Reproducible quantification of osteoclastic activity: Characterization of a biomimetic calcium phosphate assay
Salwa M. Maria
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Shriners Hospital for Children-Canada, Montreal, Quebec, Canada
Search for more papers by this authorChristiane Prukner
Institute of Materials Science and Technology, Friedrich Schiller University, Jena, Germany
Search for more papers by this authorZeeshan Sheikh
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Search for more papers by this authorFrank Mueller
Institute of Materials Science and Technology, Friedrich Schiller University, Jena, Germany
Search for more papers by this authorJake. E. Barralet
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Search for more papers by this authorCorresponding Author
Svetlana V. Komarova
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Shriners Hospital for Children-Canada, Montreal, Quebec, Canada
Correspondence to: S. V. Komarova (e-mail: [email protected])Search for more papers by this authorSalwa M. Maria
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Shriners Hospital for Children-Canada, Montreal, Quebec, Canada
Search for more papers by this authorChristiane Prukner
Institute of Materials Science and Technology, Friedrich Schiller University, Jena, Germany
Search for more papers by this authorZeeshan Sheikh
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Search for more papers by this authorFrank Mueller
Institute of Materials Science and Technology, Friedrich Schiller University, Jena, Germany
Search for more papers by this authorJake. E. Barralet
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Search for more papers by this authorCorresponding Author
Svetlana V. Komarova
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Shriners Hospital for Children-Canada, Montreal, Quebec, Canada
Correspondence to: S. V. Komarova (e-mail: [email protected])Search for more papers by this authorAbstract
Osteoclasts are responsible for bone and joint destruction in rheumatoid arthritis, periodontitis, and osteoporosis. Animal tusk slice assays are standard for evaluating the effect of therapeutics on these cells. However, in addition to batch-to-batch variability inherent to animal tusks, their use is clearly not sustainable. Our objective was to develop and characterize a biomimetic calcium phosphate assay based on the use of phase pure hydroxyapatite coated as a thin film on the surface of culture plates, to facilitate the reproducible quantification of osteoclast resorptive activity. Osteoclasts were formed from RAW 264.7 mouse monocyte cell line using a pro-resorptive cytokine RANKL (50 ng/mL). No change in substrate appearance was noted after culture with media without cells, or undifferentiated monocytes. Only in the presence of osteoclasts localized areas of calcium phosphate dissolution were observed. The total area resorbed positively correlated with the osteoclast numbers (R2 = 0.99). The resorbed area was significantly increased by the addition of RANKL, and decreased after application of known inhibitors of osteoclast resorptive activity, calcitonin (10 μM), or alendronate (100 μM). Thus, calcium phosphate coated substrates allow reliable monitoring of osteoclast resorptive activity and offer an alternative to animal tusk slice assays. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 102B: 903–912, 2014.
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