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Differentiation of osteoblasts and formation of mineralized bone in vitro

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Summary

Periostea consisting of the osteogenic layer and the fibrous layer of the periosteum were dissected from 17-day-old embryonic chick calvariae, leaving the osteoblasts behind on bone. The dissected periostea were folded with the osteogenic cells in apposition. The explants were cultured on plasma clots for up to 6 days, during which time osteodifferentiation was observed followed by osteoid formation in between the two layers. These cultures consistently mineralized in the presence of 5 or 10 mMβ-glycerophosphate. The mineralization and osteoid formation displayed many characteristics identical with those seen in vivo. Specifically, the osteoid formed was birefringent under polarized light, the central zone of osteoid became mineralized within 24 h of formation in vitro, and a clear border between mineralized and non-mineralized osteoid suggestive of a mineralization front was present. The unmineralized osteoid at the periphery was surrounded by osteoblasts. These data suggest that physiologic mineralization of osteoid produced in vitro did occur in this system by the addition of the alkaline phosphatase substrateβ-glycerophosphate.

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References

  1. Mazza, A., Felluga, B., Curci, G.: A histochemical study of the matrix of long bones of the mouse embryo grownin vitro, Metab. Bone Dis. Rel. Res.2:65–72, 1979

    Article  Google Scholar 

  2. Nijweide, P. J., Embryonic chicken periosteum in tissue culture: osteoid formation and calcium uptake, Proc. Kon. Ned. Akad. Wet.C78:410–417, 1975

    Google Scholar 

  3. Thorogood, P.:In vitro studies on skeletogenic potential of membrane bone periosteal cells, J. Embryol. Exp. Morphol.54:185–207, 1979

    CAS  PubMed  Google Scholar 

  4. Binderman, I., Duksin, D., Harrell, A., Katzir, E., Sachs, L.: Formation of bone tissue in culture from isolated bone cells, J. Cell Biol.61:427–439, 1974

    Article  CAS  PubMed  Google Scholar 

  5. Binderman, I., Greene, R. M., Pennypacker, J. P.: Calcification of differentiating skeletal mesenchymein vitro, Science206:222–224, 1979

    CAS  PubMed  Google Scholar 

  6. Fell, H. B.: The osteogenic capacityin vitro of periosteum and endosteum isolated from the limb skeleton of fowl embryos and young chicks, Anatomy66:157–180, 1932

    CAS  Google Scholar 

  7. Gaillard, P. J.: Developmental changes in the composition of the body fluids in relation to growth and differentiation of tissue cultures, Protoplasma23:145–174, 1934

    Article  Google Scholar 

  8. Goldhaber, P.: Remodelling of bone in tissue culture, J. Dent. Res.45:490–499, 1966

    Google Scholar 

  9. Ito, Y., Endo, H., Enomoto, H., Wakabayashi, K., Takamura, K.: Ossification in tissue culture, Exp. Cell Res.31:119–127, 1963

    Article  CAS  PubMed  Google Scholar 

  10. Jackson, S. F., Smith, R. H.: Studies on the biosynthesis of collagen, J. Biophys. Biochem. Cytol.3:897–911, 1957

    Article  CAS  PubMed  Google Scholar 

  11. Osdoby, P., Caplan, A. I.: The possible differentiation of osteogenic elementsin vitro from chick limb mesodermal cells, Dev. Biol.52:283–299, 1976

    Article  CAS  PubMed  Google Scholar 

  12. Osdoby, P., Caplan, A. I.: Osteogenesis in cultures of limb mesenchymal cells, Dev. Biol.73:84–102, 1979

    CAS  PubMed  Google Scholar 

  13. Marvaso, V., Bernard, G. W.: Initial intramembranous osteogenesisin vitro, Am. J. Anat.149:453–467, 1977

    Article  CAS  PubMed  Google Scholar 

  14. Nijweide, P. J., Van der Plas, A.: Studies on the uptake of calcium and strontium by isolated periosteal cells, Proc. Kon. Ned. Akad. Wet.C78:418–434, 1975

    Google Scholar 

  15. Paul, J. (ed.): Cell and Tissue Culture. Churchill-Livingstone, Edinburgh, 1975

    Google Scholar 

  16. Albritton, E. C.: Standard Values in Blood. W. B. Saunders Company, Philadelphia, 1952

    Google Scholar 

  17. Anderson, H. C.: Calcification of rachitic rat cartilagein vitro by extracellular matrix vesicles, Am. J. Pathol.79:237–245, 1975

    CAS  PubMed  Google Scholar 

  18. Anderson, H. C.: Calcification of rachitic cartilage to study matrix vesicle function, Fed. Proc.35:148–153, 1976

    CAS  PubMed  Google Scholar 

  19. Thomas, M. L., Ramp, W. K.: Effects of PTH on alkaline phosphatase activity and mineralization of cultured chick embryo tibiae, Calcif. Tiss. Int.27:137–142, 1979

    CAS  Google Scholar 

  20. Anderson, H. C.: A new method to measure45calcium accumulation by matrix vesicles, Metab. Bone Dis. Rel. Res.1:193–197, 1978

    Article  Google Scholar 

  21. Drury, R. A. B., Wallington, E. A., Cameron, R.: Carleton's Histological Technique, 4th ed., p. 150. Oxford University Press, London, 1973

    Google Scholar 

  22. Burstone, M. S.: In: Symposium on Calcification in Biological Systems, pp. 225–227. AAAS Symposium Publication, Washington, D.C., 1960

    Google Scholar 

  23. Ralis, H., Ralis, Z.: A simple method for demonstration of osteoid in paraffin sections, Med. Lab. Technol.32:203–213, 1975

    CAS  PubMed  Google Scholar 

  24. Raisz, L. G., Dietrich, J. W., Canalis, E. M.: Factors influencing bone formation in organ cultures, Isr. J. Med. Sci.12:108–114, 1976

    CAS  PubMed  Google Scholar 

  25. Harris, W. H., Heaney, R. P., Davis, L. A., Weinberg, E. H., Coutts, R. D., Schiller, A. L.: Stimulation of bone formationin vivo by phosphate supplementation, Calcif. Tissue Res.22:85–98, 1976

    Article  CAS  PubMed  Google Scholar 

  26. Hoffman, W. S.: Biochemistry of Clinical Medicine, 4th Ed., pp. 554–555. Year Book Medical Publishers, Chicago, 1970

    Google Scholar 

  27. Anderson, H. C., Reynolds, J. J.: Phosphate stimulation of calcium uptake into cultured embryonic bones, fine structure of matrix vesicles and their role in calcification, Dev. Biol.34:211–227, 1973

    Article  CAS  PubMed  Google Scholar 

  28. Ramp, W. K., Neuman, W. F.: Some factors affecting mineralization of bone in tissue culture, Am. J. Physiol.220:270–274, 1971

    CAS  PubMed  Google Scholar 

  29. Stern, P. H., Raisz, L. G.: In D. J. Simmons, A. S. Kunin, (eds.): Skeletal Research, pp. 21–59. Academic Press, New York, 1979

    Google Scholar 

  30. Ham, A. W., Cormack, D. H.: Histophysiology of Cartilage, Bone and Joints, p. 450. J. B. Lippincott Company, Philadelphia, 1979

    Google Scholar 

  31. Authors data. Manuscript submitted for publication.

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Tenenbaum, H.C., Heersche, J.N.M. Differentiation of osteoblasts and formation of mineralized bone in vitro. Calcif Tissue Int 34, 76–79 (1982). https://doi.org/10.1007/BF02411212

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