Engineered 3D Periodontal Tissue Constructs for Defining Functional Outcomes of Regenerative Processes

项目摘要。了解炎症对再生功能的影响， 牙周膜（PDL）细胞是不完整的。这限制了用于以下目的的技术的发展： 牙周再生，将保持功能性牙齿支持长期。 牙周膜再生包括多个细胞过程和一个不太了解的组成部分 其中之一是PDL细胞收缩性。细胞收缩力对细胞的生长至关重要。 排列胶原纤维，加强牙周组织并保持其功能 完整本研究的长期目标是确定调节PDL细胞的机制， 可以作为临床工具用于再生和维持结构的力学 和牙周复合体的功能。因此，本提案的目的是 证明调节PDL细胞收缩力的机制之间的联系， 促炎微环境与PDL结构和组织力学。中央 该建议的假设是炎症微环境调节PDL细胞 对PDL组织结构和力学有影响的收缩力。这一假设将是 通过鉴定调节体外PDL细胞的机制，在特定目标1中进行了测试 在单细胞水平的促炎微环境内的收缩力。西方 将使用印迹法确定肿瘤坏死因子α（TNF α）和透明质酸的作用 寡糖（oHA）对产生细胞收缩力的信号传导途径的作用，如 Rho/Rock路径为了将炎症环境和细胞信号传导与 收缩力，细胞牵引力将在有和没有炎症介质的情况下测量 和信号通路抑制剂。在特定目标2中，三维PDL构建将 开发用于将调节组织水平收缩性的信号通路与基质 结构和刚度。工程化PDL构建体将使用PDL细胞开发， 将测量胶原和原位力。PDL构建体将用刺激剂处理， Rho/Rock途径的抑制剂， 炎症这些目标的顺利实现将有助于 用于将PDL或再生组织维持在促炎性环境中的临床技术 环境未来的研究将扩大这一模型，包括骨水泥样组织和骨; 因此，这项初步研究是朝着牙周再生的未来目标迈出的第一步。 并长期保持其功能完整性。

Project Summary. Knowledge of the effects of inflammation on the regenerative functions of periodontal ligament (PDL) cells is incomplete. This limits the development of techniques for periodontal regeneration that will maintain functional tooth support over the long term. Periodontal regeneration includes multiple cellular processes and a less understood component of these processes is PDL cell contractility. Cellular contractile forces are critical to the alignment of collagen fibrils that strengthen periodontal tissue and maintain its functional integrity. The long-term goal of this research is to identify mechanisms regulating PDL cell mechanics that can be used as clinical tools for regenerating and maintaining the architecture and function of the periodontal complex over time. Thus, the objective of this proposal is to demonstrate links between mechanisms regulating PDL cell contractile forces in proinflammatory microenvironments with PDL architecture and tissue mechanics. The central hypothesis of this proposal is that the inflammatory microenvironment regulates PDL cell contractile forces with effects on PDL tissue architecture and mechanics. This hypothesis will be tested in Specific Aim 1 through identification of mechanisms that regulate in vitro PDL cell contractile forces within proinflammatory microenvironments at the single-cell level. Western blots will be used to determine effects of tumor necrosis factor alpha (TNF) and hyaluronan oligosaccharide (oHA) on signaling pathways that generate cellular contractile force, such as the Rho/Rock pathway. In order to link the inflammatory environment and cell signaling with contractility, cellular traction forces will be measured with and without inflammatory mediators and signaling pathway inhibitors. In Specific Aim 2, three-dimensional PDL constructs will be developed to link the signaling pathways that regulate tissue-level contractility with matrix architecture and stiffness. Engineered PDL constructs will be developed using PDL cells and collagen and in situ forces will be measured. PDL constructs will be treated with stimulants and inhibitors of the Rho/Rock pathway under conditions that model periodontal homeostasis and inflammation. The successful completion of these aims will contribute to the development of clinical techniques for maintaining the PDL or regenerated tissues in a proinflammatory environment. Future research will expand this model to include cementum-like tissue and bone; thus, this pilot study is an initial step toward the future goal of regenerating the periodontal complex and maintaining its functional integrity over the long-term.