Functional Repair of Elastin with Recomb. Tropoelastin

Recomb 功能性修复弹性蛋白。

• 批准号: 6794641
• 负责人: PHILLIP J STONE
• 金额: $ 24.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-25 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6794641
• 关键词: bioengineering /biomedical engineering; collagen; crosslink; elastin; extracellular matrix; fibrogenesis; intermolecular interaction; laboratory rat; newborn animals; proteoglycan; recombinant proteins; tissue /cell culture; tissue engineering; vascular smooth muscle

DESCRIPTION (provided by applicant): In disease states, such as emphysema and aneurysmal degeneration, the degradation of elastin fibers that characterizes these conditions is accompanied by repair processes that are insufficient and fail to prevent disease progression. Towards this goal our objectives are to regenerate elastin in tissue culture systems by crosslinking recombinant tropoelastin (rTE) into damaged elastin and to characterize the effectiveness of this regeneration by assessing the functional mechanical, biochemical, and ultrastructural properties. Our central hypothesis is that elastin can be regenerated in vitro and in vivo by administering rTE to damaged elastin, thereby restoring, at least in part, the pre-existing elastin fiber organization and its mechanical properties. Such repair might prevent mechanical forces from causing failure of the remodeled matrix at loci of stress concentrations. Aim 1.To identify the separate contributions of elastin, proteoglycans, collagens, and cells to the macroscopic mechanical properties of elastogenic cultures. This will be carried out in well-defined cultures by manipulating the amount of elastin, fibrillar collagen, proteoglycans or by generating cell-free matrices. This will provide essential information on the structural and mechanical properties of the components of extracellular matrices and provide the baseline information for this in vitro model system. Aim 2. To assess the changes in functional mechanical, biochemical, and ultrastructural properties of the defined elastogenic cultures from Aim 1 during and after treatment with elastases to degrade elastin and proteoglycans or with non-elastolytic enzymes to digest primarily collagen or proteoglycans. The cultures will then be monitored as the endogenous smooth muscle cells effect repair. The information will shed light on how enzymatic injury degenerates structure and mechanical function and the effectiveness of endogenous repair processes. Aim 3. To determine the mechanical, biochemical and ultrastructural properties of rTE mediated regeneration of elastase-treated matrices in the presence or absence of endogenous tropoelastin production.

描述（由申请人提供）： 在疾病状态下，例如肺气肿和动脉瘤变性，弹性蛋白纤维的降解是这些疾病的特征，伴随着修复过程，这些修复过程不足，无法预防疾病进展。为了实现这一目标，我们的目标是通过将重组型tropoelastin（RTE）交联成弹性蛋白，并通过评估功能机械，生化和超微结构特性来表征这种再生的有效性，从而再生组织培养系统中的弹性蛋白。我们的中心假设是，可以通过对损坏的弹性蛋白施用RTE来在体外和体内再生，从而至少部分恢复了先前存在的弹性蛋白纤维组织及其机械性能。这种修复可能会阻止机械力导致应力浓度基因座重塑基质的故障。目标1.确定弹性蛋白，蛋白聚糖，胶原蛋白和细胞对弹性培养物的宏观机械性能的独立贡献。通过操纵弹性蛋白，原纤维胶原蛋白，蛋白聚糖或产生无细胞的矩阵，将在明确的培养物中进行。这将提供有关细胞外矩阵组件的结构和机械性能的重要信息，并为此体外模型系统提供基线信息。目的2。评估定义的弹性培养物的功能机械，生化和超微结构特性的变化，从AIM 1和使用弹性酶处理期间和治疗后，以降解弹性蛋白和蛋白聚糖或非肾上腺素溶酶以降解主要是胶原蛋白或蛋白酶的非肾上腺素酶。然后，随着内源性平滑肌细胞的影响修复，培养物将受到监测。该信息将阐明酶促损伤如何退化结构和机械功能以及内源性修复过程的有效性。目的3。确定在存在或不存在内源性tropoelastin产生的情况下，RTE介导的弹性酶处理矩阵再生的机械，生化和超微结构特性。