质粒修饰BMSC联合纳米支架促进软骨缺损修复的相关研究

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摘要目的探究慢病毒介导聚蛋白多糖(Aggrecan)过表达质粒转染骨髓间充质干细胞联合Gelatin/PLGA纳米纤维多孔支架向软骨细胞转化及软骨缺损修复的作用效果。方法构建慢病毒聚蛋白多糖过表达载体，培养骨髓间充质干细胞，构建兔模型软骨缺损模型，实验组造模后植入Gelatin/PLGA三维支架+转染聚蛋白多糖过表达载体骨髓间充质干细胞复合物；阴性对照组造模后植入Gelatin/PLGA三维支架+未转染骨髓间充质干细胞复合物；模型组造模后加入生理盐水处理。体外实验利用HE染色和免疫组织化学染色检测骨髓间充质干细胞联合Gelatin/PLGA纳米纤维多孔支架的联合培养体系向软骨细胞转化中聚蛋白多糖和Ⅱ型胶原蛋白的表达。构建兔膝关节软骨缺损模型，HE染色和免疫组织化学分析复合支架材料对软骨缺损的修复效果。结果 a)实验组(Gelatin/PLGA纳米纤维多孔支架+转染Aggrecan过表达载体骨髓间充质干细胞复合物)中支架表面黏附的细胞与对照组(Gelatin/PLGA纳米纤维多孔支架+未转染骨髓间充质干细胞)相比，细胞数明显增多；b)动物模型大体观察结果显示，实验组的修复效果要明显优于阴性对照组；c)动物模型HE染色结果显示，阴性对照组多为纤维性修复，而实验组多为细胞性修复；d)免疫组织化学染色结果显示，实验组修复软骨组织中Aggrecan和Ⅱ型胶原含量要明显优于阴性对照组。结论 Gelatin/PLGA三维支架加转染Aggrecan过表达载体骨髓间充质干细胞复合物可以促进BMSC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，从而发挥其促进软骨缺损修复的作用。

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	郑润泉 1
	康继 2
	张贵春 1*

关键词 ： Gelatin/PLGA三维支架, 骨髓间充质干细胞, 聚蛋白多糖, 软骨缺损

Abstract：Objective To explore the effect of lentivirus-mediated Aggrecan overexpression plasmid transfection of bone marrow mesenchymal stem cells combined with Gelatin/PLGA nano-fiber porous scaffold into chondrocytes.Methods The lentivirus Aggrecan overexpression vector was constructed，bone marrow mesenchymal stem cells were cultured.A rabbit model of cartilage defect was established.Gelatin/PLGA three-dimensional scaffolds and bone marrow mesenchymal stem cell complex transfected with polyproteoglycan overexpression vector were implanted in the experimental group；Gelatin/PLGA threedimensional scaffolds and non-transfected bone marrow mesenchymal stem cell complex were implanted in the negative control group；saline was added to the model group.In vitro，HE staining and immunohistochemical staining were used to detect the expression of Aggrecan and type Ⅱ collagen in the co-culture system of bone marrow mesenchymal stem cells combined with Gelatin/PLGA nanofiber scaffold into chondrocytes.Establishment of rabbit articular cartilage defect model with HE staining and immunohistochemical study on the effect of repairing cartilage defect in animal model.Results In the experimental group (Gelatin/PLGA nanofiber porous scaffold transfected with Aggrecan over-expression vector bone marrow mesenchymal stem cell complex)，the cells attached on the scaffold surface were compared with the control group (Gelatin/PLGA nano-fiber porous scaffold did not transfect bone marrow mesenchymal stem cells).The number of cells increased obviously.The results showed that the effect of the experimental group was better than that of the negative control group.The results of HE staining showed that the negative control group was mostly fibrous repair，while the experimental group was mostly cellular repair.The results of immunohistochemical staining showed that the contents of Aggrecan and type Ⅱ collagen in the repaired cartilage in the experimental group were significantly higher than those in the negative control group.Conclusion Gelatin/PLGA three-dimensional scaffold transfection of Aggrecan overexpression vector bone marrow mesenchymal stem cell complex can promote the differentiation of BMSCs into chondrocytes and secrete more extracellular matrix containing Aggrecan and type Ⅱ collagen，which help the repair of cartilage defects.

Key words： Gelatin/PLGA three-dimensional scaffold bone marrow mesenchymal stem cells Aggrecan cartilage defect

通讯作者: 张贵春

作者简介: 郑润泉(1989－ )，男，医师，济南军区总医院骨创伤外科，250000。

引用本文:

郑润泉 1，康继 2，张贵春 1*. 质粒修饰BMSC联合纳米支架促进软骨缺损修复的相关研究[J]. 实用骨科杂志, 2019, 25(5): 428-433.
Zheng Runquan 1，Kang Ji 2，Zhang Guichun 1. Experimental Study of Lentivirus Mediated Aggrecan Overexpression Plasmid Transfection into Bone Marrow Mesenchymal Stem Cells Combined with Gelatin / PLGA Nano-fiber Porous Scaffold for Cartilage Defect. sygkzz, 2019, 25(5): 428-433.

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http://www.sygkzz.com/CN/ 或 http://www.sygkzz.com/CN/Y2019/V25/I5/428

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