This study aimed to compare the treatment effects of lentiviral vector-mediated hBMP2 which was overexpressed in the femoral bone marrow stromal cells of osteoporotic rats through genetic infection in vitro and in vivo. Comparison of the two transgenic effects may be crucial to determining the lentivirus infection method to be used. Following a comparison of the rat bone marrow stromal cells (rBMSCs) in osteoporotic (MSCs OVX) and normal (MSCs CON) groups, the lentiviral vector-mediated human bone morphogenetic protein 2 (hBMP2), which overexpressed the BMSCs of osteoporotic rats in vitro (rBMSCs in OE group), was constructed. The osteogenic ability in the overexpressed (OE) group was then compared to that of the MSCs CON. The rBMSCs in the OE group (transplants of genetic infection in vitro) and the lentivirus-containing solution (injected material of genetic infection in vivo) were injected into the femurs. The treatment effect of each group was compared via bone mineral density (BMD) and bone histomorphometry. The hBMP2-modified osteoporosis rBMSCs formed by genetic infection in vitro (n=7) had an ameliorated treatment effect on the femur as compared to that of the in vivo (n=7) (BMD: 0.315 vs. 0.19 g/cm2, P<0.01; bone histomorphometry: For bone trabeculars (Tb.Ar/T.Ar): 0.301 vs. 0.114, P<0.01; for trabecular thickness (Tb.Th): 43.54 vs. 21.39 µm, P<0.01; for trabecular separation (Tb.Sp): 115.7 vs. 304.87 µm, P<0.01). The results showed that the treatment effects of osteoporotic rBMSCs on local osteoporosis performed by genetic infection were improved in vitro as compared to those in vivo.