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Assessing Bone Formation Using Mouse Calvarial Organ Cultures

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Bone Research Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 80))

Abstract

The history of culturing bone explants goes back to the early 1920s, when Robinson reported that the enzyme alkaline phosphatase played an important role in bone mineralization based on studies of chick bone fragments. Subsequently, Reynolds used bone explants to study collagen synthesis in bone (1) and to investigate the effects of a wide variety of other agents on bone turnover such as vitamin A (2), ascorbic acid (3), calcitonin (4,5), hydrocortisone (6) and bisphosphonates (7). Others have used rodent calvarial and long bone explants to study the effects of cytokines on bone resorption (812) and bone formation (13,14). This chapter focuses on the use of cultured neonatal calvariae as an assay for agents with anabolic activity (1319) (see Note 1).

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References

  1. Reynolds, J. J. (1967) The synthesis of collagen by chick bone rudiments in vitro. Exp. Cell Res. 47, 42–48.

    Article  PubMed  CAS  Google Scholar 

  2. Reynolds, J. J. (1968) Inhibition by calcitonin of bone resorption induced in vitro by vitamin A. Proc. R. Soc. Lond. B. Biol. Sci. 170, 61–69.

    Article  PubMed  CAS  Google Scholar 

  3. Reynolds, J. J. (1966) The effect of ascorbic acid on the growth of chick bone rudiments in chemically defined medium. Exp. Cell Res. 42, 178–188.

    Article  PubMed  CAS  Google Scholar 

  4. Reynolds, J. J. and Dingle, J. T. (1968) Time course of action of calcitonin on resorbing mouse bones in vitro. Nature. 218, 1178–1179.

    Article  PubMed  CAS  Google Scholar 

  5. Minkin C., Reynolds, J. J., and Copp, D. H. (1971) Inhibitory effect of salmon and other calcitonins on calcium release from mouse bone in vitro. Can. J. Physiol. Pharmacol. 49, 263–267.

    PubMed  CAS  Google Scholar 

  6. Reynolds, J. J. (1966) The effect of hydrocortisone on the growth of chick bone rudiments in chemically defined medium. Exp. Cell Res. 41, 174–189.

    Article  PubMed  CAS  Google Scholar 

  7. Reynolds, J. J., Minkin, C., Morgan, D. B., Spycher, D., and Fleisch, H. (1972) The effect of two diphosphonates on the resorption of mouse calvaria in vitro. Calcif. Tissue Res. 10, 302–313.

    Article  PubMed  CAS  Google Scholar 

  8. Gowen, M., Wood, D. D., Ihrie, E.J., McGuire, M. K., and Russell, R. G. (1983) An interleukin 1 like factor stimulates bone resorption in vitro. Nature 306, 378–380.

    Article  PubMed  CAS  Google Scholar 

  9. Bertolini, D. R., Nedwin, G. E., Bringman, T. S., Smith, D. D., and Mundy, G. R. (1986) Stimulation of bone resorption and inhibition of bone formation in vitro by human tumor necrosis factors. Nature 319, 516–518.

    Article  PubMed  CAS  Google Scholar 

  10. Lorenzo, J. A., Sousa, S. L., and Leahy, C. L. (1990) Leukemia inhibitory factor (LIF) inhibits basal bone resorption in fetal rat long bone cultures. Cytokine 2, 266–271.

    Article  PubMed  CAS  Google Scholar 

  11. Reynolds, J. J. and Dingle, J. T. (1968) The induction and inhibition of bone resorption in vitro. Calcif. Tissue Res. (Suppl) 50–50a.

    Google Scholar 

  12. Reynolds, J. J. and Dingle, J. T. (1970) A sensitive in vitro method for studying the induction and inhibition of bone resorption. Calcif. Tissue Res. 4, 339–349.

    Article  PubMed  CAS  Google Scholar 

  13. Traianedes, K., Dallas, M. R., Garrett, I. R., Mundy, G. R., and Bonewald, L. F. (1998) 5-Lipoxygenase metabolites inhibit bone formation in vitro. Endocrinology 139, 3178–3184.

    Article  PubMed  CAS  Google Scholar 

  14. Mundy, G., Garrett, R., Harris, S, et al. (1999) Stimulation of bone formation in vitro and in rodents by statins. Science 286, 1946–1949.

    Article  PubMed  CAS  Google Scholar 

  15. Garrett, I. R., Esparza, J., Chen, D., et al. (2000) Statins mediate their effects on osteoblasts by inhibition of HMG-CoA reductase and ultimately BMP-2. J. Bone Miner. Res. 15S, 225.

    Google Scholar 

  16. Garrett, I. R., Chen, D., Zhao, M., et al. (2001) Statins mediate bone formation by enhancing BMP2 expression. Bone 28, 75.

    Google Scholar 

  17. Yin, J. L., Grubbs, B. G., Cui, Y., et al. (2000) Endothelin A receptor blockade inhibits osteoblastic metastasis. J. Bone Miner. Res. 15S, 201.

    Google Scholar 

  18. Garrett, I. R., Gutierrez, G., Chen, D., et al. (2000) Specific inhibitors of the chymotryptic component of the proteasome are potent anabolic agents in vivo. J. Bone Miner. Res. 15S, 197.

    Google Scholar 

  19. Chen, D., Garrett, I.R., Qiao, M., et al. (2001) Proteasome inhibitors stimulate osteoblast differentiation and bone formation by inhibiting Gli3 degradation and enhancing BMP-2 expression. Bone 28, 74.

    Google Scholar 

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© 2003 Humana Press Inc., Totowa, NJ

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Garrett, I.R. (2003). Assessing Bone Formation Using Mouse Calvarial Organ Cultures. In: Helfrich, M.H., Ralston, S.H. (eds) Bone Research Protocols. Methods in Molecular Medicine, vol 80. Humana Press. https://doi.org/10.1385/1-59259-366-6:183

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  • DOI: https://doi.org/10.1385/1-59259-366-6:183

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-044-1

  • Online ISBN: 978-1-59259-366-8

  • eBook Packages: Springer Protocols

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