Promotion of chondrogenesis of marrow stromal stem cells by TGF-β3 fusion protein in vivo. corrected.
10.1007/s11596-013-1182-z
- Author:
Wei WU
1
;
Yang DAN
2
;
Shu-Hua YANG
2
;
Cao YANG
2
;
Zeng-Wu SHAO
2
;
Wei-Hua XU
2
;
Jin LI
2
;
Xian-Zhe LIU
2
;
Dong ZHENG
3
Author Information
1. Department of Pediatics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
2. Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
3. Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. xhzhengdong@163.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Blotting, Western;
Bone Marrow Cells;
metabolism;
Cartilage, Articular;
pathology;
surgery;
Cell Differentiation;
genetics;
Cells, Cultured;
Chondrocytes;
metabolism;
Chondrogenesis;
genetics;
Green Fluorescent Proteins;
genetics;
metabolism;
Matrix Metalloproteinases;
genetics;
metabolism;
Mesenchymal Stem Cell Transplantation;
methods;
Mesenchymal Stromal Cells;
metabolism;
Microscopy, Fluorescence;
Molecular Sequence Data;
Osteoarthritis;
surgery;
Rats;
Rats, Sprague-Dawley;
Recombinant Fusion Proteins;
genetics;
metabolism;
Reverse Transcriptase Polymerase Chain Reaction;
Transfection;
Transforming Growth Factor beta3;
genetics;
metabolism;
Treatment Outcome
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2013;33(5):692-699
- CountryChina
- Language:English
-
Abstract:
The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.
