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Calcium hydroxide reduces lipopolysaccharide-stimulated osteoclast formation
Jin Jiang1, Jian Zuo, Shih-Hua Chen
1College of Dentistry, Department of Endodontics, University of Florida, Gainsville 32610-0436, USA. jjiang@dental.ufl.edu
Objective:
The purpose of this study was to determine the direct effects of lipopolysaccharide (LPS) on osteoclastogenesis and to assess the ability of calcium hydroxide-Ca(OH)(2)-to inhibit the osteoclast formation stimulated by LPS.
Study Design:
RAW 264.7 cells were cultured with 50 ng/mL recombinant receptor activator of NF-kappaB ligand (RANKL) for 72 hours. RANKL was then removed, and the cells were treated with 0, 1, 10, or 100 ng/mL of LPS, Ca(OH)(2)-treated LPS, or 50 ng/mL of RANKL as a positive control for an additional 48 hours. Cells were fixed and stained with fluorescein isothiocyanate-conjugated phalloidin to detect actin ring formation, and histochemistry was performed to detect multinucleated cells expressing tartrate-resistant acid phosphatase activity.
Results:
LPS induced osteoclast-like cell (OCL) formation in a dose-dependent manner when osteoclast precursor RAW 264.7 cells were pretreated for 72 hours with RANKL. Ca(OH)(2) significantly inhibited the ability of LPS to stimulate OCL formation.
Conclusion:
This study shows that LPS directly stimulates OCL formation. The detoxification of LPS by treatment with Ca(OH)(2) significantly reduced its ability to trigger the differentiation of OCLs.