Global Innovation in Wound Care Summit Series - Part 2 - Biofilm Masterclass by woundmasterclass

December 2023

Global Innovation in Wound Care Summit Series Part 2: Biofilm Masterclass Moderator Miss Negin Shamsian Consultant Plastic and Reconstructive Surgeon (Locum) London, United Kingdom

So, What is Biofilm? Biofilm is defined as one of the oldest lifeforms on Earth and is something developed by bacterial organisms. It is only within the last 40 years that Biofilm has been given this name in Science and Medicine; and it is the natural and predominant

Introduction

way that bacteria live and thrive. Bacteria will attach

themselves to a surface, which can either be liquid or mmersive Interactive Event; Taking wound care

solid, and once they’re securely attached, they secrete

into a new dimension. Hosted by Dr N. Shamsian

a polymeric matrix around themselves – This is what

in partnership with Dr M. Sanders and Dr P Bowler.

we call Biofilm. Biofilm acts as a protective layer that

Bowler has an extensive history and career within

defends the bacterial organism from hostilities within the environment or hosted organism.

wound care development and research with over

Supported by

30 years of experience in microbiology, particularly

This Biofilm layer contains exopolysaccharides – high-

focused on infections. He has been involved in the

molecular-weight polymers that are composed of

development of wound dressings alongside various

sugar residues and are secreted by a microorganism

independent consultants. Sanders has worked for

into the surrounding environment – DNA and RNA.

a diagnostic wound care company that developed

Both DNA and RNA are almost identical, are used

diagnostics for bacterial status that was sold back into

to store genetic information, but they have differing

the wound-check business. After 16 years working

base pairs and can be found in various places of the

with the company, he joined a consulting firm where

cell, but mostly within the nucleus. Due to this nature

he expanded this experimental field by developing

and structure of the Biofilm, it is easy for them to

numerous products for wound care and targeted the

adapt by altering or changing their phenotype and

anti-inflammatory stages, and infection whilst also

genotype for survival. This presents a challenge in

maintaining an interest in the fields of cancer and

the microbiology world of Science and Medicine as

brain injury through studying neuroinflammatory

it means antibiotics are ineffective in the treatment

processes. He now works in partnership with the

of bacterial infections that include such complex

consulting firm ProDevLabs who are supporting the

structures.

event discussed today.

Wound Masterclass - Vol 2 - December 2023

Global Innovation in Wound Care Summit Series Part 2: Biofilm Masterclass

“Biofilm is inextricably linked to chronic infections, leading to delayed healing and recurrence. This requires tailored treatment for wound care.” Dr M. Sanders

Treatment: Antibiotics and Initial Stages

no mention of Biofilm; during this time, Science and Medicine were aware of the existence but unsure of

Antibiotics and antiseptics are mostly ineffective

the capabilities, adaptability and extensive nature of

against Biofilm due to this shifting nature and structure

the bacteria.

that Biofilm possess, meaning the inflammatory response within a wound is increased. At this stage,

At What Point Does the Wound Become Critical

it is integral and crucial that debridement and the

During This Colonisation?

washing of the wound is used as an intervention to discover the depth and nature of the inflammatory

When we consider the critical nature of a wound, it is

process.

important to consider the comorbidities and health of the patient overall. This staging of ‘critical’ will differ

How Does the Biofilm Trigger the Inflammatory

from patient to patient, and there is no set timeframe.

Response?

Essentially, this will be when the host is no longer able to control the microbial contamination, therefore

When the Biofilm hijacks the hosts skin/wound,

requiring

additional

and

interventive

microbial

neutrophils attempt to discover the parasite or

support to manage the wound and return control to

bacterial infection where they undergo an oxidated

the host.

burst. The Biofilm itself is tolerant to the immune cells and attracts them towards the bacteria where they

Bowler states that he would not use this term

spill enzymes into the surrounding environment that

in modern society and the world of Science and

damage the hosts tissue further, which is why we can

Medicine as it has been superseded by the Biofilm

almost consider Biofilm to be a parasitic infection.

Continuum. Bacteria initially starts as a planktonic cell

As this tissue devitalises, it provides more tissue

on the surface, mostly a wound, where they attach

for the Biofilm to consume and continue to spread.

themselves and adapt rapidly. As the Biofilm layer is

Elimination of the Biofilm at this stage becomes

produced, this is the continuum; the process by which

crucial to prevent further inflammatory response and

the wound becomes colonised.

infection spread, whether this is through physical intervention, antimicrobial or new strategies within

Randy Walker, a pioneer in the field of Science and

wound care.

Medicine, recently spoke about ‘When does a wound become chronic? Does this take 30 days or longer?’

Bowler very clearly agrees that all chronic wounds

The problem with this question is that Biofilm can

have a Biofilm element due to their nature being so

quicken the process, and a wound can easily turn

disruptive and the response to antibiotic treatment.

chronic within a matter of days. Walker acknowledges

As the antibiotics are ineffective and the persistence

this and takes the approach that; if there are warning

of chronic wounds prevails, there is clear evidence

signs there, effectively treat in the early stages.

that a Biofilm layer is preventing efficient healing.

This will minimise the chance of chronic wounds,

The impact of Biofilm on the healing stages is vast

inflammation and the need for further intervention

and includes, but it not limited to; delayed healing

through new strategies and emerging technologies.

response, chronic wounds, recurrent infections, delayed closure, and impaired blood vessel formation.

Challenges: What Do Professionals Face When Treating Biofilm?

Critical Colonisation is a term that was used for around 25 years and refers to the stage when

Some of the challenges that consultants and working

bacteria colonise a wound to cause further infection

professionals face when treating wounds, those

and increased challenge to treatment. When this

that have this Biofilm layer, is the microscopic and

term was first penned by professionals, there was

hidden nature of the Biofilm itself, the variable

Wound Masterclass - Vol 2 - December 2023

Global Innovation in Wound Care Summit Series Part 2: Biofilm Masterclass

“We need a Multifactorial Blockade to healing.” Dr N. Shamsian

composition of the bacteria, sampling techniques

For immunocompromised individuals, Pseudomonas

for definitive diagnosis, laboratory techniques to

Aerginosa is a major issue as it is multi-antibiotic

extract the DNA, dynamic nature of the wound, false-

resistant and this reduces the effectiveness of

negative results and the final clinical interpretation.

treatment so further treatment and combination

From the initial stage of seeing the wound, to the

therapies must be considered.

final stage of diagnoses there are clear barriers and challenges that need to be considered. There are also

New Treatment and Strategies

further factors to consider, like the comorbidities of patients, particularly those with Diabetes or Venous

Dr Bowler and Dr Christine Murphy, specialist from

insufficiency as this can cause further issues when

Ottawa, Canada, have penned the term Granulitisas

determining the nature of the wound; and whether

the induced hyperinflammatory process of Biofilm.

the wound does have a Biofilm component.

Although developing technologies are helping to identify the nature of the wound, it is crucial to

Accurate diagnosis is vital for effective treatment of

start effective treatment and reduce the risk of

the wound as we can guide our targeted antimicrobial

occurrence of Granulitis. This treatment can include

selection for patients, and it allows room for

Photodynamic Therapy, Quorom Sensing Inhibitors,

combination therapy where appropriate. Consultants

Biofilm Disputing Enzymes, Nanoparticle based

can be more informed of the optimal wound

approaches,

management and strategies to reduce the risk of

Systems, Electrochemical Treatments, Antibiofilm

recurrence, whilst optimising the resource utilisation

Surfaces and Coatings, Biofilm Imaging Techniques,

and improving the overall treatment outcome.

Combination Therapies and Vaccines.

Some examples of specific microorganisms include:

Bowler mentions that they have been developing a

Biofilm-responsive

Drug

Delivery

3-Dimensional printing method to copy Biofilm, or •

Staphylococcusaureus

a collagen substrate, to assess antibiofilm agents in

•

Pseudomonasaeruginosa

vitro. By implanting the genetically contained and

•

Escherichia coli

grown organisms, they have transplanted this onto

•

Streptococcus species

animals for experimentation and testing. These

•

Fungi

models prove vital for representing the stages

•

Viruses specifically

and demand of resistance that we see on humans’

•

Bacteriophages

wounds. These clinical models are invasive and

•

Protozoa such as acanthamoeba and naegleria

hard to remove, and professionals find themselves

species

using fine and sharp debridement as a method for removing and disrupting Biofilm, enhancing the

And some further examples of microorganisms that

importance of an imaging device to see the definitive

we know leave Biofilm in wounds include:

nature of a wound. By using this strategy, we can visualise the location of the Biofilm and target directly with treatment.

•

Staphylococcus aureus

•

Pseudomonas aeruginosa

•

Escherichia coli

•

Candida Albicans

•

Proteus Mirabilia’s

Using a Cellink 3D BioX Printer to print methacrylate

•

Enterococcus Faecalis

collagen

3-D Printed Biofilms

loaded

aeruginosaor

with

either

Staphylococcus

GFP-Pseudomonas aureus

and

then

photo-activated the collagen with lithium phenyl-2, 4, 6-trimethylbenzoyl phosphonate (LAP) dye and

Wound Masterclass - Vol 2 - December 2023

Global Innovation in Wound Care Summit Series Part 2: Biofilm Masterclass

““Infection diagnostics are difficult in the US due to the diagnostic stages and determining the Biofilm infection.” Dr M. Sanders

polymerised with a near UV-light source. An amber

Conclusion

syringe containing a suspension of 2mls of Lifelink 200 collagen and 250 jul of an overnight culture of

Sanders states that the two main aspects of Biofilm

each microbe was loaded into the printer in a dark

in relation to wound care is to; Prevent the formation

room to avoid premature curing of the collagen

at the initial stages; Disrupt and Destroy the Biofilm.

with ambient light. After printing all the layers of the

Enhanced treatment efficacy and a personalised

collagen-microbial disk, the UV light was utilised to

treatment approach are essential for reducing the

cure the 3D print.

risk of recurrence, prevention of chronic infection and the improvement of wound healing, which, results in

Following the printing, the Biofilm disks were exposed

patient quality of life and cost savings for Science and

to natural light for 30 minutes to allow for complete

Medicine.

curing and then incubated overnight at 37C. The disks were then incubated for another 24 hours in a proprietary Biofilm binding buffer and were either used immediately or stored in tryptic soy broth with 2% sucrose for lyophilisation followed by ambient storage at -80C for future use. In these trials for 3-D printing, Bowler also states that he has found some antimicrobials are effective at reducing the exopolysaccharide matrix and others

See all Wound Masterclass MasterSeries on demand: bigmarker.com/wound-masterclass

are good for destroying Biolase. Primarily, these are ineffective once the Biofilm has occurred and that is the reason for no further development of new antibiotics within the laboratory for treatment of wounds.

Wound Masterclass - Vol 2 - December 2023