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

项目摘要。了解炎症对神经再生功能的影响 牙周膜(PDL)细胞不完整。这限制了技术的发展 牙周再生，将长期维持牙齿的功能支持。 牙周再生包括多个细胞过程和一个鲜为人知的组成部分 这些过程中包括PDL细胞的收缩能力。细胞的收缩力量对于 增强牙周组织并维持其功能的胶原纤维排列 正直。这项研究的长期目标是确定调节PDL细胞的机制 可用作再生和维护建筑的临床工具的机械 随着时间的推移，牙周复合体的功能。因此，这项提案的目标是 演示调节PDL细胞收缩力量的机制之间的联系 具有PDL架构和组织力学的促炎微环境。中环 这一理论的假设是炎症微环境调节PDL细胞 收缩力对PDL组织结构和力学的影响。这一假设将是 通过鉴定在体外调节PDL细胞的机制，在特定目标1中进行测试 在单细胞水平的促炎微环境中的收缩力量。西式 将使用印迹法来确定肿瘤坏死因子和透明质酸的作用 低聚糖(OHA)在产生细胞收缩力量的信号通路上，例如 Rho/Rock路径。为了将炎症环境和细胞信号与 在有无炎症介质的情况下，测量细胞的收缩能力和牵引力。 和信号通路抑制剂。在具体目标2中，三维PDL结构将是 开发用于将调节组织水平收缩的信号通路与基质联系起来 建筑和僵硬。设计的PDL结构将使用PDL细胞和 将测量胶原蛋白和原位作用力。PDL结构将用刺激剂和 Rho/Rock途径的抑制剂在模拟牙周动态平衡和 发炎。这些目标的成功实现将有助于 在促炎状态下维持PDL或再生组织的临床技术 环境。未来的研究将把这一模型扩展到包括牙骨质样组织和骨； 因此，这项初步研究是迈向未来再生牙周目标的第一步。 复杂，并长期保持其功能的完整性。

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.