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Background: Physical plasma is a mixture of reactive particles and electromagnetic radiation. Due to the antimicrobial, immunomodulatory, anti-inflammatory, wound-healing promoting, and antineoplastic effects of body tempered physical plasma under atmospheric pressure (cold atmospheric plasma: CAP), CAP therapy is increasingly becoming the focus of surgical and oncological disciplines. However, when applied in practice, a potential emission of harmful noxae such as toxic nitrogen oxides must be taken into account, which was investigated in the following study. Materials and Methods: MiniJet-R Ar CAP device was characterized with respect to NO X -specific spectra, ultraviolet radiation C (UVC) intensity in the range of 200-275 nm and the formation of NO X gases. Instrument-specific parameters such as gas flow, energy setting of the highfrequency generator, and flow rate of the carrier gas Ar were varied. To test the toxic properties of the NO 2 concentrations formed by CAP, SK-OV-3 human ovarian cancer cells were incubated with different NO 2 concentrations and cell growth was monitored for 120 h. Results: The operation of MiniJet-R led to the formation of NO 2 in the proximity of the CAP effluent. Synthesis of NO led to a NO-specific spectrum in the range of 100-275 nm, whereby UVC radiation produced reached intensities of up to 90 mW/m 2 . NO gas itself, however, was not detectable, as it was converted to NO 2 rapidly. Cell culture incubation experiments demonstrated that NO 2 in these concentration ranges had no influence on the cell growth of human cancer cells. Conclusion: Although no limit values were exceeded in the present study, the emission of high-energy UVC radiation and toxic NO 2 is a risk factor with regard to the legal regulations on workplace protection (operator hazard) and the approval of medical devices (patient hazard). This is important for considerations regarding treatment frequency and duration. The growth inhibitory effect of CAP treatment on human cancer cells principally suggests a medical application of the MiniJet-R device, although more extensive studies will have to follow.Physical plasma is a mixture of particles of a carrier gas and ambient air with different ionization and excitation states, as well as electromagnetic radiation of different wavelengths including ultraviolet radiation C (UVC). In medical applications, body-tempered ('cold') plasmas up to about 40˚C under normal atmospheric pressure (cold atmospheric plasma: CAP) are utilized (1, 2). CAP has antimicrobial, immunomodulating, anti-inflammatory, wound-healing promoting, and antineoplastic effects on biological tissue and is therefore tested for its applicability in various medical fields. Due to the physical properties of CAP and against the 2591

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Non-thermal atmospheric pressure plasma consists of partially ionized gas and contains a range of reactive species including biological active ROS and RNS. There are numerous future applications planned in medicine, e.g. blood coagulation, disinfection and wound care. Recently it has been shown that plasma treatment can have lethal effects on bacteria, whereas eukaryotic cells can be promoted to grow and proliferate. The aim of this study was to investigate the impact of non-thermal plasma on the stimulation of mammalian cells. The cells were treated with the atmospheric pressure plasma jet kINPen09 and it was shown that the effects were dependent on treatment time. While long treatment induced apoptosis, short time plasma treatment increased the proliferation. Therefore the underlying processes needed to be identified in order to modulate the plasmas for future applications. Applying state of the art transcriptomic as well as proteomic approaches (DNA-microarrays and HPLC-MS) we identified several hundred genes and their according proteins which were modulated following non-thermal plasma treatment. Latest software solutions (e.g. IPA, Partek) were applied to disentangle the cellular networks connecting the identified molecules. Qualitative Real-Time Polymerase Chain Reaction was used for verification and revealed up-regulation of signaling molecules (e.g. ERK, MEK and JNK) as well as growth factors and cytokines (e.g. VEGF, FGF and IL-6). Furthermore, applying phosphor-specific western blotting or ELISA techniques we also could proof that plasma treatment activated several signaling cascades and induced the secretion of various cytokines (e.g. IL-6 and IL-8). These results underline the huge potential of plasma for cell modulation with regard to signaling processes involved in cell proliferation and differentiation. ________________________________

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