骨形态发生蛋白2／血管内皮生长因子165转染骨髓间充质干细胞复合多孔纳米羟基磷灰石／聚酰胺66修复兔桡骨缺损

Repair of rabbit radius defect by using recombinant of hBMP2/hVEGF165 modified BMSCs with porous nano-hydroxyapatite/polyamide 66 composite

目的评估骨形态发生蛋白2／血管内皮生长因子165（hBMP2／hvEGF165）双基因腺病毒转染骨髓间充质干细胞（BMSCs）复合多孔纳米羟基磷灰石／聚酰胺66（n—HA／PA66）对骨缺损的修复作用。方法选取60只成年雄性新西兰大白兔，制作15mm双侧桡骨中段骨缺损模型。按完全随机法分为3组，各组20只，根据双侧桡骨制作骨缺损模型，各大组分2小组，共6组。A1组：左桡骨缺损处不植入任何材料为空白对照组；A2组：右桡骨缺损处单纯植入n—HA／PA66人工骨材料；B1组：左桡骨缺损处植入hVEGF165／BMscs／n—HA／PA66人工骨材料；B2组：右桡骨缺损处植入BMP2／BMSCs／n—HA／PA66人工骨材料；C1组：左桡骨缺损处植入BMSCs／n—HA／PA66人工骨材料；C2组：右桡骨缺损处植入hBMP2／hVEGF165／BMscs／n—HA／PA66人工骨材料。分别于2，4，8，12周行x线片、HE和Masson染色观察桡骨缺损修复情况。结果c2组术后8周x线片、HE和Masson染色显示移植部位有大量软骨形成，且在边缘有大量骨组织生成，术后12周可见组织材料被外层骨组织包裹，骨缺损修复效果优于其他组（P〈0．05）；且术后各时相点C2组血管数目均优于其他组（P〈0．05），B1组各时相点血管数均多于B2组（P〈0．05），且两组明显多于A2、c1组（P〈0．05）；A2与c1组间差异无统计学意义（P〉0．05）。结论hBMP2／hVEGF165双基因转染BMSCs复合多孔n—HA／PA66具有明显提高修复兔桡骨干缺损的作用。

Objective To assess the effect of bone defect repair using the recombinant of adeno- virus-mediated hBMP2 and hVEGF165 genes transfer of BMSCs with porous nano-hydroxyapatite/polyam- ide 66 （n-HA/PA66）. Methods Sixty male adult New Zealand rabbits were assigned to groups A, B and C according the completely random design, with 20 rabbits per group. Bone defect of 15 mm in length was made in the middle segment of bilateral radii in rabbits. In Group A, the defects were filled with nothing on the left side in blank controls （ Group A1 ） and with n-HA/PA66 material alone on the right side （Group A2）. In Group B, the defects were filled with hVEGF165/BMSCs/n-HA/PA66 on the left side （Group B1 ） and hBMP2/BMSCs/n-HA/PA66 on the right side （Group B2）. In Group C, the de- fects were filled with BMSCs/n-HA/PA66 on the left side （ Group C1 ） and hBMP2/hVEGF165/BMSCs/ n-HA/PA66 on the right side （Group C2）. Radiological analysis, HE staining, and Masson coloration were performed 2, 4, 8 and 12 weeks after operation. Results Radiographs, HE staining and Masson staining taken 8 weeks after cell transplantation showed large amount of new cartilage grown into the defect area and massive bony tissue formation around the margin in Group C2. At postoperative 12 weeks, Group C2 showed transplants were surrounded by outer bone tissues with superior bone repair effect to other groups （ P 〈 0. 05 ）. Number of vessels in Group C2 increased compared with that in other groups （ P 〈 0. 05 ）. Number of vessels was greater in Group B1 than in Group B2 （ P 〈 0.05 ） , and both were greater than those in Groups A2 and C1 （ P 〈 0. 05 ）. Moreover there was no significant difference between Groups A2 and C1 （ P 〉 0.05 ）. Conclusion hBMP2/hVEGF165 genes transferred BMSCs seeded on porous n-HA/PA66 can contribute to osteogenesis during the repair of rabbit radius defect.