壳聚糖负载原代培养自体骨髓间充质干细胞修复全层关节软骨缺损实验研究  

Primary culture of autologous bone marrow mesenchymal cells loaded with oligosaccharide repair of full-thickness articular cartilage defects with mesenchymal stem cells experimental research

作　　者：李军 张鹏 卢雪玲 刘文忠 范磊 LI Jun;ZHANG Peng;LU Xueling;LIU Wenzhong;FAN Lei(Department of Joint Surgery,Gaomi People's Hospital,Gaomi 261500,China)

机构地区：[1]高密市人民医院骨科

出　　处：《潍坊医学院学报》2019年第5期345-347,397,共4页Acta Academiae Medicinae Weifang

摘　　要：目的利用壳聚糖负载原代培养转化生长因子-β1(TGF-β1)基因修饰自体骨髓间充质干细胞(BM-MSCs)构建组织工程修复体系,观察该体系对实验体软骨缺损修复的成果。方法原代培养并通过表面抗原鉴定BM-MSCs,将pcDNA3-TGF-β1基因导入BM-MSCs制备种子细胞,30只实验兔随机分为1,2,3共3组,分别用壳聚糖、壳聚糖复合BM-MSCs和壳聚糖复合TGF-β1基因修饰BM-MSCs修复全层关节软骨缺损,于术后4周、24周用甲苯胺蓝染色法观察缺损区软骨细胞和软骨基质的形态,蛋白质印迹法检测TGF-β1和Ⅱ型胶原的表达。结果 TGF-β1基因修饰后,BM-MSCs修复软骨缺损时间缩短、新生透明软骨均匀一致,与周围正常透明软骨相容良好,效果优于其它两组;24周后修复区域软骨组织TGF-β1和Ⅱ型胶原表达明显高于其它两组。结论壳聚糖负载转化生长因子-β1基因修饰BM-MSCs构建组织工程修复软骨可用于全层关节软骨缺损治疗,其远期疗效较稳定。Objective Autologous bone marrow mesenchymal stem cells(BM-MSCs) were modified by chitosan-loaded primary cultured transforming growth factor-β1(TGF-β1) gene to construct a tissue engineering repair system,and observe the results of the system for repairing cartilage defects.Methods Generational culture BM-MSCs was identified by surface antigen,and the pcDNA3-TGF-β1 gene was introduced into BM-MSCs to prepare seed cells.Thirty experiment rabbits were divided into group 1,2,3 randomly.The rabbits suffered full-thickness articular cartilage defects,were repaired by chitosan,chitosan composite BM-MSCs and chitosan composite TGF-β1 gene modified BM-MSCs respectively.The morphology of chondrocytes and cartilage matrix in the defect area was observed by toluidine blue staining 4 and 24 weeks after operation,and the expression of TGF-β1 and type Ⅱ collagen was detected by Western blotting.Results It took lesser time for hyaline cartilage to regenerate and repair the whole defects in group 3 than group 1 and group 2.The regenerated hyaline cartilage in group 3 appeared uniformly and was compatible well with the adjacent hyaline cartilage around.The expression of TGF-β1 and type Ⅱ collagen in the repaired regional cartilage was significantly higher than the other two groups after 24 weeks.Conclusion The construction of cartilage constructed by chitosan-loaded transforming growth factor-β1 gene-modified BM-MSCs can be used for the treatment of full-thickness articular cartilage defects,and its long-term efficacy is relatively stable.

关 键 词：组织工程 转化生长因子-Β1 壳聚糖 基因修饰 骨髓间充质干细胞 

分 类 号：R684[医药卫生—骨科学]