COLLAGEN METABOLISM IN WOUND REPAIR AND KELOID

该项目的长期目标是定义和分析 细胞间相互作用及参与调控的分子机制 组织修复过程中的胶原蛋白新陈代谢。这一目标将通过以下方式实现 正常人成纤维细胞亚群的生化特征 真皮、瘢痕和瘢痕疙瘩与胶原合成、胶原酶的关系 表达与TGFβ受体数目和亲和力的关系。这些研究测试了 纤维化是选择成纤维细胞的结果的假说 能够产生更多胶原蛋白的亚群。在……里面 此外，正在进行的研究将完成，以确定 创面早期胶原生成单核细胞的观察 修理。然后，这一特定数量的细胞将从 伤口部位确定其在组织中的生理意义 修理。在其他研究中，胶原、纤维连接蛋白、 胶原酶和转化生长因子β将在正常和生物学上进行定量 受损的大鼠伤口检查转录控制机制 组织修复。此信息将与实际的 用Western印迹分析对这些蛋白的表达进行了定量 伤口液和组织提取液以及免疫组织化学 本地化。此外，将对胶原酶进行定量，以确定 胶原蛋白降解在胶原蛋白沉积净量中的作用 伤口处。胶原酶信使核糖核酸的量将与 酶活性分析酶的调节机制。瘢痕疙瘩 外植体将被放置在裸鼠身上以研究胶原代谢 在体内使用氧-18标记技术。这将使我们能够研究 瘢痕疙瘩体内发病机制的动态研究为临床提供初步数据 最终的活体人体研究。这种动物模型也应该被证明是有用的 用于评价治疗瘢痕疙瘩的药理药物。可能的角色 肥大细胞和肿瘤坏死因子(TN-F)在瘢痕疙瘩形成中的作用 在裸鼠裸鼠模型和细胞中都进行了检测 文化。这些研究的最终结果将是更好地理解 调节正常伤口修复的分子机制及其如何 在异常修复中，调节事件会发生变化。

The long-term objective of this project is to define and analyze the cellular interactions and molecular mechanisms involved in the regulation of collagen metabolism during tissue repair. This objective will be met by biochemically characterizing fibroblast subpopulations derived from normal dermis, scar, and keloids with respect to collagen synthesis, collagenase expression and TGFbeta receptor number and affinities. These studies test the hypothesis that fibrosis is the result of a selection of fibroblast subpopulations capable of producing increased amounts of collagen. In addition, ongoing studies will be completed to identify the collagen-producing monocytes observed during the early phase of wound repair. This specific population of cells will then be eliminated from the wound site to determine its physiological significance during tissue repair. In other studies, the amount of mRNA for collagens, fibronectin, collagenase and TGFbeta will be quantitated in normal and biologically compromised rat wounds to examine transcriptional control mechanisms during tissue repair. This information will be correlated with the actual expression of these proteins as quantitated using Western blot analysis of wound fluid and tissue extracts as well as immunohistochemical localization. In addition, collagenase will be quantitated to determine the role of collagen degradation in the net amount of collagen deposited in a wound site. The amount of collagenase mRNA will then be correlated with enzyme activity to analyze the mechanism of enzyme regulation. Keloid explants will be placed onto athymic nude mice to study collagen metabolism in vivo using oxygen-18 labeling techniques. This will allow a study of the dynamics of keloid pathogenesis in vivo and provide initial data for eventual in vivo human studies. This animal model should also prove useful for evaluation of pharmacologic agents to treat keloids. The probable role of the mast cell and tumor necrosis factor (TN-F) in keloid formation will be examined both in the athymic nude mouse model as well as in cell culture. The net result of these studies will be a better understanding of the molecular mechanisms which regulate normal wound repair and how these regulatory events are altered in abnormal repair.