神经肽基因转染干细胞联合自体骨移植治疗兔股骨头坏死  

Calcitonin gene -related peptide -transfected bone marrow mesenchymal stem cells combined with autologous bone grafting to treat osteonecrosis of the femoral head in rabbit model

作　　者：王义生[1] 段家晨 李劲峰[1] 王亚寒[1] 王秀利[1] 刘鸣[1] 李月白 

机构地区：[1]郑州大学第一附属医院骨科河南省高等学校临床医学重点学科开放实验室,450052 [2]郑州大学基础医学院生物化学与分子生物学系,450001

出　　处：《中华实验外科杂志》2017年第10期1731-1733,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81171776、81572216）

摘　　要：目的探讨降钙素基因相关肽转染骨髓间充质干细胞（BMSCs）联合自体骨移植治疗模型兔股骨头坏死（ONFH）的组织病理学改变。方法采用骨内液氮冷冻法成功制作出家兔ONFH动物模型。将48只ONFH模型动物随机分为3组：A组：人降钙素基因相关肽α（hCGRPα）基因转染BMSCs组：将hCGRPα基因转染BMSCs联合自体骨移植于股骨头内；B组：空质粒组：植入干细胞为空质粒BMSCs联合自体骨移植；C组：无关序列组：植入干细胞为无关基因BMSCs联合自体骨移植。于实验第3、6个月末分批处死兔，观察兔股骨头的组织病理学变化。结果实验3个月时，A、B、C组正常侧股骨头中，骨小梁面积分数分别为（40.89±1.12）%、（36.12±1.17）%、（36.33±1.53）%、（42.11±1.91）%，空骨陷窝计数分别为（11.34±1.58）%、（16.33±1.45）%、（17.21±1.23）%、（10.47±1.41）%。A组中股骨头内有大量骨细胞增殖生长，骨小梁较成熟，骨小梁面积分数高，空骨陷窝少，接近于正常侧，明显优于B、C组。而B、C组中骨小梁细小、不规则，骨小梁面积分数低，空骨陷窝多，骨修复不完整。6个月时，A组中骨小梁成熟，致密饱满，排列规则整齐，骨小梁面积分数与空骨陷窝计数均仍优于B、C组。结论hCGRPα转基因BMSCs联合自体骨移植入动物模型股骨头中，能够显著促进细胞增殖，加速成骨，增强骨组织修复，获得治疗ONFH的良好效果。ObjectiveTo investigate the histopathological changes of calcitonin peptide-transfected bone marrow mesenchymal stem cells （BMSCs） combined with bone transplantation in the treatment of osteonecrosis of the femoral head （ONFH） in rabbit model.MethodsThe osteonecrosis models on one side femoral head of rabbits were established by liquid nitrogen. Forty-eight rabbit ONFH models were randomly divided into 3 groups. In human calcitonin gene-related peptide α （hCGRPα）-transfected BMSCs group （A）, the BMSCs transfected with hCGRPα in combination with autologous bone graft were implanted into the femoral head. In empty plasmid group （B）, the BMSCs transfected with empty plasmid （only pLNCχ2 plasmid） and autologous bone graft were implanted. In irrelated sequence group （C）, BMSCs transfected with irrelated gene [only pLNCχ2-green fluorescent protein （GFP）] and autologous bone graft were implanted. Animals were sacrificed at 3rd and 6th month respectively to observe the histopathological changes in the femoral heads of rabbits.ResultsAt 3rd month, the bone trabecular area fraction respectively was （40.89±1.12）%, （36.12±1.17）%, （36.33±1.53）% and （42.11±1.91）% in groups A, B and C and normal femoral head. The percentage of empty osteocyte lacunae respectively was （11.34±1.58）%, （16.33±1.45）%, （17.21±1.23）% and （10.47±1.41）% in groups A, B and C and normal femoral head. In group A, there were proliferation and growth of a large number of osteocytes, more mature bone trabeculae and higher area fraction, less empty osteocyte lacunae, and stronger biomechanical strength in the femoral head, which were nearly close to those in the normal control group and obviously superior to those in groups B and C. In groups B and C, there were thin and irregular bone trabeculae, more empty osteocyte lacunae, lower area fraction and imperfect bone repair. At 6th month, the bone trabeculae were mature, densification and satiation, and neatly arranged rules were seen i

关 键 词：降钙素基因相关肽 骨髓间充质干细胞 股骨头坏死 模型 动物 

分 类 号：R681.8[医药卫生—骨科学]