Liu Hongchun, Guo Xiaoheng, Liu Xiao, Chen Weijian, Zhao Naru, Liao Yidong
Department of Preventive Dentistry, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University & Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China. Email:
Zhonghua Kou Qiang Yi Xue Za Zhi. 2015 Jan;50(1):38-42.
To investigate the effects of 20% fluor-hydroxyapatite (FHA) on proliferation and osteogenic differentiation of human MG63 osteosarcoma cells.
FHA was prepared by chemical precipitation method, and its structure and surface features were tested by scanning microscope, X-ray diffraction (XRD) and Fourier transform infrared spectroscopy. MG63 cells were cultured and divided into FHA, hydroxyapatite (HA) and control groups (n = 3). The proliferation of the cells was evaluated using methylthiazol tetrazolium (MTT) assay. ALP activity of the cells was assessed. Osteogenic differentiation was evaluated based on the reverse transcription PCR (RT-PCR) of differentiation-related genes, namely, collagen type I (Col I), alkaline phosphatase (ALP), osteocalcin (OCN) and core binding factor α1 (Cbfα1). The data were analyzed statistically by one-way analysis of variance using SPSS 13.0 software.
XRD test showed that the main crystalline phase of FHA was similar to that of HA. Absorptance value of cells exposed to FHA(1.87±0.06) measured by MTT was higher than that of the control(1.25±0.02) on the third day(P < 0.05), and there was no statistically significant difference between the cells exposed to FHA and HA(1.84±0.03) (P > 0.05). ALP activity of the cells exposed to FHA(4.62±0.09)was higher than that of the control (1.92 ± 0.05) (P < 0.05). RT-PCR tests showed that compared with the control, FHA up-regulated the expression of Col I, ALP and OCN mRNA, down-regulated the expression of Cbfα1 mRNA.
FHA enhances the proliferation and osteogenic differentiation-related gene expression, and has good biocompatibility.
研究20%氟羟基磷灰石（FHA）对人MG63骨肉瘤细胞增殖和成骨分化的影响。
采用化学沉淀法制备FHA，通过扫描显微镜、X射线衍射（XRD）和傅里叶变换红外光谱对其结构和表面特征进行检测。培养MG63细胞并分为FHA组、羟基磷灰石（HA）组和对照组（n = 3）。采用甲基噻唑四氮唑（MTT）法评估细胞增殖情况。检测细胞的碱性磷酸酶（ALP）活性。基于分化相关基因即Ⅰ型胶原（Col Ⅰ）、碱性磷酸酶（ALP）、骨钙素（OCN）和核心结合因子α1（Cbfα1）的逆转录聚合酶链反应（RT-PCR）评估成骨分化情况。使用SPSS 13.0软件通过单因素方差分析对数据进行统计学分析。
XRD检测显示FHA的主要晶相与HA相似。MTT法测得第3天FHA组细胞的吸光度值（1.87±0.06）高于对照组（1.25±0.02）（P < 0.05），FHA组与HA组细胞的吸光度值（1.84±0.03）比较差异无统计学意义（P > 0.05）。FHA组细胞的ALP活性（4.62±0.09）高于对照组（1.92±0.05）（P < 0.05）。RT-PCR检测显示，与对照组相比，FHA上调了Col Ⅰ、ALP和OCN mRNA的表达，下调了Cbfα1 mRNA的表达。
FHA可促进细胞增殖及成骨分化相关基因表达，具有良好的生物相容性。
