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The pathway of the biologically active molecule hydrogen peroxide (H 2 O 2 ) from the plasma generation in the gas phase by an atmospheric pressure argon plasma jet, to its transition into the liquid phase and finally to its inhibiting effect on human skin cells is investigated for different feed gas humidity settings. Gas phase diagnostics like Fourier transformed infrared spectroscopy and laser induced fluorescence spectroscopy on hydroxyl radicals ( • OH) are combined with liquid analytics such as chemical assays and electron paramagnetic resonance spectroscopy. Furthermore, the viability of human skin cells is measured by Alamar Blue ® assay. By comparing the gas phase results with chemical simulations in the far field, H 2 O 2 generation and destruction processes are clearly identified. The net production rate of H 2 O 2 in the gas phase is almost identical to the H 2 O 2 net production rate in the liquid phase. Moreover, by mimicking the H 2 O 2 generation of the plasma jet with the help of an H 2 O 2 bubbler it is concluded that the solubility of gas phase H 2 O 2 plays a major role in generating hydrogen peroxide in the liquid. Furthermore, it is shown that H 2 O 2 concentration correlates remarkably well with the cell viability. Other species in the liquid like • OH or superoxide anion radical (O •− 2 ) do not vary significantly with feed gas humidity.
This work focused on qualitative and quantitative detection of oxygen free radicals in liquids after plasma treatment with an atmospheric pressure argon plasma jet by electron paramagnetic resonance spectroscopy (EPR). For the treatment, a shielded plasma jet, where the active effluent zone is surrounded by a protective gas curtain was used.
Finding a solution for air species contamination of atmospheric pressure plasmas in plasma medical treatment is a major task for the new field of plasma medicine. Several approaches use complex climate chambers to control the surrounding atmosphere. In this paper, ambient species are excluded in plasma-human-skin-cell treatment by ensheathing the plasma jet effluent with a shielding gas. Not only does this gas curtain protect the plasma jet effluent from inflow of air species but it also, more importantly, allows controlling the effluent reactive species composition by adjusting the mixture of the shielding gas. In the present investigations, the mixture of nitrogen to oxygen within the gas curtain around an argon atmospheric pressure plasma jet (kinpen) is varied. The resulting reactive plasma components produced in the jet effluent are thus either oxygen or nitrogen dominated. With this gas curtain, the effect of reactive oxygen species (ROS) and reactive nitrogen species (RNS) on the cell viability of indirectly plasma-treated HaCaT skin cells is studied. This human keratinocyte cell line is an established standard for a skin model system. The cell viability is determined by a fluorometric assay, where metabolically active cells transform nonfluorescent resazurin to the highly fluorescent resorufin. Plasma jet and gas curtain are characterized by numerical flow simulation as well as by optical emission spectroscopy. The generation of nitrite within the used standard cell culture medium serves as a measure for generated RNS. Measurements with the leukodye dichlorodihydrofluorescein diacetate show that, despite a variation of the shielding gas mixture, the total amount of generated reactive oxygen plus nitrogen species is constant. It is shown that a plasma dominated by RNS disrupts cellular growth less than a ROSdominated plasma.Index Terms-Atmospheric pressure plasma jet, gas curtain, plasma liquid interaction, plasma medicine, reactive nitrogen species (RNS), reactive oxygen plus nitrogen species (RONS), reactive oxygen species (ROS), skin cells.
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