We proposed a novel combined gene therapy of human vascular endothelial growth factor 165 gene (hVEGF165) and human bone morphogenetic protein 2 gene (hBMP2) for bone regeneration by lentivirus-mediated co-transfection of both genes into rat bone marrow-derived mesenchymal stromal cells (MSCs). Both genes were successfully co-expressed in MSCs confirmed by real-time PCR and ELISA. And the alkaline phosphatase activity of MSCs was significantly augmented by the co-transfection with both genes than any single gene transfection (P < 0.01). These results demonstrated the feasibility of the combined gene therapy by using MSCs lentivirally co-transfected with hVEGF165 and hBMP2 for bone regeneration.