Basic fibroblast growth factor enhances the growth and expression of the osteogenic phenotype of dexamethasone-treated human bone marrow-derived bone-like cells in culture.
Basic fibroblast growth factor (bFGF) was shown to enhance rat stromal bone marrow cells in culture to produce mineralized bone-like tissue in response to dexamethasone (Dex) treatment (Pitaru et al., J Bone Miner Res 8:919; 1993). The purpose of this study was to explore the effect of bFGF on Dex-treated human stromal bone marrow cells (hSBMC) in culture. Human SBMC from 6 patients were cultured for 14 days (P0) and then subcultured and grown for 28 days in the presence of Dex (10(-8) mol/L). The effect of bFGF on cell proliferation at P0 and protein content, DNA content, alkaline phosphatase activity (ALP), osteocalcin secretion, and formation of mineralized bone-like tissue (MBT) at P1 was analyzed. bFGF treatment resulted in a 2.4-fold increase in cell number at P0 and a concentration-dependent increase in [3H]-thymidine incorporation at P1, reaching a maximum increase of 3.7-fold at a concentration of 0.3 ng/mL. Furthermore, bFGF significantly increased both DNA content (two- to threefold), protein content (five- to sixfold), and the amount of MBT (up to 20-fold) at P1 cultures. Morphological evaluation of the MBT at the electron microscope level revealed a mineralization process along collagen fibrils similar to the natural process. The osteogenic nature of the bFGF-treated cultures was further shown by their ALP activity, as well as osteocalcin secretion in response to 1,25-dihydroxyvitamin D3. In conclusion, bFGF demonstrated a stimulatory effect on the proliferation of Dex-treated hSBMC-derived osteoprogenitors while maintaining their capacity to fully differentiate and form bone-like tissue in culture.
Maurice and Gabriela Goldschleger Eye Research Institute, Maurice and Gabriela Goldschleger School of Dental Medicine, Sackler Faculty of Medicine, Tel-Aviv University, Israel.
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S
Pri-Chen
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S
Pitaru
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F
Lokiec
N
N
Savion
D000293:Adolescent; D000469:Alkaline Phosphatase / Q000032:analysis; D000880:Anthraquinones; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000648:ultrastructure; D001842:Bone and Bones / Q000187:drug effects / Q000648:ultrastructure; D002452:Cell Count / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D008854:Microscopy, Electron; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000235:genetics; D010641:Phenotype; D011506:Proteins / Q000032:analysis; D017154:Stromal Cells / Q000187:drug effects
D000880:Anthraquinones; D005938:Glucocorticoids; D011506:Proteins; D016222:Fibroblast Growth Factor 2; D015675:Osteocalcin; C004468:Alizarin Red S; D003907:Dexamethasone; D004247:DNA; D000469:Alkaline Phosphatase