摘要
背景:骨髓间充质干细胞与纤维蛋白胶联合应用可提高前交叉韧带重建后的早期腱-骨界面断裂强度与刚度,促进移植物与骨隧道早期愈合。目的:探讨骨形态发生蛋白2基因转染骨髓间充质干细胞复合纤维蛋白胶对前交叉韧带重建术后腱-骨愈合的影响。方法:将腺病毒载体Ad-GFP-骨形态发生蛋白2转染至骨髓间充质干细胞,48 h后检测GFP基因表达及骨形态发生蛋白2表达。将54只新西兰大白兔(购买于重庆市医学实验动物中心)随机分为空白组、实验组、对照组,每组18只,均建立前交韧带重建模型,实验组将基因转染48 h的骨髓间充质干细胞与纤维蛋白胶混合,注射至腱-骨界面;对照组将未转染的骨髓间充质干细胞与纤维蛋白胶混合,注射至腱-骨界面;空白组未注射任何材料。术后第4,8,12周分别获取股骨-移植肌腱-胫骨标本,进行组织学检测与生物力学检测。结果与结论:(1)转染48 h后,荧光显微镜下可见细胞表达绿色荧光,转染效率为(86.1±6.3)%;SP免疫组织化学检测细胞胞浆内骨形态发生蛋白表达阳性;Western-blot检测可见骨形态发生蛋白2阳性表达条带;(2)苏木精-伊红染色显示,实验组、对照组腱-骨愈合效果明显优于空白组,实验组腱-骨愈合效果优于对照组;(3)生物力学检测显示,实验组、对照组的最大载荷、刚度均强于空白组(P <0.05),实验组的最大载荷、刚度均强于对照组(P <0.05);(4)结果表明,骨形态发生蛋白2进一步增强了骨髓间充质干细胞复合纤维蛋白胶促进前交叉韧带重建后的腱-骨愈合效果。
BACKGROUND:The combination of bone marrow mesenchymal stem cells and fibrin glue can improve the early rupture strength and stiffness of the tendon-bone interface after anterior cruciate ligament reconstruction,and promote the early healing of graft and bone tunnel.OBJECTIVE:To explore the effect of bone morphogenetic protein 2 transfected bone marrow mesenchymal stem cells combined with fibrin glue on the tendon-bone healing after the reconstruction of the anterior cruciate ligament.METHODS:Adenovirus vector Ad-GFP-bone morphogenetic protein 2 was transfected into bone marrow mesenchymal stem cells,and the expressions of GFP gene and bone morphogenetic protein 2 were detected after 48 hours.Fifty-four New Zealand white rabbits purchased from the Medical Animal Center of Chongqing were randomly divided into blank group,experimental group and control group,with 18 rats in each group.The anterior cruciate ligament reconstruction model was established in each rat.In the experimental group,after 48 hours of transfection,the transfected bone marrow mesenchymal stem cells were mixed with fibrin glue and injected into the tendon-bone interface.In the control group,non-transfected bone marrow mesenchymal stem cells were mixed with fibrin glue and injected into the tendon-bone interface.In the blank group,no treatment was given.After 4,8,12 weeks,the femur-tendon-tibia specimens were taken for histological and biomechanical examinations.RESULTS AND CONCLUSION:(1)After transfection of 48 hours,the cells expressed green fluorescence under the fluorescence microscope,and the transfection efficiency was(86.1±6.3)%.The expression of cytoplasmic bone morphogenetic protein was positive as shown by SP immunohistochemical test,and the positive band of bone morphogenetic protein 2 was detected by western blot.(2)Findings from the hematoxylin-eosin staining showed best better tendon-bone healing in the experimental and control group than the blank group,and the tendon-bone healing was further better in the experimental group than i
作者
朱乐全
卢剑华
冉俊岭
杨卫保
王辉
Zhu Lequan;Lu Jianhua;Ran Junling;Yang Weibao;Wang Hui(Department of Orthopedics,Qianjiang Central Hospital of Chongqing(Qianjiang Hospital of Jishou University),Chongqing 409000,China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第1期30-34,共5页
Chinese Journal of Tissue Engineering Research
