力学负载通过骨髓微环境外泌体调控破骨细胞增殖分化及迁移的研究

Activation of osteoclast induced bone absorption contributed to mechanisms of osteoporosis and prosthetic loosening after surgery. Proliferation and differentiation of osteoclast play an important role in development of osteoporosis. This project proposed a hypothesis that myeloid microenvironment derived exosome after mechanical loading play an important role in the proliferation and differentiation of osteoclast, following the previous study of bone absorption after fatigue loading in vivo. In this project, we will explore the function of exosome in regulation of osteoclast proliferation, differentiation and migration. Firstly, the exosome would be extract form ovariectomized (OVX) rats, and then co-culture with mononuclear macrophage to confirm the function of exosome toward osteoclast proliferation and differentiation. Then, we will perform the transcriptome using high-throughput sequencing. Divergence analysis was also used to explore the target pathway, and verified by cell experiment. Finally, explore the function of exosome on osteoclast proliferation, and analyze the pathway associated with focal adhesion. This project will systematically analyze the function of myeloid microenvironment derived exosome exert on osteoclast.

破骨细胞活化在骨质疏松症及术后假体松动中起重要作用。破骨细胞增殖及分化的调控在骨质疏松症发展中居核心作用。本研究在活体疲劳负载后骨吸收的研究基础上，提出力学负载后，骨髓微环境中外泌体功能改变，影响破骨细胞的增殖、分化及迁移，拟从分子-细胞-动物实验，整体探讨骨髓微环境外泌体调控破骨细胞的机制。首先通过动物活体疲劳负载实验获取外泌体，将其与单核巨噬-破骨细胞分化体系共培养，证明其调控作用；再通过转录组高通量测序及差异性分析寻找外泌体调控破骨细胞分化的靶通路，进而在细胞系中验证；最后，观察外泌体在单核-巨噬细胞及破骨前体迁移过程中的作用，分析粘附斑形成相关通路分子的表达。系统论证骨髓外泌体对破骨细胞的整体作用。