Original Article

Evaluation of cold atmospheric-pressure plasma against burn wound infections and gene silencing


Background and Objectives: Non-thermal atmospheric-pressure plasma or cold plasma is defined as an ionized gas. This study aimed to investigate the effect of cold plasma on Pseudomonas aeruginosa strains. Also, the expression level of the alp virulence gene before and after treatment with cold plasma was compared with the Housekeeping gene gyrA.
Materials and Methods: P. aeruginosa isolates recovered from hospitalized burn patients at Shahid Motahari Burns Hospital, Tehran, Iran. The Kirby Bauer disk diffusion method was used to determine the antimicrobial susceptibility test. Then, the antibacterial effect of atmospheric non-thermal plasma was evaluated on P. aeruginosa in as in vitro and in vivo studies at different times on Muller Hinton agar and in mouse model (treated by plasma every day/ 90 sec). The histopathological study was evaluated by Hematoxylin-Eosin staining. Data were analyzed using SPSS software by the Chi-square test and Pvalues less than 0.05 considered as statistically significant.
Results: Results indicated that non-thermal atmospheric plasma inhibited the growth of P. aeruginosa. The non-thermal helium plasma accelerates wound healing for 6 days. Results showed that cold plasma decreased virulence gene expression alp after treatment. Therefore, cold plasma can be suggested as a complementary therapeutic protocol to reduce bacterial infection and accelerate wound healing and reduce the expression of virulence genes of pathogens.
Conclusion: Cold plasma showed pathogen inhibitory properties of P. aeruginosa and virulence alkaline protease and wound healing properties in animal models, so this inexpensive and suitable method can be presented to the medical community to disinfect burn wounds and improve wound healing.

1. Mah TF, Pitts B, Pellock B, Walker GC, Stewart PS, O'Toole GA. A genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance. Nature 2003; 426:306-310.
2. Klausen M, Heydorn A, Ragas P, Lambertsen L, Ages‐Jørgensen A, Molin S, et al. Biofilm formation by Pseudomonas aeruginosa wild type, flagella, and type IV pili mutants. Mol Microbiol 2003; 48:1511-1524.
3. Meskini M, Esmaeili D. The study of formulated Zoush ointment against wound infection and gene expression of virulence factors Pseudomonas aeruginosa. BMC Complement Altern Med 2018; 18:185.
4. Döring G, Conway SP, Heijerman HG, Hodson ME, Høiby N, Smyth A, et al. Antibiotic therapy against Pseudomonas aeruginosa in cystic fibrosis: a European consensus. Eur Respir J 2000; 16:749-767.
5. Smith EE, Buckley DG, Wu Z, Saenphimmachak C, HoffmanLR, D’Argenio DA, et al. Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients. Proc Natl Acad Sci U S A 2006; 103:8487-8492.
6. Sadeghi-Nejad B, Shiravi F, Ghanbari S, Alinejadi M, Zarin M. Antifungal activity of Satureja khuzestanica (Jamzad) leaves extracts. Jundishapur J Microbiol 2010; 3:36-40.
7. Yang Y, Guo J, Zhou X, Liu Z, Wang C, Wang K, et al. A novel cold atmospheric pressure air plasma jet for peri-implantitis treatment: an in vitro study. Dent Mater J 2018; 37:157-166.
8. ChurchD, Elsayed S, Reid O, Winston B, Lindsay A. Burn wound infection. Clin Microbiol Rev 2006; 19:403-434.
9. Filipić A , Primc G , Zaplotnik R , Mehle N , Gutierrez-Aguirre I , Ravnikar M , et al. Cold atmospheric plasma as a novel method for inactivation of potato virus Y in water samples. Food Environ Virol 2019; 11:220-228.
10. Stratmann B, Costea TC, Nolte C, Hiller J, Schmidt J, Reindel J, et al. Effect of cold atmospheric plasma therapy vs standard therapy placebo on wound healing in patients with diabetic foot ulcers: arandomized clinical trial. JAMA Netw Open 2020; 3(7):e2010411.
11. Niedzwieds I, Wasko A, PawlatJ, Polak-Berecka M. The State of research on antimicrobial activity of cold plasma. Pol J Microbiol 2019; 68:153-164.
12. Keidar M, Shashurin A, Volotskova O, Stepp MA,Srinivasan P, Sandler A, Trink B. Cold atmospheric plasma in cancer therapy. Phys Plasma 2013; 20:057101.
13. Cordaro L, de Masi G, Fassina A, Gareri C, Pimazzoni A, Desideri D, et al. The role of thermal effects in plasma medical applications: biological and calorimetric analysis. Appl Sci 2019; 9:5560.
14. Heslin C, Boehm D, MilosavljevicV, Laycock M, Cullen PJ, Bourke P. Quantitative assessment of blood coagulation by cold atmospheric plasma. Plasma Med 2014; 4:153-163.
15. Semmler ML, Bekeschus S, Schäfer M, Bernhardt T, Fischer T, Witzke K, et al. Molecular mechanisms of the efficacy of cold atmospheric pressure plasma (CAP) in cancer treatment. Cancers (Basel) 2020; 12:269.
16. Dai X, Bazaka K, Thompson EW,Ostrikov KK. Cold atmospheric plasma: apromising controller of cancer cell states. Cancers (Basel) 2020; 12:3360.
17. Pankaj SK, Wan Z, Keener KM. Effects of cold plasma on food quality: areview. Foods 2018; 7:4.
18. Terefinko D, Dzimitrowicz A, Bielawska-PohlA, Klimczak A, Pohl P, Jamroz P. The influence of cold atmospheric pressure plasma-treated media on the cell viability, motility, and induction of apoptosis in human non-metastatic (MCF7) and metastatic (MDA-MB-231) breast cancer cell lines. Int J Mol Sci 2021; 22:3855.
19. Nazar Namini Y, Heidarzadeh S, Khaledi A, Abbasi E, Abbasi A, Esmaeili D. Study on the killing effect of cold atmospheric pressure plasma on MRSA Staphylococcus aureus in vitro and in vivo infection model. Malaysian J Microbiol 2019; 15:394-399.
20. Tajadod Y, Jangravi Z, Mahmoudabadi AZ, Esmaeili D, Dadseresht S, Bahadoran H, Korani M. Comparative study of the effects of Zoush ointment as a natural product and sliver sulfadiazine on the second-degree burn wounds healing in mice: role of antioxidants and the gene expression of matrix metalloproteinase-9. Bull Pharm Sci 2021; 44:149-160.
21. Betancourt-Ángeles M, Peña-Eguiluz R, López-Callejas R, Domínguez-Cadena NA, Mercado-Cabrera A, Muñoz-Infante J, et al. Treatment in the healing of burns with a cold plasma source. Int J Burns Trauma 2017; 7:142-146.
22. Pei X, Lu X, Liu J, Liu D, Yang Y, Ostrikov K, et al. Inactivation of a 25.5 µm Enterococcus faecalis biofilm by a room-temperature, battery-operated, handheld air plasma jet. J Phys D: Appl Phys 2012; 45:165205.
23. Isbary G, Zimmermann JL, Shimizu T, Li YF, Morfill GE, Thomas HM, et al. Non-thermal plasma—more than five years of clinical experience. Clin Plasma Med 2013; 1:19-23.
24. Brehmer F, Haenssle H, Daeschlein G, Ahmed R, Pfeiffer S, Görlitz A, et al. Alleviation of chronic venous leg ulcers with a hand‐held dielectric barrier discharge plasma generator (PlasmaDerm(®) VU‐2010): results of a monocentric, two‐armed, open, prospective, randomized and controlled trial (NCT01415622). J Eur Acad Dermatol Venereol 2015; 29:148-155.
25. Alkawareek MY, Algwari QT, Gorman SP, Graham WG, O'Connell D, Gilmore BF. Application of atmospheric pressure nonthermal plasma for the in vitro eradication of bacterial biofilms. FEMS Immunol Med Microbiol 2012; 65:381-384.
26. Mai-Prochnow A, Murphy AB, McLean KM, Kong MG, Ostrikov KK. Atmospheric pressure plasmas: infection control and bacterial responses. Int J Antimicrob Agents 2014; 43:508-517.
27. Deben JA, Zago CE, Tyhovych N, Duarte S, Vergani CE. Effect of atmospheric-pressure cold plasma on pathogenic oral biofilms and in vitro reconstituted oral epithelium. PLoS One 2016;11(5):e0155427.
28. Chatraie M, Torkaman G, Khani M, Salehi H, Shokri B. In vivo study of non-invasive effects of non-thermal plasma in pressure ulcer treatment. Sci Rep 2018; 8:5621.
29. Haertel B, von Woedtke T, Weltmann KD, Lindequist U. Non-thermal atmospheric-pressure plasma possible application in wound healing. Biomol Ther (Seoul) 2014; 22:477-490.
IssueVol 13 No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v13i4.6982
Cold plasma; Pseudomonas aeruginosa; Burn; Wound; Alp gene; Real-time reverse transcription polymerase chain reaction

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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Abbasi E, Fallah Mehrabadi J, Nourani M, Nazar Namini Y, Mohammadi S, Esmaeili D, Abbasi A. Evaluation of cold atmospheric-pressure plasma against burn wound infections and gene silencing. Iran J Microbiol. 2021;13(4):544-552.