携载FGF2基因的改性壳聚糖微球促进兔桡骨骨缺损修复的研究  

Modified Chitosan Microspheres Loaded FGF2 Gene Promote the Repair of Radial Bone Defect in Rabbits

作　　者：陈胤贤 蒋健一 赵宇[1] 孙军[1] CHEN Yinxian;JIANG Jianyi;ZHAO Yu;SUN Jun(Department of Orthopedics,Anhui Provincial Children's Hospital Affiliated to Anhui Medical University,Hefei 200011,China.)

机构地区：[1]安徽医科大学附属省儿童医院骨科,安徽省合肥市230041

出　　处：《组织工程与重建外科杂志》2020年第2期81-86,共6页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金(81171829);安徽省医疗卫生重点专科建设项目。

摘　　要：目的研究携载FGF2基因的改性壳聚糖微球在体内促进兔桡骨临界骨缺损愈合过程中的作用。方法以羟丁基壳聚糖(HBC)、巯基烷基化壳聚糖(TACS)与pFGF2质粒制备HBC@TACS-pFGF2微球,体外与BMSCs共培养。Western-blot检测FGF2蛋白表达水平,CCK8检测细胞增殖情况,PCR检测ALP mRNA和CD31 mRNA的表达情况。动物实验选用15只新西兰兔,手术建立双侧桡骨中段18 mm骨缺损模型,左侧植入明胶海绵和HBC@TACS-pFGF2微球(实验组,n=15),右侧不作植入(对照组,n=15)。移植术后第4周、8周、12周收集标本分别进行大体观察、X线检查和Masson染色。结果HBC@TACS-pFGF2微球体外可促进BMSCs增殖,并可显著提高ALP和CD31的表达。动物实验显示,术后4周、8周、12周时,实验组兔桡骨缺损处的断端桥接情况、骨密度以及新生骨矿化程度均优于对照组。结论含pFGF2质粒的HBC@TACS-pFGF2微球可在体内促进兔桡骨临界骨缺损修复。Objective To explore the effect of modified chitosan microspheres loaded FGF2 gene in promoting the healing of critical radial bone defect in rabbits.Methods HBC@TACS-pFGF2 microspheres were prepared by HBC,TACS and pFGF2 plasmid,and co-cultured with bone marrow mesenchymal stem cells in vitro.The expression of FGF2 protein was detected by western-blot.The cell viability of BMSCs was assessed by CCK8.The gene expressions of ALP and CD31 were analyzed by Real time RT-PCR.In the animal experiment,15 New Zealand rabbits were selected,and an 18 mm bone defect model of the bilateral middle radius was established.Gelfoam and HBC@TACS-pFGF2 microspheres were implanted on the left side(experimental group,n=15),while no implant was performed on the right side(control group,n=15).At the 4th,8th and 12th weeks after the transplant,specimens were collected for general observation,X-ray examination and Masson staining.Results The HBC@TACS-pFGF2 particles can promote the proliferation of BMSCs in vitro and significantly increase the expression of ALP and CD31 genes.According to the results of gross observation,X-ray and Masson staining,at 4 weeks,8 weeks and 12 weeks after operation,the bridging,bone density and mineralization of new bone in the radius defect of the experimental group were all better than those in the control group.Conclusion The HBC@TACS-pFGF2 microspheres which containing pFGF2 plasmids can promote the repair of critical radial bone defect in rabbits in vivo.

关 键 词：壳聚糖 基因治疗 骨缺损 碱性成纤维细胞生长因子 

分 类 号：R683.41[医药卫生—骨科学]