Effect of hydroxyapatite bioceramics on the growth of osteoblasts and HIF-α/VEGF signal axis in partial hypoxia environment in vitro.

ABSTRACT

BACKGROUND: Hydroxyapatite bioceramic is a kind of bone implant commonly used in oral clinic treatment. In the early stage of tissue repair, cells will suffer hypoxic due to the interruption of blood supply. OBJECTIVE: Studying the expression of osteoblasts in hypoxic environment will help us to understand the expression and response mechanism of osteoblasts at the implantation site of hydroxyapatite in the early stage of hypoxia. METHODS: MG63 osteoblast cell line was used in this study. The cells of normal group were incubated under normal oxygen and hydroxyapatite ceramics condition. The cells of hypoxia group were incubated under hypoxia (37∘C, 8% CO2, 8% O2, 86% N2) and hydroxyapatite ceramics condition. Cell proliferation was measured by CCK8 assay. Apoptosis was measured by flow cytometry. Serum alkaline phosphatase (ALP) activity was measured by ALP kit. Hypoxia inducible factor (HIF-α) and vascular endothelial growth factor (VEGF) were detected by Western blot. RESULTS: Compared to the normal group, the cells of hypoxia group showed a dramatically higher proliferation ability, especially at 48 h (P< 0.05). Due to hypoxia, cell apoptosis was induced, but there is no difference between these two groups. Interestingly, the ALP activity of hypoxia group was higher than that of normal group at 24 h and 48 h (P< 0.05). Mechanically, western blot result showed that the protein level of both HIF-α and VEGF were up-regulated in hypoxia group. CONCLUSIONS: Under hypoxia condition, hydroxyapatite bioceramics can promote the proliferation of MG63 osteoblasts, elevate the activity of alkaline phosphatase and upregulate HIF-α and VEGF expression without effect on apoptosis.