IGF-1过表达质粒转染BMSCs联合纳米纤维多孔支架修复兔软骨缺损效果

EFFECT OF BONE MARROW MESENCHYMAL STEM CELL TRANSFECTION WITH IGF-1 OVEREXPRESSION PLASMIDS COMBINED WITH NANOFIBER POROUS SCAFFOLD IN TREATMENT OF RABBITS WITH CARTILAGE DEFECT

目的探讨慢病毒介导胰岛素样生长因子1(IGF-1)过表达质粒转染骨髓间充质干细胞(BMSCs)联合Gelatin/PLA纳米纤维多孔支架修复兔软骨缺损的效果。方法构建慢病毒IGF-1过表达载体,分离、培养并鉴定BMSCs,构建纳米纤维多孔支架+转染IGF-1过表达载体BMSCs复合物。构建兔膝关节软骨缺损模型,采用苏木精-伊红染色和免疫组织化学方法观察纳米纤维多孔支架+转染IGF-1过表达载体BMSCs复合物对兔软骨缺损的修复效果。结果纳米纤维多孔支架+转染IGF-1过表达载体BMSCs复合物支架表面黏附的细胞数较纳米纤维多孔支架+未转染BMSCs复合物明显增多。大体观察显示,实验组(造模后植入纳米纤维多孔支架+转染IGF-1过表达载体BMSCs复合物)兔软骨缺损的修复效果要明显优于对照组(造模后植入纳米纤维多孔支架+未转染BMSCs复合物)。苏木精-伊红染色显示,对照组兔软骨缺损多为纤维性修复,而实验组多为细胞性修复。免疫组织化学染色显示,实验组修复软骨组织中Aggrecan和Ⅱ型胶原的含量要明显高于对照组(F=15.342、18.928,P<0.05)。结论纳米纤维多孔支架+转染IGF-1过表达载体BMSCs复合物可以促进BMSCs向软骨细胞的分化,并且分泌更多的含Aggrecan和Ⅱ型胶原成分的细胞外基质,从而发挥促进软骨缺损修复的作用。

Objective To investigate the effect of bone marrow mesenchymal stem cell (BMSC) transfection with lentivirus-mediated insulin-like growth factor-1 (IGF-1) overexpression plasmids combined with Gelatin/PLA nanofiber porous scaffold in the treatment of rabbits with cartilage defect. Methods The lentivirus IGF-1 overexpression vector was constructed, BMSCs were isolated, cultured, and identified, and the complex of nanofiber porous scaffold+BMSCs transfected with IGF-1 overexpression vector was constructed. A rabbit model of knee cartilage defect was established, and hematoxylin and eosin (HE) staining and immunohistochemical staining were used to evaluate the effect of the complex of nanofiber porous scaffold+BMSCs transfected with IGF-1 overexpression vector in the repair of cartilage defect in rabbits. Results The complex of nanofiber porous scaffold+BMSCs transfected with IGF-1 overexpression vector had a significantly higher number of cells attached on the scaffold surface than the complex of nanofiber porous scaffold+BMSCs not transfected with IGF-1 overexpression vector. Gross observation showed that the experimental group (with the transplantation of the complex of nanofiber porous scaffold+BMSCs transfected with IGF-1 overexpression vector after modeling) had significantly better repair of cartilage defect than the control group (with the transplantation of the complex of nanofiber porous scaffold+BMSCs not transfected with IGF-1 overexpression vector after modeling). HE staining showed mainly fibrous repair in the control group and cellular repair in the experimental group. Immunohistochemical staining showed that the experimental group had significantly higher content of Aggrecan and type Ⅱ collagen in the repaired cartilage than the control group ( F =15.342 and 18.928, P <0.05). Conclusion The complex of nanofiber porous scaffold+BMSCs transfec- ted with IGF-1 overexpression vector can promote the differentiation of BMSCs into chondrocytes, increase the secretion of extracellular matrix containing more Aggrecan and type Ⅱ collagen, and thus promote the repair of cartilage defect.