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-Plasma）是一种相对较新的基于物理的技术。虽然关于该技术在生物科学中的应用的报道很少，但已知nt -血浆主要通过激活活性氧和氮（ROS/RNS）信号通路来影响细胞功能。在与Drexel等离子体研究所（美国等离子体物理领域最重要的专家）的合作中，我们建议使用nt等离子体作为一种工具来特异性地操纵细胞氧化还原，以促进MSC的承诺和分化。这项研究是基于最近的观察，即nt -血浆系统促进活性氧的产生，促进胚胎结构的发育，并启动许多与细胞分化相关的基因的表达。第一个具体目标是描述nt -血浆产生的ROS/RNS促进MSC增殖、承诺和分化的机制；同时开发用于此应用的NT-Plasma设备。我们将检验nt -血浆促进干细胞沿软骨、内皮和成骨谱系分化的假设。此外，这种作用是通过ROS/RNS依赖的信号通路介导的，这些信号通路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 -等离子体系统调制处理的长度，处理的次数和振幅和放电参数。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，以及反应性信号通路。第二个特定目的是确定nt -血浆如何促进两种模型系统（软骨内成骨系统和组织工程血管组织异体移植）中祖细胞的分化。我们建议在体内验证nt -血浆积极影响干细胞承诺和分化的假设。每个系统都提供了一个独特的机会来评估NT血浆治疗促进组织愈合和替代的潜力和可行性，同时获得对所产生的信号网络的有价值的理解。如果这项应用的目标成功实现，那么在NT-Plasma技术发展中获得的知识将具有变革性的科学和临床重要性。nt -血浆扩增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.