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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 threedimensional 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.
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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[J]. sygkzz, 2019, 25(5): 428-433.
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| http://www.sygkzz.com/EN/ OR http://www.sygkzz.com/EN/Y2019/V25/I5/428 |
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