title: In-vitro analysis of rhBMP-2 effects in human osteogenic cells.
abstract: This study evaluated the in vitro expression of bone-related proteins by osteoblasts in the presence of different concentrations of human recombinant bone morphogenetic protein-2 (rhBMP-2). Immortalized human fetal osteoblastic cell line 1.19 (hFOB) were exposed to different concentrations of rhBMP-2 (10, 50, or 100 ng/mL) for 72 h. Cell proliferation and viability (MTT assay), as well as the expression of fibronectin, osteonectin, and osteopontin were assessed by indirect immunofluorescence and Western blot. Neither of the 3 concentrations of rhBMP-2 caused statistically significant alterations in cell proliferation and viability, although the concentration of 100 ng/mL showed lower values for both assays after both 48 and 72 h of exposure. There was no alteration in the expression of noncollagenous proteins, as analyzed by immunofluorescence, when compared with the control group. Furthermore, in the Western blot assay we observed a statistically significant decrease in fibronectin and osteonectin at 100 ng rhBMP-2/mL (p < 0.05) by comparison with the medium alone. The expression of osteopontin decreased slightly in all 3 concentrations of rhBMP-2 tested; however, the change was not statistically significant (p > 0.05). In this in-vitro study, the tested concentrations of rhBMP-2 appeared to decrease the expression of important bone-related molecules in pre-osteoblast cells.
authors.affiliation: a Laboratory of Immunology and Molecular Biology, São Leopoldo Mandic Institute and Research Center, Campinas, São Paulo, Brazil.
authors.forename: Eder M F
authors.initials: EM
authors.lastname: de Oliveira
authors.forename: Elizabeth F
authors.initials: EF
authors.lastname: Martinez
authors.forename: Jeruza P
authors.initials: JP
authors.lastname: Bossonaro
authors.forename: Renato C
authors.initials: RC
authors.lastname: Ribeiro
authors.forename: Vera C
authors.initials: VC
authors.lastname: de Araújo
authors.forename: Marcelo Henrique
authors.initials: MH
authors.lastname: Napimoga
mesh_terms: D000328:Adult; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000032:analysis* / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis; D005455:Fluorescent Antibody Technique; D006801:Humans; D010006:Osteoblasts / Q000378:metabolism*; D010012:Osteogenesis / Q000502:physiology; D015676:Osteonectin / Q000096:biosynthesis; D053495:Osteopontin / Q000096:biosynthesis; D011247:Pregnancy; D011994:Recombinant Proteins / Q000032:analysis; D013778:Tetrazolium Salts; D013844:Thiazoles; D014404:Tubulin / Q000096:biosynthesis
chemical_list: D055396:Bone Morphogenetic Protein 2; D004396:Coloring Agents; D005353:Fibronectins; D015676:Osteonectin; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014404:Tubulin; D053495:Osteopontin; C022616:thiazolyl blue