Cellular toxicity, cell adhesion and proliferation, and alkaline phosphatase (ALP) activity were investigated for an artificial bone substitute composed of heated carbonate apatite (CAp) and Type I atelocollagen (AtCol) extracted from bovine tail skins (88/12 in %wt/wt). To enhance the intramolecular crosslinking between collagen molecules, the CAp-AtCol substitutes were irradiated by ultraviolet rays (wave length 254 nm) at 4°C for 4 h or vacuum dried at 150°C for 2 h. Cytotoxicity tests by a direct contact method and an extract dilution method revealed that the CAp-AtCol substitutes were cytocompatible for balb 3T3 fibroblasts. Osteoblast adhesion studies demonstrated that the substitute disks composed of 980°C-heated CAp and AtCol were significantly more adhesive for osteoblasts than those of 1,200°C-sintered CAp and AtCol (p < 0.05). Proliferation studies showed that the number of osteoblasts grown in the media containing substitutes of 980°C-heated CAp and AtCol was statistically higher than grown in those of 1,200°C-sintered CAp and AtCol after 5 days (p < 0.05). It was found that osteoblasts grown in the substitutes of 980°C-heated CAp and AtCol only expressed similar ALP activity to the controls. These results suggested that the substitutes consisting of 980°C-heated CAp and AtCol show more favorable interactions with osteoblasts than those of 1,200°C-sintered CAp and AtCol.