RECOMBINANT HUMAN COLLAGEN & CTGF FOR TISSUE ENGINEERING

能够影响再生的生长因子的组合 指具有适合作为该组织的支架的基质的组织 是组织工程学的核心原则。我们建议对此进行测试 开发原型性组织工程生物材料的假说 体内和体外概念验证研究。结缔组织生长 结缔组织生长因子(CTGF)是一种重要的纤维化生长因子，作用于转化生长因子-2的下游。 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，在 软骨、皮肤和骨骼的发育和分化。使用 CTGF在组织工程中的应用尚未被探索。力量， 持久性和生物兼容性是独一无二的固有属性 使这些蛋白质非常适合作为支架的胶原蛋白 学习。尽管动物来源的胶原蛋白是可用的，而且大多数 由于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。在已知的20种胶原蛋白中， 只有1型胶原蛋白被评估为生物材料，同样是因为 它是唯一经济地从组织中提取的胶原蛋白。直到 近年来，重组人胶原蛋白的表达一直比较困难。 由于需要进行重大的翻译后修改， 功能。我们在胶原蛋白表达技术方面的成功现在提供了 一种产生和探索更罕见的组织特异性人类的用途的方法 胶原蛋白，如II型(软骨占优势)和III型(突出 在船只上)。 重组人胶原蛋白和CTGF作为组织成分的评价 工程疗法我们具体将：1)生产重组 商业上具有成本效益的人I型、II型和III型胶原蛋白 活性酵母表达系统。这将涉及到表达， 提纯、制备和表征；2)评价 重组人胶原蛋白与组织的生物相容性比较- 衍生材料；3)体外研究间充质细胞如何匹配 细胞类型和胶原基质影响细胞的附着、分化， 迁徙、扩散和生存，也决定了 在这些支架中包括CTGF；以及，4)评估这些CTGF增强 支架材料在组织工程中的应用 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开发这些 能够解决重大问题的新型治疗原型 医疗需要。此外，进一步的研究和研究途径 基于新型细胞因子组合和工程技术的开发 和/或组织特异性胶原蛋白，将被鉴定出来。

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.