Vitamin D receptor expression in human bone tissue and dose-dependent activation in resorbing osteoclasts

Abstract

The effects of vitamin D on osteoblast mineralization are well documented. Reports of the effects of vitamin D on osteoclasts, however, are conflicting, showing both inhibition and stimulation. Finding that resorbing osteoclasts in human bone express vitamin D receptor (VDR), we examined their response to different concentrations of 25-hydroxy vitamin D₃ [25(OH)D₃] (100 or 500 nmol·L⁻¹) and 1,25-dihydroxy vitamin D₃ [1,25(OH)₂D₃] (0.1 or 0.5 nmol·L⁻¹) metabolites in cell cultures. Specifically, CD14+ monocytes were cultured in charcoal-stripped serum in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Tartrate-resistant acid phosphatase (TRAP) histochemical staining assays and dentine resorption analysis were used to identify the size and number of osteoclast cells, number of nuclei per cell and resorption activity. The expression of VDR was detected in human bone tissue (ex vivo) by immunohistochemistry and in vitro cell cultures by western blotting. Quantitative reverse transcription-PCR (qRT-PCR) was used to determine the level of expression of vitamin D-related genes in response to vitamin D metabolites. VDR-related genes during osteoclastogenesis, shown by qRT-PCR, was stimulated in response to 500 nmol·L⁻¹ of 25(OH)D₃ and 0.1–0.5 nmol·L⁻¹ of 1,25(OH)₂D₃, upregulating cytochrome P450 family 27 subfamily B member 1 (CYP27B1) and cytochrome P450 family 24 subfamily A member 1 (CYP24A1). Osteoclast fusion transcripts transmembrane 7 subfamily member 4 (tm7sf4) and nuclear factor of activated T-cell cytoplasmic 1 (nfatc1) where downregulated in response to vitamin D metabolites. Osteoclast number and resorption activity were also increased. Both 25(OH)D₃ and 1,25(OH)₂D₃ reduced osteoclast size and number when co-treated with RANKL and M-CSF. The evidence for VDR expression in resorbing osteoclasts in vivo and low-dose effects of 1,25(OH)₂D₃ on osteoclasts in vitro may therefore provide insight into the effects of clinical vitamin D treatments, further providing a counterpoint to the high-dose effects reported from in vitro experiments.

Vitamin D: Bone resorption boosted by vitamin D derivatives

Derivatives of vitamin D can increase the number and the resorption activity of the osteoclast cells that break down bone tissue. A team led by Cameron Brown from the University of Oxford, UK, showed that osteoclasts from human bone tissue and from cell cultures express the receptor that is activated by vitamin D. The researchers stimulated the receptor with various concentrations of two vitamin D derivatives and saw an increase in osteoclast number and activity. They also observed a decrease in the levels of two proteins that cause fusion between osteoclasts, resulting in more but smaller cells. The findings suggest that the increase in bone mass density seen in people with osteoporosis who take vitamin D supplements may not be because of the suppression of osteoclasts as previously believed.

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