Impact of Oxygen Tension on Human Meniscus Tissue Engineering

项目概述半月板功能丧失可导致关节软骨退变和骨关节炎。这对年龄太小的患者进行全膝关节置换术提出了临床挑战。为了满足临床对半月板缺陷更有效治疗的需求，该培训项目旨在促进长期的职业目标，即创建一个模拟天然半月板生物学和功能的间充质干细胞（MSC）种子半月板支架结构。本项目将评估缺氧、HIF-1a信号和微粒氧生成系统对人类间充质干细胞在多孔异体移植人类半月板上分化的影响。将利用定量PCR、共聚焦显微镜和蛋白多糖合成分析等方法评估培养的支架的生物学特性，并与正常半月板组织的特性进行比较。HIF-1a信号将被上调和下调，利用化学和分子技术阐明HIF-1a在缺氧培养环境中介导纤维软骨细胞稳态和分化中的作用。该建议是一个长期项目的一部分，旨在创建一个基于生物学的半月板替代结构，与同种异体半月板移植相比，其长期效果更好。本课题的研究将：1)提高我们对正常成人半月板组织生物学的认识，以及HIF-1a转录介导缺氧对半月板细胞稳态和表型的影响。2)更好地理解干细胞向纤维软骨细胞表型分化的程序，以及调节缺氧作为纤维软骨组织工程生物反应器变量的作用。3)研究缺氧和颗粒氧生成系统对组织工程半月板替代结构蛋白聚糖生成的影响。综上所述，从这些研究中收集的信息将为医生更好地了解正常半月板生物学、体内平衡和分化机制提供帮助。组织工程领域的成功可能会导致与半月板病理相关的膝关节疼痛的替代和扩展治疗。作为一名内科科学家，这项培训资助将帮助我进一步提高我的分子生物学技能和研究培训，通过从基于机制的分子方法接近组织工程策略，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.