Elsevier

Bone

Volume 11, Issue 2, 1990, Pages 115-119
Bone

Original article
Oxygen derived free radicals in osteoclasts: The specificity and location of the nitroblue tetrazolium reaction

https://doi.org/10.1016/8756-3282(90)90058-7Get rights and content

Abstract

Oxygen derived free radicals are generated by osteoclasts. In a novel culture system, isolated rat osteoclasts were stained when nitroblue tetrazolium (NBT) was reduced by cellular oxidants to formazan, an insoluble precipitate. Superoxide dismutase (SOD) inhibited the accumulation of formazan by the isolated osteoclasts. Osteoclasts in mouse calvarial organ cultures also reduced NBT to formazan. The reaction products were localized to the area of the osteoclast-bone interface. At the light microscopic level, the formazan granules appeared to be concentrated within the cytoplasm. Formazan accumulation was significantly inhibited by calcitonin (hCT). The inhibition of NBT reduction by SOD indicates that the isolated osteoclasts were capable of producing superoxide. The localization of the formazan granules between the external osteoclastic membrane and the bone, and the inhibition of this reaction during hCT exposure suggests that oxygen derived free radicals may contribute to bone resorption.

References (16)

  • M.C. Seeds et al.

    Independent stimulation of membrane potential changes and the oxidative metabolic burst in polymorphonuclear leukocytes

    Blood

    (1985)
  • B.M. Babior

    The respiratory burst of phagocytes

    J. Clin. Invest.

    (1984)
  • R.L. Baehner et al.

    Quantitative nitroblue tetrazolium test in chronic granulomatous disease

    N. Engl. J. Med.

    (1968)
  • C.J. Beard et al.

    Neutrophil defect associated with malignant osteopetrosis

    J. Lab. Clin. Med.

    (1986)
  • R.T. Briggs et al.

    Hydrogen peroxide production in chronic granulomatous disease

    J. Clin. Invest.

    (1977)
  • J.T. Curnutte et al.

    Manganese-dependent NADPH oxidation by granulocytes

    J. Clin. Invest.

    (1976)
  • M.D. Fallon et al.

    The oxidative metabolism of isolated osteoclasts is regulated by calcitropic agents

    J. Bone Min. Res.

    (1986)
  • I.R. Garrett et al.

    Osteoblastic bone resorption in vitro is dependent on oxygen-derived radical production

    J. Bone Min. Res.

    (1987)
There are more references available in the full text version of this article.

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