关节镜下髓芯减压基因转染骨髓间质干细胞修复兔股骨头缺血性坏死

Treament of osteonecrosis of femoral head by core decompression with autograft of hBMP-2 gene modified bone marrow stromal cells using arthroscope

目的研究髓芯减压关节镜监视下基因转染骨髓间质干细胞（BMSCs）治疗兔股骨头缺血性坏死的效果。方法在无菌条件、戊巴比妥钠麻醉下，获取成年新西兰兔骨髓，体外培养。倒置显微镜下观察细胞形态和生长情况，以腺相关病毒介导的hBMP-2转染兔BMSCs，感染倍率（MOI）为100，继续培养。应用Western blot检测转染后的BMSCs的hBMP-2基因表达。建立兔右侧股骨头缺血性坏死模型，并分为A组单纯髓芯减压组，B组髓芯减压并植入BMSCs组，C组髓芯减压并植入hBMP-2转染的BMSCs组。关节镜监视并对坏死骨清除后植入BMSCs或hBMP-2转染的BMSCs，术后4周和8周对股骨头行组织学检查。结果hBMP-2基因转染的兔BMSCs可有效表达hBMP-2。组织学观察显示B组周边有一定量骨形成，骨小梁表面新骨形成；C组周边坏死骨小梁表面新骨带明显增宽，新生骨小梁局部毛细血管丰富，表面有较多成骨细胞。术后4、8周C组血管计数及修复区新骨面积比分别与A、B组比较，差异有统计学意义。结论关节镜监视下髓芯减压并植入hBMP-2转染的BMSCs可促进股骨头缺血．性坏死的修复。

Objective To observe the therapeutic effect on the treatment of earliy osteonecrosis of femoral head by core decompression with autograft of bone marrow stromal cells （BMSCs） using arthroscope. Methods By aseptic conditions, bone marrow was obtained from an adult male New Zealand white rabbit under sodium pentobarbital anesthesia. Rabbit BMSCs were isolated by density the cluture media. The cellular histological characteristics and growth aspects were observed under phase contrast microscope, hBMP-2 plasmids were extracted and were transfected into rabbit marrow stromal stem cells, hBMP-2 mediated by adeno-assoeiated virus（AAV） transfected BMSCs. MOI was 100. The transcription and expression of hBMP-2 protein expression were investigated by Western blot. Established the animal models of femoral head necrosis in 36 rabbits by delivery. Establish the model of earliy osteonecrosis of rabbit femoral head on right leg. Three groups were divided, A group was single core decompression group, B group was core decompression and autografi of mesenchymal stem cells. C group was decompression with rAAV-hBMP-2/BMSCs group. Necrosis bone was cleared out, then BMSCs was implanted on arthroscope. The femoral head was inspected by histology postoperative 4-8 weeks. Results The expression of hBMP-2 gene could be found distinctly in the transfected rabbit BMSCs and hBMP-2 protein in the supematants of transfected cell cultures possesses highly biological activity, which could promote rabbit osteoblast increased and augment cytoactive effective. New bone was evident in the implantation field of C group. The area of new bone repair and the number of vascular in group C were significant larger than those in group A and B postoperative 4-8 weeks. Conclusion The treatment of core decompression with autograft of BMSCs is safe and effective, and it deserves to application in early necrosis of femoral head.