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

用于定义再生过程功能结果的工程 3D 牙周组织结构

• 批准号: 10189554
• 负责人: TRACY E POPOWICS
• 金额: $ 17.65万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189554
• 关键词: 3-Dimensional; Actins; Affect; Age; Age-Years; Architecture; Biomedical Engineering; CD44 gene; Catabolic Process; Cell physiology; Cells; Clinical; Collagen; Collagen Fibril; Complex; Dental Cementum; Dental Plaque; Engineering; Environment; Future; Gene Expression; Goals; Harvest; Health; Homeostasis; Hyaluronan; Hyaluronidase; In Situ; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Knowledge; Laboratories; Link; Longevity; Manuals; Measures; Mechanics; Mediating; Modeling; Myosin ATPase; Natural regeneration; Oligosaccharides; Oral; Outcome; Pathway interactions; Periodontal Diseases; Periodontal Ligament; Periodontitis; Physiological; Pilot Projects; Play; Population; Process; Regulation; Research; Risk; Role; Signal Pathway; Signal Transduction; Silicones; Systemic disease; TLR4 gene; TNF gene; Techniques; Testing; Time; Tissues; Tooth structure; Traction; Western Blotting; Work; age related; aging population; bone; clinical development; dexterity; enzyme activity; functional outcomes; inflammatory milieu; inhibitor/antagonist; novel; oral care; preservation; receptor; receptor binding; regeneration function; regenerative; rho; technique development; tissue regeneration; tool

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.

项目摘要了解炎症对再生功能的影响。 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功能性牙齿支撑。 牙周再生包括多个细胞过程和一个不太了解的组成部分 这些过程中 PDL 细胞的收缩力对于细胞的收缩至关重要。 胶原纤维排列，强化牙周组织并维持其功能 这项研究的长期目标是确定调节 PDL 细胞的机制。 可用作再生和维护架构的临床工具的力学 牙周复合体随时间的变化和功能因此，该提案的目标是 证明调节 PDL 细胞收缩力的机制之间的联系 具有 PDL 结构和组织力学的促炎微环境。 该提议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将是这一假设。 通过鉴定体外调节 PDL 细胞的机制，在特定目标 1 中进行测试 单细胞水平上促炎微环境中的收缩力。 印迹将用于确定肿瘤坏死因子 α (TNFα) 和透明质酸的作用 寡糖 (oHA) 作用于产生细胞收缩力的信号通路，例如 Rho/Rock 通路，以将炎症环境和细胞信号传导联系起来。 将在有或没有炎症介质的情况下测量收缩性、细胞牵引力 在具体目标 2 中，将构建三维 PDL 结构。 开发用于将调节组织水平收缩性的信号通路与基质连接起来 工程 PDL 结构将使用 PDL 细胞和进行开发。 将测量胶原蛋白和原位力，并用兴奋剂进行处理。 在模拟牙周稳态的条件下，Rho/Rock 通路的抑制剂和 这些目标的成功完成将有助于炎症的发展。 在促炎状态下维持 PDL 或再生组织的临床技术 未来的研究将扩展该模型以包括牙骨质样组织和骨骼； 因此，这项试点研究是朝着牙周再生的未来目标迈出的第一步。 复杂并长期保持其功能完整性。

项目成果

Black dots: High-yield traction force microscopy reveals structural factors contributing to platelet forces.

• DOI: 10.1016/j.actbio.2021.11.013
• 发表时间: 2023-06
• 期刊: ACTA BIOMATERIALIA
• 影响因子: 9.7
• 作者: Beussman, Kevin M.;Mollica, Molly Y.;Leonarda, Andrea;Milesc, Jeffrey;Hocterd, John;Songe, Zizhen;Stollac, Moritz;Hang, Sangyoon J.;Emerya, Ashley;Thomasb, Wendy E.;Sniadecki, Nathan J.
• 通讯作者: Sniadecki, Nathan J.