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BMC Musculoskeletal Disorders
Published in: BMC Musculoskeletal Disorders 1/2010

Open Access 01-12-2010 | Research article

Screening of protein kinase inhibitors identifies PKC inhibitors as inhibitors of osteoclastic acid secretion and bone resorption

Authors: Mette G Sørensen, Morten A Karsdal, Morten H Dziegiel, Jean A Boutin, Olivier Nosjean, Kim Henriksen

Published in: BMC Musculoskeletal Disorders | Issue 1/2010

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Abstract

Background

Bone resorption is initiated by osteoclastic acidification of the resorption lacunae. This process is mediated by secretion of protons through the V-ATPase and chloride through the chloride antiporter ClC-7. To shed light on the intracellular signalling controlling extracellular acidification, we screened a protein kinase inhibitor library in human osteoclasts.

Methods

Human osteoclasts were generated from CD14+ monocytes. The effect of different kinase inhibitors on lysosomal acidification in human osteoclasts was investigated using acridine orange for different incubation times (45 minutes, 4 and 24 hours). The inhibitors were tested in an acid influx assay using microsomes isolated from human osteoclasts. Bone resorption by human osteoclasts on bone slices was measured by calcium release. Cell viability was measured using AlamarBlue.

Results

Of the 51 compounds investigated only few inhibitors were positive in both acidification and resorption assays. Rottlerin, GF109203X, Hypericin and Ro31-8220 inhibited acid influx in microsomes and bone resorption, while Sphingosine and Palmitoyl-DL-carnitine-Cl showed low levels of inhibition. Rottlerin inhibited lysosomal acidification in human osteoclasts potently.

Conclusions

In conclusion, a group of inhibitors all indicated to inhibit PKC reduced acidification in human osteoclasts, and thereby bone resorption, indicating that acid secretion by osteoclasts may be specifically regulated by PKC in osteoclasts.
Appendix
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Metadata
Title
Screening of protein kinase inhibitors identifies PKC inhibitors as inhibitors of osteoclastic acid secretion and bone resorption
Authors
Mette G Sørensen
Morten A Karsdal
Morten H Dziegiel
Jean A Boutin
Olivier Nosjean
Kim Henriksen
Publication date
01-12-2010
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2010
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/1471-2474-11-250

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