“Biofilm formation can make bacteria up to 1000 times more resistant to antibiotics, antimicrobial agents, disinfectants and the host immune system and are acknowledged to be one of the main contributors to the “antibiotic resistance crisis”
– Singh S, Singh SK, Chowdhury I, Singh R. Understanding the Mechanism of Bacterial Biofilms Resistance to Antimicrobial Agents. The Open Microbiology Journal. 2017;11:53-62.
With 81 patents and patents pending, Kane Biotech is a leader in biofilm research. Kane has laboratory and clinical evidence that these technologies have the potential to significantly improve the ability to prevent and destroy biofilms.
Kane Biotech has a growing pipeline of technologies based on their ongoing research on biofilm formation and how this process can be disrupted. Kane is committed to developing products to meet the demand for safe and effective anti-biofilm compounds for a variety of industries and applications.
Biofilms are estimated to be responsible for 80% of all human infections and cost industry, cities and hospitals in excess of $500 billion each year.
What Are Biofilms
Biofilms are pervasive and represent the most prevalent bacterial mode of growth. They form on living and non-living surfaces and can be found in natural, industrial, and healthcare settings.
Biofilms are formed when bacteria and/or fungi adhere to surfaces and excrete a glue-like substance that acts as an anchor and provides protection from the environment. Biofilm formation can make bacteria up to 1000 times more resistant to antibiotics, antimicrobial agents, disinfectants and the host immune system and are acknowledged to be one of the main contributors to the “antibiotic resistance crisis”xvi.
Biofilms can form on both living and non-living surfaces including teeth (plaque and tartar), skin (wounds and diseases like seborrheic dermatitis), medical devices (catheters and endoscopes), kitchen sinks and counter tops, food and food processing equipment, hospital surfaces, pipes and filters in water treatment plants and oil, gas and petrochemical process control facilities.
Biofilm related infections are difficult to treat and they commonly manifest themselves as chronic or recurrent in nature. According to an estimate by the National Institute of Health (NIH, USA), approximately 80% of all human bacterial infections are caused by biofilmsxvii. These structures are implicated in a range of health concerns such as periodontal disease, the healing of chronic wounds, medical device associated infection, inflammatory skin conditions, Hospital-Acquired Infections (HAIs) and food safety.
It was not until the 1990s that the elaborate organization of attached bacteria was identified as a biofilmxviii. Research on biofilms has only progressed rapidly in the last decade, leading to a greater understanding of the role biofilms play in infection and antimicrobial resistance. New understandings of how biofilms develop and propagate will suggest ideas for preventing and eliminating them.
These efforts have led to the current definition of microbial biofilms as “a structured community of microbial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.”
Biofilms can be comprised of single or multiple microbial species. Although mixed-species biofilms predominate in most environments, single-species biofilms exist in a variety of infections and on the surface of medical implants.
Biofilms exhibit a mode of growth that allows survival in a hostile environment. The structures biofilms form contain channels in which nutrients can circulate, and cells in different regions of biofilms exhibit different patterns of gene expression. These biofilm communities can rapidly multiply and disperse. In this light, it is not surprising a considerable number of chronic bacterial infections involve biofilms.
New understandings of how biofilms develop and propagate suggest ideas for preventing and eliminating them. Standard antibiotics often fail because they do not penetrate biofilms fully or do not kill bacteria of all species or metabolic states when protected in a biofilm.
Kane Biotech is a leader in biofilm intellectual property, protected technologies based on molecular mechanisms of biofilm formation and dispersal, and methods for finding components that inhibit or disrupt biofilms.
– Marc Edwards, CEO
Development of Biofilms
Stage 1. Initial attachment:
Colonizing bacteria anchor to a surface through basic adhesion techniques. This is when the biofilm is weakest, so many makers of medical devices, such as catheters, design their equipment in a way that attempts to disrupt initial adhesion.
Stage 2. Irreversible attachment:
After the cells aggregate they form micro colonies and excrete EPS or “slime” to form an irreversible attachment that can weather shear forces and maintain a steadfast grip on the surface.
Stage 3. Maturation I:
The biofilm is fully formed. As it matures the biofilm becomes a multi-layered cluster.
Stage 4. Maturation II:
The biofilm continues to grow and become three-dimensional. As the biofilm matures it is able to provide protection against the host immune system, anti-microbials, disinfectants and antibiotics.
Stage 5. Dispersion:
The biofilm reaches its critical mass and releases planktonic bacteria to continue colonizing other surfaces.
Kane Patented Technologies
Kane Biotech’s patented coactiv+™ technology is specifically formulated to destabilize biofilm and create an environment for fast wound healing. This multi-functional and gentle formulation makes it a perfect companion treatment to DispersinB® Hydrogel, also part of the Kane Biotech antibiofilm wound care portfolio.
coactiv+™ is a biofilm destabilizing formula with continuous activity. The key ingredients are recognized as safe by the FDA and have been purposefully selected to provide support throughout the entire wound healing cascade.
EDTA: ethylenediaminetetraacetic acid (EDTA) is a chelating (binding) agent that sequesters the metal ions present in the wound and needed for bacterial growth, function and ultimately, biofilm organization. Thus, once the metal ions are bound to EDTA, bacterial growth is inhibited and biofilm is destabilized.
Sodium citrate/citric acid: Elevated wound pH is a characteristic of hard to heal chronic wounds which are often inflamed and infected. Sodium citrate/citric acid acts as a buffering agent that helps to reduce elevated pH and/or maintain a lower pH which is conducive to wound healing. Similar to EDTA, sodium citrate is also a metal ion chelator, thus aids in microbial growth inhibition and biofilm destabilization.
The combination of metal ion sequestering and pH lowering activity of coactive+ provide an environment for effective biofilm destabilization. In addition, this activity has been shown to reduce overactive proteolytic function within wounds. Elevated levels of proteases are associated with chronic wounds and are known to cause tissue damage, inflammation and delayed healing.
DispersinB® is a naturally occurring enzyme that cleaves the bacterial surface polysaccharide poly-b-1, 6-N-acetylglucosamine (PNAG). This polysaccharide is produced by a wide range of bacteria and fungi and is a key component in biofilm formation. DispersinB cleaves PNAG, inhibiting bacterial adhesion and disperses the biofilm. This is especially useful for treating wounds and otic infections, which can become chronic due to the persistent nature of the bacterial biofilms. Once the biofilm is dispersed the bacteria can be eradicated and the infection can be remedied.
Kane Biotech Inc. has an exclusive worldwide license agreement with the University of Medicine and Dentistry of New Jersey, NJ, USA (now part of Rutgers University) to commercialize the DispersinB technology for human, animal, agricultural and industrial applications. The US patent on this technology has already been issued (U.S. Pat. No.7,294,497) and Kane Biotech holds 7 other patents in this area. Full biocompatibility, toxicity and stability testing has been conducted on this enzyme. Kane Biotech has a number of DispersinB products in development including formulations with antibiotics and the antibiofilm enzyme β-N-Acetylglucosaminidase. Kane Biotech has developed an antibiofilm technology in combination with Gentamycin and formulated an Enzyme-Gentamycin wound gel spray containing a thermo reversible gelling agent that makes the liquid spray become a gel when applied at body temperature.
Glossary of Terms
Biological catalysts that increase the rate or velocity of a chemical reaction without itself being changed in the overall process.
The whole range of biochemical processes that occur within us and in all living organisms. Metabolism consists of anabolism (the buildup of substances) and catabolism (the breakdown of substances). The term is commonly used to refer to the breakdown of food and its transformation into energy.
A colony of a few microscopic cells. For example, a minute colony of bacteria growing under suboptimal conditions.
Deficient in active properties; especially: lacking a usual or anticipated chemical or biological action.
A biomolecule is any organic molecule that is produced by a living organism, including large polymeric molecules such as proteins, polysaccharides, and nucleic acids as well as small molecules such as primary metabolites, secondary metabolites, and natural products.
A branch of biology that deals with the molecular structure and function of genes, with gene behavior in the context of a cell or organism, with patterns of inheritance from parent to offspring, and with gene distribution, variation and change in populations.
A branch of biology that deals with the molecular basis of biological activity. This field overlaps with other areas of biology and chemistry, particularly genetics and biochemistry. Molecular biology chiefly concerns itself with understanding and the interactions between the various systems of a cell, including the interactions between the different types of DNA, RNA and protein biosynthesis as well as learning how these interactions are regulated.
i Singh S, Singh SK, Chowdhury I, Singh R. Understanding the Mechanism of Bacterial Biofilms Resistance to Antimicrobial Agents. The Open Microbiology Journal. 2017;11:53-62. doi:10.2174/1874285801711010053.
ii Marshall MD, Wallis CV, Milella L, Colyer A, Tweedie AD, Harris S. A longitudinal assessment of periodontal disease in 52 miniature schnauzers. BMC Veterinary Research. 2014;10:166. doi:10.1186/1746-6148-10-166.
v Production effects related to mastitis and mastitis economics in dairy cattle herds. Henri Seegers, Christine Fourichon and François Beaudeau, Vet. Res., 34 5 (2003) 475-491 DOI: https://doi.org/10.1051/vetres:2003027
vi Lee S-I, Kim J, Han Y, Ahn K. A proposal: Atopic Dermatitis Organizer (ADO) guideline for children. Asia Pacific Allergy. 2011;1(2):53-63. doi:10.5415/apallergy.2011.1.2.53.
vii Del Rosso JQ. Adult Seborrheic Dermatitis: A Status Report on Practical Topical Management. The Journal of Clinical and Aesthetic Dermatology. 2011;4(5):32-38.
viii Crawford F. Athlete’s foot. BMJ Clinical Evidence. 2009;2009:1712.
ix Attinger C, Wolcott R. Clinically Addressing Biofilm in Chronic Wounds. Advances in Wound Care. 2012;1(3):127-132. doi:10.1089/wound.2011.0333.
x Sen CK, Gordillo GM, Roy S, et al. Human Skin Wounds: A Major and Snowballing Threat to Public Health and the Economy. Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society. 2009;17(6):763-771. doi:10.1111/j.1524-475X.2009.00543.x.
xi Peleg AY, Hooper DC. Hospital-Acquired Infections Due to Gram-Negative Bacteria. The New England journal of medicine. 2010;362(19):1804-1813. doi:10.1056/NEJMra0904124.
xii Bryers JD. Medical Biofilms. Biotechnology and bioengineering. 2008;100(1):1-18. doi:10.1002/bit.21838.
xiii Finch, J.E., Prince J., and Hawksworth, M (1978) A bacteriological survey of the domestic environment. Journal of Applied Bacteriology 45, 357-364.
xiv Scott, E., Bloomfield, S.F., and Barlow, C.G. (1984) Evaluation of disinfectants in the domestic environment under ‘in use’ conditions. Journal of Hygiene, Cambridge 92, 193-203.
xv Josephson, K., Rubino, J. and Pepper, I. (1997), Characterization and quantification of bacterial pathogens and indicator organisms inhousehold kitchens with and without the use of a disinfectant cleaner. Journal of Applied Microbiology, 83: 737-750. doi:10.1046/j.1365-2672.1997.00308.x
xviSingh S, Singh SK, Chowdhury I, Singh R. Understanding the Mechanism of Bacterial Biofilms Resistance to Antimicrobial Agents. The Open Microbiology Journal. 2017;11:53-62. doi:10.2174/1874285801711010053.
xvii NIH RESEARCH ON MICROBIAL BIOFILMS. Available online: http://grants.nih.gov/grants/guide/pa-files/PA-03–047.html.
xviii Donlan RM. Biofilms: Microbial Life on Surfaces. Emerging Infectious Diseases. 2002;8(9):881-890. doi:10.3201/eid0809.020063.
xix Wenzel R.P. (2007) Health care-associated infections: Major issues in the early years of the 21st century. Clinical Infectious Diseases 45 (Suppl 1), S85–S88.
xx Stone PW. Economic burden of healthcare-associated infections: an American perspective. Expert review of pharmacoeconomics & outcomes research. 2009;9(5):417-422. doi:10.1586/erp.09.53.
xxi Klevens RM, Morrison MA, Nadle J, et al. Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA. 2007;298:1763–71.
xxii Tilton, D. (2003). Nosocomial infections: diseases from within our doors. Retrieved May 15, 2005 from http://www.nursingceu.com/NCEU/courses/nosocomial/.
xxiii Bryers JD. Medical Biofilms. Biotechnology and bioengineering. 2008;100(1):1-18. doi:10.1002/bit.21838.
xxiv Dancer SJ. (2008) Importance of the environment in methicillin-resistant Staphylococcus aureus acquisition: the case for hospital cleaning. Lancet Infect Dis;8:101e113.
xxv Boyce JM. (2007) Environmental contamination makes an important contribution to hospital infection. J Hosp Infect;65(Suppl. 2): 50e54.
xxvi Carling PC, Bartley JM. (2010) Evaluating hygienic cleaning in health care settings: what you do not know can harm your patients. Am J Infect Control;38(5 Suppl. 1). S41e50.
xxvii Dancer SJ, White LF, Lamb J, Girvan EK, Robertson C. (2009) Measuring the effect of enhanced cleaning in a UK hospital: a prospective cross-over study. BMC Medicine;7:28.
xxviii Hota B, Blom DW, Lyle EA, Weinstein RA, Hayden MK.(2009) Interventional evaluation of environmental contamination by vancomycin resistant enterococci: failure of personnel, product, or procedure? J Hosp Infect;71:123e131.
xxix Marsh PD. Dental plaque as a biofilm and a microbial community – implications for health and disease. BMC Oral Health. 2006;6(Suppl 1):S14. doi:10.1186/1472-6831-6-S1-S14.
xxx Weidlich, Patrícia, Cimões, Renata, Pannuti, Claudio Mendes, & Oppermann, Rui Vicente. (2008). Association between periodontal diseases and systemic diseases. Brazilian Oral Research, 22(Suppl. 1), 32-43.
xxxi Nazir MA. Prevalence of periodontal disease, its association with systemic diseases and prevention. International Journal of Health Sciences. 2017;11(2):72-80.
xxxii Sen CK, Gordillo GM, Roy S, Kirsner R, Lambert L, Hunt TK, et al. Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen. 2009
xxxiii Attinger C, Wolcott R. Clinically Addressing Biofilm in Chronic Wounds. Advances in Wound Care. 2012;1(3):127-132. doi:10.1089/wound.2011.0333.
xxxiv Borda LJ, Wikramanayake TC. Seborrheic Dermatitis and Dandruff: A Comprehensive Review. J Clin Investig Dermatol. 2015;3 doi: 10.13188/2373-1044.1000019.
xxxvOkokon EO, Verbeek JH, Ruotsalainen JH, Ojo OA, Bakhoya VN. Topical antifungals for seborrhoeic dermatitis. Cochrane Database Syst Rev. 2015;5:CD008138.
xxxvi Gupta AK, Batra R, Bluhm R, et al. Skin diseases associated with Malassezia species. J Am Acad Dermatol. 2004;51:785–798
xxxvii Ghannoum MA, Rice, LB; Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance. Clin Microbiol Rev. 1999 Oct; 12(4):501-17.
xxxviii Figueredo, L. A., Cafarchia, C., & amp; Otranto, D. (2013). Antifungal susceptibility of Malassezia pachydermatis biofilm. Medical mycology, 51(8), 863-867.
xxxix Krakowski AC, Eichenfield LF, Dohil MA. Management of atopic dermatitis in the pediatric population. Pediatrics. 2008 Oct;122(4):812-24.
xl Akiyama H, Yamasaki O, Tada J, et al. Adherence characteristics and susceptibility to antimicrobial agents of Staphylococcus aureus strains isolated from skin infections and atopic dermatitis. J Dermatol Sci. 2000 Aug;23(3):155-60.
xli Ikezawa Z, Komori J, Ikezawa Y, et al. A role of Staphylococcus aureus, interleukin-18, nerve growth factor and semaphorin 3A, an axon guidance molecule, in pathogenesis and treatment of atopic dermatitis. Allergy Asthma Immunol Res. 2010 Oct;2(4):235-46.
xlii Katsuyama M, Ichikawa H, Ogawa S, et al. A novel method to control the balance of skin microflora. Part 1. Attack on biofilm of Staphylococcus aureus without antibiotics. J Dermatol Sci. 2005 Jun;38(3):197-205.
xliii Auger, P., Marquis, G., Joly, J., and Attye, A. 1993. Epidemiology of tinea pedis in marathon runners: prevalence of occult athlete’s foot. Mycoses. 36(1-2): 35-41.
xliv Costa-Orlandi, C. B. et al. In vitro characterization of Trichophyton rubrum and T. mentagrophytes biofilms. Biofouling. Abingdon: Taylor & Francis Ltd, v. 30, n. 6, p. 719-727, 2014.
xlvi Chmielewski, R.A.N, and Frank, J.F. (2003) Biofilm formation and control in food processing facilities. Comprehensive Reviews in Food Science and Food Safety 2, 22-32.
xlvii Keskinen, L.A., Todd, E.C.D., and Ryser, E. (2008) Transfer of surface dried Listeria monocytogenes from stainless steel knife blades to roast turkey breast. Journal of Food Protection 71, 176-181.
xlviii Lee Wong, A.C. (1998). Biofilms in food processing environments. Journal of Dairy Science 81, 2765-2770.
xlix Pui, C.F., Wong, W.C., Chai, L.C., Lee, H.Y., Tang, J.Y.H., Noorlis, A., Farinazleen, M.G., Cheah, Y.K., and Son. R. (2011) Biofilm formation by Salmonella Typhi and Salmonella Typhimurium on plastic cutting board and its transfer to dragon fruit. International Food Research Journal 18, 31-38.
l Reuter, M., Mallett, A., Pearson, B.M., and van Vliet, A.H.M. (2010) Biofilm formation by Campylobacter jejuni is increased under aerobic conditions. Applied and Environmental Microbiology 76, 2122-2128.
li Finch, J.E., Prince J., and Hawksworth, M (1978) A bacteriological survey of the domestic environment. Journal of Applied Bacteriology 45, 357-364.
lii Scott, E., Bloomfield, S.F., and Barlow, C.G. (1984) Evaluation of disinfectants in the domestic environment under ‘in use’ conditions. Journal of Hygiene, Cambridge 92, 193-203.
liii Furuhata, K., Ishizaki, N., and Fukuyama, M. (2010) Characterization of heterotrophic bacteria isolated from the biofilm of a kitchen sink. Biocontrol Science 15, 21-25.
liv Neth, K., Girard, D., and Albrecht, J. (2008) Determination of biofilm on plastic cutting boards. RURALS: Review of Undergraduate Research in Agricultural and Life Sciences, University of Nebraska-Lincoln, 3, Article 5.
1. James GA, Swogger E, Wolcott R et al. Biofilms in chronic wounds. Wound Repair Regen 2008; 16(1): 37–44.
2. Percival S, McCarty S and Lipsky B. Biofilms and wounds: An overview of the evidence Advances in Wound Care 2015; 4(7):373–381.
3. Cooper R, Bjarnsholt T and Alhede M. Biofilms in wounds: A review of present knowledge. Journal of Wound Care 2014 23(11): 570–582.
4. Thomson CH. Biofilms: do they affect wound healing? Int W J 2011; Feb 8(1):63–7. doi: 10.1111/j.1742-481X.2010.00749.x. Epub 2010 Dec
5. Phillips PL, Wolcott RD, Fletcher J, Schultz GS. Biofilms Made Easy. Wounds International 2010; 1(3): Available from http:// www.woundsinternational.com/