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目的 :观察氟化钠对体外培养的人牙周膜细胞增殖及矿化能力的影响,为氟添加入牙周组织工程药物中的应用提供依据。方法:原代培养并鉴定人牙周膜细胞,应用CCK8检测不同浓度Na F对h PDLCs增殖的影响,并筛选出4个浓度用于矿化实验。矿化条件下,将0、1×10~(-5)、5×10~(-4)和1×10~(-3) mol/L的Na F作用h PDLCs后,通过碱性磷酸酶(ALP)染色、茜素红染色和实时荧光定量PCR检测矿化能力及成骨相关基因的表达。采用SPSS20.0软件包对数据进行单因素方差分析。结果:5×10~(-5)、1×10~(-4)、5×10~(-4) mol/L的Na F均能促进h PDLCs增殖,且以5×10~(-4) mol/L效果最佳(P<0.05)。而1×10~(-5) mol/L的Na F碱性磷酸酶染色阳性面积最大、茜素红染色矿化结节数量最多(P<0.05)。RT-PCR结果根据时间、指标变化程度较大。结论:5×10~(-5)、1×10~(-4)、5×10~(-4) mol/L的Na F能促进h PDLCs的增殖能力,1×10~(-5) mol/L的Na F能提高h PDLCs的碱性磷酸酶活性及钙结节形成。
OBJECTIVE: To observe the effect of sodium fluoride on the proliferation and mineralization ability of cultured human periodontal ligament cells in vitro, and provide the basis for the application of fluoride addition to periodontal tissue engineering drugs. Methods: Primary human periodontal ligament cells were cultured and identified. The effects of different concentrations of NaF on the proliferation of h PDLCs were detected by CCK8, and four concentrations were selected for mineralization experiments. Under the conditions of mineralization, hPLCs were treated with 0,1 × 10 -5, 5 × 10 -4 and 1 × 10 -3 mol / L Na F, and then treated with alkaline phosphatase (ALP) staining, alizarin red staining and real-time fluorescence quantitative PCR were used to detect the mineralization ability and the expression of osteogenesis-related genes. Data were analyzed by one-way ANOVA using SPSS20.0 software package. Results: Na F at 5 × 10 ~ (-5), 1 × 10 ~ (-4) and 5 × 10 ~ (-4) mol / L all promoted the proliferation of h PDLCs. ) mol / L was the best (P <0.05). The positive area of NaF alkaline phosphatase staining was the largest at 1 × 10 ~ (-5) mol / L, and the number of mineralized nodules by alizarin red staining was the highest (P <0.05). RT-PCR results based on time, a larger degree of change indicators. CONCLUSION: Na F at 5 × 10 -5, 1 × 10 -4 and 5 × 10 -4 mol / L can promote the proliferation of h PDLCs, NaF at mol / L increased alkaline phosphatase activity and calcium nodule formation in h PDLCs.