9/EPO双基因共转染ADSCs复合nHAC/PLA支架修复兔下颌

作者：王韵壹、李英、熊慧卿

摘要： 目的：探讨腺病毒介导BMP-9/EPO双基因共转染脂肪干细胞（ADSCs）复合nHAC/PLA支架材料促进体内成骨的可行性。方法：分离培养兔ADSCs，利用重组腺病毒构建携带BMP-9、EPO、BMP-9/EPO基因的表达载体，并转染至兔ADSCs。14 d后，在荧光显微镜下观察各组细胞荧光表达情况，计算病毒转染效率，采用Western免疫印迹检测各组细胞 BMP-9及EPO蛋白的表达。50只大白兔随机分为5组，A组为空白组，B组为单纯材料组，C组为BMP-9/nHAC/PLA组，D组为EPO/nHAC/PLA组，E组为BMP-9/EPO/nHAC/PLA组。制备兔双侧下颌骨缺损模型，按照实验分组，将转染组细胞分别与nHAC/PLA支架复合培养后植入兔下颌骨缺损处，于4、8、12周在缺损处取材进行影像学、组织学检测及扫描电镜观察，比较各组动物下颌骨缺损修复情况。采用 SPSS17.0软件包对结果进行统计学分析。结果：荧光显微镜下观察，BMP-9、EPO及BMP-9/EPO重组腺病毒均能稳定转染 ADSCs，转染效率为 80%～93%。转染后，BMP-9、EPO及BMP-9/EPO基因和蛋白均能稳定表达。转染后的ADSCs与nHAC/PLA支架复合，其骨缺损区修复情况优于对照组，且BMP-9/EPO/nHAC/PLA组成骨情况优于其他各组。扫描电镜下见转染组骨缺损与材料结合修复情况优于对照组，BMP-9/EPO/nHAC/PLA组修复最明显。结论：BMP-9、EPO均可转染ADSCs并稳定表达。与nHAC/PLA支架材料复合后，可显著促进下颌骨缺损的修复，为骨组织修复工程提供了新的选择。

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张广德，靳霞，李荣亮，杨世茂，郭延伟，房殿吉. BMP-9/EPO双基因共转染ADSCs复合nHAC/PLA支架修复兔下颌骨缺损的实验研究[J]. 中国口腔颌面外科杂志, 2017, 15(3): 202-208.

ZHANG Guang-de, JIN Xia, LI Rong-liang, YANG Shi-mao, GUO Yan-wei, FANG Dian-ji.. Repair of critical sized mandibular bone defect with nHAC / PLA scaffold combing with rabbit adipose tissue derived mesenchymal stem cells transfected with BMP-9 / EPO gene[J]. China Journal of Oral and Maxillofacial Surgery, 2017, 15(3): 202-208.

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