figures (B) Cell uptake (SW480)

(A) TEM Images DAPI

Nile Red

Transmitted light

Nile Red

Merge

Development of nanocarriers polyelectrolytes as antimicrobial agents for dual use as drugs and bioimaging probes in human health 1

A. Mira-Carrió , R.Vázquez, Z. Kahveci , C.R. Mateo, A. Falcó, R. Mallavia. 1 Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Elche (a.mira@alu.umh.es) Abstract The combination of polymer science with nanomedicine and nanobiotechnology research fields is a consortium with the potential of addressing complex societal challenges such as the search of new efficient antimicrobial compounds for multidrug-resistance pathogens. In combination with nanotechnology, the development of funtionalized polyelectrolytes (PEs) for diagnostic and therapeutic applications is an interesting approach for the new generation of monitoring tools and treatments in medicine. Thus, the application of this tecnology to obtain more effective diagnostic and therapeutic systems than current ones would increase human wealthfare standards. From this premise, we developed new formulations of PE- based nanocarriers with antimicrobial activity against important bacterial pathogens. Alternatively, within this main objective, it will be also assessed the potential of this compounds as bioimaging probes for the rapid detection of bacterial infections in patients. Regarding the synthesis of PEs, they normally emit blue light and, although researchers in the area have made significant efforts to tune color to longer wavelengths, bright red-emitting PEs are still rare, even if at present they are of great interest for in vivo fluorescence imaging studies because of minimum photodamage to biological samples, deep tissue penetration, and minimum interference from background autofluorescence by biomolecules in the living systems. In this sense, our group has made successful advances and recently described the synthesis of + this kind of molecules (PFNT ) and characterized them and their interaction with two different types of bioactive molecules such as human serum albumin (HSA) and lipid vesicles. Moreover, more successful advances for controlled released purposes have been achieved recently in our laboratory by using a commercial copolymer derived from maleic acid and vinyl ether (PMV) to encapsulate the PEs or components either as nanofibers by electrospinning or as nanoparticles by nanoprecipitation solvent displacement method. Till the date, the development, characterization and optimization of PE-based nanostructured + + systems give us nanostructures containing PFTN or PFP that will be efficient delivery systems for their controlled release in topical (with nanofibers) and/or systemic (with nanoparticles) clinical applications. Some other results are their non toxicity in eukariyotic cells (MTT essays) and antimicrobial activity against an array of important bacterial pathogens for human health. Also, PEs have been tested as bacteria markers with proven affinity for bacterial membranes and some structure cell markers like lipids structures. These achievements would be of great pharmacological and commercial interest.

References [1] Coates AR, Halls G, Hu Y., Br J Pharmacol. 2011 May;163(1):184-94. [2] Merkel O., Material Matters, 2013, 8 (2) 74-77. [3] R Materials & Interface, 2016, 8, 1958í1969.

MJ, Mallavia R., Mateo C, ACS Applied

[5] Chakraborty S., Liao IC, Adler A, Leong KW., Advance Drug Delivery Rev, 2009. 61 (12):1043-54.

Mammalian Cell Viability on Hydrophobic and Superhydrophobic Fabrics 1,2

M. Carmen Morán , Guillem, Ruano , Francesca Cirisano , Michele Ferrari 1

Departament de Bioquímica i Fisiologia, Secció de Fisiologia; Facultat de Farmàcia, Universitat de Barcelona, Avda. Joan XXIII s/n, 08028-Barcelona, Spain 2 Institut de Nanociència i Nanotecnologia-IN2UB, Universitat de Barcelona, Avda. Joan XXIII s/n, 08028-Barcelona, Spain 3 CNR-,&0$7( ,VWLWXWR GL &KLPLFD GHOOD 0DWHULD &RQGHQVDWD H GL 7HFQRORJLH SHU O¶(QHUJLa, via De Marini, 6, 1649-Genova, Italy mcmoranb@ub.edu

Abstract Highly water repellent coatings with wettability properties known as superhydrophobicity (SH) are related to surfaces with contact angle above 150° and a very small hysteresis. The small area available for these surfaces when in contact with water address to be exploited in many applications where interactions with aqueous environment are usually strongly to be avoided. The combination of low surface energy and the existence of a specific surface morphology (micro-nano roughness) coexist, lead to significantly less wettable surfaces [1]. Cell adhesion, the binding of a cell to the extracellular matrix, other cells, or a specific surface, is essential for the growth and survival of the cell and also its communication with other cells. The hydrophobicity and roughness of the surface affect the cell attachment and growth [2]. In this work the influence of coatings at different degree of hydrophobicity has been investigated. Polyester fabrics have been coated by mixed organic-inorganic coating resulting in moderated to highly water repellence. In vitro experiments have been performed in order to establish the influence of surface modification on adhesion of representative model mammalian cell lines such as 3T3 fibroblasts, HaCaT keratinocytes and HeLa epithelial carcinoma cells. The obtained results suggested that the chemistry and the geometry of the resulting coatings are important parameters on the final cell viabilities. References [1] M. Ferrari, A. Benedetti, E. Santini, F. Ravera, L. Liggieri, E. Guzman, F. Cirisano, Colloids and Surfaces A, 480 (2015) 369. [2] F. Dongb, M. Zhangb, W. Huangb, L. Zhoub, MS. Wongb, Y.i Wanga, Colloids and Surfaces A, 482 (2015) 718.

Multichannel Optical Waveguide Lightmode Spectroscopy: Sensor and Instrument M. Moreno, N. Darwish, M. Giménez-Conejo, I. Bernat, J.Bosch, A. Romano-Rodríguez Department of Electronics, Universitat de Barcelona, Martí i Franquès 1, Barcelona, Spain mmoreno@el.ub.edu Abstract Since the development of the first sensors in 1989 [1], the Spectroscopy of Optical Waveguide modes has been fruitful for label-free affinity-based detection of biomolecules. The two independent TE and TM polarization modes provide a very unique feature: the thickness and optical density of an adsorbed molecular layer are measured directly, providing results based on evidence instead of a model. It's not necessary to deepen into the mechanics of the molecule adsorption or the structure of the formed biolayers to measure the concentration of a target molecule, and many different biosensing techniques can do it accurately [2]. Nevertheless, when the process itself is the object of study, the optical waveguide spectroscopy is possibly one of best suited transduction principles [3]. In 2010 we demonstrated a multichannel waveguide spectroscopy sensor with integrated on-line reference [4]. By measuring optical coupling resonances in the angular space we obtained sensitivities 2 around 9deg./RIU, or a 4ng/cm as a limit of detection for molecular adsorption. Using the dual-grating configuration [5] and a passive channel as a robust reference, several channels could be analyzed at the same time. Here we present a rotary in-coupling grating coupler [6] with similar capabilities (fig.1). For this purpose it was necessary to develop the appropriate instrumentation to detect resonances for different channels simultaneously [7]. Because the detection of coupling resonances doesn't need a particularly high sensitivity, a 512 pixel CMOS array detected the coupled beams at the side border of the waveguide device. As different sensor arrays can be used, the number of measurement channels provided by the instrument is only limited by practical constraints, as the width of the fluidic channels and the spatial resolution of the functionalized areas. The optical sensor consist on several diffraction gratings with a period of /=500nm fabricated on Si3N4 waveguides of 90nm thickness over glass substrates. The sensitivity obtained is 8.69º/RIU. References [1] Tiefenthaler, K.; Lukosz, W. J. Opt. Soc. Am. B, 6 (2), 1989. 209-220. [2] Fan, X.; White, I.M.; Shopova S.I.; Zhu-H.; Suter J.D.; Sun Y. Analytica Chimica Acta, 620, 2008. 8-26. [3] Vörös, J. Biophysical Journal, 87(1), 2004. 553-561. [4] Darwish, N.; Caballero, D.; Moreno M.; Errachid A.; Samitier J. Sensors and Actuators B: Chemical, 144(2), 2010. 413-417. [5] Clerc, D., & Lukosz, W. Sensors and Actuators B: Chemical, 40 (1), 1997. 53-58. [6] Adányi, N.; Levkovets, I. A.; Rodríguez-Gil S.; Ronald A.; Váradi M.;Szendro I., Biosensors and Bioelectronics, 22(6), 2007. 797-802 [7] García-Conejo, M.A. Master thesis, Universitat de Barcelona, 2016. 45-46

Figures

512 pixels array

Left: Layout of a multi-grating coupler sensor Center: Coupled beam for the reference channel in the center. Right: Output frame showing two out-coupling beams

Influence of stabilization agents on the properties of biocompatible magnetite nanoparticles C. Nadejde1, E.-L. Ursu2 and G. Ababei3 1³$OH[DQGru

Ioan Cuza´ University, Interdisciplinary Research Department ± Field Science, L. Catargi Str. 54, Iasi, Romania 2³3HWUX 3RQL´ ,QVWLWXWH RI 0DFURPROHFXODU &KHPLVWU\ RI 5RPDQLDQ $FDGHP\, Centre of Advanced Research in Bionanoconjugates and Biopolymers, Gr. Ghica Voda 41A, Iasi, Romania 3National Institute of Research and Development for Technical Physics, D. Mangeron Bd. 47, Iasi, Romania claudia.nadejde@uaic.ro

Abstract The aim of the present investigation was to assess the influence of various surfactants on the structural and magnetic properties of magnetite (Fe3O4) nanoparticles (MNPs) [1,2]. In this respect, surface modified Fe3O4 nanoparticles have been synthesized using the co-precipitation method [3,4], in the presence of four different biocompatible capping agents (citric acid, gallic acid, chitosan and polyethylene glycol) [5±9]. The choice of a non-hazardous surfactant is crucial in the stabilization of magnetic cores, since it is not only protecting the MNPs against oxidation and agglomeration, but is also maintaining their long-term dispersion in aqueous medium. The size of the magnetite cores was tuned by varying the temperature and pH during the reaction. Lower temperature (70 °C), pH 11 and fast agitation were the favorable conditions to yield nano-sized particles below 30 nm. Nevertheless, during the preparation protocol, the type of stabilization agent, also plays a major role on particle size and sample stability. By adding the surfactant during or post-synthesis procedure, the nanoparticle size could be tuned-up. The relationship between nanoparticle crystallinity, size and stability was evaluated using standard characterization techniques. X-Ray Diffraction (XRD), High-Resolution Transmission Electron Microscopy (HR-TEM), and Vibrating Sample Magnetometry (VSM) were used to analyze the structural, morphological and, respectively, the magnetic properties of the as-synthesized MNPs; Fourier Transform-Infrared Spectroscopy (FT-IR) was employed in order to confirm the binding of the surfactant onto magnetite, while Dynamic Light Scattering (DLS) measurements in the diluted aqueous dispersions provided information regarding the stability of the samples. The average crystallite size of cubic spinel magnetic phase ranged from 13 to 27 nm (as resulted following XRD analyses), being also dependent on the type of surfactant and the moment of its addition into reaction. The presence of surfactant led to decrease of the size of formed nanoparticles during the precipitation step. Smaller particle sizes resulted when citric or gallic acids were used as surfactant as compared to the polymer encapsulated MNPs. TEM data revealed the nano-sized particles of quasi-spherical shape in all samples, with average sizes that are in agreement with the XRD results. All samples exhibited superparamagnetic behavior at room temperature; the magnetic properties were found to be dependent on both grain size and surfactant type. The maximum value of saturation magnetization of 74.65 emu/g was obtained for uncoated magnetite. Additionally, the produced coated MNPs were found to be well dispersed in water, exhibiting excellent stability as confirmed by the zeta potential values following DLS measurements. The results showed that the obtained stabilized MNPs possess a great potential for biomedical and environmental applications. References [1] A.-H. Lu, E.L. Salabas, F. Schuth, Angew. Chem. Int. Ed., 46 (2007) 1222. [2] B.I. Kharisov, H.V. Rasika Dias, O.V. Kharissova, A. Vázquez, Y. Peña, I. Gómez, RSC Adv., 4 (2014) 45354. [3] R. Massart, IEEE Trans. Magn., MAG-17 (1981) 1247. [4] F.A. Tourinho, R. Franck, R. Massart, J. Mater. Sci., 25 (1990) 3249. [5] E. Tombácz, M. Szekeres, A. Hajdú, I.Y. Tóth, R.A. Bauer, D. Nesztor, E. Illés, I. Zupkó, L. Vékás, Per. Pol. Chem. Eng., 58(Sup) (2014) 3. [6] Z. Sohrabijam, A. Zamanian, M. Saidifar, A. Nouri, Procedia Mater. Sci., 11 (2015) 282. [7] D.H. Kim, S.H. Lee, K.H. Im, K.N. Kim, K.M. Kim, I.B. Shim , M.H. Lee , Y.-K. Lee, Curr. Appl. Phys., 6S1 (2006) e242. [8] E. Illés, M. Szekeres, E. Kupcsik, I.Y. Tóth, K. Farkas, A. Jedlovszky-Hajdú, E. Tombácz, Colloids and Surfaces A: Physicochemical and Engineering Aspects, 460 (2014) 429. [9] M. Khalkhali, K. Rostamizadeh, S. Sadighian, F. Khoeini, M. Naghibi, M. Hamidi, DARU J. Pharm. Sci., 23(1) (2015) 45.

Stimulation of Dermal Fibroblasts with Calcium-Phosphate Nanoparticles for the Design of Novel Bioactive Dressings C Navarro-Requena 1

1, 2

, S Pérez-Amodio

1, 2

, J Martí-Muñoz

, O Castaño

1, 2

, E Engel

1, 2,3

Institute for Bioengineering of Catalonia, Barcelona, Spain. CIBER de Bioingeniería, Biomateriales y 3 Nanomedicina, Barcelona, Spain. Dpt. Material Science and Metallurgical Engineering, Technical University of Catalonia, Barcelona, Spain. cnavarro@ibecbarcelona.eu Abstract 1

Hard-to-heal wounds are a major socio-economic problem in developed countries . In the recent years, bioactive dressings have been developed, mainly based in the addition of biological factors. However, they present significant limitations, such as high cost of production or regulatory challenges. Extracellular calcium could be considered as an alternative biostimulator, since it has been probed to 2-3 influence the healing process of skin injuries . However, a better understanding of its effect, together with a system to obtain a controlled release is needed. In this study we tested whether the ionic dissolution of calcium-releasing nanoparticles may exert a similar stimulating effect as extracellular calcium on dermal fibroblast. Rat dermal fibroblasts were cultured in the presence of medium containing 2+ different calcium concentrations, normally ranging from 0.1 to 3.5mM Ca . Several relevant aspects of the wound healing process were assessed by different in vitro assays, including cell viability and proliferation, migration, collagen synthesis, matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) activity, contraction capacity and expression of relevant genes for the wound healing process. For most 2+ 2+ of the assays significant differences were observed among the Ca concentrations tested. Higher Ca concentrations of CaCl2 (3.5mM) increased metabolic activity, in vitro wound closure (Fig. 1), MMP activity, collagen synthesis and cytokine expression. Furthermore, extracellular calcium reduced cell contraction capacity. Interestingly, the levels of migration and contraction measured followed a calcium concentration-dependent behavior. Media conditioned with CaP stimulated the same activities as media conditioned with CaCl2, but effects such as inflammatory factor expression and MMP activity were 2+ reduced compared to the equivalent CaCl2 concentration. Overall, the concentration of 3.5mM Ca from CaP was able to stimulate relevant activities that might promote a faster wound healing while avoiding undesired effects, such as increased MMP activity or excessive cell contraction. The differences observed between the two different calcium sources arise from the fact that the particles used release other ions such as phosphates, which might have other effects on the cells. A better understanding of how other cell types involved in the healing process respond to a range of concentrations of calcium-phosphate particles may lead to a wiser design of novel growth factor/cellfree bioactive dressings for skin wound healing.

References: 1

Figure 1: The wound scratch assay was used to study cell 2+ migration in different CaCl2 concentrations and Ca released from the CaP. The reduction of the wound was measured at different time points.

100

(a fte r 2 4 h )

Figures

% w o u n d a re a

CK Sen et al., Wound Repair Reg 17(6) (2009) 763±771. JW Doyle et al., J Biomed Mater Res, 32 3 (1996) 561. J Belmin et al., J Am Geriatr Soc, 50 (2002) 269. 80 60 40 20

-C

m .5 3

-C M

2 .5 2

Acknowledgements:

m .5

m 5

.5 0

The authors would like to thank the Spanish MINECO (FPU grant ref. AP-2012-5310) and MAT201238793 for providing financial support to this project.

Inhibiting Notch Activity in Breast Cancer Stem Cells by Glucose Functionalized Nanoparticles Carrying Č&#x2013;-secretase Inhibitors

Niemi Rasmus1,2, Mamaeva Veronika1,2,3, Beck Michaela4, Ă&#x2013;zliseli Ezgi1,5, Desai Diti5, Landor Sebastian1,2,6, GrĂśnroos Tove7,8, Kronqvist Pauliina9, Pettersen Ina KN10, McCormack Emmet3,11, Rosenholm Jessica M5, LindĂŠn Mika4, Sahlgren Cecilia1,2,12 1Turku

Centre for Biotechnology, University of Turku and Ă&#x2026;bo Akademi University, Turku, Finland of Science and Engineering, Ă&#x2026;bo Akademi University, Turku, Finland 3Department of Clinical Science, University of Bergen, Bergen, Norway 4Inorganic Chemistry II, Ulm University, Ulm, Germany 5Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Ă&#x2026;bo Akademi University, Turku, Finland 6Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden 7Turku PET Centre, University of Turku, Turku, Finland 8Medicity Research Laboratories, University of Turku, Turku, Finland 9Department of Pathology, University of Turku, Turku, Finland 10Department of Biomedicine, University of Bergen, Bergen, Norway 11Department of Medicine, Haematology Section, Haukeland University Hospital, Bergen, Norway 12Department of Biomedical Engineering, Technical University of Eindhoven, Institute for Complex Molecular Systems, Eindhoven, the Netherlands 2Faculty

Abstract Cancer stem cells (CSCs) RU ÂłWXPRXU LQLWLDWLQJ FHOOVÂ´ are a challenge in cancer treatment due to their therapy resistance. The CSC population has also been linked to tumour initiation, metastasis and recurrence[1]. The Notch signalling pathway, an important regulator of cell fate decisions, is crucial for maintenance of regular stem cells, is often deregulated in cancer, and has been shown to be elevated in CSCs[2],[3] in various types of cancer[4],[5]. We demonstrated that enhanced Notch signaling in breast cancer promotes self-renewal of CSCs that display high glycolytic activity [6] and aggressive hormone-independent tumor growth in vivo. We took advantage of the glycolytic phenotype and the dependence on Notch activity of the CSCs and designed nanoparticles to target the CSCs. Mesoporous silica nanoparticles were functionalized with glucose moieties and loaded with a Č&#x2013;-secretase inhibitor, a potent interceptor of Notch signaling. Cancer cells and CSCs in vitro and in vivo efficiently internalized these particles, and particle uptake correlated with the glycolytic profile of the cells. Nanoparticle treatment of breast cancer transplants on chick embryo chorioallantoic membranes efficiently reduced the cancer stem cell population of the tumor. Our data reveal that specific CSC characteristics can be utilized in nanoparticle design to improve CSC-targeted drug delivery and therapy.

References [1] Dean M, Fojo T, Bates S, Nat Rev Cancer, 5 (2005), 275-284. [2] Pannuti A, Foreman K, Rizzo P, Osipo C, Golde, T Osborne B et al, Clin Cancer Res 16 (2010), 3141-3152. [3] 5L]]R 3 0LDR + 'Âś6RX]D * 2VLSR & 6RQJ // <XQ - et al, Cancer Res, 68 (2008), 5226-5235. [4@ 2Âś%ULHQ &$ 3ROOHWW $ *DOOLQJHU 6 'LFN -( 1DWXUH 445 (2007), 106-110. [5] Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF, Proc Natl Acad Sci USA 100 (2003), 3983-3988. [6] Landor SK, Mutvei AP, Mamaeva V, Jin S, Busk M, Borra R et al, Proc Nat Acad Sci USA 108 (2011), 18814-18819.

NANOLIPOSOMES TARGETING MYELOID CELLS TO DELIVER HIV-1 REACTIVATION COMPOUNDS 1,+

2,+

Jon Ander Nieto-Garai , Susana Benet , Eneritz Bilbao , Itziar Erkizia , Julia G. Prado , 2,3 2,* 1,* Javier Martinez-Picado , Nuria Izquierdo-Useros , Maier Lorizate +

Equal contribution

Instituto Biofisika (UPV/EHU, CSIC) and Department of Biochemistry and Molecular Biology 2 (UPV/EHU), Barrio Sarriena s/n, 48940, Leioa, Spain AIDS Research Institute IrsiCaixa, 3 Badalona, Spain ICREA, Barcelona, Spain

*Corresponding authors: nizquierdo@irsicaixa.es; maier.lorizate@ehu.eus

Abstract Antiretroviral therapy is capable of suppressing HIV-1 replication, but it does not hamper the establishment of viral reservoirs within latently infected cells. A major drawback for eradication is how to reactivate latently infected cells and purge HIV-1 reservoirs from anatomical sanctuaries [1]. Hence, viral eradication demands innovative combined strategies designed not only to reactivate latent viral reservoirs, but also to promote antiviral immune control. Indeed, prior antigen-specific stimulation of CD8+ T-cells will be likely required for an effective clearance of HIV-1 reservoirs [2]. Mature dendritic cells (mDCs) are potent antigen presenting cells that constantly interact with T-cells to initiate immune responses. However, HIV-1 has evolved strategies to subvert mDC antiviral immunity and promote infection of CD4+ T-cells throughout a mechanism that involves binding and uptake of HIV-1 by mDCs, traffic of internalized virus, and its final release allowing for productive infection of CD4+ T-cells. We identified the molecular mechanism underlying this highly infectious route, which relies on the expression of the membrane receptor Siglec-1 on mDCs [3] that recognizes sialyllactose exposed on viral membrane gangliosides [4]. This novel pathway offers the unique opportunity to achieve HIV-1 eradication by targeting engineered nanoparticles to Siglec-1 expressed on mDCs and promote i) the reactivation of latently infected cells and, ii) the specific killing of these cells by antiviral cytotoxic responses. We intend to exploit the infectious HIV-1 transmission route mediated by Siglec-1 on mDCs and use it to transfer reactivation compounds within nanocarriers designed to mimic the size and external lipid composition of HIV-1. These nanoparticles would not only be efficiently captured by Siglec-1 expressed on mDCs or other myeloid cells such as monocytes [3, 5], but would specifically deliver reactivation molecules and antiretroviral compounds to CD4+ T-cells and viral peptides to CD8+ T-cells. By targeting Siglec-1 with nanoparticles we could reach antigen presenting cells scattered throughout secondary lymphoid tissues [6]. Indeed, targeting liposomes to Siglec-1 has proven efficacy in inducing antigen presentation in vivo [7]. Fluorescent PEGylated nanoliposomes were prepared with distinct sialyllactosecontaining gangliosides (GM). Siglec-1+ primary myeloid cells were exposed to the same concentration of nanoliposomes and capture was measured by FACS and confocal microscopy. Statistical differences were assessed with a paired t test. Nanoliposomes were loaded with the latency reactivators romidepsin and bryostatin-1. Reactivation assays were carried out using the latently infected J-lat cell line. When HIV-1 reactivation occurs GFP is expressed, which can be measured by FACS.

Fluorescent nanoliposomes containing GM1 or GM3 specifically targeted mDCs and activated monocytes with equal efficiency as fluorescent HIV-1 viral like particles. An increase in the ganglioside content in the liposome formulation led to an improved capture efficiency in a dose dependent manner (figure 1). Nanoliposome specificity for Siglec-1 was demonstrated by blocking capture with two specific mAbs against Siglec-1 (P RU ZLWK VROXEOH sialyllactose (P ILJXUH 7KH DGGLWLRQ RI 3(*-lipids to the nanoliposomes conferred a robust stability in up to 50% of serum (figure 3A) but preserved Siglec-1 targeting capability (figure 3B). Nanoliposomes bearing romidepsin and bryostatin-1 efficiently reactivated latently infected J-lat cells (figure 4). We have developed a stable nanoliposome formulation with a potent and specific capacity to target Siglec-1 expressing myeloid cells. This technology could be crucial for delivering small therapeutic molecules and purging latent viral reservoirs within lymphoid tissues.

References [1] Deeks, S.G., Autran, B., Berkhout, B., Benkirane, M., et al., Nat Rev Immunol, 12 (2012) 607-614. [2] Shan, L., Deng, K., Shroff, N.S., Durang, C. M., et al., Immunity, 36 (2012) 491-501. [3] Izquierdo-Useros, N., Lorizate, M., Puertas, M.C., Rodriguez-Plata, M.T., et al., PLoS Biology, 10 (2012) e1001448. [4] Izquierdo-Useros, N., Lorizate, M., Contreras, F.X., Rodriguez-Plata, M.T., et al., PLoS Biology, 10 (2012) e1001315. [5] Pino, M., Erkizia, I., Benet, S., Erikson, E., et al., Retrovirology, 12 (2015) 37. [6] Steinman, R.M. & Banchereau, J., Nature, 449 (2007) 419-426. [7] Kawasaki, N., Vela, J.L., Nycholat, C.M., Rademacher, C., et al., Proc Natl Acad Sci USA, 110 (2013), 7826-7831. Figures

Fig. 1 ± Nanoliposomes prepared with increasing concentrations of GM led to an improved capture on myeloid cells.

Fig. 2 ± Specificity for Siglec-1 was demonstrated by blocking capture with two monoclonal antibodies against Siglec-1 and with sialyllactose.

Fig. 3 ± A) Incorporation of PEG-lipids conferred a robust stability in up to 50% of serum. B) PEG-lipid containing nanoliposomes preserved Siglec-1 targeting capability.

Fig. 4 ± Nanoliposomes bearing romidepsin and bryostatin-1 efficiently reactivated latently infected cells.

Paper-based gold-modified electroanalytical devices for glucose and inorganic arsenic sensing 1

Estefanía Nunez-Bajo , M. Carmen Blanco-López, Agustín Costa-García, M. Teresa Fernández2

Abedul

Departamento de Química Física y Analítica. Universidad de Oviedo 33006 Oviedo (Asturias) Spain. Tel.: +34 (9)85102968. FAX: + 34 (9)85103125. 1,2

e-mail: nunezestefania@uniovi.es ; mtfernandeza@uniovi.es

Abstract Lab-on-paper sensing represents promising advances to portable and disposable devices for diagnostic testing as well as food and environmental analysis, in which highly sensitive methods must be combined with inexpensive, rapid, and simple fabrication and operation. In this way, integration of electrochemical detection does not greatly increase the size, price or complexity and allows both portability and disposability. Moreover, paper-based electrochemical devices allow easy immobilization of biological reagents (e.g. enzymes) due to the porosity of the substrate and results very convenient for performing bioassays. On the other hand, advantages of electrode nanostructuration are well known and this step can be easily introduced in analytical procedures developed on paper substrates. In this work, we aim to design simple electroanalytical microfluidic platforms based on paper for the determination of analytes of interest. A paper-based gold working electrode (Au-PWE) can be easily fabricated to proportionate conductivity and improve the adsorption of biological materials on paper sample zones. This is made in two different ways: i) through the deposition of thin gold films on the substrate surface by sputtering under low vacuum or ii) with the electrogeneration of gold nanoparticles (AuNPs) on a conductive layer (carbon ink) that was previously deposited on the surface of cellulose fibers. As a proof-of-concept, in this work we apply these simple and inexpensive electrochemical platforms for the enzymatic determination of glucose by chronoamperometry and for the amperometric stripping determination of arsenic. A biosensor will help to find glucose levels that should be regulated to prevent metabolic disorders and a point-of-use nanostructured device will allow avoiding harmful arsenic effects in human health employing a calibration-free and fast analytical methodology. Electrochemical stability and reproducibility of both platforms is achieved coupling reusable reference and counter electrodes (metallic wires or screen-printed electrodes) to disposable gold-sputtered and gold-nanostructured PWEs (Figure 1). References [1] A.C. Glavan, D. Christodouleas, B. Mosadegh, H.D. Yu, B. Smith, J. Lessing, M.T. FernándezAbedul, G.M. Whitesides, Analytical Chemistry, 86 (2014) 11999-12007. [2] M.M. Hamedi, A. Ainla, F. Guder, D.C. Christodouleas, M.T. Fernández-Abedul, G.M. Whitesides, Advanced Materials, 28 (25) (2016) 5054-5063.

Figure 1. Scheme of the electrochemical platforms with main enzymatic and electrochemical reactions that take place in the Au-PWEs. A) gold thin-film electrodes and wires employed for the construction of glucose enzymatic sensors where enzymes and mediator are immobilized on Au-PWE. B) gold nanostructured carbon PWEs coupled to screen-printed carbon electrodes for arsenic sensing.

Acknowledgements This work has been supported by the Spanish Ministry of Economy and Competitiveness under projects MINECO-15-CTQ2014-58826-R and the Asturias Regional Government through the project FC-15GRUPIN14-021

Self-Assembled Up-standing Chiral Architectures for the Construction of Functional Surfaces Ani Ozcelik, José Lorenzo Alonso Gómez, Raquel Pereira Cameselle University of Vigo, Campus Universitario Lagoas-Marcosende, 36310, Vigo, Spain ozcelik@uvigo.es

Chiral two-dimensional (2D) surfaces are of particularly interest due to the potential applications in stereoselective chemical synthesis, chiroptical sensing and optoelectronic devices.

[1]

In this

context, a versatile and efficient approach is provided by the selfassembly of optically active molecules onto achiral substrates. Recently, we have shown that Chiral Frameworks (CFs) bearing chiral axes are capable of forming surface-confined Up-Standing Chiral Architectures (UCAs) orthogonal to the metallic substrate. Nevertheless, Scanning Tunneling Microscopy (STM) studies revealed that these first examples were only stable at very low temperature hampering their applicability.

[2]

The stability of a chiral molecule-substrate pair plays a vital role for tailoring surface properties in a well-controlled fashion. In this regard, the strategy of Self-Assembled Monolayers (SAMs) offers a great possibility to construct stable self-assembled systems by the incorporation of thiols as the most employed head group.

[3]

Additionally, axially chiral diethynylallenes (DEAs) could serve as

appropriate building blocks for the construction of novel systems with remarkable chiroptical responses.

[4]

We herein report thiol-derivatized allenic CFs to improve the stability of the previously

reported UCAs.

Synthesis of (±)-CF-1 and (±)-CF-2 was successfully achieved through Cu-free Sonogashira reaction followed by enantiomeric resolution and absolute configuration assignment. Furthermore, photo- and thermal-stability studies of the enantiopure chiral frameworks were performed in solution through Electronic Circular Dichroism (ECD) spectroscopy. Regarding to the results obtained, both molecules

presented stability at room temperature (in the dark) and slow photoismerization. On the other hand, the time scale for the photoisomerization of both molecules was similar and sufficiently slow to study the self-assembly processes. The synthesized CFs were employed in order to form SAMs onto semi-transparent Au substrates. Subsequent

spectroscopic

ellipsometry

and

X-ray

Photoelectron

Spectroscopy

(XPS)

characterizations have proved the formation of stable Up-standing Chiral Architectures (UCAs) at room temperature. The analysis of the chiroptical responses of these novel functionalized surfaces will be discussed.

References [1]

Y. Mastai, Chem. Soc. Rev. 2009, 38, 772±780.

[2]

Y.-Q. Zhang, M. A. Öner, I. R. Lahoz, B. Cirera, C.-A. Palma, S. Castro-Fernández, S. MíguezLago, M. M. Cid, J. V. Barth, J. L. Alonso-Gómez, et al., Chem. Commun. 2014, 50, 15022± 15025.

[3]

C. Vericat, M. E. Vela, G. Benitez, P. Carro, R. C. Salvarezza, Chem. Soc. Rev. 2010, 39, 1805±1834.

[4]

S. Castro-Fernández, M. M. Cid, C. S. López, J. L. Alonso-Gómez, J. Phys. Chem. A 2015, 119, 1747±1753.

Silver nanoparticles restore bactericidal activity of inactive antibiotics against multiresistant bacteria a

Ales Panacek , Monika Smekalova , Renata Vecerova , Milan Kolar , Marketa Havrdova , Radek a a Zboril and Libor Kvitek a

Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacky University in Olomouc, 17.listopadu 12, 771 46 Olomouc, Czech Republic ales.panacek@upol.cz b Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Hnevotinska 5, 775 15 Olomouc, Czech Republic c Regional Centre of Advanced Technologies and Materials, Department of Experimental Physics, Faculty of Science, Palacky University in Olomouc, 17.listopadu 12, 771 46 Olomouc, Czech Republic

Abstract Bacterial resistance to conventional antibiotics is currently one of the most important healthcare issues, and has serious negative impacts on medical practice [1]. Strong synergistic effect of antibiotics combined with silver nanoparticles (silver NPs) presents a potential solution to this problem. Silver NPs inhibit bacterial growth via a multilevel mode of antibacterial action at concentrations ranging from a few ppm to tens of ppm [2]. Silver NPs strongly enhance antibacterial activity against multiresistant, betalactamase and carbapenemase-producing Enterobacteriaceae when combined with the following antibiotics: cefotaxime, ceftazidime, meropenem, ciprofloxacin and gentamicin. All the antibiotics, when combined with silver NPs, showed enhanced antibacterial activity at concentrations far below the minimum inhibitory concentrations (tenths to hundredths of one ppm) of individual antibiotics and silver NPs. The enhanced activity of antibiotics combined with silver NPs, especially meropenem, was weaker against non-resistant bacteria than against resistant bacteria. Low silver concentrations were required for effective enhancement of antibacterial activity against multiresistant bacteria. These low silver concentrations showed no cytotoxic effect towards mammalian cells, an important feature for potential medical applications. Acknowledgement The authors gratefully acknowledge the support provided by project LO1305 of the Ministry of Education, Youth and Sports of the Czech Republic, the Czech Science Foundation (project No.1522248S), the Ministry of Health of the Czech Republic (AZV VES 15-27726A) and the Internal Grants of Palacky University in Olomouc (IGA PrF_2015_022 and IGA_LF_2015_035). References [1] D.M. Livermore, Int. J. Antimicrob. Agents 39 (2012) 283. [2] A. Panacek, L. Kvitek, R. Prucek, M. Kolar, R. Vecerova, N. Pizurova, V.K.Sharma, T. Nevecna, R. Zboril, J. Phys. Chem. B 110 (2006) 16248.

CeO2NPs Time-evolution stability 1

1,2,3

A.P. Guillamon , I.Salvo , V.F. Puntes . (1)Institut de Recerca Vall d´Hebron (VHIR). Pg. Vall d'Hebron, 119-129 08035 Barcelona (Spain) (2)Institució Catalana de Recerca i Estudis Avançats(ICREA) Pg. Lluís Companys, 23, 08010 BCN (3)Institut Català de Nanociencia i Nanotecnologia (ICN2), Campus de la UAB, Av. Serragalliners, s/n, 08193 BCN. ariadna.peral.ing@gmail.com

Abstract The use of nanoparticles (NPs) in medical applications is an emerging field. NPs have the potential to offer novel methods, strategies and therapies, for example, in cancer detection, diagnostics and 1 treatment. One of the common steps to pathologic processes is deregulated inflammation, deriving oxidative stress. In the last years there has been an increasing interest in Cerium oxide nanoparticles (CeO2NPs) 2 due to their regenerative antioxidant properties. 3+ 4+ Cerium oxide can have two oxidation states Ce and Ce due the oxygen vacancies in the lattice, thus having attractive redox properties. At nanoscale, CeO2NPs have improved oxidant capacity from bulk material. However, the same novel properties that arise at the nanoscale, may also lead to unexpected toxicity of nanomaterials. In order to produce safe nanomaterials it is fundamental to study their physicochemical properties, to take into account, for example, size, shape, catalytic activity and 3 persistency. In this particular case, is critical to understand the tendency to form aggregates, which are directly related with the surface area, thus, with the antioxidant capacity. The purpose of this research is to study the time-evolution stability of CeO2NPs in order to determine their physical-chemical properties. We focus on the size distribution profile (Dynamic light scattering), surface charge (Z-Potential) and wavelength absorption (Spectroscopy). Our results can contribute to a better understanding of CeO2NPs, advancing towards their use in medical applications.

Figure 1. CeO2NPs reaction schematic: Defect sites of the nanoparticle contain dangling bonds in the +3 oxidation state. Cerium reacts with singlet oxygen to form an oxygen trimer (top). The Ce 3+ scavenging of ROS results in the generation of Ce 4+. The Ce 3 + state is regenerated through several mechanisms, including superoxide reduction and regeneration of defect states on the surface lattice.4

1. 2. 3. 4.

Alexandra M. N. Hauser-Kawaguchi, L. G. L. Nanomedicine²Nanoparticles in Cancer Imaging and Therapy. (2014). doi:10.1007/978-3319-12136-9_10 Das, S. et al. Cerium oxide nanoparticles฀ Nanomedicine 8, 1483 1508 (2013). Reichenbach, V., Casals, G., González, B. & Presa, D. Cerium oxide nanoparticles reduce steatosis, portal hypertension and display antiinflammatory properties in rats with liver fibrosis. J. Hepatol. 64, 691 698 (2015). Boghossian, A. A. et al. Application of Nanoparticle Antioxidants to Enable Hyperstable Chloroplasts for Solar Energy Harvesting. Adv. Energy Mater. 1 13 (2013). doi:10.1002/aenm.201201014

Fluorescence signal enhancement and background suppression in autofluorescence microscopy imaging by using two-dimensional nanostructured glass substrates 1

P.A. Postigo , C. Heeschen , S. Alcalรก , J. Llorens , R. Serna , I. Camps , H. Y. Feng , F. 5 Luo

IMM-Instituto de Microelectrรณnica de Madrid (CNM-CSIC), Isaac Newton 8, PTM, E-28760 Tres Cantos, Madrid, Spain

Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK. 3 Department of Preventive Medicine, Public Health and Microbiology, Universidad Autรณnoma, Madrid, Spain 4 Laser Processing Group, Instituto de ร ptica, CSIC, C/Serrano 121, 28006, Madrid, Spain 5 IMDEA Nanociencia, Faraday 9, Ciudad Universitaria de Cantoblanco, 28049, Madrid, Spain pabloaitor.postigo@imm.cnm.csic.es Abstract: We have recently identified an intrinsic autofluorescent phenotype in cancer stem cells, which are rare cells, found in tumors and bear exclusively tumorigenic and metastatic capacity. This distinct inherent cancer stem cell property represents a novel biological feature that is traceable in real time and provides unprecedented robustness and power for the identification and purification of cancer stem cells without the use of antibodies nor any kind of manipulation, thus drastically reducing experimental errors and artifacts. Autofluorescence was due to riboflavin accumulation in membrane-bounded cytoplasmic structures, which results in a distinct, but rather weak fluorescent signal upon excitation with a 488 blue laser (emission max at 532nm), even in the presence of high riboflavin concentrations. Two-dimensional nanostructures fabricated on glass have been used as substrates for advancing cell culture and fluorescence imaging. Pancreatic tumor-derived human primary cancer cells were enriched for autofluorescent cancer stem cells using anchorage-independent conditions for 7 days. Cells were exposed to 5uM riboflavin for 24 hours prior to seeding the cells on top of the nanopatterns, and images were obtained by confocal microscopy (Leica SP5) after the cells had adhered. The recorded images of autofluorescent cells were processed to obtain the image histograms and pixel-depth plots (Figure 1). An enhancement of the fluorescence (5-10%) in the normal direction to the substrate glass was observed along a strong inhibition of the background fluorescence inside all of the nanopatterns, being almost 100% for some of them. The inhibition and enhancement of the fluorescence signal can be well rationalized by the optical properties of the nanopatterns.

Figure 1. Pixel depth in log scale for autofluorescence images of cells cultured in 12 nanopatterns fabricated on glass

Optofluidic chips with nanochannels for dynamic molecular detection using enhanced fluorescence 1

P.A. Postigo , R. Alvaro, A. Juarros and S. Merino 1

IMM-Instituto de MicroelectrĂłnica de Madrid, IMM-CNM-CSIC, Isaac Newton 8, PTM E-28760 Tres Cantos, Madrid, Spain 2 Micro and Nanofabrication Unit, IK4-Tekniker, 20600. Eibar, Spain pabloaitor.postigo@imm.cnm.csic.es

Abstract The fabrication of a novel optofluidic chip using nanochannels optimized for DNAstretched molecules and optical detection by enhanced fluorescence is reported (Fig.1). The chips are composed of a series of microchannels that allow the transport of molecules in the femto-liter per second inside a fluid or gas. The nanochannels are surrounded by a photonic crystal structure to enhance the emission of fluorescent light from the molecules, which can travel along the nanochannel, allowing for enhanced optical detection of the molecules in motion. The photonic crystal structure provides an enhancement up to 2.5 times in the light emitted from fluorescent molecules inside the nanochannels which increases to around 250 when normalized to the area of the nanochannels emitting fluorescence. The results may help to the detection of fluorescent molecules (like marked-DNA) in series by speeding it and allowing the use of less sophisticated equipment. [1] References [1] P. A. Postigo, R. Alvaro, A. Juarros, and S. Merino, Biomed. Opt. Express, 7(9), 3289â&#x20AC;&#x201C;3298, (2016).

Figure 1. Scanning electron microscopy (SEM) images of the chips fabricated by FIB etching showing greater magnifictions of the areas marked by the white boxes (a,b and c) . The images in (d) is an atomic force microscopy (AFM) image of the area marked by the white box in c).

Photosensitizing flavoproteins: improved bio-tags for nanoscale imaging with correlative microscopy a

Alberto Rodríguez-Pulido, Aitziber L. Cortajarena, Joaquim Torra, Rubén Ruiz-González, Santi c a Nonell and Cristina Flors a

Madrid Institute for Advanced Studies in Nanoscience (IMDEA Nanoscience) and Nanobiotechnology Unit Associated to the National Center of Biotechnology (CSIC), C/ Faraday, 9 ± 28049 Madrid, Spain b CIC biomaGUNE, Paseo de Miramón, 182 ± 20009 Donostia-San Sebastián, Spain c Institut Químic de Sarrià, Universitat Ramon Llull ± 08017 Barcelona, Spain alberto.rpulido@imdea.org

Abstract Correlative light and electron microscopy (CLEM) is a powerful technology for observing structure and function of proteins at high spatial resolution. A popular strategy for CLEM is based on oxidation of diaminobenzidine (DAB) by enzymatic or photoinduced reactions to form an insoluble osmiophilic [1] polymer, which provides contrast in EM after staining with osmium tetroxide. Synthetic photosensitizers have been traditionally used in CLEM to produce reactive oxygen species (ROS), in particular singlet oxygen, upon light irradiation that in turn oxidize DAB. Lately, photosensitizing proteins have gained great interest as genetically-encoded tags for CLEM because they can site-specifically induce DAB polymerization in the cells. However, in spite of their potential applicability, current genetically-encoded photosensitizers are limited by their low yield of ROS production and/or unspecific generation of a kind of ROS (singlet oxygen, superoxide anion and/or hydroxyl radical) that can affect to the expected efficiency of DAB polymerization. With the aim of overcoming current limitations of genetically-encoded photosensitizers, we have studied [2] variants from miniSOG (mini Singlet Oxygen generator), a fluorescent flavin-binding protein (flavoprotein), by introducing mutations in strategic positions that are expected to improve their photosensitizing properties. We have screened the ability of these mutants to photoxidize DAB as well [3] as their fluorescence photobleaching properties in order to predict their performance in CLEM. We compare the DAB photo-oxidation data to direct measurements of singlet oxygen photosensitation efficiency using time-resolved detection of its phosphorescence in the NIR, as well as to their relative [3] generation of other ROS. Our results help understanding the requirements for optimal CLEM tags and reveal, a complex relation between fluorescence photobleaching and the production of different ROS. [3] Importantly, we identify one novel improved miniSOG mutant that induces more efficiently DAB polymerization and generates singlet oxygen with higher efficiency than others reported to date, which has great potential for future CLEM experiments. References [1] C. Meisslitzer-Ruppitsch, C. Rohrl, J. Neumuller, M. Pavelka and A. Ellinger; J. Microsc., 235 (2009) 322-335. [2] X. Shu, V. Lev-Ram, T. J. Deerinck, Y. Qi, E. B. Ramko, M. W. Davidson, Y. Jin, M. H. Ellisman and R. Y. Tsien; PLoS Biol., 9 (2011) e1001041. [3] A. Rodríguez-Pulido, A. L. Cortajarena, J. Torra, R. Ruiz-González, S. Nonell and C. Flors; ChemComm, 52 (2016) 8405-8408.

Control synthesis of metal oxide nanoparticles for medical applications 1

Ignacio Salvo Ibáñez . Víctor Franco Puntés

1, 2, 3

(1)Institut de Recerca Vall d´Hebron (VHIR), Pg Vall d´Hebron 119-129, 08035, Barcelona (Spain). (2) Institució Catalana de Recerca i Estudis Avançats (ICREA) Pg. Lluís Companys, 23, 08010 Barcelona. (3) Institut Català de Nanociencia i Nanotecnologia (ICN2), Campus de la UAB, Av. Serragalliners, s/n, 08193 Barcelona. Nanobiotechnology is growing the last years, and the use of nanoparticles in medicine is increasing, specially metal oxide nanoparticles. These kinds of particles have interesting properties which they would not have if we had only the metal. Their use has applications such as design of new materials, development of sensors, catalysis, biological fields, environmental [1] remediation . It is important to know and control the synthesis of nanoparticles. Most of metal oxide nanoparticles are synthesized following the basification of a metal precursor in aqueous solution. In general, the metal hydroxide is synthesized and, later it is dehydrated, losing a molecule of water and forming the metal oxide nanoparticle. Depending on the kind and concentration of the base, we can do nanoparticles with different sizes. We employ this technique because it is more precise to control size and polidispersity of nanoparticles. Our aim consists in improving the synthesis of these nanoparticles because, in this way, we can benefit better from their properties and apply them for different medical applications. Metal oxide nanoparticles have characteristics that allow them to act as active substances. For these reasons, we focus on the synthesis of silica (SiO2), iron oxide (Fe3O4) and titanium oxide (TiO2) nanoparticles. Titanium oxide can be an efficient photocatalyst, useful in [2] [3] photodynamic therapy for cancer . Iron oxide can be used as ions releaser for anemia and [4] silica can do an important task promoting activity and differentiation of bone , helping in the treatment against osteoporosis.

Figure: TiO2 nanoparticles 10nm in the left. SiO2 nanoparticles 250nm in the center. Fe3O4 nanoparticles 7nm in the right.

[1] Gerko Osman, Journal of sol-gel Science and technology, 2006, 37, 161-163. [2] Zi Fei Yin, Long Wu, Hua Gui Yang and Yong Hua Su, Physical Chemistry Chemical Physics, 2013, 15, 4819-4831. [3] David C Ford, Naomi V Dahl, William E Strauss, Charles F Barish, David J Hetzel, Kristine Bernard, Zhu Li, Lee F Allen, Clin exp gastroenterol, 2016, 9, 151-162. [4] M. Neale Weitzmann, Ph.D, Shin-Woo H, Tatyana Vikulina, Susanne Roser-Page, Jin-Kyu Lee, George R. Beck Jr.Ph.D, Nanomedicine, 2015, 11, 959-967.

A Comparative Study of Folate Receptor-Targeted Doxorubicin Delivery Systems: Dosing Regimens and Therapeutic Index 1

Anna Scomparin , Stefano Salmaso , Anat Eldar-Boock , Dikla Ben-Shushan , Shiran Ferber , 1 3 4 4 2 GaliaTiram , Hilary Shmeeda , Natalie Landa-Rouben , Jonathan Leor , Paolo Caliceti , Alberto 3 1 Gabizon and Ronit Satchi-Fainaro 1

Department of Physiology and Pharmacology, Sackler School of Medicine, Tel Aviv University, Tel Aviv 2 69978, Israel. Department of Pharmaceutical and Pharmacological Sciences, University of Padova, 3 Padova 35131, Italy. Department of Oncology, Shaare Zedek Medical Center and Hebrew University4 School of Medicine, Jerusalem 91031, Israel. Neufeld Cardiac Research Institute, Sheba Medical Center, Tel-Aviv University, Tel-Hashomer, Israel anna.scomparin@gmail.com

Abstract Supramolecular drug delivery nanosystems are designed to accumulate in tumors by extravasation-dependent targeting through the leaky vessels via the enhanced permeability and 1 retention (EPR) effect . The conjugation of targeting moieties results in receptor-mediated drug delivery. Folic acid (FA) is a low molecular weight ligand of the folate receptor (FR), widely used as targeting 2 agent due to overexpression of FR on many tumor types . In this study, we compared the selectivity, safety and activity of doxorubicin (Dox) entrapped in PEGylated liposomes (PLD) versus Dox 3 4 conjugated to polymeric Pullulan nanocarriers (Pull-Dox), targeted with FA (PLD-FA and FA-Pull-Dox ). Both receptor-targeted nanosystems were shown to interact in vitro specifically with cells via the FA. Treatment of FR-overexpressing human cervical carcinoma KB tumor-bearing mice with threeweekly injections resulted in enhanced anticancer activity of FR-targeted Dox-loaded PEGylated liposomes (PLD-FA) and Dox-loaded PEGylated liposomes (PLD), with slight advantage for PLD-FA. Under the same regimen, there was no significant reduction of tumor size with both pullulan-based conjugates (Panel A). When the nanosystems were administered intravenously every other day, the FA-Pull-Dox and the Pull-Dox displayed similar and low antitumor activity as compared to free Dox. At this dosing regimen, the liposome-based formulations displayed again enhanced antitumor activity with a slight advantage for PLD (Panel B). However, both liposomal formulations caused toxicity that was reversible following treatment discontinuation. Using a daily dosing schedule, the FA-Pull-Dox conjugate strongly inhibited tumor growth, and was significantly more active than the Pull-Dox conjugate, while free Dox was toxic at this regimen (Panel C). For polymeric constructs, increasing dose intensity and cumulative dose strongly affects the therapeutic index and reveals a major therapeutic advantage for the FR-targeted formulation. Liposome-based nanosystems require longer dose intervals to prevent toxicity. All nanosystems were able to abrogate Dox-induced cardiotoxicity. This study reports the first side-by-side comparison of two receptor-targeted systems, namely polymer therapeutics versus nanoparticulate systems, that were evaluated in the same mouse tumor model at several dosing 5 regimens . References 1.Matsumura, Y.; Cancer Res 46 (1986), 6387-92. 2.Gabizon, A., In Targeted Drug Strategies for Cancer and Inflammation., Jackman, A. L.; Leamon, C. P.; vol. Springer-Verlag. Heidelberg, G., Eds. Springer-Verlag. Heidelberg, Germany; (2012), 217-248. 3.Gabizon, A. Bioconjugate chemistry 10 (1999), 289-98. 4.Scomparin, A.,Eur J Pharm Sci 42 (2011), 547-58. 5.Scomparin, A., et al.. J control release 208 (2015), 106-20. Figures

In vitro study of gold nanoparticles, nanorods and microparticles, as potential carriers for photodynamic therapy Albert SerrĂ 1,3, MarĂa E. Alea-Reyes2,3, Mafalda Rodrigues2,3, Margarita Mora4, Maria L. SagristĂĄ4, Asensio GonzĂĄlez2, Sara Duran5, Marta Duch5, JosĂŠ Antonio Plaza5, Elisa VallĂŠs1,3, David A. Russell6, and LluĂŻsa PĂŠrez-GarcĂa2,3 1

Ge-CPN, Departament de CiĂ¨ncia de Materials i QuĂmica FĂsica, Universitat de Barcelona, C/ MartĂ i FranquĂ¨s 1, 08028 Barcelona, Spain. 2Departament de Farmacologia, Toxicologia i QuĂmica TerapĂ¨utica, Universitat de Barcelona, Avda. Joan XXIII 27-31, 08028 Barcelona, Spain. 3Institut de NanociĂ¨ncia i NanotecnologĂa UB (IN2UB), Universitat de Barcelona, 08028 Barcelona, Spain. 4 Departament de BioquĂmica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 643, 08028 Barcelona, Spain. 5Instituto de MicroelectrĂłnica de Barcelona (IMB-CNM (CSIC)), Campus UAB, 08193, Cerdanyola, Barcelona, Spain. 6School of Chemistry, University of East Anglia, Norwich Research Park, Norwich, Norfolk, NR4 7TJ, UK. a.serra@ub.edu Abstract Photodynamic therapy (PDT) is a treatment used in cancer therapy. It requires a photoactive drug (photosensitizer), from which porphyrins are among the most commonly used [1]. However, many photosensitizers have low specific bio-distribution, which results in adverse side effects. To overcome this issue, the photosensitizers can be incorporated into nanostructured vehicles, to improve their distribution and targeting to the specific site of action. We report a comparative study of three different types of gold-based nanostructured vehicles, with different sizes and morphologies, loaded with porphyrins: spherical gold nanoparticles (GNP, Fig. 1A), hexahedral silicon-gold microparticles (ÂľP, Fig. 1B) [2] and cylindrical gold nanorods with a magnetic core (NR, Fig. 1C) [3]. The evaluation of the influence of the type of vehicle in the phototoxic effect of the porphyrin, as well as the internalization of the porphyrin-loaded vehicles, was studied using HeLa cells. Independently of the vehicle, the zinc-containing porphyrin is more photocytotoxic. GNP loaded with the zinc-containing porphyrin showed the highest photocytotoxic effect and the best internalization in the cells (Fig. 1D), followed by the microparticles (Fig. 1E) and finally the nanorods (Fig. 1F). Porphyrin-loaded ÂľP present a lower internalization when compared to the GNP, but also showed high cytotoxicity, probably due to the high amount of porphyrin present per vehicle unit. Further analysis of the reactive oxygen species (ROS) produced by the porphyrin upon irradiation showed that the porphyrin immobilized on the GNP had a higher ROS production when compared with the porphyrin free in solution, suggesting a synergistic effect derived from the immobilization in that vehicle. The synthesized porphyrin-loaded GNP therefore appears as a promising delivery system for PDT. References [1] Agostinis, P.; Berg, K.; Cengel, K. A.; Foster, T. H.; Girotti, A. W.; Golinick, S. O.; M., H. S.; Hamblin, M. R.; Juzeniene, A.; Kessel, D.; Korbelik, M.; Moan, J.; Mroz, P.; Nowis, D.; Piette, J.; Wilson, B. C.; Golab, J., CA Cancer J. Clin. 61 (2011) 250Âą281. [2] PatiĂąo, T.; Soriano, J.; Amirthalingam, E.; DurĂĄn, S.; GonzĂĄlez-Campo, A.; Duch, M.; IbĂĄĂąez, E.; Barrios, L.; Plaza, J. A.; PĂŠrez-GarcĂa, L.; NoguĂŠs, C., Nanoscale. 8 (2016) 8773Âą8783. [3] Gispert, C.; SerrĂ , A.; Alea, M. E.; Rodrigues, M.; GĂłmez, E.; Mora, M.; SagristĂĄ, M. L.; PĂŠrezGarcĂa, L.; VallĂŠs, E., Electrochem. commun. 63 (2016) 18Âą21. Figures

Fig. 1. TEM micrograph of porphyrin-GNP (A), fluorescence microscopy images of porphyrin-ÂľP (B) and porphyrinNR (C); Confocal images of HeLa cells with: porphyrin-GNP (D), porphyrin-ÂľP (E) and porphyrin-NR (F). HeLa cells ZHUH DOVR ORDGHG ZLWK WKH IOXRURSKRUH &HOO 7UDFNHUÂ&#x152; *UHHQ &0)'$ WR PDUN WKH F\WRSODVP LQ JUHHQ DQG WKH porphyrin-loaded carriers present red color. Acknowledgements: This work was supported by the EU ERDF (FEDER) funds and the Spanish Government grant TEC2014-51940-C2. The LED source for PDT treatment was purchased with a grant from the Spanish Government (CTQ2013-48767-C3-1-R).

Chitosan-TPP nanoparticles as a DNA carrier to silence gene expression in vivo 1

3,4

Jonás I. Silva-Marrero , Juan D. González , Albert Caballero-Solares , Ivan Viegas , Felipe 2 5 5 5 3 Fernández , Montserrat Miñarro , Anna Fàbregas , Josep R. Ticó , John G. Jones , Isabel V. 1 1 Baanante , Isidoro Metón 1

Departament de Bioquímica i Fisiologia. Secció de Bioquímica i Biologia Molecular, Facultat de Farmàcia L &LqQFLHV GH O¶$OLPHQWDFLy, Universitat de Barcelona, Joan XXIII 27, 08028 Barcelona, Spain 2 Departament de Biologia Evolutiva, Ecologia i Ciències Ambientals, Facultat de Biologia, Universitat de Barcelona, Diagonal 645, 08028 Barcelona, Spain 3 Center for Neuroscience and Cell Biology, University of Coimbra, Largo Marquês de Pombal, 3004-517 Coimbra, Portugal 4 Center for Functional Ecology, Department Life Sciences, University of Coimbra, Calçada Martins de Freitas 3000-456 Coimbra, Portugal 5 Departament de Farmàcia, Tecnologia Farmacèutica i Fisicoquímica, Facultat de Farmàcia i Ciències GH O¶$OLPHQWDFLy, Universitat de Barcelona, Joan XXIII 27, 08028 Barcelona, Spain imeton@ub.edu Chitosan is a linear cationic polymer composed of glucosamine and N-acetylglucosamine derived from chitin through a process of deacetylation. An increasing amount of research is being conducted to use chitosan nanoparticles as a vehicle for drug delivery and transport of peptides, proteins and DNA. In the present study we obtained stable chitosan-TPP (tripolyphosphate) nanoparticles complexed with a DNA plasmid expressing an shRNA to knockdown the expression of cytosolic alanine aminotransferase (cALT) in the liver of gilthead sea bream (Sparus aurata). Several siRNAs were designed to silence cALT expression in vitro. The siRNA producing higher silencing effect was subsequently used to construct an shRNA expression plasmid. Using ionic gelation, chitosan-TPP-DNA nanoparticles carrying the shRNA expression plasmid were obtained through spontaneous complexing of chitosan with TPPDNA. Optimum formulation for obtaining chitosan-TPP nanoparticles included analysis of pH on particle size and zeta potential. Physical and chemical properties of the chitosan-TPP-DNA nanoparticles revealed that the particles were small, spherical and smooth. Particle size and surface charge were determined by atomic force microscopy and laser Doppler microelectrophoresis, respectively, indicating a mean diameter size of approximately 200-250 nm and a Z potential of between 14 and 35 mV. Intraperitoneal administration of chitosan-TPP-DNA nanoparticles allowed us study the effect of expressing an shRNA to knockdown cALT in the liver of S. aurata. To this end, the hepatic expression of cALT isoforms was monitored 24, 48 and 72 h after the treatment with chitosan-TPP (control without DNA), chitosan-TPP-scramble (expressing a control scramble sequence with no homology with known sequences) and chitosan-TPP-DNA (expressing an shRNA to silence cALT expression). Administration of chitosan-TPP-DNA nanoparticles produced maximal silencing effects 72 h following the treatment, leading to significantly decreased cALT1 mRNA levels, immunodetectable ALT and ALT enzyme activity in the liver of S. aurata. This study was supported by the BIO2009-07589 (MCI, Spain) and AGL2012-33305 (MEC, Spain, cofunded by the European Regional Development Fund, ERDF, EC) grants.

Magnetic hyperthermia of MgxFe3-xO4 ferrofluid synthesized by hydrothermal procedure. 1

Vojislav Spasojevic , Irena Spasojevic , Biljana Dojcinovic and Bratislav Antic

Institute RI 1XFOHDU 6FLHQFHV 9LQĂža, University of Belgrade, P.O. Box 522, Belgrade Serbia Universitat Rovira i Virgili, Campus Sescelades, Department of Analytical and Organic Chemistry C/MarcellĂ, Domingo s/n, Tarragona, Spain 3 Institute of Chemistry, Technology DQG 0HWDOOXUJ\ 8QLYHUVLW\ RI %HOJUDGH 1HJRĂŁHYD %HOJUDGH 6HUELD * on live University of Belgrade, Faculty of Physical Chemistry, Studentski trg 12-16, Belgrade, Serbia 1

vojas@vinca.rs Abstract Set of ferrofluids, consisting of MgxFe3-xO4 QDQRSDUWLFOHV Â&#x201D;[Â&#x201D; ZKLFK DUH FRDWHG ZLWK FLWULF DFLG and dispersed in water, were synthesized by using hydrothermal method. The main objective was to measure their heating efficiency (HE) and to compare these values with the undoped Fe3O4 ferrofluid. Since several main factor influence HE, like structural and magnetic properties of magnetic nanoparticles (MNPs) as well as amplitude and frequency of applied field, complete structural and magnetic characterization were done. Formation of single phase was evident in all as prepared dried samples in the whole range of concentrations x. XRD line broadening showed that the particle diameter decreases from 18 nm for x=0 to 13 nm for x=0.6, and TEM photographs showed that the particles are spherical with log-normal distribution of particle sizes (Fig.1). All samples are superparamagnetic at room temperature, while at low temperatures super-spinglass state is evident (Fig.2 insets). Saturation magnetization is decreasing with the concentration increase (Fig.2). SPA values of ferrofluid samples are increasing with concentration x and applied frequency (Fig.3) showing improvement of heating efficiency comparing to pure magnetite. Measured SPA values are comparable or higher with so far known commercial ferrofluids [1]. [1] M. Kallumadil, M. Tada, T. Nakagawa, M. Abe, P. Southern, Q.A. Pankhurst, J. Mag.Mag.Mat. 321(21) (2009) 3650-3651.

100 60 40

f = 472 kHz f = 728 kHz 600

-40

Mg0.6Fe2.4O4

H=1000 Oe

SPA [W/g]

M(emu/g)

700

-20

-60

f = 323 kHz

800

x=0.6

-1500 -1000 -500

500

1000 1500

H(Oe)

-20

M(emu/g)

5K 300K

60 40

x=0.1

X=0.6

-40 -60

Tf=43.6K ZFC

500 400 300

200 -80

100

150

200

250

300

T (K)

-100 -60000

100 -40000

-20000

H(Oe)

20000

40000

60000

0.00

0.05

0.10

mol Mg

0.15

0.20

Improved formulation of cationic solid lipid nanoparticles displays cellular uptake and biological activity of nucleic acids

1,2

1,3

Marc Suñé-Pou , Silvia Prieto , Anna Fàbregas , Isaac Nofrerias-Roig , Anna Nardi , Encarna Garcia1,3

Montoya , Pilar Pérez-Lozano , Josep R. Ticó , Montserrat Miñarro , Cristina Hernández-Muniain , 1,3

Josep M. Suñé-Negre , and Carlos Suñé

Department of Pharmacy and Pharmaceutical Technology and Physical Chemistry, University of Barcelona, Spain.

Department of Molecular Biology and Department of Cell Biology and Immunology, Instituto de ParasitoloJtD \ %LRPHGLFLQD ³/ySH]-1H\UD´ (IPBLN-CSIC), PTS Granada 18016, Spain 3

Group of Pharmacotherapy, Pharmacogenetics and Pharmaceutical Technology of the Bellvitge Biomedical Research Institute (IDIBELL) e-mail: msunepou@gmail.com

Abstract

In recent decades, the transfection of nucleic acids using nanostructured non-viral delivery systems has become a powerful strategy in Nanomedicine to achieve gene expression regulation for the treatment of diseases, thus representing a promising new avenue for gene therapy. Cationic solid lipid nanoparticles (cSLNs) are one of the most promising nanoparticle systems for nucleic acid delivery for therapeutic usage. In a previous work, we showed that cSLNs are capable of forming complexes with DNA plasmids with low toxicity when assayed in human cells [1]. Formulation of these cSLNs consists in only three components: matrix lipid (stearic acid), cationic lipid (octadecylamine), and surfactant (Poloxamer 188). We obtained cSLNs with a mean diameter of approximately 120-130 nm [1]. Here, we present data concerning the functional characterization of cSLNs-nucleic acids lipoplexes. We found that these lipoplexes efficiently transfect circular DNA and linear RNA molecules into cells using fluorescence- and luminescence-based methodologies. We also observed that they deliver nucleic acid molecules into cells where they display full bioactivity at nontoxic concentrations. In the case of DNA, our results indicate that upon cSLN-complexation, the nucleic acid is protected against nuclease degradation, internalized into cells, released in the cell, transcribed to RNA in the nucleus, exported to the cytoplasm, and translated to protein, displaying full bioactivity. Hence, we established a novel and simple cSLN formulation as a powerful tool for future therapeutic use. References

[1] Fàbregas, A., Sánchez-Hernández, N., Ticó, JR., García-Montoya, E., Pérez-Lozano, P., SuñéNegre, JM., Hernández-Muniain, C., Suñé, C., Miñarro, M., Int J Pharm 473 (2014) 270.

Importance of calcium ions at bioinspired integrative implant surfaces for tissue regeneration Ricardo Tejero, Eduardo Anitua, Miguel Ángel Pacha, Mar Zalduendo, María Luisa González BTI Biotechnology Institute, C/Leonardo da Vinci 14B, Miñano, Spain ricardo@minin.es Abstract The calcium ion (Ca) is centrally involved in a multitude of biological processes, among which the entire lifecycle of bone from formation to repair [1]. Ca mediates platelet activation and provisional matrix formation, binds acidic-rich proteins and creates supersaturating conditions for bone-mineral nucleation [2]. Elevated extracellular Ca levels have been associated with enhanced osteogenic cell activity and increased osteoclast apoptosis [3]. At implant surfaces, Ca enables the electrostatic binding of several cell types and biomolecules relevant for biomineralization [4]. Also, Ca has been identified as the principal constituent of the thin layer formed between bone and osseointegrated implants [5]. However, once implanted, blood calcium has a limited accessibility to the surfaces. Therefore we hypothesize that implant surfaces modified previously with Ca may be more effective to advance and improve healing around integrative implants. Here we present a comprehensive characterization of Ca ion modified titanium implant surfaces. We used Time of Flight Secondary Ion-Mass Spectrometry (ToF-SIMS) to analyze the ion exchange process in air. We employed goniometry to measure the surface free energy and inductively coupled plasma optical emission spectrometry (ICP-OES) to quantify the release of Ca ions in solution. Further, we combined absorbance and confocal innmunofluorescence to evaluate surface-mediated platelet activation and coagulation. Finally, these interfaces were functionally tested in terms of their tissue integrative potency with mammalian and bacterial cell experiments and in vivo integration tests. Unlike standard implant surfaces, ToF-SIMS depth profiles revealed absence of hydrocarbons at the titanium oxide interface, which together with the presence of Ca ions conveys superhydrophilic properties to these surfaces. In water, roughly ¾ of the Ca is released/dissolved almost instantly followed by a progressive decay over more than 80 days. However, in the presence of blood plasma, Ca induces platelet activation and the formation of a fibrin clot bound to the surface. Ca surfaces significantly increased Osteoblast Primary Cell adhesion, proliferation and osteopontin expression and reduced significantly the initial bacterial adhesion of three bacterial strains. Finally, the in vivo tests showed increased levels of integration around Ca-modified implants. In summary, we have outlined the importance of Ca ions in the initial and long-term healing events around implants, particularly when delivered from the surface. Relevant for clinical applications, these results highlight the importance of bioinspired multifunctional surface designs to modulate the wide variety of complex processes that govern tissue-regeneration around implants. References [1] Habibovic P & Barralet JE Acta Biomater, 7 (2011) 3013±26 [2] R. Tejero, E. Anitua, G. Orive, Prog Polym Sci, 39 (2014) 1406±47 [3] Dvorak MM, et al. PNAS USA, 101 (2004) 5140±5 [4] Mann S. Oxford University Press (2001) 89-124 [5] Davies JE, J Dent Educ, 67 (2003) 593±6

Computer simulations of nanosized biomaterials Antonio Tilocca Department of Chemistry, University College London, 20 Gordon Street, London WC1H 0AJ, UK a.tilocca@ucl.ac.uk Abstract %LRDFWLYH JODVVHV %*V LQVSLUHG WR WKH RULJLQDO +HQFKÂśV 6 IRUPXODWLRQ DUH VXFFHVVIXOO\ HPSOR\HG LQ biomedical applications, for instance as bone defect fillers, where they are used in compact or particulate form. These materials also exhibit great potential in regenerative medicine, especially in their various nanosized forms such as nanoparticles (NPs) and nanorods, which appear to offer enhanced biological activity compared to their larger-size particulate and compact counterparts [1]. This promising potential has not translated into practical applications thus far, due to the limited fundamental understanding of the origin of the superior properties of BG nanosystems is required: in this context, computer simulations are playing an increasingly important role [2-3]. Previous Molecular Dynamics (MD) simulations show how the reduced size may affect structural and dynamical features of a BG nanoparticle, in a way which could in turn enhance the bioreactivity of these systems [4]. We present MD simulation data showing in detail how, compared to the corresponding bulk systems or to the NP core, reducing the particle size may lead to a reduction in the already low silicate connectivity of these + 2+ materials, as well as to a higher Na /Ca ratio and a higher mobility of Na cations, in the outer regions of the nanoparticle directly exposed to the biological environment. We discuss how these nanosizeinduced effects can give rise to - and thus be further controlled to achieve - higher bioreactivity in BG nanoparticles. References [1] S. Labbaf et al., Biomaterials 32 (2011) 1010. [2] A. Tilocca, J. Mater. Chem. 20 (2010) 6848. [3] E. Berardo et al. RSC Advances 4 (2014) 36425. [4] A. Tilocca, J. Mater. Chem. 21 (2011) 12660.

Suspended silicon micro-photodiodes for single cell stimulation Carolina Vargas-Estevez1*, Lilian Enríquez Barreto2, M. Duch1, F.J. Del Campo1, Gonzalo Murillo1, Nuria Torras1, Carlos A. Saura2, José Antonio Plaza Plaza1, Jaume Esteve1* 1*

Micro and Nano-tools, Instituto de Microelectrónica de Barcelona IMB-CNM (CSIC), Barcelona, Spain. 2 Institut de Neurociències (INc), Universitat Autònoma de Barcelona, Spain.

Abstract Electric stimulation has been widely used to treat medical conditions of excitable cells [1]. The main problem with the technology used for this stimulation is that besides reducing the size of the devices, there are still long terms adverse effects related with the hardware [2], and even non-invasive approaches have still much work to accomplish a focalized stimulation on a targeted group of cells [3]. Taking the advantages of microtechnology in reduce the scale, we have developed suspended silicon micro-photodiodes to electrically stimulate single cells. These microstructures were electrically characterized in substrate, measuring its photovoltaic behavior. Once they were suspended, they were also electrochemically characterized to probe their performance in liquid by a redox reaction quantified by spectroscopy. Then as a proof-of-concept, an in vitro test with mice neurons was performed activating the micro-photodiodes with pulses of an intense white lamp. The effect of stimulation was studied morphologically by scanning electron microscope (SEM) imaging and fluorescence images. The combination of these techniques revealed a visible effect on the neurons compared with cell control. The branching length seemed to be enlarged by effect of these stimuli plus morphological changes in the branches itself were observed. The tests carried in this study showed the feasibility of these suspended micro-photodiodes to electrically stimulate single cells in a wireless less invasive way.

References

[1] D. R. Merrill, M. Bikson and J. G. Jefferys, "Electrical stimulation of excitable tissue: design of efficacious and safe protocols," Journal of neuroscience methods, vol. 141, no. 2, pp. 171-198, 2005. [2] M. Y. Oh, A. Abosch, S. H. Kim, A. E. Lang and A. M. Lozano, "Long-term hardware-related complications of deep brain stimulation," Neurosurgery, vol. 50, no. 6, pp. 1268-1276, 2002. [3] P. M. Rossini et al., "Non-invasive electrical and magnetic stimulation of the brain, spinal cord and roots: basic principles and procedures for routine clinical application. Report of an IFCN committee.," Electroencephalography and clinical neurophysiology, vol. 91, no. 2, pp. 79-92, 1994.

TiO2 nanostructures as UV blocking components in sunscreens Bogdan Stefan VASILE, Roxana DUTU, Ecaterina ANDRONESCU, Adrian Vasile SURDU, Roxana TRUSCA, Ovidiu OPREA, Otilia Ruxandra VASILE University POLITEHNICA from Bucharest, National research Centre for Food Safety, No. 313 Splaiul Independentei Street, Bucharest, Romania. bogdan.vasile@upb.ro Abstract TiO2 nanoparticles were synthesized using sol-gel method, starting from organic precursors: titanium n-butoxide and titanium isopropoxide. The nanoparticles obtained have been used as UV blocking component in the preparation of sunscreen creams. The synthesized nanostructures were characterized by means of powder X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM). The obtained products, sunscreen containing small amounts of TiO2 nanostructures were analysed using UV-VIS spectroscopy. References [1] Celline Botta, Jerome Labille, Melanie Auffan, Daniel Borschneck, Helene Miche, Martiane Cabie, Armand Masion, Jerome Rose, Jean-Yves Bottero, Environmental Pollution, 159, 2011, 1543-1550 [2] L.Kubac, J. Akram, K.Keijlova, H. Bendova, K. Klanova, Z. Hladikova, P. Pikal, L. Kavorikova, L. Kasparova, D. Jirova, International Journal of Pharmaceutics, 481, 2015, 91-96. Figures 35

(110) Rutile

10000 8000 6000 4000 2000 0 20

(200) Anatase

(101) Rutile (112) Anatase (200) Rutile (111) Rutile (210) Rutile

12000

(202) Rutile

(301) Rutile, (116) Anatase (112) Rutile

(202) Rutile, (301) Anatase

14000

(220) Rutile

16000

TiO2 but. 450oC

(101) Anatase

Counts (a.u.)

18000

(311) Rutile

2000 0 20000

(200) Anatase

4000

(211) Rutile, (105) Anatase

6000

(101) Anatase

8000

(112) Anatase (200) Rutile (111) Rutile (210) Rutile

10000

TiO2 Ip. 450 C

(220) Rutile

(101) Rutile

14000 12000

(301) Rutile, (116) Anatase (112) Rutile

16000

(211) Rutile, (105) Anatase

Counts (a.u.)

18000

(310) Rutile, (204) Anatase (221) Rutile

(110) Rutile

20 20000

2theta

1.2

10% but 15% but 20% but

1.0

Abs (a.u.)

10% ip 15% ip 20% ip

1.0

Abs (a.u.)

1.2

0.8

0.6 0.6

0.4 0.4

0.2 0.2 200

300

400

500

600

Wavelength (nm)

700

800

200

300

400

500

600

Wavelength (nm)

700

800

Pentacyclic triterpenes with anti-inflammatory activity formulated in topical nanoemulsion.

Karla Ximena Vázquez-Prada1,2, Paola Bustos-Salgado3, Berenice Andrade Carrera2, Beatriz Clares-Naveros4, Helen L. Alvarado3, Ana Calpena-Campmany3 and María Luisa Garduño-Ramírez2. 1

Facultad de Ciencias Biológicas, Universidad Autónoma del Estado de Morelos. Centro de Investigaciones Químicas, Instituto de Investigación en Ciencias Básicas y Aplicadas, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, 62209, Cuernavaca, Morelos, México 2 Department of Pharmacy, Pharmaceutical Technology and Physical Chemical, Faculty of Pharmacy and Food Sciences, University of Barcelona, Av. Sant Joan s/n, Barcelona, Spain. 4 Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of Granada, Granada, Spain. lgarduno@uaem.mx 2

Introduction: The nanoemulsions are colloidal dispersions of immiscible liquids translucent stabilized 1,2 by a surfactant, with a droplet size between 50 and 1000 nm. These formulations must be monophasic. The aim of the project is nanostructured formulations with three different pentacyclic triterpenes: obtusilinic acid (1), acid oleanolic-acid urseolic (2) and oleanolic acid (3), isolated from Plumeria obtusa used as anti-inflammatory in traditional medicine and one reference Indomethacin (4); subsequently the droplet size and polydispersity of such formulations was measured. The NEs1 and 3 NEs4 were evaluated in mouse ear oedema induced by TPA. Materials and methods: Obtusilinic acid was isolated from methanol extract of the aerial parts of P. obtusa, which was identified by spectroscopic techniques such as 1D and 2D NMR. Tritepene (1) and Indomethacin (4) were prepared in formulations nanostructured (nanoemulsion) to improve 3 bioavailability as anti-inflammatory agents above have already been evaluated in vivo (TPA Model). 3 The nanoemulsions were prepared under the methodology of V. Dominguez-Villegas. Subsequent to preparing the formulations were characterized and determinated the droplet size with ZetaSizer NanoSeries. Finally the nanoemulsions (NEs1 and NEs4) were evaluated in mouse ear oedema induced by TPA. Results and discussion: NEs1, NEs4 and NEs (free drug) were monophasic, transparent and translucent systems. The nanoemulsions Z-Average was determined with Z-Sizer NanoSeries and NEs has a droplet size 537.8 ± 154.2 nm; NEs1 700.1 ± 150.7 nm and NEs4 622.1 ± 122.0 nm. The topical anti-inflammatory activities of triterpene (1) and Indomenthacin (4) (free drug and encapsulated) were determined by assessing the effect of the drugs on mouse ear oedema induced by TPA. Triterpene (1) and Indomethacin (4) anti-inflammatory activities were 68.63 ± 0.40 and 90.1 ± 0.40 %, respectively. The anti-inflammatory activity nanoemulsions (NEs1 and NEs4) were 78.75 ± 2.16 and 85.00 ± 3.75 %. Conclusions: The nanoemulsion prepared were monophasic formulations achieved are within the nanometer range, since one of them is greater than 1 m. Indomethacin anti-inflammatory activity did not show a significant difference between Indomethacin free and nanoemulsion (NEs4); howerever, the values to triterpene (1) free and the formulations (NEs1) showed an increase the anti-inflammatory activity about 10 % in the formulation. The nanoemulsions Z-Average did not show a significant difference in two formulations compared between them but in the anti-inflammatory effect was it; this effect could be probably due to the obtusilinic acid lipophilicity. Acknowledgments: The authors would like to thank Gattefossé, the LANEM Laboratory, UAEM and the Ministry of Science and Innovation of Spain with MAT201459134R project. References [1] G. Guglemini. Dermato Cloin. 26 (4) (2008) 341. [2] T.P.U. Ravi, T. Padma. Res. Biotechnol. 2 (3) (2011) 1. [3] V. Dominguez-Villegas, B. Clares-Naveros, M. L. Garcia-Lopez, A.C. Calpena-Campmany, P. Bustos-Zagal, M. L. Garduño-Ramirez. Colloids and Surfaces B: Biointerface. 116 (2014) 183.

Exosome-like nanoparticles for microRNA selective transport to tumoral cells 1

Vázquez-Ríos AJ , Bouzo BL , Abuín C , Grijalvo S , Avinyó A , Eritja R , López-López R , 1 de la Fuente M . 1 Nano-oncologicals Lab, Translational Medical Oncology Group, Health Research Institute of Santiago (IDIS), SERGAS, Santiago de Compostela, Spain. 2 Nucleic Acid Chemistry Group, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Barcelona, Spain. abivazquezrios@hotmail.com Abstract Exosomes are extracellular membrane vesicles (40-100 nm) secreted by most cell types in healthy and pathological conditions. Given their role in cell-to-cell communication, carrying genetic material and proteins selectively, exosomes have increasingly gained much attention as 1 the “ideal drug delivery vehicle” . However, cells release relatively low amounts of exosomes 2 and purification and manipulation is cumbersome . Furthermore, their use in human therapy requires much deeper knowledge of its composition and function to ensure the safety of this approach. In order to avoid the mentioned drawbacks and making use of nanotechnology, we have developed a nanoplatform that simulate natural exosomes in structure and functionality but with a controlled composition and much higher yield of production, exosome-like nanoparticles (ELN). Besides, our ELN are based on liposome technology with an easily 3 scalable preparation method (injection of ethanol in water ) and highly reproducible. Our ELN are stable in different biological media, no toxic, and most importantly, efficiently internalized by tumor cells. To demonstrate their potential in drug delivery, ELN were loaded with different therapeutic cargoes (microRNAs, hydrophobically modified nucleic acids and curcumine), and compared with exosomes isolated from cell cultures of cancer cells. In all cases, higher encapsulation efficiencies were reported for ELN. The functional significance of lost microRNAs has been reported in several human 4 malignancies ; however, lack of an effective tumor-specific delivery system remains an unmet clinical challenge for successful translation of microRNAs. In this study, the tumor suppressor microRNA-145 was associated to our nanosystems and efficiently transported to tumoral cells, achieving a high transfection efficacy. Finally, we have also proved that ELN can be successfully functionalized with proteins that have been previously identified in exosomes, and reported for their organotropic properties. We believe that this strategy will favour the accumulation of ELN in the tumor microenvironment and hence the delivery of the associated therapeutic microRNAs. Current experiments are being designed to prove this hypothesis. In conclusion, we have succesfully achieved the preparation of novel nanosystems completely inspired by exosomes, with a versatile load capacity, and an efficient interaction with tumoral cells, thefore holding a great potencial in cancer therapy. References [1] Record M, et al. Biochim Biophys Acta 1841 (2014) 108. [2] Durán JDG, et al. J Pharm Sci 97 (2008) 2948. [3] Holgado MA, et al. Int J Pharm 358 (2008) 27. [4] Setua S, et al. J Gastrointest Surg (2016) 1. Figures exosome

Exosome

Exosome-like N

D N"

% EE

)*+,-./$011+2.0,.+/$

Exosomes

Liposomes

siRNA

53 ± 3

53 ± 1

DNA-CH

75 ± 2

87 ± 8

Curcumin

33 ± 4

87 ± 6

!"#$%&’$($ -3/$

1/$

Figure. A) TEM images of natural exosomes and exosome-like nanoparticles showing a similar size and morphology. B) Encapsulation efficiencies of different therapeutic molecules in exosomes and ELN, higher EE were achieved in ELN than in exosomes. C) Confocal image of ELN carriung miR145 inside tumoral cell (A549 cell line). ELN-NBD. miR145-Cy5, Nuclei-DAPI. D) Western Blot showing the protein association of the isolated ELN compared to the supernatant (isolation by ultracentrifugation at 35000 rpm, 1h, 15ºC).

Understanding the Kinetics of Protein-Nanoparticle Corona Formation Oriol Vilanova, Judith J. Mittag, Philip M. Kelly, Silvia Milani, Kenneth A. Dawson, Joachim O. Rädler, Giancarlo Franzese Secció de Física Estadística i Interdisciplinària ± Departament de Física de la Matèria Condensada, Facultat de Física, Universitat de Barcelona Institut de Nanociència i Nanotecnologia, Universitat de Barcelona ovilanova@ffn.ub.edu Abstract When a pristine nanoparticle (NP) encounters a biological fluid, biomolecules spontaneously form adsorption layers around the NP, called ``protein corona''. The corona composition depends on the time-dependent environmental conditions and determines the NP's fate within living organisms. Understanding how the corona evolves is fundamental in nanotoxicology as well as medical applications. However, the process of corona formation, is challenging due to the large number of molecules involved and to the large span of relevant time-scales ranging from 100 ȝs, hard to probe in experiments, to hours, out of reach of all-atoms simulations. Here we combine experiments, -3 3 simulations, and theory to study (i) the corona kinetics (over 10 to 10 s) and (ii) its final composition for silica NPs in a model plasma made of three blood proteins (human serum albumin, transferrin, and fibrinogen). When computer simulations are calibrated by experimental protein-NP binding affinities measured in single-protein solutions, the theoretical model correctly reproduces competitive protein replacement as proven by independent experiments. When we change the order of administration of the three proteins, we observe a memory effect in the final corona composition that we can explain within our model. Our combined experimental and computational approach is the first step towards the development of systematic prediction and control of protein-NP corona composition based on a hierarchy of equilibrium protein binding constants. References [1] Monopoli, M. P.; Aberg, C.; Salvati, A.; Dawson, K. A.; Aberg, C. Nature nanotechnology 7, (2012) 779±86. adler, J.; Baldelli Bombelli, F. ACS nano 6, (2012) 2532±41. [3] Vilaseca, P.; Dawson, K. A.; Franzese, G. Soft Matter 9, (2013) 6978. [4] Salvati, A.; Pitek, A. S.; Monopoli, M. P.; Prapainop, K.; Bombelli, F. B.; Hristov, D. R.; Kelly, P. M.; Aberg, C.; Mahon, E.; Dawson, K. A. Nature nanotechnology, 8, (2013) 137±43. Figures

Titania coating of mesoporous silica nanoparticles- Protective effects on vasodilator function, ex vivo. Debra Whitehead, May Azzawi, Asima Farooq, Peter Kelly, Lubomira Tosheva, Manchester Metropolitan University, Chester Street, Manchester, M1 5GD, UK d.whitehead@mmu.ac.uk Abstract: Titania (TiO2) nanoparticles encompass unique properties that allow them to be used in numerous biomedical applications. Our previous research studies have suggested that the hydroxyl groups on silica surface lead to the attenuation in vascular function through the possible production of reactive oxygen species (ROS). Furthermore, we have previously demonstrated that ceria coating of silica [1] nanoparticles significantly improved the magnitude of vasodilation . Mesoporous silica nanoparticles (MSNs) are widely used for drug delivery applications owing to their well-ordered pore architecture 2 giving them very high surface area to volume ratios (>1100 m /g). The MSNs particles were synthesized using a liquid crystal templating method and characterised using electron microscopy for size and pore structure. The nanoparticle size and morphology were analysed using transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. The MSNs were further characterised by Zeta potential, nitrogen adsorption measurements and infra-red spectroscopy. The MSNs utilised were monodispersive and spherical with a diameter of ~100 nm and the interior pores had an average diameter of ~3 nm. We have loaded these pores with endothelial-independent vasodilator, sodium nitroprusside (SNP) and demonstrated real time drug delivery by the MSNs on by assessing the degree [2] of aortic vessel dilation, in real time, using an organ bath system, ex vivo . Vessel exposure to unloaded MSNs resulted in a small attenuation in constriction that occurred after approximately 1 h. In contrast, MSNs loaded with SNP led to a rapid relaxation of aortic vessels that was sustained over a 3 h period (p<0.001). Thus, our aim was to determine the influence of TiO 2 coating on improving the vasodilator function of aortic vessels exposed to mesoporous silica nanoparticles (MSNs) by testing vasodilation response, ex vivo. The SNP drug loaded MSNs were coated with titania (Ti) by magnetron sputtering, a smooth coating was observed by TEM (Figure 1). The drug loading and drug release was determined before and after the titania coating in high potassium physiological salt solution using dialysis and inductively coupled plasma atomic emission spectroscopy. MSNs loaded with SNP (with or without surface titania 12 -1 11 -1 coating) were incubated with 3 mm aortic vessel rings (at 1.96Ă&#x2014;10 NP mL and 1.31Ă&#x2014;10 NP mL ) over a 3 h time period. The effects of both MSNs and TiMSNs on endothelial-dependent [acetylcholine (ACh); 0.01 M-1.0 mM] and independent [sodium nitroprusside (SNP); 0.01-10 M] dilator responses were investigated. The TiO2 coating the MSNs lead to the significant improvement in vasodilation, suggesting that the titania reduces the ROS produced by silica. Our results suggest that MSNsTi have great implications for future biomedical applications in the treating of conditions where attenuated dilation occurs. These MSNs are promising drug delivery platforms for the targeted delivery of drugs to the vasculature. References Farooq, A., Mohamed, T., Whitehead, D. and Azzawi, M., Journal of Nanomedicine & [1]

[2]

Nanotechnology, 2014. Farooq, A., Tosheva, L., Azzawi, M. and Whitehead, D., Journal of Colloid and Interface Science, 2016, 478, pp.127-135.

Figures:

Transmission electron microscope image of MSNs and (left) and TiMSNs (right).

ISBN 978-84-617-6662-8 Calle Alfonso Gomez 17 Planta 2 - Loft 16 28037 Madrid (Spain) info@phantomsnet.net www.phantomsnet.net