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在HET-MN模型中研究冷氩等离子体射流的致突变性。

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

作者信息

Kluge Susanne, Bekeschus Sander, Bender Claudia, Benkhai Hicham, Sckell Axel, Below Harald, Stope Matthias B, Kramer Axel

机构信息

Institute of Hygiene and Environmental Medicine, University Medicine Greifswald, Walther-Rathenau-Str. 49a, 17485 Greifswald, Germany.

Leibniz-Institute for Plasma Science and Technology, ZIK plasmatis, Felix-Hausdorff-Str. 2, 17489 Greifswald, Germany.

出版信息

PLoS One. 2016 Sep 1;11(9):e0160667. doi: 10.1371/journal.pone.0160667. eCollection 2016.

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

目的

用于生物医学应用的所谓冷物理等离子体可产生活性氧和氮物种,而后者在高浓度时可引发DNA损伤。因此,评估了一种经过认证的大气压氩等离子体射流(kINPen MED)及其前身型号(kINPen 09)的致突变风险。

方法

受精鸡蛋的内卵膜接受单次处理,处理设备为kINPen 09(处理1.5、2.0或2.5分钟)或kINPen MED(处理3、4、5或10分钟)。孵化三天后,制作血涂片(全视野May-Grünwald-Giemsa染色),对每个鸡蛋的1000个红细胞进行评估,以确定是否存在多色和正色核染色以及核畸变和双核细胞(鸡蛋微核诱导试验,HET-MN)。同时,记录胚胎死亡率。每个实验均设置阳性对照(环磷酰胺和甲氨蝶呤)和阴性对照(NaCl溶液、氩气)。此外,处理后通过抗坏血酸氧化测定法评估血浆的抗氧化潜力。

结果

对于两种等离子体源,均未发现遗传毒性证据,尽管在最长的10分钟等离子体暴露时间时,胚胎死亡率超过了40%。暴露于冷等离子体后,鸡蛋血浆中的抗氧化潜力在即刻(p = 0.32)或1小时后(p = 0.19)均未显著降低。

结论

使用kINPen MED的最长等离子体处理时间比临床治疗慢性伤口的推荐时间上限高5至10倍。使用kINPen 09或kINPen MED在任何等离子体处理时间下均未发现致突变作用。本研究提供的数据似乎证实了缺乏遗传毒性潜力,表明这些氩等离子体射流在兽医或临床应用中不会带来致突变风险。

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