Culture of osteoblasts on bio-derived bones.
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 microcentrifugable 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.
Lan
Xu
X
Division of Stem Cell and Tissue Engineering, Key Laboratory of Biotherapy of Human Diseases of Ministry of Education, West China Hospital, Sichuan University, Chengdu 610041, China. lanxu2000@163.net
Yang
Zhi-ming
ZM
Ge
Bao-feng
BF
Liu
Xue-mei
XM
D000818:Animals; D018786:Bone Substitutes; D002102:Cadaver; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D006801:Humans; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis; D011817:Rabbits; D012886:Skull / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*
D018786:Bone Substitutes