FIBRIN GEL CONTRACTION & MATRIX SYNTHESIS BY FIBROBLASTS

纤维蛋白凝胶收缩

• 批准号: 2080030
• 负责人: TAI-LAN TUAN
• 金额: $ 9.29万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2080030
• 关键词: affinity chromatography; cell adhesion; collagen; crosslink; epidermal growth factor; extracellular matrix proteins; fibrin; fibrinogen receptors; fibrinopeptide; fibroblast growth factor; fibroblasts; fibronectins; fibrosis; gel electrophoresis; growth factor receptors; human tissue; immunofluorescence technique; platelet derived growth factor; radiotracer; receptor binding; transforming growth factors; western blottings

The process of repair and regeneration following traumatic or surgical injuries represents a fundamental protection of an organism against its environment. Thus, cutaneous wound healing is not only one of the most important research subjects, but also a challenging one due to the complex nature of skin. The general features of wound healing have been described (Peacock, E. E. 1984.), however, the exact mechanisms of healing remain undefined. The long term objective of the research plan is to understand the cellular and biochemical mechanisms underlying fibroplasia in wound repair. The information gained from this study will uncover important details of biological events essential in fibroblast-fibrin interactions and possibly lead to the treatment of keloid and hypertrophic scars and other pathologic skin conditions. During the five years of the proposed project, research will focus on the interactions between skin fibroblasts and fibrin and their influence on matrix remodeling using an in vitro 3-dimensional fibrin gel culture system. The following specific aims will be systematically studied. I. Characterization and purification of fibrinogen receptor on fibroblasts. We will study the binding characteristic of fibrin to fibroblasts, and use antibodies and adhesion peptides to investigate the adhesion function of fibroblasts on fibrinogen substratum and in fibrin gels and use the methods of chemical crosslinking and affinity chromatography to isolate the receptor. I. Study the effect of fibroblast types, age, and density on fibroblast contraction of fibrin gels. We will culture papillary and reticular fibroblasts separately from dermis and study their interactions with fibrin gels in gel contraction and degradation. III. Study the effect of TGF-beta on fibroblast contraction of fibrin gels. A preliminary study has indicated that TGF-beta enhanced gel contraction and prevented fibrinolysis under serum free condition. We will further investigate the mechanism of the effect regarding fibrin matrix contraction and degradation by fibroblasts. IV. Study collagen synthesis by fibroblasts cultured in fibrin gels and the effect of TGF-beta on the process. First, we will study the collagen synthesized by fibroblasts in fibrin gels and compare to monolayer and collagen gel cultures; second, the effect of fibrinolysis on collagen synthesis and the enzyme that involved in the lytic process; and third, the effect of TGF-beta on collagen synthesis and examine its regulatory mechanisms on both fibrin degradation and collagen synthesis by fibroblasts. V. To study the behavior of fibroblasts isolated from keloids and hypertrophic scars in fibrin gel contraction and collagen production and the effect of TGF-beta on the process. We will use fibroblasts isolated from papillary and reticular dermis separately in the studies and investigate the possible pathologic mechanism involved in the tissue fibrosis.

创伤或手术后的修复和再生过程 伤害是生物体抵御其 环境因此，皮肤伤口愈合不仅是最重要的治疗之一， 重要的研究课题，但也是一个具有挑战性的，由于复杂的 皮肤的性质。伤口愈合的一般特征已被描述 （Peacock，E. E. 1984年），然而，确切的愈合机制仍然存在， 未定义。研究计划的长期目标是了解 伤口纤维增生的细胞和生化机制 修复.从这项研究中获得的信息将揭示重要的 成纤维细胞-纤维蛋白相互作用中重要的生物学事件的详细信息 并可能导致瘢痕疙瘩和增生性瘢痕的治疗， 其他病理性皮肤病。 在拟议项目的五年期间，研究将集中在 皮肤成纤维细胞与纤维蛋白的相互作用及其对 使用体外三维纤维蛋白凝胶培养物进行基质重塑 系统将系统地研究以下具体目标。 I.成纤维细胞纤维蛋白原受体的鉴定与纯化。 我们将研究纤维蛋白与成纤维细胞的结合特性， 抗体和粘附肽，以研究粘附功能， 纤维蛋白原基质上和纤维蛋白凝胶中的成纤维细胞和使用该方法 化学交联和亲和色谱分离， 受体的 I.研究成纤维细胞类型、年龄、密度对成纤维细胞增殖的影响 纤维蛋白凝胶的收缩。我们将培养乳头状和网状 分离真皮成纤维细胞，并研究其与纤维蛋白的相互作用 凝胶在凝胶收缩和降解。 三.研究TGF-β对纤维蛋白凝胶成纤维细胞收缩的影响。 初步研究表明，TGF-β增强凝胶收缩， 并在无血清条件下阻止纤溶。我们将进一步 探讨纤维蛋白基质收缩作用的机制 和被成纤维细胞降解。 四.研究纤维蛋白凝胶中培养的成纤维细胞的胶原合成， TGF-β在这个过程中的作用。首先，我们将研究胶原蛋白 由纤维蛋白凝胶中的成纤维细胞合成，并与单层和 胶原凝胶培养;第二，纤维蛋白溶解对胶原的影响 合成和参与裂解过程的酶;第三， TGF-β对胶原合成影响及其调节作用的研究 纤维蛋白降解和胶原合成的机制， 成纤维细胞 诉研究瘢痕疙瘩成纤维细胞的行为， 纤维蛋白凝胶收缩和胶原蛋白生成中的增生性瘢痕， TGF-β在这个过程中的作用。我们将使用分离的成纤维细胞 在研究中分别从乳头状和网状真皮中提取， 探讨组织中可能的病理机制 纤维化