Objective: To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts. Methods: Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion. In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifugale tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 48 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials. Results: Osteoblasts attached to the bio-derived bone materials in a time-dependent manner. There were significantly (P< 0.05) more attached cells after 180 minutes than after 15 and 90 minutes of incubations (P< 0.05). Osteoblasts were proliferated in a large amount on the surface and in the materials. Osteoblasts seeded onto 100 mg bio-derived bones resulted in significantly (P< 0.05) more measurable proliferation than those seeded onto 10 mg bones. Osteoblasts appeared round as they attached to the materials, then flattened and spread over with time passing. Conclusions: Bio-derived bones can provide a good environment for the attachment and proliferation of osteoblasts. Objective: To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts. Methods: Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifuge tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with the liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 4 8 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials. Results: Osteoblasts attached to the bio-derived bone materials in a time-dependent Osteoblasts were proliferated in a large amount on the surface and in the materials. Osteoblasts seeded onto 100 mg (P <0.05) more attached cells after 180 minutes than after 15 and 90 minutes of incubations Bio-derived bones resulted in significantly (P <0.05) more measurable proliferation than those seeded onto 10 mg bones. Osteoblasts appeared as they attached to the materials, then flattened and spread over with time passing. Conclusions: Bio-derived bones can provide a good environment for the attachment and proliferation of osteoblasts.