中药骨康含药血清培养脂肪干细胞移植促进骨质疏松性骨折愈合过程中cbfa1和OCN的表达变化

目的 探讨中药骨康含药血清培养脂肪干细胞移植促进骨折愈合过程中骨折端cbfa1和OCN的动态变化。方法 制备中药骨康含药血清并进行脂肪干细胞培养，移植脂肪干细胞到骨折部位，用免疫组化观察骨折部cbfa1和OCN的动态变化。结果 第3 代脂肪干细胞流式鉴定CD90阳性，CD34、CD45低表达。免疫组化观察骨折部cbfa1和OCN的动态变化：第一周干预组Cbfal和OCN组均可见血肿、肉芽组织及纤维性骨痂组织中出现阳性细胞和基质阳性信号，阳性较强，对照组可见阳性细胞和基质阳性信号，阳性较弱，空白组基本上无阳性表达；第二周Cbfal和OCN组均出现软骨岛和新生骨小梁，胶原纤维排列整齐，大量阳性细胞和基质阳性信号，强阳性表达, 对照组骨小梁和软骨岛均出现，但比干预组少，亦出现阳性细胞和基质阳性信号，信号比干预组弱，空白组少见软骨岛和骨小梁，胶原纤维排列混乱，阳性细胞和基质阳性信号亦可见，但比前两组要弱；第四周干预组Cbfal组阳性表达减弱，OCN组继续强阳性表达，对照组两指标均阳性表达，空白组阳性细胞和基质阳性信号存在，阳性程度比以往稍强。结论 通过免疫组化观察骨折部cbfa1和OCN的动态变化，表明脂肪干细胞移植可促进骨质疏松性骨折愈合，而中药骨康含药血清培养脂肪干细胞移植效果更理想。

Expression of Cbfa1 and OCN in osteoporotic fracture healing process promoted by transplantation of Gukang-containing serum cultivated adipose-derived stem cells

Objective To investigate the dynamic changes of the expression of Cbfa1 and OCN in osteoporotic fracture healing progress promoted by the transplantation of adipose-derived stem cells (ADSCs), which were cultivated in Gukang-containing serum. Methods Gukang-containing serum was prepared and used for the culturing of ADSCs. And then the ADSCs were transplanted to the fracture site. The dynamic changes of Cbfa1 and OCN were observed using immunohistochemistry. Results The third passage of ADSCs were identified as CD90 positive, CD34 and CD45 low expression. The dynamic changes of Cbfa1 and OCN could be observed using immunohistochemistry. At the first week, hematoma, granulation tissue, Cbfa1 and OCN-positive cells, and matrix positive signal in fibrous callus could be observed in intervention group, while positive cells and matrix positive signal in control group could be observed, but not as strong as in intervention group. And no positive expression was observed in blank group. At the second week, cartilage islands and new trabecular bone were observed in intervention group, and collagen fibers arranged in neat rows. Large number of Cbfa1 and OCN-positive cells and matrix positive signal could be observed, showing a strong positive expression. Trabecular bone and cartilage islands were also observed in control group, but in less extent than that in intervention group. Less strong positive expression of positive cells and matrix positive signal was observed in control group. While rare cartilage islands and trabecular were observed in blank group, chaotic arrangement of collagen fibers, least positive cells, and weakest matrix positive signal in 3 groups were also observed. At the fourth week, the expression of Cbfa1 decreased, while the strong expression of OCN continued in intervention group. In control group, the two indicators showed positive expression. In blank group, positive cells and matrix positive signal showed much stronger than those previously. Conclusion Through the observation of dynamic changes of the Cbfa1 and OCN expression using immunohistochemistry in the fracture sites, we find that the transplantation of ADSCs can promote osteoporotic fracture healing process. The transplantation of ADSCs cultivated in Gukang-containing serum has better effect.