创伤和骨肿瘤切除等可造成大段骨缺损，应用失活同种异体骨并配合自体骨髓间充质干细胞（MSC）移植修复是目前主要治疗方法。但移植物上MSC细胞增殖低下，粘附效率低，MSC成骨分化能力低下，治疗效果不佳。前期研究，我们建立了MSC体外培养系统（Jag1/Notch2 MSC)，可以使MSC迅速扩增，并维持干细胞特性；通过温度反应性培养皿生成的细胞膜片技术进行移植，提高MSC体内的粘附性与生存率；根据临床用,经验，筛选出一批可以促进MSC成骨分化的补肾中药及其有效单体。在此基础上，本研究拟优化并联合利用Jag1/Notch2 MSC培养系统、细胞膜片技术和补肾中药干预方案。体外制备时通过激活Notch信号通路促进MSC增殖、抑制分化，移植时提高细胞的粘附性与生存率；移植后通过抑制Notch信号通路，促进MSC成骨分化。该研究可有效促进骨修复，并阐释疗效机制，为补肾中药联合治疗骨缺损提供实验依据。

Large bone defect caused by wound and tumor resection can be treated with allogenic bone together with autologous bone marrow mesenchymal stem cell (MSC) transplantation. But the low ability of proliferation, adhesion and differentiation of MSC onallograft remains problems to be resolved..In previous study, we have established the primary MSC in vitro culture system (Jag1/Notch2 MSC) through activating Notch signaling pathway. This culture system has been proved to stimulate MSC proliferation and maintein MSC properties. We also improved the transplanted MSC viability with MSC sheet technology. Acoording to our clinical practice, kidney torifying Chinese medicine and their effective components have been sceened and proved to promote MSC differentiation both in vitro and in vivo. .On the basis of previous study, in this study, the Jag1/Notch2 MSC cultrue system, the MSC sheet technology and the kidney torifying Chinese medicine will be optimized and combined together to promote MSC proliferation through activation of Notch signaling pathway when MSCs were prepared in votro, and to enhance the cell viability when transplanted, and also to promote MSC differentaiton through inhibition of Notch signaling pathway. The combination and healing of allogenic bone and host bone will be observed. This study will effectively 批romote bone healing, and clarfy the mechanisms, and provide the experimental evidence for the combined treatment with kidney torifying Chinese medicine on bone defects.

创伤和骨肿瘤切除等可造成大段骨缺损，应用失活同种异体骨并配合自体骨髓间充质干细胞（MSC）移植修复是目前主要治疗方法。但移植物上MSC细胞增殖低下，粘附效率低，MSC成骨分化能力低下，治疗效果不佳。我们首先采用温度敏感技术培养小鼠MSCs，形成的细胞膜层包裹移植骨进行移植，可以显著加速骨修复；进一步采用重组Jag1蛋白处理人源MSCs，激活细胞中Notch信号通路，显著延缓MSCs细胞周期和细胞衰老；形成的细胞膜层应用于骨移植可以促进骨痂形成，提高修复骨的生物力学性能。我们还发现补肾中药有效组分齐墩果酸可以剂量依赖性的抑制Notch信号通路活性及其靶基因Hes1mRNA的表达，促进MSCs的早期成骨分化，并增强BMP2诱导的成骨分化和骨形成作用。为Jag1干细胞体外增殖成膜技术和OA在骨修复的临床应用提供了基础研究依据。