Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect
10.3969/j.issn.1003-0034.2015.05.011
- VernacularTitle:慢病毒介导的Sox9基因在兔骨髓间充质干细胞的过表达促进软骨损伤修复
- Author:
Zhen WANG
1
;
Chuan Da LIANG
;
Yu Jie BAI
;
Ning KANG
;
Yu Jun FENG
;
Quan Zi YANG
Author Information
1. 山西医科大学第二医院骨科
- Keywords:
Sox9 gene;
Bone marrow mesenchymal stem cells(BMSCs);
Transfection;
Cartilage;
Rabbit
- From:
China Journal of Orthopaedics and Traumatology
2015;(5):433-440
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for re?pairing articular cartilage injury in vivo. Methods:Rabbit bone marrow mesenchymal stem cells(BMSCs)were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treat?ment. After animals anesthesia,a full thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile,the transfected cells were implanted to repair the rabbit model with full thickness cartilage defects. Cartilage defects tissue was observed with light microscope,electron microscope,HE and im?munohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation. Re?sults:At 3 days after the transfection,Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection,the expression of collagen typeⅡbegan and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox 9 gene. Histologi?cal observation showed that at 6 weeks after the operation,the defects in the experimental group was filled with hyaline like cartilage tissue,12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively,the defects were filled with fibrous tissues in control groups. Meanwhile,immunohistochemical stain?ing of sections with typeⅡcollagen antibodies showed the proteins in the regenerated tissue stained positive for typeⅡcolla?gen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances. Conclusion:Overexpression of Sox9 gene on rab? bit bone marrow mesenchymal stem cells(BMSCs)promote the repair of cartilage defect.
