MECHANISMS OF FETAL WOUND REPAIR--CELLULAR AND MOLECULAR

描述：(改编自申请人的摘要)长期 该项目的目标是确定负责的独特机制 用于胎儿伤口愈合。胎儿的伤口通过以下过程愈合 与那些参与成人伤口愈合的人有显著不同。那里 是没有急性炎症、收缩和纤维化，导致 很少或没有疤痕形成。胎儿组织基质丰富 透明质酸的含量被认为是关键因素之一 负责最佳胎儿伤口愈合反应。这项建议 将确定透明质酸调节细胞外基质的机制 真皮成纤维细胞沉积。这将包括测量影响 透明质酸对I型和III型胶原、纤维连接蛋白、 基质金属蛋白酶、透明质酸及其受体。蛋白质含量 合成将通过凝胶电泳和放射自显影进行量化 脉冲标记的蛋白质。基质的功能活性 金属蛋白酶(胶原酶和明胶酶)将使用 研究人员建立的放射性标记胶原底物测定法 实验室。此外，这项提案还将定义信号 透明质酸调节基质基因表达的转导机制。 透明质酸受体的表达将通过定量测定 RNA和蛋白质水平。测定透明质酸能力的几种方法 还将使用与配体结合的受体。信号通路将是 通过在存在或不存在的情况下测量蛋白质磷酸化来识别 特定途径的抑制物。这些研究将集中在正常皮肤上 以及从胎兔和成年兔分离的伤口成纤维细胞。在……里面 此外，还设计了具体的研究来检验在 在胎兔和成年兔体内发现细胞培养。活体内 研究将使用原位杂交进行转录分析和 蛋白质的表达将通过免疫组织化学来完成。 这些研究产生的信息将提供更好的理解 胎儿的生理和发育。这些基本的生物信息将 为治疗许多人类伤口愈合提供了更好的临床依据 问题，如瘢痕疙瘩，增生性烧伤疤痕紧缩，腹膜 粘连和狭窄以及慢性不可愈合的伤口，如 压力性、糖尿病和静脉淤积性溃疡。这些临床问题 导致严重的发病率和医疗费用。

DESCRIPTION: (Adapted from the applicant's abstract) The long term objective of this project is to determine the unique mechanisms responsible for fetal wound healing. Wounds in the fetus heal by processes which are significantly different than those involved in adult wound healing. There is an absence of acute inflammation, contraction, and fibrosis resulting in minimal or no scar formation. The fetal tissue matrix has abundant quantities of hyaluronan which is thought to be one of the critical factors responsible for the optimal fetal wound healing response. This proposal will define the mechanisms whereby hyaluronan modulates extracellular matrix deposition by dermal fibroblasts. This will include measuring the effects of hyaluronan on the expression of collagen types I and III, fibronectin, matrix metalloproteinases, hyaluronan and its receptors. Rates of protein synthesis will be quantified by gel electrophoresis and autoradiography of pulse-labeled proteins. Functional activity of the matrix metalloproteinases (collagenases and gelatenases) will be determined using a radiolabeled collagen substrate assay developed in the investigators laboratory. In addition, this proposal will also define the signal transduction mechanism whereby hyaluronan modulates matrix gene expression. Expression of hyaluronan receptors will be determined by quantitation at the RNA and protein levels. Assays determining the ability of hyaluronan receptors to bind to ligand will also be used. Signaling pathways will be identified by measuring protein phosphorylation in the presence or absence of pathway-specific inhibitors. These studies will focus on normal dermal and wound fibroblasts isolated from both fetal and adult rabbits. In addition, specific studies are designed to test the observations made in cell culture to in vivo findings in fetal and adult rabbits. The in vivo studies will employ in situ hybridization for transcript analysis and protein expression will be accomplished using immunohistochemistry. Information generated by these studies will provide a better understanding of fetal physiology and development. This basic biological information will lead to better clinical rationales for treating the many human wound healing problems such as keloid, hypertrophic burn scar contractures, peritoneal adhesions and strictures as well as chronic non-healing wounds such as pressure, diabetic, and venous stasis ulcers. These clinical problems result in major morbidity and health care costs.