VEGF过表达的骨髓间充质干细胞对大鼠牙槽骨缺损修复的影响

The effects of bone marrow mesenchymal stem cells with VEGF overexpression on alveolar bone repair in rats

目的:探讨VEGF过表达的BMMSCs对大鼠牙槽骨损伤修复的影响。方法:分离培养及鉴定BMMSCs;VEGF过表达载体转染BMMSCs;制备BMMSCs成骨膜片并鉴定;构建大鼠牙槽骨损伤模型并植入成骨膜片;术后2、4、6、8周Micro-CT扫描牙槽骨并计算骨体积分数。结果:分离后的BMMSCs形态多为菱形;VEGF过表达的BMMSCs荧光镜下可见明显绿色荧光,且VEGF表达提高(83.1±4.8)%(P<0.05)。Vonkossa染色发现,BMMSCs成骨膜片矿化结节染色阳性,胞外局部有黑色钙结节。植入BMMSCs膜片后,第4和8周模型大鼠牙槽骨骨体积分数增加(P<0.05);与BMMSCs膜片相比,过表达VEGF的BMMSCs膜片在第4、6和8周骨体积分数增加(P<0.05)。结论:VEGF过表达的BMMSCs能增强大鼠牙槽骨损伤的修复。

Objective: To investigate the effects of BMMSCs with VEGF overexpression on the repair of alveolar bone defect in rats. Methods: BMMSCs were isolated,cultured and identified. The VEGF overexpression vector was transfected into BMMSCs. The BMMSCs osteogenic membrane was prepared and identified. The rat model of alveolar bone defect was constructed and the osteogenic membrane was implanted into the alveolar bone defect of the rats. The alveolar bone was scanned by Micro-CT and the bone volume fraction was calculated 2,4,6 and 8 weeks after implantation. Results: The morphology of isolated BMMSCs was mostly rhomboid, BMMSCs overexpressing VEGF showed obvious green fluorescence under fluoroscopy,and the expression of VEGF increased by ( 83. 1 ± 4. 8)%( P < 0. 05). vonkossa staining showed that BMMSCs were positive for mineralized nodule in osteogenic membrane and black calcium nodules were extracellular. After implantation of BMMSCs,the volume of alveolar bone volume increased in the 4th and 8th week( P < 0. 05). BMMSCs with VEGF overexpression increased bone volume fraction at 4,6 and 8 weeks compared with BMMSCs( P < 0. 05). Conclusion: BMMSCs with VEGF overexpression can enhance the repair of rat alveolar bone defect.