Non-Thermal Plasma in Biomedicine: A New Paradigm for Redox Cell Activation

描述（由申请人提供）：非热介质阻挡放电等离子体（NT-等离子体）是一种相对较新的基于物理的技术。尽管有关该技术在生物科学中应用的报道很少，但众所周知，NT-血浆主要通过激活活性氧和氮物质（ROS/RNS）信号通路来影响细胞功能。与德雷克塞尔等离子体研究所合作，可以说是美国等离子体物理学领域最重要的专家，我们建议使用NT-等离子体作为一种工具，专门操纵细胞氧化还原，以促进MSC的承诺和分化。这项拟议的研究是基于最近的观察，即NT-血浆系统促进活性氧的产生，增强胚胎结构的发育，并启动与细胞分化相关的许多基因的表达。第一个具体目标是描述NT-血浆产生的ROS/RNS促进MSC增殖、定型和分化的机制;同时开发用于该应用的NT-血浆装置。我们将检验NT-血浆增强干细胞分化沿着软骨，内皮和成骨谱系的假设。此外，这种作用是通过ROS/RNS依赖性信号通路介导的，该信号通路用于影响细胞的氧化状态。为了验证这一假设，首先，我们将定义允许NT-血浆调节细胞氧化状态的条件。同时，我们将微调NT-等离子体系统，调节治疗长度、治疗次数以及振幅和放电参数。我们将测量ROS和RNS，细胞的氧化还原状态，抗氧化蛋白的表达和活性，以及响应信号通路。第二个具体目标是确定NT-血浆如何在两个模型系统中促进祖细胞分化，即软骨内骨化系统和组织工程血管组织同种异体移植物。我们建议测试的假设，NT-血浆积极影响干细胞的承诺和分化在体内。每个系统都提供了一个独特的机会来评估NT等离子体治疗的潜力和可行性，以增强组织愈合和替换，同时获得对所产生的信号网络的有价值的理解。如果成功实现了本申请的目标，那么在NT-Plasma技术开发过程中获得的知识将具有变革性的科学和临床重要性。NT-Plasma放大干细胞功能的能力将成为组织工程和再生医学的宝贵工具，并将为干细胞生物学中ROS信号传导的基础生物学提供新的见解。

DESCRIPTION (provided by applicant): Non-thermal dielectric barrier discharge plasma (NT-Plasma) is a relatively new physics-based technology. Although there are few reports concerning the application of this technology to biological sciences, it is known that NT-plasma influences cell function mainly through activation of reactive oxygen and nitrogen species (ROS/RNS) signaling pathways. In collaboration with the Drexel Plasma Institute, arguably the foremost experts in the field of plasma physics in the United States, we propose to use NT-plasma as a tool to specifically manipulate cellular redox to promote MSC commitment and differentiation. The proposed study is based on recent observations that the NT-Plasma system promotes reactive oxygen species generation enhances development of embryonic structures and initiates the expression of many genes linked to cell differentiation. The first Specific Aim is to delineate the mechanism by which NT-Plasma generated ROS/RNS promotes MSC proliferation, commitment and differentiation; while simultaneously developing the NT-Plasma device for this application. We will test the hypothesis that NT-Plasma enhances stem cell differentiation along chondrogenic, endothelial and osteogenic lineages. Moreover, that this effect is mediated via ROS/RNS dependent signaling pathways that serve to influence the cell's oxidative state. To test this hypothesis, first, we will define the conditions that permit NT-Plasma to regulate the oxidative state of the cell. At the same time, we will fine tune the NT-Plasma system modulating the length of treatment, times of treatment and amplitude and discharge parameters. We will measure ROS and RNS, the redox status of the cells, the expression and activity of antioxidant proteins, as well as responsive signaling pathways. The second Specific Aim is to determine how NT-Plasma advances differentiation of progenitor cells in two model systems, an endochondral ossification system and a tissue engineered vascular tissue allograft. We propose to test the hypothesis that NT-Plasma positively influences stem cell commitment and differentiation in vivo. Each system provides a unique opportunity to evaluate the potential and feasibility of NT plasma treatment to enhance tissue healing and replacement, while at the same time gaining valuable understanding of the resulting signaling networks. If the goals of this application are successfully achieved, then the knowledge gained in development of NT-Plasma technology will be of transformative scientific and clinical importance. NT-Plasma's ability to amplify stem cell function will be an invaluable tool for tissue engineering and regenerative medicine in general, and will provide new insights into the basic biology of ROS signaling in stem cell biology.