COLLAGEN METABOLISM IN WOUND REPAIR AND KELOID

描述：(改编自申请人的摘要)虽然是动物 模型和体外系统在过去一直非常有用，以获得 对细胞、生化和分子事件的洞察 组织修复，这些系统在解决独特的 与人类伤口愈合相关的临床问题。人类 响应包括广泛的治愈现象，包括 一端过度愈合和纤维化，一端形成慢性溃疡 另一头。动物不会出现过度的疤痕问题，也不会 明显存在愈合不足的问题。因此，这个项目是 旨在描述和分析正常伤口愈合反应 在人类中以及在相反的两端看到的病理反应 光谱。人的瘢痕疙瘩代表纤维化的情况 而慢性无法愈合的糖尿病溃疡将成为 治疗不力。所有研究都将在人类身上进行，使用 通常可获得的伤口渗出物、活组织检查和切除的组织 按照正常计划对这些患者进行手术治疗。 此外，一种可植入管，由扩展形式的 特氟龙，被称为Impra，已被证明对收集新的伤口很有用 用于分析的组织。这个实验室现在已经发展和适应了 研究人类伤口愈合的最新技术和方法。 因此，这些策略将被用来分析炎症， 细胞因子的产生，基质的合成和降解，血管生成和 正常人伤口、瘢痕疙瘩和慢性不可愈合的收缩 溃疡。实验室方法包括常规组织学， 免疫染色、酶联免疫吸附试验、原位杂交、核糖核酸酶保护 一种新的胶原蛋白放射性底物定量分析方法 胶原酶和明胶酶活力及羟脯氨酸的高效液相色谱定量 和亮氨酸沉积，以及蛋白质印迹分析。这些方法有 已被修改为适用于分析少量组织 以及可以从这些人类来源获得的渗出物。 这些研究将比较和对比正常的机制 负责修复健康组织中的这些机制 负责瘢痕疙瘩的过度愈合和慢性瘢痕疙瘩的愈合不足 溃疡。

DESCRIPTION: (Adapted from the applicant's abstract) Although animal models and in vitro systems have been very useful in the past to gain insight into the cellular, biochemical and molecular events during tissue repair, these systems have limitations when addressing the unique clinical problems associated with human wound healing. The human response encompasses a wide spectrum of healing phenomena with overhealing and fibrosis on one end and chronic ulcer formation on the other end. Animals do not develop excessive scar issue nor do they manifest problems with deficient healing. Therefore, this project is designed to characterize and analyze the normal wound healing response in humans as well as the pathologic responses seen at opposite ends of the spectrum. The human keloid will represent conditions of fibrosis whereas chronic non-healing diabetic ulcers will serve as a model of deficient healing. All studies will be done in humans using the normally available wound exudates, biopsies, and excised tissue following normally scheduled surgical treatments for these patients. In addition, an implantable tube consisting of an expanded form of Teflon, called Impra, has proven to be useful to collect new wound tissue for analysis. This laboratory has now developed and adapted state-of-the-art technologies and methods to study human wound healing. Therefore, these strategies will be employed to analyze inflammation, cytokine production, matrix synthesis and degradation, angiogenesis and contraction in normal human wounds, keloids and chronic non- healing ulcers. The laboratory methods include routine histology, immunostaining, ELISA assays, in situ hybridization, RNase protection assays, a new collagen radiolabeled substrate assay to quantify collagenase and gelatinase activity, HPLC quantitation of hydroxyproline and leucine deposition, and Western blot analysis. These methods have been modified to be applicable to analyze the small amounts of tissue and exudate material that can be obtained from these human sources. These studies will compare and contrast the normal mechanisms responsible for healing in healthy tissue to those mechanisms responsible for overhealing in keloids and underhealing in chronic ulcers.