Impact of Oxygen Tension on Human Meniscus Tissue Engineering

描述(申请人提供)：项目摘要半月板功能丧失可导致关节软骨变性和骨关节炎。这对太年轻而不能进行全膝关节置换术的患者构成了临床上的挑战。为了满足临床对半月板缺乏症更有效治疗的需求，该培训项目旨在促进创造一种模拟本地半月板生物学和功能的骨髓间充质干细胞(MSC)种植的半月板支架结构的长期职业目标。该项目将评估低氧、HIF-1a信号和颗粒制氧系统对种植在多孔性同种异体半月板上的人间充质干细胞分化的影响。通过定量聚合酶链式反应、共聚焦显微镜和蛋白多糖合成分析，对培养的支架的生物学特性进行评估，并与正常半月板组织的特性进行比较。利用化学和分子技术上调和下调HIF-1a信号，以阐明HIF-1a在低氧培养环境中调节纤维软骨细胞动态平衡和分化中的作用。这项建议是一个长期项目的一部分，该项目旨在创造一种基于生物的半月板替代结构，与同种异体半月板移植相比，具有更好的长期效果。这项研究将：1)加深我们对正常成人半月板组织生物学以及转录HIF-1a在低氧对半月板细胞动态平衡和表型影响中的作用的理解。2)更好地理解干细胞向纤维软骨细胞表型分化的程序，以及作为纤维软骨组织工程生物反应变量的调节缺氧的作用。3)研究缺氧和颗粒制氧系统对组织工程半月板置换结构蛋白多糖合成的影响。综上所述，从这些研究中收集的信息将使医生更好地了解正常的半月板生物学、内稳态和分化机制。组织工程领域的成功可能会导致半月板病理相关的膝关节疼痛的替代和扩展治疗方法。作为一名内科科学家，这笔培训资金将帮助我进一步提高我的分子生物学技能和研究培训，从基于机制的分子方法来探讨组织工程策略，并为验证未来组织工程技术在临床领域的有效性奠定基础。

DESCRIPTION (provided by applicant): Project Summary Loss of meniscus function can lead to articular cartilage degeneration and osteoarthritis. This poses a clinical challenge in patients who are too young for total knee arthroplasty. To address the clinical need for a more effective treatment for meniscus deficiency, this training project is designed to facilitate a long-term career goal of creating a mesenchymal stem cell (MSC) seeded meniscus scaffold construct that emulates native meniscus biology and function. This project will evaluate the effect of hypoxia, HIF-1a signaling, and particulate oxygen generating systems on the differentiation of human mesenchymal stem cells seeded on a porous allograft derived human meniscus. The biologic characteristics of the cultivated scaffolds will be evaluated and compared with the properties of normal meniscus tissue using quantitative PCR, confocal microscopy, and proteoglycan synthesis analysis. HIF-1a signaling will be upregulated and downregulated using chemical and molecular techniques to elucidate the role of HIF-1a in mediating fibrochondrocyte homeostasis and differentiation in a hypoxia culture environment. This proposal is part of a long term project to create a biologically based meniscus replacement construct with improved long term results compared with allograft meniscus transplant. The studies described in this proposal will: 1) Improve our understanding of the biology of normal adult human meniscus tissue and the role of the transcription HIF-1a in mediating the effect of hypoxia on meniscus cell homeostasis and phenotype. 2) Result in a better understanding of the program of stem cell differentiation toward the fibrochondrocyte phenotype and the role of regulated hypoxia as a bioreactor variable for fibrocartilage tissue engineering. 3) Investigate the impact of hypoxia and particulate oxygen generating systems on the proteoglycan production of tissue engineered meniscus replacement constructs. Taken together, the information gathered from these studies will provide physicians with a better understanding of normal meniscus biology, homeostasis, and differentiation mechanisms. Success in this realm of tissue engineering could lead to alternative and expanded treatments for knee pain related to meniscus pathology. As a physician scientist, this training grant will help me further advance my molecular biology skills and research training by approaching tissue engineering strategies from a mechanism based molecular approach and lay the groundwork for validating the effectiveness of future tissue engineering technologies in the clinical arena.