The aim of the current study was to observe the effects of Baimai ointment (MPBM) medicated plasma for stimulation of osteogenesis in mesenchymal stem cells (MSCs) in vitro. This study tested both rat plasma and medicated plasma (BM to stimulate MSCs). CCK-8 assays were employed to detect proliferation levels of MSCs. This study also applied alkaline phosphatase (ALP) staining and Alizarin Red S staining to detect reactive osteogenic differentiation of MSCs. Moreover, mRNA levels of ALP, OCN, Runx2, Col1a1, Col1a2, OPN, and OPG were quantified using RT-qPCR. Protein levels of Akt and Wnt/beta-catenin signaling pathways were measured using Western blotting. Compared with the control group, there were significant increases in proliferation of MSCs, as well as increased mRNA expression of ALP, OCN, Runx2, Col1A1, ColA2, OPN, and OPG, in the MPBM group (P < 0.05). Accordingly, phosphorylated Akt (Ser473, Thr308) and beta-catenin signals were strongly activated in the MPBM group (P < 0.05). Results indicate that MPBM enhanced proliferation and osteoblastic differentiation of MSCs via Akt and Wnt/beta catenin signaling pathways, suggesting BM as a promising therapeutic drug for improved fracture healing.