Objective To explore the role and possible mechanism of a disintegrin and metalloproteinase 10 (ADAM10) in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and tibial fracture union. Methods BMSCs of SD rats were cultured, stably transfected with a negative control (NC) shRNA pGFP-V-RS vector, and divided into sh-NC and sh-NC+DMSO groups. BMSCs were stably transfected with an ADAM10 shRNA pGFP-V-RS vector and divided into sh-ADAM10, sh-ADAM10+DMSO, and sh-ADAM10+valproic acid (VPA) groups. After 14 days of osteogenic induction, absorbance at 405 nm after alizarin red staining, ALP activity, and ADAM10, OCN, Runx2, CoLI, NICD, and Hes1 expression levels were analyzed. A tibial fracture model was established in SD rats, and NC shRNA or ADAM10 shRNA pCMV5. 0 vectors were injected locally. Fracture healing and gene expression were then observed for 4 weeks. Results The ADAM10 expression level in BMSCs of the sh-ADAM10 group was lower than that in the sh-NC group. After osteogenic induction, the absorbance value of alizarin red staining at 405 nm, ALP activity, and OCN, Runx2, CoL-I, NICD, and Hes1 expression levels of the sh-ADAM10 group were higher than those of the sh-NC group (P< 0. 05). The absorbance value of alizarin red staining at 405 nm, ALP activity, and OCN, Runx2, and Col-I expression levels of the sh-ADAM10+VPA group after osteogenic induction were lower than those in the sh-ADAM+DMSO group (P< 0. 05). Healing of the tibial fracture in the sh-ADAM10 group was better than that in the sh-NC group, and OCN, Runx2, CoL-I, NICD, and Hes1 expression levels were higher than those in the sh-NC group (P<0. 05). Conclusions Knocking down ADAM10 expression promotes osteogenic differentiation of BMSCs and tibial fracture healing, which is related to inhibition of the Notch1 pathway.