【摘要】 目的:研究低浓度的H2O2对人牙周膜细胞(human periodontal ligament cells,h PDLCs)增殖及成骨分化的影响。方法:采用组织块法培养h PDLCs,免疫细胞化学染色鉴定其来源;采用不同浓度的H2O2处理h PDLCs,CCK-8法检测细胞的增殖活性;并对处理后的h PDLCs进行成骨诱导,碱性磷酸酶检测试剂盒测定细胞的碱性磷酸酶(ALP)活性,茜素红染色法(ARS)观察细胞矿化结节的形成情况。结果:采用组织块法成功培养出h PDLCs,细胞抗波形丝蛋白染色阳性,抗角蛋白染色阴性; CCK-8结果显示:低浓度的H2O2(20～300μM)可提高h PDLCs的增殖活性,更低浓度(0～10μM)和较高浓度(500μM)对细胞增殖活性无影响,而更高浓度(700μM)则抑制细胞的增殖活性; ALP活性及ARS染色半定量结果分析显示:适当浓度范围的H2O2(10～80μM)可提高细胞ALP活性及增加矿化结节形成量,而当浓度上升至100～300 m M时,细胞的ALP活性降低,矿化结节形成量减少。结论:H2O2对h PDLCs的增殖及分化的影响存在剂量效应,一定低浓度范围的H2O2可提高h PDLCs的增殖活性并促进其成骨分化。更多还原

【Abstract】 Objective: To observe the effect of low concentrations of hydrogen peroxide（ H₂O₂） of proliferation and osteogenic differentiation on human periodontal ligament cells（ h PDLCs）. Methods: The primary human periodontal ligament cells were cultured from periodontal ligament tissues. The immunocytochemical staining was used to identify cell sources. The CCK-8 assay was used to determine the proliferation curve of h PDLCs treated by different concentrations of H₂O₂. Alkaline phosphatase（ ALP） activity kit and Alizarin Red S staining（ ARS） were performed to assess osteogenic differentiation administrated H₂O₂. Results: The primary h PDLCs were obtained,and the immunocytochemical staining showed that the anti-vimentin staining of h PDLCs was positive while the anti-keratin staining was negative. The CCK-8 assay manifested that low concentrations（ 20 ～ 300 μM） of H₂O₂ could improve the proliferation of h PDLCs,while lower concentrations（ 0 ～ 10 μM） and high concentration（ 50 μM） of H₂O₂ had no influence on the proliferation of h PDLCs,and higher concentration（ 700 μM） decreased the proliferation of h PDLCs. The ALP activity assay results indicated that the ALP activity was enhanced when the concentrations of H₂O₂ were between 10 and 80 μM,and the degree of mineralization was also enhanced within these concentrations according to the ARS staining. But when the concentrations of H₂O₂ were between 100 and 300 μM,both the ALP activity and the degree of mineralization were dropped. Conclusion: H₂O₂ had a dose effect on the proliferation and osteogenic differentiation of h PDLCs,and the certain low concentrations of H₂O₂ could enhance both the proliferation activity and osteogenic differentiation of h PDLCs.更多还原