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Mouse embryo-derived NIH3T3 fibroblasts adopt an osteoblast-like phenotype when treated with 1α,25-dihydroxyvitamin D3 and dexamethasone in vitro

Authors

  • Chaoxiang Shui,

    1. Department of Human Metabolism and Clinical Biochemistry, University of Sheffield Medical School, Sheffield, United Kingdom
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  • Andrew M. Scutt

    Corresponding author
    1. Department of Human Metabolism and Clinical Biochemistry, University of Sheffield Medical School, Sheffield, United Kingdom
    2. Institute of Child Health, University of Sheffield Medical School, Sheffield, United Kingdom
    • Tissue Engineering Group, Department of Engineering Materials, Sir Robert Hadfield Building, University of Sheffield Medical School, Mappin Street, Sheffield S1 3JD, UK.
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Abstract

This study examines the capability of NIH3T3 fibroblasts to express osteoblastic markers following stimulation with a number of hormones and growth factors in vitro. Of the agents tested, 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) dose-dependently induced alkaline phosphatase (ALP) activity in NIH3T3 cells, and this effect was enhanced by the addition of dexamethasone (Dex), which when administered alone caused no detectable ALP expression. The combined use of 1,25(OH)2D3 and Dex also stimulated the synthesis of osteocalcin, and osteopontin. Furthermore, cells treated with the both hormones, in the presence of β-glycerophosphate and l-ascorbic acid, formed mineralized plaques, indicating an osteoblast (OB) phenotype. By contrast, the differentiation induced by 1,25(OH)2D3 or 1,25(OH)2D3 plus Dex was significantly antagonized by transforming growth factor-β1 and all trans-retinoic acid. These data indicate that NIH3T3 cells have the potential to adopt an OB-like phenotype and may prove to be a convenient model for studying the early events of osteogenic differentiation and the specific interactions of 1,25(OH)2D3 with glucocorticoids in controlling this process in vitro. © 2002 Wiley-Liss, Inc.

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