Alteration of osteocalcin mRNA expression in ovine osteoblasts in dependence of sodium fluoride and sodium selenite medium supplementation

Abstract

Objective of this study was to assess the quantification of osteocalcin (OCN) expression by ovine osteoblasts cultured with different concentrations of sodium fluoride (F) and sodium selenite (Se) to evaluate the interaction of these agents on OCN expression in vitro . We wanted to demonstrate a possible protective effect of selenium on the toxic effect of fluoride. Osteoblasts were isolated by complete trypsin and collagenase digestion from ovine calvarial bone and cultured in DMEM supplemented with 15% FBS at 37 degrees C in a humidified 5% CO 2 incubator. Identified osteoblasts were divided into one control group (C) and eight experimental groups, which were exposed to different concentrations of sodium fluoride (F; 0, 0.5, 1 mM) sodium selenite (Se; 0, 0.1, 1 microM). At different time points after treatment total RNA was extracted and reverse transcribed into first-strand cDNA. OCN mRNA was indirectly measured by real-time fluorescent quantitative PCR (qPCR). OCN mRNA expression in F 1 mM with Se 1 microM group was found to have a high peak at day seven and was lower before and afterwards. Expression of OCN mRNA in all groups except control could be promoted by F and/or Se showing a general upregulation. Furthermore, the toxicity from excessive exposure of osteoblast with F could be circumvented by usage of moderate concentration of Se. Osteoblasts cultured in vitro may have stressful responses to F and Se at the first few days. Low concentrations of Se inhibit the toxic effects of high concentrations of F. Therefore, F and Se could be used as antagonistic factors, which could regulate osteocalcin expression.