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In Vitro effects of zinc on markers of bone formation

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Abstract

Zinc deficiency is associated with a reduced rate of bone formation that can be corrected by supplementation of the deficient diet with adequate amounts of zinc. This study was conducted to examine the effects of zinc on bone cell parameters associated with bone formation.

Tibiae were removed from 19-d-old chicken embryos and incubated for 48 h in Dulbecco’s modified Eagle’s medium supplemented with antibiotics, bovine serum albumin, and HEPES. The addition of zinc (25–200 Μg/dL) to tibial cultures resulted in a concentrationdependent increase in alkaline phosphatase activity, an increase in the incorporation of proline into bone protein and an increase in the posttranslational oxidation of proline to peptidyl hydroxyproline. These effects of zinc were all diminished by the addition of 2,6-pyridine dicarboxylic acid, a chelator of zinc. The addition of either cycloheximide (10-5M), dactinomycin (10-8M), or hydroxyurea (10-3M) to tibial cultures also attenuated the effects of zinc.

The effect of zinc on bone cell DNA synthesis was measured by following the incorporation of3H-thymidine into DNA and by fluorometric measurement of cellular DNA content. These methods revealed that the addition of zinc to cultured tibiae resulted in a concentration-dependent increase in tibial DNA content and synthesis rate. The magnitude of the zinc-induced DNA increase was similar to the magnitude of the zinc-induced increases in alkaline phosphatase activity, proline incorporation, and hydroxyproline synthesis. Normalization of these latter responses to tibial DNA content yield data indicating that the effect of zinc on bone formation results from a zinc-induced increase in bone cell proliferation.

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References

  1. M. J. Jackson, Physiology of zinc: general aspects, inZinc in Human Biology, C. E Miller, ed., Springer-Verlag, Berlin, pp. 1–14 (1989).

    Google Scholar 

  2. L. S. Hurley, J. Gowan, and G. Milhaud, Calcium metabolism in manganese-deficient and zinc-deficient rats,Proc. Soc. Exp. Biol. Med. 130, 856–860 (1969).

    PubMed  CAS  Google Scholar 

  3. Z. K. Roughead and M. E. Kunkel, Effect of diet on bone matrix constituents,J. Amer. Coll. Nutr. 10, 242–246 (1991).

    CAS  Google Scholar 

  4. D. Chen, L. C. Waite, and W. M. Pierce, Jr., In vitro bone resorption is dependent on physiological concentrations of zinc,Biol. Trace Element Res. 61, 9–18 (1998).

    Article  CAS  Google Scholar 

  5. G. Oner, B. Bhaumick, and R. M. Bala, Effect of zinc deficiency on serum somatomidin levels and skeletal growth in young rats,Endocrinology 114, 1860–1863 (1984).

    Article  PubMed  CAS  Google Scholar 

  6. A. S. Prasad and D. Aberleas, Changes in activities of zinc-dependent enzymes in zinc-deficient tissues of rats,J. Appl. Physiol. 31, 842–846 (1971).

    PubMed  CAS  Google Scholar 

  7. A. Suwarnasarn, J. C. Wallwork, G. I. Lykhen, F. N. Lou, and H. H. Sandstead, Epiphyseal plate development in the zinc-deficient rat,J. Nutr. 112, 1320–1328 (1982).

    PubMed  CAS  Google Scholar 

  8. M. Yamaguchi and T. Sakashita, Enhancement of vitamin D3 effect on bone metabolism in weanling rats by orally administered zinc sulfate,Acta Endocrinol. 11, 285–288 (1986).

    Google Scholar 

  9. B. Bergman, U. Freberg, and S. Lohmander; The importance of zinc to cell proliferation in endochondral growth sites in the white rat,Scand. J. Dent. Res. 80, 486–492 (1972).

    PubMed  CAS  Google Scholar 

  10. B. C. Starcher, C. H. Hill, and J. G. Madaras, Effect of zinc deficiency on bone collagenase and collagen turnover,J. Nutr. 110, 2095–2102 (1980).

    PubMed  CAS  Google Scholar 

  11. M. Yamaguchi, H. Oishi, and Y. Sukata, Stimulatory effect of zinc on bone formation in tissue culture,Biochem. Pharmacol. 36, 4007–4012 (1987).

    Article  PubMed  CAS  Google Scholar 

  12. T. Kaji, R. Kawatani, M. Takata, T. Hoshino, T. Miyahara, H. Kozuka, et al., The effects of cadmium, copper or zinc on formation of embryonic chick bone in tissue culture,Toxicology 50, 303–316 (1988).

    Article  PubMed  CAS  Google Scholar 

  13. M. Yamaguchi, A. Mochizuki, and S. Okada, Stimulatory effect of zinc in bone growth in weanling rats,J. Pharm. Dyn. 5, 619–626 (1982).

    CAS  Google Scholar 

  14. L. R. DeChatelet and M. R. Cooper, A modified procedure for the determination of leukocyte alkaline phosphatase,Biochem. Med. 4, 61–68 (1970).

    Article  PubMed  CAS  Google Scholar 

  15. J. E. Puzas and D. B. P. Goodman, A rapid assay for cellular deoxyribonucleic acid,Anal. Biochem. 86, 50–55 (1978).

    Article  PubMed  CAS  Google Scholar 

  16. W. R. Todd, C. A. Elvehjim, and E. B. Hart, Zinc in the nutrition of the rat,Am. J. Physiol. 107, 146–156 (1934).

    CAS  Google Scholar 

  17. A. Togari, S. Orakawa, M. Arai, and S. Matsumoto, Alterations of in vitro bone metabolism and tooth formation by zinc,Gen. Pharmacol. 24, 1133–1140 (1993).

    PubMed  CAS  Google Scholar 

  18. M. Yamaguchi and R. Yamaguchi, Action of zinc on bone metabolism in rats. Increases in alkaline phosphatase activity and DNA content,Biochem. Pharmacol. 35, 773–777 (1986).

    Article  PubMed  CAS  Google Scholar 

  19. T. Kaje, Y Fujiwara, C. Yamamoto, M. Sakamoto, and H. Kozuka, Stimulation by zinc of cultured endothelial cell proliferation: possible involvement of endogenous basic fibroblast growth factor,Life Sci. 55, 1781–1787 (1994).

    Article  Google Scholar 

  20. E. B. Trujillo, Effects of nutritional status on wound healing,J. Vasc. Nurs. 11, 12–18 (1995).

    Google Scholar 

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Chen, D., Waite, L.C. & Pierce, W.M. In Vitro effects of zinc on markers of bone formation. Biol Trace Elem Res 68, 225–234 (1999). https://doi.org/10.1007/BF02783905

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  • DOI: https://doi.org/10.1007/BF02783905

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