RECOMBINANT HUMAN COLLAGEN & CTGF FOR TISSUE ENGINEERING

重组人类胶原蛋白

• 批准号: 6171712
• 负责人: JAMES W POLAREK
• 金额: $ 28.8万
• 依托单位: FIBROGEN, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-28 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6171712
• 关键词: SCID mouse; biomaterial compatibility; biomaterial development /preparation; cell adhesion; cell growth regulation; cell migration; cell type; clinical research; collagen; growth factor; human subject; laboratory mouse; recombinant proteins; regeneration; tissue engineering

The combination of a growth factor capable of effecting the regeneration of a tissue with a matrix suitable to act as a scaffold for that tissue is a central tenet in tissue engineering. We propose to test this hypothesis by developing prototype tissue engineering biomaterials for in vivo and in vitro proof of concept studies. Connective tissue growth factor (CTGF), a key fibrogenic growth factor, acts downstream of TGF- beta to promote mesenchymal cell proliferation and to stimulate extracellular matrix deposition. Our recent data suggest a role in development and differentiation of cartilage, skin and bone. Uses of CTGF in tissue engineering applications are unexplored. Strength, persistence and biocompatibility are unique and inherent properties of collagens that make these proteins well suited as scaffolds for these studies. Although animal-derived collagens are available and most frequently used as biomaterials, there are increasing concerns about their purity, safety and immunogenicity. Of the 20 known collagens, only type 1 collagen has been evaluated as a biomaterial, again because it is the only collagen economically extracted from tissue. Until recently, recombinant expression of human collagens has been difficult due to the significant post translational modifications needed for function. Our success with collagen expression technology now provides a means to generate and explore uses of the rarer, tissue-specific human collagens such as types II (predominant in cartilage) and III (prominent in vessels). To evaluate recombinant human collagens and CTGF as components of tissue engineering therapeutics we specifically will: 1) produce recombinant human collagen types I, II, and III in a cost effective, commercially viable yeast expression system. This will involve expression, purification, formulation and characterization; 2) evaluate biocompatibility of the recombinant human collagens compared to tissue- derived materials; 3) investigate in vitro how matching mesenchymal cell type and collagen matrix affects cell attachment, differentiation, migration, proliferation and survival and also determine the effects of including CTGF in these scaffolds; and, 4) evaluate these CTGF augmented scaffolds in tissue engineering applications using in vivo models of tissue repair and regeneration that focus on cartilage, bone and skin. The momentum of our ongoing programs and commitments in related areas will leverage additional support to expedite the development of these novel therapeutic prototypes that are capable of addressing significant medical needs. In addition, further avenues for research and development, based on novel combinations of cytokines and engineered and/or tissue-specific collagens, will have been identified.

能够影响再生的生长因子的组合 具有适合充当该组织支架的基质的组织 是组织工程的核心原则。 我们建议对此进行测试 通过开发原型组织工程生物材料来提出假设 体内和体外概念验证研究。 结缔组织生长 因子 (CTGF) 是一种关键的纤维生长因子，作用于 TGF-下游 β促进间充质细胞增殖并刺激 细胞外基质沉积。 我们最近的数据表明， 软骨、皮肤和骨骼的发育和分化。用途 CTGF 在组织工程中的应用尚未被探索。力量， 持久性和生物相容性是独特且固有的特性 胶原蛋白使这些蛋白质非常适合作为这些蛋白质的支架 研究。 尽管动物源性胶原蛋白是可用的，而且大多数 经常被用作生物材料，人们越来越关注 它们的纯度、安全性和免疫原性。 在 20 种已知的胶原蛋白中， 只有 1 型胶原蛋白被评估为生物材料，同样是因为 它是唯一从组织中经济地提取的胶原蛋白。 直到 近年来，人类胶原蛋白的重组表达一直很困难 由于需要进行大量的翻译后修改 功能。 我们在胶原蛋白表达技术方面的成功现在提供了 一种生成和探索更稀有的、特定组织的人类的用途的方法 胶原蛋白，例如 II 型（主要是软骨）和 III 型（主要是软骨） 在船只中）。 评估重组人胶原蛋白和 CTGF 作为组织成分 工程疗法我们具体将：1）生产重组 人类 I、II 和 III 型胶原蛋白具有成本效益、商业化 可行的酵母表达系统。 这将涉及表达， 纯化、配制和表征； 2）评估 与组织相比，重组人胶原蛋白的生物相容性 衍生材料； 3）研究体外如何匹配间充质 细胞类型和胶原基质影响细胞附着、分化、 迁移、增殖和生存，也决定了影响 在这些支架中包括CTGF；以及，4) 评估这些 CTGF 增强 使用体内模型的组织工程应用中的支架 专注于软骨、骨骼和皮肤的组织修复和再生。 我们在相关领域正在进行的计划和承诺的势头 将利用额外的支持来加快这些项目的开发 能够解决重大问题的新颖治疗原型 医疗需求。 此外，进一步的研究和 基于细胞因子和工程改造的新型组合的开发 和/或组织特异性胶原蛋白，将被鉴定。