Mesoporous bioactive glass (MBG) has been demonstrated to play a vital role in bone tissue engineering due to its bioactivity, biocompatibility, and osteoinduction properties. Here, we report that MBG grafted with an amino group (MBG-NH2) and MBG-NH2 adsorbed epidermal growth factor (EGF) (MBG-NH2/EGF) sustained-release EGF, and MBG-NH2/EGF could accelerate osteoblast differentiation and mineralization in MC3T3-E1 cells. We found that MBG-NH2 could promote bone-like deposit formation and Ca deposition in vitro. Intriguingly, we observed that MBG-NH2/EGF enhanced MC3T3-E1 cell adhesion. We also showed that extracellular signal-regulated kinase 1/2 (ERK1/2) was phosphorylated when MC3T3-E1 cells were cultured on MBG-NH2/EGF. Interestingly, the transcription factor Runx2, important for osteoblast differentiation, was also activated when MC3T3-E1 cells were cultured on MBG-NH2/EGF. We showed that MC3T3-E1 cells cultured on MBG-NH2/EGF activating Runx2 was through ERK1/2 phosphorylation. Consistent with this survey, we observed that MC3T3-E1 cells cultured on MBG-NH2/EGF accelerated osteoblastic marker gene expressions, including osteopontin (Opn) and osteocalcin (Ocn). Taken together, we conclude that the osteoblast differentiation and mineralization were accelerated in MC3T3-E1 cells cultured on MBG-NH2/EGF through ERK-activated Runx2 pathway. These findings support the idea that MBG-NH2/EGF is a potential biomaterial for bone tissue repair in bone defect-related diseases.