Deciphering non thermal plasma - human cell interaction using the proteomics approach

Non thermal plasmas are promising medical tools, e.g. for skin or wound treatment. To identify and basically understand the plasma - cell interaction and to guide plasma tuning efforts a deep insight into plasma - cell interaction is necessary. The cellular proteome covers the complete protein composition of a biological cell at a given time. It can be called the toolbox of a cell - besides regulating cell processes proteins are needed for structural, metabolic or defense mechanisms. Therefore, the snapshot of a cellular proteome enables in-depth findings on the cellular state. We established a protocol to obtain time (hours past treatment) and/or space (localization within a cellular compartment) resolved protein abundance data in a bottomup approach using a human keratinocytes cell line (HaCaT). As plasma source an argon based plasma jet was used (kinpen, Neoplas Tools). With an ABSciex TripleTOF 5600 high resolution mass spectrometer and nano-flow liquid chromatography, we were able to identify 3000+ human proteins within different cellular compartments. Proteins detected cover a wide range of molecular functions (e.g. antioxidant activities) and a broad spectrum of biological processes (e.g. regulation of apoptosis; DNA replication). Measured protein abundances differ between argon jet plasma treated cells and controls (argon gas only) in a specific manner. Among the most regulated proteins after plasma treatment are regulatory/enzymatic active proteins involved in reactive oxygen/reactive nitrogen (RONS) metabolism and cell division, indicating an oxidative influence on the DNA. We also measured the activation of protective and pro-proliferative signal molecules indicating specific cell activation after plasma treatment and encourage a possible use in wound care. Hand in hand with transcriptomics data the presented overview of HaCaT proteome after non-thermal plasma treatment helps streamlining the identification of both biological and physical key players and the conclusion of further research topics.