Inhibition of Biofilms by Non-Thermal Plasma Treated Novel Solutions

Utku K. Ercan¹, Siddharth S. Joshi¹, Adam Yost¹, Natalie Gogotsi¹, Sean O’Toole¹, Michelle Paff¹, Eric Melchior¹, Suresh G. Joshi^1,2*

¹Department of Surgery, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA.
²Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA.

DOI: 10.4236/aim.2014.416128                

Abstract

Biofilms act as a reservoir of infection, and periodically release cells in vicinity that are capable of developing new biofilm colonies and disseminate infection. Many chronic bacterial infections are serious that are associated with biofilms and have high morbidity and mortality, partly due to their higher resistance to antimicrobial agents, and partly due to lack of strong biocides which can efficiently treat and inhibit biofilm formation. We recently demonstrated that nonequilibrium non-thermal dielectric-barrier discharge plasma (Plasma) can also be applied to control pathogens via applying treated-liquids, and these liquids acquire broad-spectrum antimicrobial properties. In present studies we demonstrated a range of plasma-activated simple chemical solutions which significantly inhibited biofilm formation by multidrug-resistant bacterial pathogens. Plasma-activated methionine solution exhibited strong inhibitory activity against the biofilms of car-bapenem-resistant Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, metal-lo-β-lactamase (NDM1)-positive Klebsiella pneumoniae, and Enterococcus faecalis, and prevented the formation of biofilms by about 70% as compared to untreated controls in single exposure. In addition to inhibition of biofilm formation, a complete inactivation of biofilm-embedded bacterial cells was observed in less than 30 minute’s exposure to candidate plasma-activated methionine solution. These findings suggest that plasma-activated solutions have a potential to prevent biofilm formation, and as biofilm inhibitor.

Keywords

Acinetobacter baumannii, Antibacterial Solution, Antibiofilm Agent, Biofilm, MRSA, Nonthermal Plasma

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Costerton, J.W., Cheng, K.J., Geesey, G.G., Ladd, T.I., Nickel, J.C., Dasgupta, M. and Marrie, T.J. (1987) Bacterial Biofilms in Nautre and Disease. Annual Review of Microbiology, 41, 435-464. http://dx.doi.org/10.1146/annurev.mi.41.100187.002251
[2]	Conrad, J.C. (2012) Physics of Bacterial Near-Surface Motility Using Flagella and Type IV pili: Implications for Biofilm Formation. Research in Microbiology, 163, 619-629. http://dx.doi.org/10.1016/j.resmic.2012.10.016
[3]	Donlan, R.M. and Costerton, J.W. (2002) Biofilms: Survival Mechanisms of Clinically Relevant Microorganisms. Clinical Microbiology Reviews, 15, 167-193. http://dx.doi.org/10.1128/CMR.15.2.167-193.2002
[4]	Wolcott, R.D. and Ehrlich, G.D. (2008) Biofilms and Chronic Infections. JAMA: The Journal of the American Medical Association, 299, 2682-2684. http://dx.doi.org/10.1001/jama.299.22.2682
[5]	Leung, C.Y., Chan, Y.C., Samaranayake, L.P. and Seneviratne, C.J. (2012) Biocide Resistance of Candida and Escherichia coli Biofilms Is Associated with Higher Antioxidative Capacities. The Journal of Hospital Infection, 81, 79-86. http://dx.doi.org/10.1016/j.jhin.2011.09.014
[6]	Smith, K. and Hunter, I.S. (2008) Efficacy of Common Hospital Biocides with Biofilms of Multi-Drug Resistant Clinical Isolates. Journal of Medical Microbiology, 57, 966-973. http://dx.doi.org/10.1099/jmm.0.47668-0
[7]	O’Toole, G.A. (2002) Microbiology: A Resistance Switch. Nature, 416, 695-696. http://dx.doi.org/10.1038/416695a
[8]	Joshi, S.G., Paff, M., Friedman, G., Fridman, G., Fridman, A. and Brooks, A.D. (2010) Control of Methicillin-Resistant Staphylococcus aureus in Planktonic Form and Biofilms: A Biocidal Efficacy Study of Nonthermal Dielectric-Barrier Discharge Plasma. American Journal of Infection Control, 38, 293-301. http://dx.doi.org/10.1016/j.ajic.2009.11.002
[9]	Ercan, U.K., Wang, H., Ji, H.F., Fridman, G., Brooks, A.D. and Joshi, S.G. (2013) Nonequilibrium Plasma-Activated Antimicrobial Solutions Are Broad-Spectrum and Retain Their Efficacies for Extended Period of Time. Plasma Processes and Polymers, 10, 544-555. http://dx.doi.org/10.1002/ppap.201200104
[10]	Poor, A.E., Ercan, U.K., Yost, A., Brooks, A.D. and Joshi, S.G. (2014) Microbial and Cellular Responses to Nonthermal Plasma-Treated Alginate Wound Dressing: Control of Multidrug-Resistant Pathogens. Surgical Infections, 15, 233-243. http://dx.doi.org/10.1089/sur.2013.050
[11]	Telang, N.V., Satpute, M.G., Niphadkar, K.B. and Joshi, S.G. (2010) An Increased Incidence of Biofilm-Producing Multidrug-Resistant Methicillin-Resistant Staphylococcus aureus in a Tertiary Care Hospital from India: A 2-Year Study. American Journal of Infection Control, 38, 165-166. http://dx.doi.org/10.1016/j.ajic.2009.07.007
[12]	Standar, K., Kreikemeyer, B., Redanz, S., Munter, W.L., Laue, M. and Podbielski, A. (2010) Setup of an in Vitro Test System for Basic Studies on Biofilm Behavior of Mixed-Species Cultures with Dental and Periodontal Pathogens. PLoS ONE, 5, e13135. http://dx.doi.org/10.1371/journal.pone.0013135
[13]	Gobor, T., Corol, G., Ferreira, L.E.N., Rymovicz, A.U.M., Rosa, R.T., Campelo, P.M.S. and Rosa, E.A.R. (2011) Proposal of Protocols Using D-glutamine to Optimize the 2,3-Bis(2-methoxy-4-nitro-5-sulfophenly)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) Assay for Indirect Estimation of Microbial Loads in Biofilms of Medical Importance. Journal of Microbiological Methods, 84, 299-306. http://dx.doi.org/10.1016/j.mimet.2010.12.018
[14]	Fridman, G., Friedman, G., Gutsol, A., Shekhter, A.B., Vasilets, V.N. and Fridman, A. (2008) Applied Plasma Medicine. Plasma Processes and Polymers, 5, 503-533
[15]	Thiyagarajan, M. and Waldbeser, L. (2012) Portable Plasma Medical Device for infection Treatment. Studies in Health Technology and Informatics, 173, 518-520.
[16]	De Geyter, N. and Morent, R. (2012) Nonthermal Plasma Sterilization of Living and Nonliving Surfaces. Annual Review of Biomedical Engineering, 14, 255-274. http://dx.doi.org/10.1146/annurev-bioeng-071811-150110
[17]	Alkawareek, M.Y., Algwari, Q.T., Gorman, S.P., Graham, W.G., O’Connell, D. and Gilmore, B.F. (2012) Application of Atmospheric Pressure Nonthermal Plasma for the in Vitro Eradication of Bacterial Biofilms. FEMS Immunology and Medical Microbiology, 65, 381-384. http://dx.doi.org/10.1111/j.1574-695X.2012.00942.x
[18]	Zelaya, A.J., Stough, G., Rad, N., Vandervoort, K. and Brelles-Marino, G. (2010) Pseudomonas aeruginosa Biofilm Inactivation: Decreased Cell Culturability, Adhesiveness to Surfaces, and Biofilm Thickness upon High-Pressure Nonthermal Plasma Treatment. IEEE Transactions on Plasma Science, 38, 3398-3403. http://dx.doi.org/10.1109/TPS.2010.2082570
[19]	Schaudinn, C., Jaramillo, D., Freire, M.O., Sedghizadeh, P.P., Nguyen, A., Webster, P., Costerton, J.W. and Jiang, C. (2013) Evaluation of a Nonthermal Plasma Needle to Eliminate ex Vivo Biofilms in Root Canals of Extracted Human Teeth. International Endodontic Journal, 46, 930-937. http://dx.doi.org/10.1111/iej.12083
[20]	Sladek, R.E.J., Filoche, S.K., Sissons, C.H. and Stoffels, E. (2007) Treatment of Streptococcus mutans Biofilms with a Nonthermal Atmospheric Plasma. Letters in Applied Microbiology, 45, 318-323. http://dx.doi.org/10.1111/j.1472-765X.2007.02194.x
[21]	Kamgang, J.O., Briandet, R., Herry, J.M., Brisset, J.L. and Naitali, M. (2007) Destruction of Planktonic, Adherent and Biofilm Cells of Staphylococcus epidermidis Using a Gliding Discharge in Humid Air. Journal of Applied Microbiology, 103, 621-628. http://dx.doi.org/10.1111/j.1365-2672.2007.03286.x
[22]	Vercaigne, L.M., Sitar, D.S., Penner, S.B., Bernstein, K., Wang, G.Q. and Burczynski, F.J. (2000) Antibiotic-Heparin lock: In Vitro Antibiotic Stability Combined with Heparin in a Central Venous Catheter. Pharmacotherapy, 20, 394-399. http://dx.doi.org/10.1592/phco.20.5.394.35063
[23]	Henrickson, K.J., Powell, K.R. and Schwartz, C.L. (1988) A Dilute Solution of Vancomycin and Heparin Retains Antibacterial and Anticoagulant Activities. The Journal of Infectious Diseases, 157, 600-601. http://dx.doi.org/10.1093/infdis/157.3.600
[24]	Henrickson, K.J. and Dunne Jr., W.M. (1992) Modification of Central Venous Catheter Flush Solution Improves in Vitro Antimicrobial Activity. The Journal of Infectious Diseases, 166, 944-946. http://dx.doi.org/10.1093/infdis/166.4.944
[25]	Ercan, U.K. and Joshi, S.G. (2013) Mechanisms Underlying Antimicrobial Efficacies of Non-Thermal Dielectric Barrier Discharge (DBD) Plasma-Treated N-Acetyl Cysteine (NAC) Solution. Ph.D. Thesis, Drexel University, Philadelphia. http://www.biomed.drexel.edu/new04/Content/news_events/display_event.cfm?EVENT_ID=969
[26]	Joshi, S.G., Cooper, M., Yost, A., Paff, M., Ercan, U.K., Fridman, G., Friedman, G., Fridman, A. and Brooks, A.D. (2011) Nonthermal Dielectric-Barrier Discharge Plasma-Induced Inactivation Involves Oxidative DNA Damage and Membrane Lipid Peroxidation in Escherichia coli. Antimicrobial Agents and Chemotherapy, 55, 1053-1062. http://dx.doi.org/10.1128/AAC.01002-10
[27]	Gilbert, P., Allison, D.G. and McBain, A.J. (2002) Biofilms in Vitro and in Vivo: Do Singular Mechanisms Imply Cross-Resistance? Journal of Applied Microbiology, 92, 98S-110S. http://dx.doi.org/10.1046/j.1365-2672.92.5s1.5.x
[28]	Cooper, M., Fridman, G., Fridman, A. and Joshi, S.G. (2010) Biological Responses of Bacillus stratosphericus to Floating Electrode-Dielectric Barrier Discharge Plasma Treatment. Journal of Applied Microbiology, 109, 2039-2048. http://dx.doi.org/10.1111/j.1365-2672.2010.04834.x