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Effects of epidermal growth factor on osteoblastic cellsin vitro

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Summary

The effect of epidermal growth factor (EGF) on clone MC3T3-El cells that have osteoblastic activity was examined by phase-contrast microscopy and electron microscopy; hydroxyproline content, collagen synthesis, collagen pattern, and alkaline phosphatase (ALP) activity were also determined. We found that EGF (0.4 ng/ml) transformed the cells from their normal polygonal shape to a spindle-like morphology by 8 h. This hormone also caused dose-related suppression of hydroxyproline content and ALP activity which was detectable 2 days and 1 day, respectively, after EGF addition. Indomethacin did not affect hydroxyproline content and ALP activity, suggesting that the effect of EGF on the cells may not be mediated by prostaglandins. Epidermal growth factor at concentrations of 2 to 50 ng/ml significantly decreased collagen synthesis in the cells, whereas protein synthesis was stimulated. Electron microscopy demonstrated that collagen fiber formation was also reduced by EGF; an immature type of fibril was observed compared with the typical cross-striated one in the controls. Moreover, the hormone treatment also resulted in the appearance of type III collagen in addition to the type I already present in the cells. These suppressive effects of EGF on MC3T3-El cellsin vitro suggest that this hormone may be involved in bone remodellingin vivo as well.

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References

  1. Luben RA, Wong GL, Cohen DV (1976) Biochemical characterization with parathormone and calcitonin of isolated bone cells: provisional identification of osteoclasts and osteoblasts. Endocrinology 99:526–534

    Article  CAS  PubMed  Google Scholar 

  2. Wong GL, Luben RA, Cohen DV (1979) 1,25 dihydroxycholecalciferol and parathormone: effects on isolated osteoclast-like and osteoblast-like cells. Science 197:663–665

    Google Scholar 

  3. Wiestner M, Fischer S, Dessau M, Müller PK (1981) Collagen types synthesized by isolated calvarium cells. Exp Cell Res 133:115–125

    Article  CAS  PubMed  Google Scholar 

  4. Kodama H, Amagai Y, Sudo H, Kasai S, Yamamoto S (1982) Establishment of a clonal osteogenic cell line from newborn mouse calvaria. Jap J Oral Biol 23:899–901

    Google Scholar 

  5. Sudo H, Kodama H, Amagai Y, Yamamoto S, Kasai S (in press) Differentiation and calcification of a new clonal osteogenic cell line derived from newborn mouse calvaria in vitro. J Cell Biol

  6. Chen LB, Gudor RC, Sun T-T, Chen AB, Mosesson MW (1977) Control of a cell surface major glycoprotein by epidermal growth factor. Science 197:776–778

    CAS  PubMed  Google Scholar 

  7. Goodman R, Slater E, Herschman HR (1980) Epidermal growth factor induces tyrosine hydroxylase in a clonal phenochromocytoma cell line, PC-G2. J Cell Biol 84:495–500

    Article  CAS  PubMed  Google Scholar 

  8. Kumegawa M, Yajima T, Hiramatsu M, Ikeda E, Hatakeyama K, Namba M (1981) Epidermal growth factor stimulates collagen synthesis in liver-derived epithelial cells. Biochim Biophys Acta 675:305–308

    CAS  PubMed  Google Scholar 

  9. Kumegawa M, Hiramatsu M, Yajima T, Hatakeyama K, Hosoda S, Namba M (1982) Effect of epidermal growth factor on collagen formation in liver-derived epithelial clone cells. Endocrinology 110:607–612

    CAS  PubMed  Google Scholar 

  10. Tashijian AH Jr, Levine L (1978) Epidermal growth factor stimulates prostaglandin production and bone resorption in cultured mouse calvaria. Biochem Biophys Res Commun 85:966–975

    Article  Google Scholar 

  11. Canalis E, Raisz LG (1979) Effect of epidermal growth factor on bone formation in vitro. Endocrinology 104:862–869

    CAS  PubMed  Google Scholar 

  12. Raisz LG, Simmons HA, Sandberg AL, Canalis E (1980) Direct stimulation of bone resorption by epidermal growth factor. Endocrinology 107:270–273

    CAS  PubMed  Google Scholar 

  13. Shupnik MA, Ip NY, Tashijian AH Jr (1980) Characterization and regulation of receptors for epidermal growth factor in mouse calvaria. Endocrinology 107:1738–1746

    CAS  PubMed  Google Scholar 

  14. Shupnik MA, Tashjian AH Jr (1981) Functional receptors for epidermal growth factor on human osteosarcoma cells. J Cell Physiol 109:403–410

    Article  CAS  PubMed  Google Scholar 

  15. Savage CR Jr, Cohen S (1972) Epidermal growth factor and a new derivative: rapid isolation procedure and biological and chemical characterization. J Biol Chem 247:7609–7611

    CAS  PubMed  Google Scholar 

  16. Cohen S (1962) Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the newborn animal. J Biol Chem 237:1555–1562

    CAS  PubMed  Google Scholar 

  17. Lowry OH, Roberts NR, Wn ML, Hixon WS, Crawford EJ (1954) The quantitative histochemistry of brain. II. Enzyme measurements. J Biol Chem 207:19–37

    CAS  PubMed  Google Scholar 

  18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin-phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  19. Woessner JF (1961) The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys 93:440–447

    Article  CAS  PubMed  Google Scholar 

  20. Peterkofsky B, Diegelmann R (1971) Use of a mixture of proteinase-free collagenase for the specific assay of radioactive collagen in the presence of other proteins. Biochem 10:988–994

    Article  CAS  Google Scholar 

  21. Yaoita H, Foidart JM, Katz SI (1978) Localization of the collagenous component in skin basement membrane. J Invest Dermatol 70:191–193

    Article  CAS  PubMed  Google Scholar 

  22. Rose SP, Pruss RM, Herschman HR (1975) Initiation of 3T3 fibroblast cell division by epidermal growth factor. J Cell Physiol 86:593–598

    Article  PubMed  Google Scholar 

  23. Carpenter G, Cohen S (1976) Human epidermal growth factor and the proliferation of human fibroblasts. J Cell Physiol 88:227–237

    Article  CAS  PubMed  Google Scholar 

  24. Cherington RV, Pardee AB (1980) Synergistic effects of epidermal growth factor and thrombin on the growth stimulation of dipoloid Chinese hamster fibroblasts. J Cell Physiol 105:25–32

    Article  CAS  PubMed  Google Scholar 

  25. Takigawa M, Ishida H, Takano T, Suzuki F (1980) Polyamine and differentiation: induction of ornithine decarboxylase by parathyroid hormone is a good marker of differentiated chondrocytes. Proc Natl Acad Sci USA. 77:1481–1485

    CAS  PubMed  Google Scholar 

  26. Beaulieu JF, Calvert R (1981) The effect of epidermal growth factor (EGF) on the differentiation of the rough endoplasmic reticulum in fetal mouse small intestine in organ culture. J Histochem Cytochem 29:765–770

    CAS  PubMed  Google Scholar 

  27. Byyny RL, Orth DN, Cohen S, Doyne ES (1974) Epidermal growth factor: effects of androgens and adrenergic agents. Endocrinology 95:776–782

    Article  CAS  PubMed  Google Scholar 

  28. Carpenter G (1978) The regulation of cell proliferation: advances in the biology and mechanism of action of epidermal growth factor. J Invest Dermatol 71:283–288

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Oral Anatomy, Josai Dental University, 350-02, Sakado, Saitama

    M. Kumegawa & T. Yajima

  2. Pharmacology, Josai Dental University, 350-02, Sakado, Saitama

    M. Hiramatsu & K. Hatakeyama

  3. Department of Physiology, Tohoku Dental School, 963, Koriyama, Fukushima

    H. Kodama

  4. Department of Oral Anatomy, Kyushu University, 812, Fukuoka, Japan

    T. Osaki & K. Kurisu

Authors
  1. M. Kumegawa
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  2. M. Hiramatsu
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  3. K. Hatakeyama
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  4. T. Yajima
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  5. H. Kodama
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  6. T. Osaki
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  7. K. Kurisu
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Kumegawa, M., Hiramatsu, M., Hatakeyama, K. et al. Effects of epidermal growth factor on osteoblastic cellsin vitro . Calcif Tissue Int 35, 542–548 (1983). https://doi.org/10.1007/BF02405091

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