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

描述(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。尽管将这项技术应用于生物科学的报道很少，但已知NT-血浆主要通过激活ROS/RNS信号通路来影响细胞功能。在与德雷克塞尔等离子体研究所的合作下，我们建议使用NT-等离子体作为一种工具，专门操纵细胞的氧化还原，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结果，即NT-等离子体系统促进了活性氧的产生，促进了胚胎结构的发育，并启动了许多与细胞分化有关的基因的表达。第一个具体目标是描述NT-等离子体产生的ROS/RNS促进MSC增殖、承诺和分化的机制；同时开发用于这一应用的NT-等离子体设备。我们将检验这一假说，即NT-血浆可促进干细胞沿软骨、内皮和成骨方向分化。此外，这种作用是通过ROS/RNS依赖的信号通路来调节的，这些信号通路有助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-等离子体系统的治疗时间，治疗次数和幅度和放电参数。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的分化，这两个模型系统是软骨内成骨系统和组织工程血管组织同种异体移植。我们建议测试假设，即NT-血浆积极影响体内干细胞的承诺和分化。每个系统都提供了一个独特的机会来评估NT等离子体治疗促进组织愈合和替代的潜力和可行性，同时获得对由此产生的信号网络的有价值的了解。如果这项应用的目标成功实现，那么在NT-等离子体技术开发中获得的知识将具有变革性的科学和临床重要性。NT-血浆放大干细胞功能的能力将是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信号的基础生物学提供新的见解。 公共卫生相关性：该项目的目的是探索和开发非热血浆在组织工程和再生医学中的生物应用。我们的初步研究和其他已发表的报告表明，这项新技术可以显著影响成体干细胞的命运，并促进它们的增殖、承诺和分化。拟议研究的结果将为组织修复和再生提供改进的方法，直接适用于临床研究和外科手术。

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. PUBLIC HEALTH RELEVANCE: The purpose of this project is to explore and develop biological applications of Non-Thermal Plasma for use in tissue engineering and regenerative medicine. Our preliminary studies and other published reports indicate that this new technology can significantly influence the fate of adult stem cells and promote their proliferation, commitment and differentiation. Results generated from the proposed studies will provide improved methods for tissue repair and regeneration with direct applicability to both clinical research and surgical procedures.