Reactivation of inhibited bone acid phosphatase and its significance in bone histomorphometry

C. C. Liu, D. J. Sherrard, N. A. Maloney, Guy Howard

Research output: Contribution to journalArticle

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Abstract

Despite biochemical demonstration of acid phosphatase (AcP) activation or reactivation in bone, few attempts have been made to show similar effects histochemically. Bones from growing rats, when fixed in 4% buffered formaldehyde at room temperature and demineralized in 5% formic acid, exhibited expected inactivation of AcP. The inhibited AcP, however, was reactivated by pre-incubation of sections for 1 hr at 37°C in the following buffers: 0.2 M Tris, 0.2 M glycine, 0.2 M NaHCO3, or 0.1 M borax, as well as in alkaline water, but not in 0.2 M Na2HPO4 (all at pH 9). The reactivation was (a) site-specific (e.g., osteoclasts, osteoblasts, osteocytes, and cement lines), (b) temperature- and pH-dependent, (c) unaffected by OH- or SH--binding agents or by an alkaline phosphatase inhibitor, and (d) inhibited completely by 10mM Na2HPO4. The reactivation process, much simplified and/or more effective than with the methods previously reported, was observed in all 83 human biopsy bones embedded in methyl methacrylate and in human bones stored in cold buffered formaldehyde for 7 monts. This study demonstrates a unique method for reactivating and thus localizing the inhibited AcP in bones, and suggests possible applications in bone histomorphometry.

Original languageEnglish
Pages (from-to)1355-1363
Number of pages9
JournalJournal of Histochemistry and Cytochemistry
Volume35
Issue number12
StatePublished - Dec 1 1987
Externally publishedYes

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Acid Phosphatase
Bone and Bones
formic acid
Formaldehyde
Osteocytes
Temperature
Methacrylates
Osteoclasts
Osteoblasts
Glycine
Alkaline Phosphatase
Buffers
Biopsy
Water

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Reactivation of inhibited bone acid phosphatase and its significance in bone histomorphometry. / Liu, C. C.; Sherrard, D. J.; Maloney, N. A.; Howard, Guy.

In: Journal of Histochemistry and Cytochemistry, Vol. 35, No. 12, 01.12.1987, p. 1355-1363.

Research output: Contribution to journalArticle

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