PLASNOW – Qualitative and Quantitative Detection of plasma-generated ROS/RNS/RCS in solution and their Impact on Biomolecules Relevant for Wound Healing

本项目研究常压低温等离子体产生的溶液中的化学物种，目的是阐明等离子体产生的与伤口愈合相关的反应物种的生物学后果。该项目将与PlatNOW财团内的几个合作伙伴合作进行。我们在这个项目中的贡献是鉴定等离子体处理形成的溶液中的活性物种，根据等离子体的组成和通量对它们进行量化，并在模型氨基酸和多肽中追踪它们的起源和去向。为了量化溶液中的物种(盐水、缓冲溶液、细胞培养液和真实伤口液)，我们将适当使用比色分析和直接光谱技术，如原位EPR光谱分析和核磁共振光谱，以及大分子的色谱技术和质谱学。我们将使用同位素标记技术(15N和18O)来验证血浆修饰的氨基酸和肽衍生物是通过与等离子体产生的活性氧或氮物种(ROS或RNS)的质量转移形成的，还是通过能量转移间接形成的。我们还建议(首次)调查活性氯物种(RC)，这些物种很可能形成于含氯的缓冲液中，但到目前为止还没有得到太多关注。该项目通过我们的合作伙伴及其对等离子体成分和气相物种的知识(Awakowicz和Schulz-von der Gaten，均在RUB)获得两个不同的、特性良好的等离子体源而得以实施。它还与生物医学合作伙伴对接，在那里通过动物实验研究和量化血浆对伤口愈合的影响。本文描述的贡献是在分子水平上描述和量化溶液中的相关活性物种，以及在成纤维细胞培养中。

This project investigates the chemical species in solution which are produced by non-thermal atmospheric pressure plasma, with the aim to elucidate the biological consequences of plasma-generated reactive species relevant for wound healing. This project will be carried out in collaboration with several partners within the PlasNOW consortium. Our contribution in this project is to identify reactive species in solution formed by plasma treatment, quantify them in dependence of the plasma composition and fluence, and trace their origin and fate in model amino acids and peptides. To quantify species in solution (saline, buffered solutions, cell culture media and authentic wound liquid) we will use colorimetric assays and direct spectroscopic techniques such as in situ EPR spectrocopy and NMR spectroscopy as appropriate, as well as chromatographic techniques and mass spectrometry for larger molecules. We will use isotopic labelling techniques (15N and 18O) to verify whether plasma-modified amino acid and peptide derivatives are formed by mass transfer with plasma-generated reactive oxygen or nitrogen species (ROS or RNS), or indirectly through energy transfer. We also propose to investigate (for the first time) reactive chlorine species (RCS) which are likely formed in chloride-containing buffers but have received very little attention so far. This project is enabled by access to two different well-characterized plasma sources through our cooperation partners and their knowledge of plasma compositions and gas-phase species (Awakowicz and Schulz-von der Gathen, both at RUB). It also interfaces to a biomedical cooperation partner where plasma effects on wound healing are investigated and quantified in animal eperiments. The contribution described herein is in describing and quantifying the relevant reactive species in solution on a molecular level, and in fibroblast cell culture.