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Tissue Engineering Scaffolds for Organ Regeneration
Tissue Engineering Scaffolds for Organ Regeneration Adaeze Eze, Naznin Sultana, and Dennis E. Daniels Undergraduate Medical Academy, Prairie View A & M University, Prairie View, TX
Abstract Background: Regenerative medicine plays a major role in restoring and repairing damaged or diseased tissue. Tissue engineering (TE) is a form of regenerative medicine. TE scaffold, a component of TE, is the idea of using biomaterials that are degradable, to aid in restoring, repairing, and treating damaged tissue. The aim of the tissue engineering scaffold technique is to insert a non-toxic component into the body that the cells can attach, migrate, differentiate, and proliferate through. Objective: The objective of this project was to fabricate and investigate the properties of the combined component of a natural, biodegradable and biocompatible polymer, chitosan, and a natural product, pectin, for TE scaffolds application.
Introduction The tissue engineering approach is an application derived from the idea of repairing damaged or diseased tissue by regenerating the tissue. In recent decades, there has been a sharp increase in the use of resorbable tissue implants, which in turn has spurred extensive research in the field of tissue engineering. Tissue engineering scaffold is a component of tissue engineering. In the methodology of producing TE scaffold, a biodegradable polymers that provide secure structural foundation for cell attachment and tissue regeneration. The study of tissue engineering scaffold is an application that researchers have done to determine if it is a less invasive and effective treatment for regenerative medicine. Materials/ Methods
Chitosan and Pectin. Chitosan was purchased from sigma with medium molecular weight. Pectin from citrus peel (Pc, galacturonic acid content of 80.2%, methoxylation degree of 7.6% and Mv of 45 kDa) was obtained from Sigma-Aldrich. Different concentration of Pectin will be used.
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Preparation of composite scaffolds. Initially, an acetic acid solution of 2% (v/v) was prepared as a solvent of which 20 ml was added to each polymer and pectin content in a beaker, for making a solution with the desired concentration of chitosan (CS) and Pectin. In this case, 1.15 g chitosan and 0.1 g pectin were weighed and dissolved in 20 ml 2% acetic acid. The solutions were magnetically stirred in a beaker until completely dissolved. Then the solutions were transferred in a freezer at - 18C and then freeze dried using a Labconco Freeze-dryer.
Quality control and physical characterization. We characterized the physical properties of the scaffolds and compared them with previously fabricated scaffolds to ensure consistency with our previous lots. Results 3-D scaffolds were successfully fabricated using Freeze drying technique. The scaffolds had good handling properties. Each scaffolds had a length of 4 cm and diameter of 1 cm. Scanning Electron Microscope (SEM) image confirmed the porous structure of the scaffolds. The pores were several micro-meter size and were interconnected. EDX spectrum confirmed the presence of C and O.
Conclusion Preliminary results shows that the scaffolds could be potential for tissue engineering applications. Further study on cell culture is needed to confirm the cytotoxicity of the materials.
References Chan, B.P.; et al. Scaffolding in tissue engineering:
General approaches and tissue-specific considerations, 2008. Sultana, N; et al. Chitosan-Based Nanocomposite
Scaffolds for Tissue Engineering Applications, 2015.
Acknowledgements R&I’s Office of Undergraduate Research (OUR and Undergraduate Medical Academy, Prairie View A&M University.
Introduction
Regenerative medicine plays a significant role in restoring and repairing damaged or diseased tissue. Tissue engineering (TE) is a form of regenerative medicine. TE scaffold, a component of TE, is the idea of using degradable biomaterials, to aid in restoring, repairing, and treating damaged tissue. (Sultana, 2015) The tissue engineering scaffold technique aims to insert a non-toxic component into the body that the cells can attach, migrate, differentiate, and proliferate through. (Chan, 2008)
Objective
This project’s objective was to fabricate and investigate the properties of the combined component of a natural, biodegradable and biocompatible polymer, chitosan, and a natural product, pectin (De Souza et al., 2019), for TE scaffolds application.
Materials and Methods
Chitosan and Pectin. Chitosan with medium molecular weight and pectin from a citrus peel (Pc, the galacturonic acid content of 80.2%, methoxylation degree of 7.6%, and M, of 45kDa) were purchased from Sigma-Aldrich.
Preparation of composite scaffolds. In preparation of the solvent, 1mL of an acetic acid solution of 2% (v/v) was added with Milli-Q water (20mL) to each polymer and pectin content in a beaker to produce a solution with the desired concentration of chitosan (CS) and pectin. The polymer solution was made by adding 0.1 g pectin and 1.15 g chitosan into the acidic solvent. Figure 2 (a) shows the solutions were magnetically stirred in a beaker for 5-10 minutes until completely dissolved. As seen in Figure 2 (b), the beaker’s polymer solution was transferred into four vials in the fume hood and then transferred in a freezer at 18°C to cool and mold for two days. Figure 2 (c) shows that on the third day, the vials were placed into a Labconco Freeze-dryer to begin the solidification process. Procedures and methods were established by Sultana (2015). After the freeze-drying step, the samples in the vials were removed and cut into smaller sections for Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray (EDX) testing.
Quality control and physical characteristics: The scaffolds’ physical properties were characterized and compared with previously fabricated scaffolds to ensure consistency with previous lots.
Fig. 1 --- flow diagram for preparation of composite scaffolds
Results and Discussion
(a) (b) Fig. 2 --- (a) general appearance; (b) sample preparation for characterization
(a) (b) Fig. 3 --- (a) SEM of the scaffold; (b) EDX of scaffold sample
3-D scaffolds were successfully fabricated using the Freezedrying technique. The scaffolds had good handling properties. Each scaffold had a length of 4 cm and a diameter of 1 cm. The SEM image confirmed the porous structure of the scaffolds. The pores were several micrometers in size and were interconnected. EDX spectrum confirmed the presence of C and O.
Conclusion
References
Sultana, N; et al. Chitosan-Based Nanocomposite Scaffolds for Tissue Engineering Applications, J. Nanomater. 2015: 1-7. Chan, B.P.; et al. Scaffolding in tissue engineering: general approaches and tissue specific considerations, Eur. Spine J. 2008: 467-479. De Souza, Bombaldi, Fernanda Carla; Bombaldi de Souza, Renata Francielle; Drouin, Bernard; Mantovani, Diego; Moraes, Ângela Maria. Comparative study on complexes formed by chitosan and different polyanions: Potential of chitosan-pectin biomaterials as scaffolds in tissue engineering, Int. J. Biomacromol. 2019. 132:178-189
Adaeze Eze is a junior, majoring in Biology with a Chemistry minor. Dr. Naznin Sultana is a Research Scientist with a research interest in Tissue Engineering.