title: Culture of osteoblasts on bio-derived bones.
abstract: 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.
authors.affiliation: 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
authors.forename: Xu
authors.initials: X
authors.lastname: Lan
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Yang
authors.forename: Bao-feng
authors.initials: BF
authors.lastname: Ge
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: Liu
mesh_terms: 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*
chemical_list: D018786:Bone Substitutes