Synergistic effects of recombinant Lentiviral-mediated BMP2 and TGF-beta3 on the osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro

•LV carrying hBMP2 and/or hTGF-beta3 genes were constructed and transduced rat BMSCs.•BMP2 and/or TGF-beta3 genes can be successfully overexpressed in BMSCs.•The expression of OPN, OCN, and OPG increased significantly in double genes groups.•Overexpression of BMP2 and /or TGF-beta3 could upregulate ALP activity.•Both BMP-2 and TGF-beta-3 had synergistic effects on the osteogenic differentiation. Bone marrow mesenchymal stem cells (BMSCs) are considered good candidates for seed cells in bone engineering. The study aim to investigate the synergistic effects of human bone morphogenetic protein 2 (hBMP2) and transforming growth factor beta3 (hTGF-beta3) modified BMSCs on inducing osteogenic differentiation in vitro. Lentivirus (LV) carrying hBMP2 and/or hTGF-beta3 genes were constructed and used to transduce rat BMSCs. The expression of osteogenic molecules was detected by qRT-PCR and western blotting. Targeted genes were PCR-amplified and confirmed by DNA sequencing and BLAST analysis. BMSCs infected by vectors effectively resulted in the overexpressions of hBMP2 and hTGF-beta3 and higher levels of hBMP2 and hTGF-beta3 in the culture supernatant. The co-transduction of hBMP2 and hTGF-beta3 induced BMSCs osteogenic differentiation more effectively than the transduction of hBMP2 or hTGF-beta3 individually. The expression levels of osteopontin (OPN), osteocalcin (OCN), and osteoprotegerin (OPG) in LV-hBMP2 + LV-hTGF-beta3 group (BMSCs transfected by vectors respectively carrying hBMP-2 gene and hTGF-beta3 gene) and LV-hBMP2-hTGF-beta3 group (BMSCs transfected by vector carrying hBMP2 and hTGF-beta3 fusion gene) were significantly higher than in LV-BMP2 (BMSCs transfected by vector carrying hBMP2 gene) and LV-TGF-beta3 (BMSCs transfected by vector carrying hTGF-beta3 gene) groups (P