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

该项目研究了非热大气压等离子体产生的溶液中的化学物质，旨在阐明等离子体产生的与伤口愈合相关的活性物质的生物学后果。该项目将与PlasNOW联合体内的几个伙伴合作执行。我们在这个项目中的贡献是确定反应物种在溶液中形成的等离子体处理，量化他们的血浆成分和流量的依赖，并跟踪其起源和命运的模型氨基酸和肽。为了定量溶液（生理盐水、缓冲溶液、细胞培养基和真实伤口液体）中的物质，我们将使用比色测定和直接光谱技术，如原位EPR光谱和NMR光谱（如适用），以及色谱技术和质谱法（用于较大分子）。我们将使用同位素标记技术（15 N和18 O）来验证等离子体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质（ROS或RNS）的质量转移形成的，还是间接通过能量转移形成的。我们还建议调查（第一次）反应性氯物种（RCS），这是可能形成的含氯化物的缓冲液，但迄今为止很少受到关注。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质的了解（Awakowicz和Schulz-von der Gathen，均在RUB）获得两种不同的表征良好的等离子体源。它还与一个生物医学合作伙伴建立了接口，在那里研究了等离子体对伤口愈合的影响，并在动物实验中进行了量化。本文所述的贡献是在分子水平上描述和定量溶液中和成纤维细胞培养物中的相关反应性物质。

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.