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为研究氟化钠对体外培养的大鼠成骨细胞增殖和分化的影响,用混合酶消化法分离24h内新生清洁级SD大鼠颅盖骨成骨细胞,进行原代培养。采用倒置相差显微镜观察成骨细胞的形态,采用Gomori改良钙钴法对成骨细胞进行碱性磷酸酶(ALP)染色。向1×103个/孔成骨细胞中接种加入0.0、2.5、5.0、10.0、20.0、40.0、60.0、80.0、100.0、120.0mg/L的氟化钠,培养1～3d,采用噻唑蓝(MTT)检测增殖活力。向1×103个/孔成骨细胞中接种加入0.0、2.5、5.0、10.0、20.0、40.0、60.0、80.0、100.0、120.0mg/L的氟化钠,培养3、5、7、9d时,采用PNPP法检测碱性磷酸酶活力。结果显示,氟化钠作用于成骨细胞第2天时,2.5、5.0mg/L的氟化钠对成骨细胞增殖活力有明显的促进作用(P<0.05);当氟化钠≥80.0mg/L时,可明显抑制成骨细胞的增殖(P<0.05)。染毒第9天,氟化钠≥40.0mg/L时对成骨细胞ALP活力有明显的抑制作用,差异均有统计学意义(P<0.05)。提示低剂量氟化钠有轻微促进成骨细胞增殖的作用,高剂量氟化钠则能抑制成骨细胞增殖和ALP的活力。
In order to study the effect of sodium fluoride on proliferation and differentiation of cultured rat osteoblasts in vitro, the calvarial osteoblasts of neonatal cleansing grade SD rats were isolated by mixed enzyme digestion and cultured in primary culture. Osteoblast morphology was observed by inverted phase contrast microscope. Alkaline phosphatase (ALP) staining of osteoblasts was performed by Gomori modified calcium and cobalt method. To 1 × 103 osteoblasts were inoculated with 0.0,2.5,5.0,10.0,20.0,40.0,60.0,80.0,100.0,120.0mg / L of sodium fluoride, cultured 1 ~ 3d, using thiazolyl blue (MTT ) To detect proliferative activity. To 1 × 103 osteoblasts were inoculated with 0.0,2.5,5.0,10.0,20.0,40.0,60.0,80.0,100.0,120.0mg / L of sodium fluoride, cultured 3,5,7,9 d, Alkaline phosphatase activity was detected by PNPP method. The results showed that when sodium fluoride was added to osteoblasts on day 2, sodium fluoride at concentrations of 2.5 and 5.0 mg / L significantly promoted the proliferation activity of osteoblasts (P <0.05); when sodium fluoride was more than or equal to 80.0 mg / L significantly inhibited the proliferation of osteoblasts (P <0.05). On the 9th day of exposure, the ALP activity of osteoblasts was significantly inhibited when sodium fluoride≥40.0mg / L, the difference was statistically significant (P <0.05). Tip low-dose sodium fluoride slightly promote osteoblast proliferation, high-dose sodium fluoride can inhibit osteoblast proliferation and ALP vitality.