High concentration of zoledronate inhibits osteogenesis differentiation and promotes apoptosis of human osteoblasts

Abstract

Abstract: PURPOSE:To investigate the influence of zoledronate on osteoblasts isolated from human jaw in vitro. METHODS: Osteoblasts were isolated from health human mandible. Third passage of osteoblasts were exposed to zoledronate with different concentrations 0 μmol/L,1 μmol/L,5 μmol/L in vitro. Cell proliferation ability was detected using CCK-8 kit and flow cytometry method was applied to identity cell apoptosis. ALP staining and quantitative analysis were used to evaluate the osteogenesis differentiation of osteoblasts. Real-time PCR were used to measure the gene expression of ALP,OPN and Runx2 and western blotWB was used to detect OPN expression.The difference between each group was analyzed with SPSS 16.0 software package. RESULTS: Cell growth curve indicated that zoledronate treatment groups showed slower growth compared with control group and flow cytometry results indicated higher proportion of apoptosis cells in zoledronate treated groups. ALP staining and quantitative analysis both revealed decreased osteogenesis differentiation ability in zoledronate treated groups. These effect were more obvious in 5 μmol/L treated group than 1μmol/L treated group. Significant down-regulation of of ALP, Runx2 and OPN gene expression were also found in zoledronate treated groups.WB results revealed the decreased expression of OPN in zoledronate treated groups. CONCLUSION: High concentration of zoledronate inhibits osteogenesis differentiation and proliferation ability of human osteoblasts and facilitate their apoptosis.

Key words: Zoledronate, Osteoblast, Osteonecrosis of the jaw, Apoptosis, Osteogenesis differentiation

HU Long-wei, HAN Jing, PAN Hong-ya, XU Li-qun. High concentration of zoledronate inhibits osteogenesis differentiation and promotes apoptosis of human osteoblasts[J]. Shanghai Journal of Stomatology, 2015, 24(5): 389-394.

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