Combination Gene Transfection of TGF-β3 and BMP-2 Mediated by Lentivirus Induces Chondrogenic Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells

Cartilage restoration continues to present a huge clinical problem due to a limited supply of vessels, nerves and lymph, but tissue engineering has produced many efficient methods that have been tested in animal models. Compared with exogenous cytokines, which remain a vital difficulty in feeding plenty of cytokines to the implanted therapeutic cells, gene transfection into the therapeutic cells could solve this challenging problem by overexpression of some cytokines in cartilage tissue engineering. Therefore, this study was conducted to investigate transforming the growth factor-beta 3 (TGF-beta 3) gene and TGF-beta 3/bone morphogenetic protein-2 (BMP-2) combination gene, transfected into rabbit bone marrow mesenchymal stem cells (BMSCs) and mediated by recombinant lentivirus (rLV), to induce BMSCs to become chondrocytes. Reverse-transcription PCR (RT-PCR), Western blot, immunocytochemistry and toluidine blue staining were performed to detect the expression of SOX-9, collagen type II (COL2) and aggrecan (ACAN). The results suggest that these chondrogenic markers were upregulated at higher levels in the TGF-beta 3/BMP-2 combined gene group than in those in the TGF-beta 3 group. Consequently, we conclude that the TGF-beta 3/BMP-2 combination gene could not only induce the chondrogenic differentiation of BMSCs but also be more efficient than single gene induction. This assay may provide an alternative mentality or method for tissue engineering to repair cartilage damage.