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Updated: Jun 10, 2026

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro
Published on: June 16, 2022
Passive and active in vitro resorption of calcium and magnesium phosphate cements by osteoclastic cells
Christian Grossardt1, Andrea Ewald, Liam M Grover
1Department for Functional Materials in Medicine and Dentistry, University of Würzburg, Würzburg, Germany.
Abstract:
Biocements are clinically applied materials for bone replacement in non-load-bearing defects. Depending on their final composition, cements can be either resorbed or remain stable at the implantation site. Degradation can occur by two different mechanisms, by simple dissolution (passive) or after osteoclastic bone remodeling (active). This study investigated both the passive and active in vitro resorption behavior of brushite (CaHPO₄ · 2H₂O), monetite (CaHPO₄), calcium-deficient hydroxyapatite (CDHA; Ca₉(PO₄)₅HPO₄OH), and struvite (MgNH₄PO₄ · 6H₂O) cements. Passive resorption was measured by incubating the cement samples in a cell culture medium, whereas active resorption was determined during the surface culture of multinuclear osteoclastic cells derived from RAW 264.7 macrophages. Osteoclast formation was confirmed by showing tartrate resistant acid phosphatase (TRAP) activity on CDHA, brushite, and monetite surfaces, as well as by measuring calcitonin receptor (CT-R) expression as an osteoclast-specific protein by Western blot analysis for struvite ceramics. An absence of passive degradation and only marginally active degradation of <0.01% were found for CDHA matrices. For the secondary calcium phosphates brushite and monetite, active degradation was predominant with a cumulative Ca²+ release of 2.02 (1.20) μmol during 13 days, whereas passive degradation released only 0.788 (0.04) μmol calcium ions into the medium. The struvite cement was the most degradable with a passive (active) release of 9.26 (2.92) Mg²+ ions and a total weight loss of 4.7% over 13 days of the study.
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