Non-thermal dielectric-barrier discharge plasma damages human keratinocytes by inducing oxidative stress

Abstract

The aim of this study was to identify the mechanisms through which dielectric-barrier discharge plasma damages human keratinocytes (HaCaT cells) through the induction of oxidative stress. For this purpose, the cells were exposed to surface dielectric-barrier discharge plasma in 70% oxygen and 30% argon. We noted that cell viability was decreased following exposure of the cells to plasma in a time-dependent manner, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The levels of intracellular reactive oxygen species (ROS) were determined using 2′,7′-dichlorodihydro-fluorescein diacetate and dihydroethidium was used to monitor superoxide anion production. Plasma induced the generation of ROS, including superoxide anions, hydrogen peroxide and hydroxyl radicals. N-acetyl cysteine, which is an antioxidant, prevented the decrease in cell viability caused by exposure to plasma. ROS generated by exposure to plasma resulted in damage to various cellular components, including lipid membrane peroxidation, DNA breaks and protein carbonylation, which was detected by measuring the levels of 8-isoprostane and diphenyl-1-pyrenylphosphine assay, comet assay and protein carbonyl formation. These results suggest that plasma exerts cytotoxic effects by causing oxidative stress-induced damage to cellular components.

Exposure of keratinocytes to non-thermal dielectric barrier discharge plasma increases the level of 8-oxoguanine via inhibition of its repair enzyme

“…lipid membrane peroxidation, DNA breaks and protein carboxylation (19). These findings suggested that DBD plasma exerts cytotoxic effects via oxidative stress-induced damage to these components.…”

“…Cells were grown in Dulbecco's modified Eagle's medium (Thermo Fisher Scientific Inc., Waltham, MA, USA) containing 10% heat-inactivated fetal calf serum (Thermo Fisher Scientific Inc.), 100 µg/ml streptomycin and 100 U/ml penicillin. Non-thermal DBD plasma was applied as described previously (19). The cell suspension was adjusted to a concentration of 2x10 5 cells/ml and 11 ml was placed into 60 mm dishes.…”

Non-thermal gas plasma-induced endoplasmic reticulum stress mediates apoptosis in human colon cancer cells

“…Plasma treatment. Non-thermal DBD was used as the plasma source, as previously described (19). For DBD plasma treatment, cells were trypsinized and counted to adjust the density to 2x10 5 cells/ml, and 11 ml of cell suspension was placed in a 60-mm cell culture dish.…”

“…For this study, we used a non-thermal dielecteic barrier discharge (DBD) plasma source with a gas consisting of 70% oxygen and 30% argon at atmospheric pressure. Previously, we showed that a DBD plasma system can generate ROS and induce apoptosis in human keratinocytes (19), and we used the same experimental settings for this study.…”

A Comparison of Floating-Electrode DBD and kINPen Jet: Plasma Parameters to Achieve Similar Growth Reduction in Colon Cancer Cells Under Standardized Conditions

“…Plasma medical research inevitably involves experiments on reactive species because these were found to be central effectors in a number of biological targets exposed to plasma, such as skin cells [13][14][15], immune cells [16][17][18], and cancer cells . In many instances, hydrogen peroxide (H 2 O 2 ) was an important mediator in these in vitro experiments [53][54][55].…”