The in vitro osteogenetic characteristics of primary osteoblastic cells from a rabbit calvarium.
Previously, we showed that recombinant human bone morphogenetic protein-2 (rhBMP-2) increased bone augmentation beyond the skeletal envelope within a titanium cap in a rabbit calvarium; many cuboidal osteoblastic cells were observed histologically. These results suggested that the new osteoblastic cells might have differentiated and matured via stimulation by rhBMP-2. To date, however, no studies have reported the characteristics of osteoblastic cells derived from adult rabbit calvarium, after addition of rhBMP-2. To determine the effects of rhBMP-2 on osteoblastic cells, we observed morphological characteristics and alkaline phosphatase activity of osteoblastic cells from an adult rabbit calvarium. The expression of proteins in the BMP signaling pathway and extracellular matrix were analyzed, and mineralized nodule formation was assessed. The alkaline phosphatase activity increased significantly after rhBMP-2 stimulation. The protein levels of phosphorylated-Smad1, Runx2, osteocalcin, osteopontin, and type I collagen were augmented by rhBMP-2 stimulation using Western blotting or ELISA; rhBMP-2 also stimulated mineralized nodule formation with alizarin red staining. The results suggest that primary osteoblastic cells derived from a rabbit calvarium have osteogenetic characteristics in vitro, underscoring the potential use of these cells as a model for studying bone formation. These cells may play an important role in in vivo bone augmentation in a rabbit experimental model.
Hasegawa
Yasunori
Y
Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry, Tokyo, Japan.
Shimada
Koichi
K
Suzuki
Naoto
N
Takayama
Tadahiro
T
Kato
Takashi
T
Iizuka
Tetsuya
T
Sato
Shuichi
S
Ito
Koichi
K
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D000880:Anthraquinones; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002113:Calcification, Physiologic / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D024042:Collagen Type I / Q000032:analysis; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006801:Humans; D008297:Male; D023421:Models, Animal; D010006:Osteoblasts / Q000187:drug effects* / Q000502:physiology; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000187:drug effects* / Q000502:physiology; D053495:Osteopontin / Q000032:analysis; D010766:Phosphorylation; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology; D012886:Skull / Q000166:cytology* / Q000187:drug effects*; D051898:Smad1 Protein / Q000032:analysis; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D000880:Anthraquinones; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit; D016326:Extracellular Matrix Proteins; D011994:Recombinant Proteins; C493716:Runx2 protein, mouse; D051898:Smad1 Protein; C497401:Smad1 protein, mouse; C506896:Spp1 protein, mouse; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D015675:Osteocalcin; D053495:Osteopontin; C010078:alizarin; D000469:Alkaline Phosphatase