MECHANISMS OF FETAL WOUND REPAIR--CELLULAR AND MOLECULAR

描述：（改编自申请人的摘要）长期 本项目的目标是确定负责的独特机制 用于胎儿伤口愈合 胎儿的伤口愈合过程 与成人伤口愈合中所涉及的那些显著不同。 那里 没有急性炎症、收缩和纤维化， 最小或无疤痕形成。 胎儿组织基质具有丰富的 大量的透明质酸被认为是 负责最佳的胎儿伤口愈合反应。 这项建议 将定义透明质酸调节细胞外基质的机制 真皮成纤维细胞沉积。 这将包括衡量影响 透明质酸对I型和III型胶原蛋白，纤连蛋白， 基质金属蛋白酶、透明质酸及其受体。 蛋白质比率 合成将通过凝胶电泳和放射自显影定量， 脉冲标记蛋白质。 基质的功能活性 金属蛋白酶（胶原酶和明胶酶）将使用 研究者开发的放射性标记胶原底物测定法 实验室 此外，这份提案还将对信号进行定义 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。 测定透明质酸的能力的试验 也可以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下测量蛋白质磷酸化来鉴定 特定途径抑制剂的作用。 这些研究将集中在正常皮肤 以及从胎兔和成年兔分离的伤口成纤维细胞。 在 此外，还设计了具体的研究，以检验 细胞培养与胎兔和成年兔体内结果的比较。 体内 研究将采用原位杂交进行转录物分析， 蛋白质表达将使用免疫组织化学完成。 这些研究所产生的信息将有助于更好地了解 胎儿的生理和发育 这些基本的生物信息将 导致更好的临床理据来治疗许多人类伤口愈合 瘢痕疙瘩、增生性烧伤瘢痕挛缩、腹膜炎等问题 粘连和狭窄以及慢性不愈合伤口， 压迫性、糖尿病性和静脉淤滞性溃疡。 这些临床问题 导致严重的发病率和卫生保健费用。

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.