转染BMP-2基因的骨髓MSC复合多孔磷酸钙骨水泥构建组织工程化骨对兔骨缺损的修复作用研究  

Bone marrow MSC transfected with BMP-2 gene and porous calcium phosphate cement to construct tissue-engineered bone for repairing bone defects

作　　者：陈波[1] 郭祥[1] 张寿[1] 钟海波 CHEN Bo;GUO Xiang;ZHANG Shou(Center of Orthopaedics,Haikou Hospital,Xiangya Medical College,Zhongnan University,Haikou Hainan 571000,China.)

机构地区：[1]中南大学湘雅医学院附属海口医院骨科中心

出　　处：《临床和实验医学杂志》2019年第17期1815-1818,共4页Journal of Clinical and Experimental Medicine

基　　金：海南省科技厅重点科技计划项目(编号:20170047)

摘　　要：目的研究骨髓间充质干细胞(MSC)经骨形态发生蛋白-2(BMP-2)基因转染后与多孔磷酸钙骨水泥(CPC)进行复合而建立组织工程化骨,分析其修复骨缺损的效果。方法取新西兰白兔32只,构建双侧股骨髁缺损的动物模型,双侧均植入骨髓MSC复合CPC组织工程化骨,其中右侧植入未转染BMP-2基因为对照组,左侧植入BMP-2基因为转染组。分别于植入后1、3个月,取两组双侧股骨髁,通过Van-Gieson染色法观察骨组织形态学变化,计算新骨形成面积在总缺损区中的占比;于植入后3个月,对两组新骨形成情况进行免疫荧光标记检测。结果植入后1个月,对照组仅于CPC外周出现少量新骨组织,转染组出现诸多新骨组织生成,以CPC外周孔隙为多见。植入后3个月,对照组植入区出现诸多新骨组织向CPC周围至中心区生长,但支架材料吸收效果并不明显;转染组人工骨内部出现诸多新骨组织生成,外周编织骨进展为成熟的小梁骨,外周区可见支架材料吸收较明显,部分被新骨组织所取代。相比植入后1个月,两组植入后3个月新骨面积在总缺损区的比率均明显提高(P <0. 01);相比对照组,转染组植入后1、3个月新骨面积在总缺损区的比率均明显提高(P <0. 01)。植入后3个月,转染组的组织工程化骨成骨速率为(5. 73±1. 24) m/d,较对照组的(3. 09±0. 98) m/d显著提高(P <0. 01)。结论通过基因转染的方法,将BMP-2基因转染至骨髓MSC,使之复合CPC材料,可明显提高新骨形成速率,从而可有效增强组织工程化骨对骨缺损的修复作用。Objective To study the effects of bone marrow mesenchymal stem cells( MSC) transfected with bone morphogenetic protein-2( BMP-2) gene and porous calcium phosphate cement( CPC) on the establishment of tissue-engineered bone for repairing bone defects.Methods 32 New Zealand white rabbits were selected to establish bilateral femoral condylar defect animal models. MSC and CPC tissue engineered bone were implanted on both sides. BMP-2 gene was implanted on the right side as the control group,and BMP-2 gene was implanted on the left side as the transfection group. At 1 and 3 months after implantation,bilateral femoral condyles of the two groups were harvested. Bone histomorphological changes were observed by Van-Gieson staining,and the proportion of new bone formation area in the total defect area was calculated. At 3 months after implantation,the new bone formation of the two groups was detected by immunofluorescence labeling. Results One month after implantation,only a small amount of new bone tissue appeared in the control group,and many new bone tissue appeared in the transfection group,especially in the peripheral pore of CPC. Three months after implantation,many new bone tissues grew from the implanted area to the central area of CPC in the control group,but the effect of scaffold material absorption was not obvious. In the transfected group,many new bone tissues were generated inside the artificial bone,and the peripheral woven bone developed into mature trabecular bone. In the peripheral area,scaffold material absorption was obvious,and some of them were replaced by new bone tissue. Compared with one month after implantation,the proportion of new bone area in total defect area increased significantly in both groups at 3 months after implantation( P < 0. 01). Compared with the control group,the proportion of new bone area in total defect area increased significantly in the transfection group at 1 and 3 months after implantation( P < 0. 01). Three months after implantation,the osteogenesis rate of tissue engineer

关 键 词：兔 骨缺损 骨髓间充质干细胞 骨形态发生蛋白-2 多孔磷酸钙骨水泥 基因转染 

分 类 号：R318.08[医药卫生—生物医学工程]