摘要
为探讨氟化物对反刍动物骨骼代谢的影响及对成骨细胞的作用机理,以初生山羊股骨骨膜为材料,采用半消化组织块培养法分离成骨细胞,进行传代培养,并通过形态观察、扫描电镜、碱性磷酸酶(AKP)和矿化结节染色进行鉴定.通过噻唑蓝(MTT)染色、AKP活性测定和钙化结节计数,研究不同浓度的氟对山羊成骨细胞增殖、分化及钙化的影响.结果表明,加氟48 h和96 h后,1.0×10-7～1.0×10-5 mol/L氟可促进成骨细胞增殖,其中1.0×10-7及1.0×10-6 mol/L氟与对照组差异极显著(P<0.01),高浓度氟(1.0×10-3 mol/L)则抑制成骨细胞增殖(P<0.01);而1.0×10-4及5.0×10-4 mol/L氟作用48 h对成骨细胞增殖无影响,作用96 h转而抑制成骨细胞增殖(P<0.01).1.0×10-7～1.0×10-5 mol/L氟能显著增强成骨细胞AKP活性及钙化能力(P<0.01),但氟浓度高于5.0×10-4 mol/L时,AKP活性及钙化能力明显下降(P<0.01).证实,低剂量氟能促进体外培养的山羊成骨细胞增殖分化及钙化,高剂量的氟则表现抑制作用.
The present study was designed to analyze regulations of fluoride on ruminant bone metabolism, and to elucidate processes and mechanisms of fluoride affecting caprine osteoblasts, which were isolated by means of tissue culture. On such a basis, osteoblasts were subcultured until they had come semiconfluent, and then identified by morphological observation, scanning electron microscopy, AKP detection and mineralized nodule stain. The regulations of different doses of fluoride on proliferation, differentiation and calcification of osteoblasts were measured respectively by the tetrazolium salt test, AKP activity detection and mineralized nodule count. After 48 or 96 h of exposure to fluoride at, 1.0×10^-7-1. 0×10^-5 mol/L of concentrations promoted osteoblast proliferation, and the role of promoting osteoblast proliferation was obvious with 1.0×10^7 and 1.0×10^-8 mol/L of fluoride（P〈0. 01） compared with the control. Higher concentration of fluoride with 1.0×10^-3 mol/L suppressed cell proliferation （P〈0.01） significantly. 1.0×10^-4 and 5.0× 10^-4 mol/L of fluoride had no effects on osteoblast proliferation after 48 h of exposure, but significant suppression was observed after 96 h（P〈0.01）. AKP activity and mineralization ability were increased obviously with 1.0×10^-7-1.0×10^-5 mol/L of fluoride（P〈0.01）, but decreased obviously with concentrations higher than 5.0×10^-4 mol/L of dose. It was concluded that lower dosage of fluoride can promote proliferation, differentiation and calcification, while higher dosage of fluoride had reverse effects on caprine osteoblasts cultured in vitro.
出处
《中国兽医科技》
CSCD
北大核心
2005年第11期895-899,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家自然科学基金项目(3010013530371065)
关键词
氟化钠
成骨细胞
增殖分化
钙化
山羊
sodium fluoride
osteoblast
proliferation and differentiation, calcification
caprine
