Investigating the Mutagenicity of a Cold Argon-Plasma Jet in an HET-MN Model

• Published in: PloS one 2016-09, Vol.11 (9), p.e0160667
• Main Authors: Kluge, Susanne, Bekeschus, Sander, Bender, Claudia, Benkhai, Hicham, Sckell, Axel, Below, Harald, Stope, Matthias B, Kramer, Axel
• Format: Article
• Language: English
• Subjects: Aberration; Analysis; Animals; Antioxidants; Apoptosis; Argon; Argon plasma; Ascorbic acid; Atmospheric pressure; Biochemistry; Biology and Life Sciences; Biomedical materials; Blood; Blood plasma; Chick Embryo; Cyclophosphamide; Data processing; Deoxyribonucleic acid; DNA; DNA damage; DNA repair; Eggs; Eggs (Food); Embryos; Environmental health; Erythrocytes; Exposure; Genotoxicity; Hygiene; Incubation; Lipid peroxidation; Medicine; Medicine and Health Sciences; Membranes; Methotrexate; Micronuclei; Micronucleus Tests; Mortality; Mutagenicity; Oxidation; Oxidative stress; Oxygen; Pathogens; Physical Sciences; Plasma; Plasma Gases; Plasma jets; Plasma physics; Reactive nitrogen species; Reactive oxygen species; Sodium chloride; Studies; Veterinary medicine; Wounds
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0160667

So-called cold physical plasmas for biomedical applications generate reactive oxygen and nitrogen species and the latter can trigger DNA damage at high concentrations. Therefore, the mutagenic risks of a certified atmospheric pressure argon plasma jet (kINPen MED) and its predecessor model (kINPen 09) were assessed. Inner egg membranes of fertilized chicken eggs received a single treatment with either the kINPen 09 (1.5, 2.0, or 2.5 min) or the kINPen MED (3, 4, 5, or 10 min). After three days of incubation, blood smears (panoptic May-Grünwald-Giemsa stain) were performed, and 1000 erythrocytes per egg were evaluated for the presence of polychromatic and normochromic nuclear staining as well as nuclear aberrations and binucleated cells (hen's egg test for micronuclei induction, HET-MN). At the same time, the embryo mortality was documented. For each experiment, positive controls (cyclophosphamide and methotrexate) and negative controls (NaCl-solution, argon gas) were included. Additionally, the antioxidant potential of the blood plasma was assessed by ascorbic acid oxidation assay after treatment. For both plasma sources, there was no evidence of genotoxicity, although at the longest plasma exposure time of 10 min the mortality of the embryos exceeded 40%. The antioxidant potential in the egg's blood plasma was not significantly reduced immediately (p = 0.32) or 1 h (p = 0.19) post exposure to cold plasma. The longest plasma treatment time with the kINPen MED was 5-10 fold above the recommended limit for treatment of chronic wounds in clinics. We did not find mutagenic effects for any plasma treatment time using the either kINPen 09 or kINPen MED. The data provided with the current study seem to confirm the lack of a genotoxic potential suggesting that a veterinary or clinical application of these argon plasma jets does not pose mutagenic risks.