Valvular interstitial cells (VICs) are integral to the progression of calcific aortic valve disease (CAVD). Dentin matrix protein-1 (DMP-1), a member of the Sibling family protein, is implicated in the calcification process. This study aims to investigate the role and mechanisms of DMP-1 in the osteogenic differentiation of VICs. Between April 2018 and December 2018, aortic valve tissues were collected from 14 patients undergoing aortic valve replacement or heart transplantation. DMP-1 expression was quantified in calcified valves versus normal controls. An in vitro model of VICs' osteogenic differentiation was established to study the regulatory mechanism of DMP-1 on valvular calcification using immunoblot, immunohistochemistry, immunofluorescence, etc. The expression of DMP-1 was significantly increased in the calcified aortic valves patients (P < 0.01). DMP-1, both short and long arginine-glycine-aspartic acid (RGD) peptides, induced the osteogenic differentiation in VICs, an effect that was inhibited by an integrin αvβ3 antagonist (P < 0.05). Furthermore, the expression levels of RAF, RAS, MEK, and phosphorylated ERK1/2 were significantly elevated in VICs upon stimulation of DMP-1 (P < 0.05). DMP-1 is involved in the progression of valvular calcification and promotes the osteogenic differentiation of VICs via the integrin αvβ3 receptor. The combination of DMP-1 and integrin αvβ3 via its RGD domain activates the MAPK signaling pathway, leading to enhanced osteogenic gene expression in VICs. Clinical trial number: not applicable.