腺病毒介导hBMP-2转染BMSCs复合DBM修复兔缺血性股骨头坏死的实验研究

Rehabilitation effect of BMSCs that was transfected with h BMP-2 through adenovirus combination with DBM on rabbit osteonecrosis in femoral head

目的评价人骨形态发生蛋白(h BMP)-2/骨髓间充质干细胞(BMSCs)/脱钙松质骨(DBM)对兔股骨头坏死的修复作用,探索临床治疗股骨头坏死的新途径。方法髓芯减压联合液氮冰冻法制备兔股骨头坏死模型。将造模成功兔随机分为A、B、C、D 4组(n=12),A组不植入材料,为对照组,B、C、D分别植入DBM、DBM/BMSCs、h BMP-2/BMSCs/DBM。术后4、8及12周各组分别处死4只,运用X线技术、大体标本观察、HE染色技术评判股骨头坏死修复情况。结果 X线示A组股骨头塌陷,无明显成骨;B、C、D组股骨头缺损区有骨再生现象,但D组再生情况明显优于B、C组。Lane-Sandhu X线评分A组<B、C组<D组(P<0.05),B和C组差异无统计学意义。大体观示A组股骨头塌陷,钻孔存在;B、C组股骨头未塌陷,钻孔存在;D组股骨头未塌陷,钻孔消失。HE染色示A组骨小梁坏死、碎裂,大量空骨陷窝;B、C组可见成骨细胞及新生幼稚骨小梁;D组大量骨细胞,新生骨小梁与正常骨小梁无异。空骨陷窝率A组>B、C组>D组(P<0.05),B和C组差异无统计学意义。结论 h BMP-2/BMSCs/DBM植入体内后能够诱导BMSCs向成骨方向分化,对兔股骨头坏死具有较好的修复效果。

Objective To evaluate the effect of human bone morphogenetic protein 2 （hBMP-2）/Bone Mesenchymal Stem Cells （BMSCs）/demineralized bone matrix（DBM） on repairing rabbits’femoral head after necrosis and to explore the new treatments for femoral head necrosis. Methods Femoral head necrosis models was established by clinical core decom-pression combined with liquid nitrogen frozen. Then, animals were randomly devided into 4 groups （n=12 per group）：Group A were not implanted anything as control group, Group B were implanted with DBM. Group C were implanted with hBMP-2/DBM. Group D were implanted with hBMP-2/BMSCs/DBM. Four rabbits from each group were sacrificed at 4,8 and 12 weeks after surgery to evaluate the the repairing effect of Osteonecrosis of the femoral head （ONFH） through X-ray examina-tion, observation of the specimen and HE staining. Results X-ray revealed defect of femoral head in Group A without clear bone formation. There is a little fibrous hyperplasia and no obvious osteogenic response. By contrast, the femoral head defect areas became fuzzy in group B, group C and group D with new bone trabeculars. And the regenerate phenomenons of group D were significantly better than that of group B and group C of the same time point. As to the Lane-Sandhu X Ray scores, it is lower in group A than that in group B;It is lower in group C than that in group D（P〈0.05）. There is no statistical difference between Group B and Group C. General observation of the specimen revealed that the femoral head of group A collapsed with drilling holes. The femoral heads of group B and group C showed no collapse but the drilling holes existed. Femoral head in group D was not collapsed and the drilling holes disappeared. HE staining showed that bone trabeculars became ne-crotic and fragmented in Group A with a lot of air trapped cells. There were newborn immature bone trabeculars and osteo- blasts in group B and group C. Group D were of large number of bone cells, fat cells, and newborn mature bone trabeculars. The ratio of empty lacuna is higher in Group A than that in Group B;it is higher in Group C than that in Group D（P〈0.05）. Conclusion hBMP-2/BMSCs/DBM can induce BMSCs differentiation into osteoblasts after being implanted. It has good re-pairing effect on ONFH with good application prospect.