Development of an In Vitro Wound Healing Model

描述(申请人提供)：纤维结缔组织为身体的其他组织提供机械支撑和框架。I型胶原是纤维结缔组织的主要蛋白质成分。成纤维细胞是主要负责胶原生物合成和重塑的细胞类型。作为机械重塑的结果，胶原和纤维结缔组织的其他成分拉伸、滑动并经历稳定的重组。这种重塑涉及正常生理和病理的多个方面，包括伤口修复、纤维化、瘢痕形成、肿瘤发生和衰老。基质重塑也是组织工程的一个重要设计特征。我们研究的前提是，我们可以使用3D胶原基质来分析和剖析组织状环境中成纤维细胞-基质相互作用的结构、功能和力学特征。了解这些特征将有助于发现促进伤口修复的干预措施，并促进组织工程领域的发展。事实上，3D矩阵模型在加速广泛的翻译研究工作方面的普遍有用性已经越来越被认识到。在目前的方案中，我们计划分析人成纤维细胞在嵌套胶原基质和纤维蛋白基质中的迁移和胶原流动。我们还将分析人成纤维细胞在胶原基质上的迁移和聚集，作为生长因子环境的函数，并确定在适当的生长因子刺激下，肌成纤维细胞是否是迁移细胞。在其他研究中，将评估细胞-细胞黏附连接在细胞聚集和细胞迁移中的作用。关于生长因子环境的工作将特别集中在鞘氨醇-1-磷酸(S1P)上，它可能起到双重调节作用--对成纤维细胞迁移不利，对收缩积极。我们将分析S1P功能的重要受体，并分析S1P和其他迁移/收缩因子在急性人体创面液中的存在。最后，我们将测试盘状结构域受体2，一种结合胶原的细胞表面酪氨酸激酶受体，是否在成纤维细胞-胶原基质相互作用中发挥特殊作用。公共卫生相关性：成纤维细胞对纤维结缔组织的重塑涉及正常生理和病理的多个方面，包括伤口修复、纤维化、瘢痕形成、肿瘤发生和衰老。基质重塑也是组织工程的一个重要设计特征。我们研究的前提是，我们可以使用含有人成纤维细胞的3D胶原基质来分析和解剖组织样环境中结缔组织重塑的结构、功能和力学特征。了解这些特征将有助于发现促进伤口修复的干预措施，并促进组织工程领域的发展。事实上，3D矩阵模型在加速广泛的翻译研究工作方面的普遍有用性已经越来越被认识到。

DESCRIPTION (provided by applicant): Fibrous connective tissue provides mechanical support and frameworks for the other tissues of the body. Type 1 collagen is the major protein component of fibrous connective tissue. Fibroblasts are the cell type primarily responsible for collagen biosynthesis and remodeling. As a result of mechanical remodeling, collagen and other components of fibrous connective tissue stretch, slip, and undergo stable reorganization. Such remodeling has been implicated in diverse aspects of normal physiology and pathology including wound repair, fibrosis, scar formation, tumorigenesis, and aging. Matrix remodeling also is an important design feature in tissue engineering. Underlying our research is the premise that we can use 3D collagen matrices to analyze and dissect the structural, functional and mechanical features of fibroblast-matrix interactions in a tissue-like environment. Understanding these features should facilitate discovery of interventions to promote wound repair and enhance development of the field of tissue engineering. Indeed, the general usefulness of 3D matrix models to accelerate a wide range of translational research work increasingly has been recognized. In the current proposal, we plan to analyze human fibroblast migration and collagen flow in nested collagen matrices and in fibrin matrices. We also will analyze human fibroblast migration and clustering on collagen matrices as a function of the growth factor environment and determine if myofibroblasts are migratory cells given appropriate growth factor stimulation. In other studies, the role of cell-cell adherens junctions in cell clustering and cell migration will be assessed. Work on the growth factor environment will focus especially on sphingosine-1-phosphate (S1P), which potentially plays a dual regulatory role -- negative for fibroblast migration and positive for contraction. We will analyze the receptors important for S1P function and analyze the presence of S1P and other promigratory/procontractile factors in acute human wound fluid. Finally, we will test if discoidin domain receptor 2, a cell surface tyrosine kinase receptor that binds collagen, plays a specific role in fibroblast-collagen matrix interactions. PUBLIC HEALTH RELEVANCE: Remodeling of fibrous connective tissue by fibroblasts has been implicated in diverse aspects of normal physiology and pathology including wound repair, fibrosis, scar formation, tumorigenesis, and aging. Matrix remodeling also is an important design feature in tissue engineering. Underlying our research is the premise that we can use 3D collagen matrices containing human fibroblasts to analyze and dissect the structural, functional and mechanical features of connective tissue remodeling in a tissue-like environment. Understanding these features should facilitate discovery of interventions to promote wound repair and enhance development of the field of tissue engineering. Indeed, the general usefulness of 3D matrix models to accelerate a wide range of translational research work increasingly has been recognized.