Osteoblast migration as well as adhesion is important aspect of bone formation. While the significance of extracellular matrix protein cell interactions are generally appreciated, the specific relationship osteoblast interactions with adhesive extracellular matrix proteins to cell motility is not well defined. We investigated effects of adhesive extracellular proteins on the motility of ROS 17/2.8 cells. Recombinant fibronectin domain 4 (+RGD) and type I collagen promoted cell motility in a dose-dependent manner. Recombinant fibronectin domain 4 - dependent motility was inhibited by anti-1 integrin antibody and GRGDSK peptide. Type I collagen - dependent motility was inhibited anti-2 integrin antibody, demonstrating that the migration supported by these proteins is specifically regulated by their receptors. Surprisingly, vitronectin, a major adhesive protein among serum components, repressed motility. This study demonstrates that diverse mechanisms of osteoblast migration on extracellular matrices exist.