DEVELOPMENT OF AN IN VITRO WOUND HEALING MODEL

这项研究的长期目标是了解细胞生物学 伤口愈合，特别是肉芽组织的形成及其 被新的真皮替代，伤口痉挛，和重新上皮化。 在接下来的四年里，我们计划系统地调查几个 体外模型系统，并开展相关的体内研究。这个 人成纤维细胞对水合胶原凝胶的重组 研究了解血清和细胞分泌因子的同一性 发生凝胶重组所必需的。此外，外源的影响 成纤维细胞迁移和凝胶重组的基质成分将是 下定决心。人角质形成细胞迁移分化的实验研究 重组的胶原蛋白将被分析；我们将确定 纤维连接蛋白受体的功能是在这些培养条件下启动的。 胶原蛋白凝胶中成纤维细胞和角质形成细胞的能力 将测量吞噬胶原和纤维蛋白碎片。一种体外模型 通过培养角质形成细胞来建立模拟皮肤创伤 在含有成纤维细胞的水合胶原凝胶上，放置一个 血浆凝块变成物理缺陷，在凝胶中制备。移民的迁移 将确定与“伤口”界面有关的细胞，并且 将分析形成肉芽组织的可能性 研究细胞的形态和生物合成特征。贯穿始终 在上述研究中，我们将使用抗纤维连接蛋白抗体和纤维连接蛋白 细胞结合区--确定纤维连接蛋白功能的多肽片段。 最后，将进行体内实验，以延长体外 调查结果。特别是，我们将尝试评估纤维连接蛋白在 肉芽组织形成和再上皮化与纤维连接蛋白受体 迁移角质形成细胞的启动及其可能的差异 新鲜分离和培养的角质形成细胞的形成能力 全层创面上的移植。

The long term goal of this research is to understand the cell biology of wound healing, particularly granulation tissue formation and its replacement by neodermis, wound contracture, and re-epithelialization. During the next four years, we plan to systematically investigate several in vitro model systems and to carry out related in vivo studies. The reorganization of hydrated collagen gels by human fibroblasts will be studied to learn the identity of the serum and cell-secreted factors necessary for gel reorganization to occur. Also, the effects of exogenous matrix components on fibroblast migration and gel reorganization will be determined. Human keratinocyte migration and differentiation on reorganized collagen will be analyzed; and we will determine whether fibronectin receptor function is initiated under these culture conditions. The ability of fibroblasts and keratinocytes in the collagen gels to phagocytize collagen and fibrin debris will be measured. An in vitro model mimicking cutaneous wounds will be established by culturing keratinocytes on top of hydrated collagen gels that contain fibroblasts, and placing a plasma clot into a physical defect prepared in the gels. The migration of cells with respect to the "wound" interface will be determined, and the possibility that a granulation-like tissue is formed will be analyzed by studying morphological and biosynthetic features of the cells. Throughout the above studies, we will use anti-fibronectin antibodies and fibronectin cell binding region-peptide fragments to determine fibronectin function. Finally, in vivo experiments will be carried out to extend the in vitro findings. In particular, we will try to assess fibronectin function in granulation tissue formation and re-epithelialization, fibronectin receptor initiation in migrating keratinocytes, and possible differences between freshly isolated and cultured keratinocytes in their abilities to form grafts on full-thickness wounds.