不同固化模式下的自粘接型树脂水门汀体外细胞毒性研究

Cytotoxic effects of two self-adhesive resin cements under different curing modes in vitro

目的评价自粘接型树脂水门汀在双重或化学固化模式下的浸提液对人牙周膜成纤维细胞的毒性作用。方法制作2种自粘接型树脂水门汀(Biscem和Rely X U200)圆盘状试件,经过双重或化学固化后,避光保存24 h。使用改良Eagle完全培养基制取试件的浸提液,滤菌后按照1∶5、1∶10、1∶20、1∶40、1∶80(体积分数)进行梯度稀释作为实验组,未含浸提液的改良Eagle完全培养基组作为对照组。各组浸提液与人牙周膜成纤维细胞共培养24 h后,使用倒置显微镜观察细胞形态及数量变化,通过Cell Counting Kit-8试剂盒检测各组的细胞增殖率,利用膜联蛋白V/碘化丙啶双染流式细胞术对细胞的凋亡/坏死情况进行检测。使用单因素方差分析对实验数据进行统计学分析。结果在不同的稀释浓度下,2种树脂水门汀化学固化下的细胞增殖率均显著低于双重固化,其中Biscem化学固化的细胞增殖率显著低于Rely X U200的(P<0.05);而在双重固化下两者间无显著差异(P>0.05)。2种树脂水门汀化学固化下的细胞凋亡/坏死率均显著高于其双重固化下的,其中Biscem双重和化学固化下的细胞凋亡/坏死率均显著高于Rely X U200(P<0.05)。结论自粘接型树脂水门汀种类及其固化模式能够影响人牙周膜成纤维细胞的增殖率及凋亡率。

Objective To evaluate the potential cytotoxic effects of two self-adhesive resin cements（ Biscem and Rely X U200） on human periodontal ligament fibroblasts（ HPDLFs） under dual or chemical curing mode. Methods Disc-shaped specimens of two selfadhesive resin cements were prepared under dual or chemical curing mode and incubated for 24 hours in the dark. Then specimens were immersed in complete DMEM（ Dulbecco＇s Modified Eagle Medium,supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin） for 24 hours（ 37 ℃ and 5% CO2）. The extract solution was diluted accordingly to 1 ∶ 5,1 ∶ 10,1 ∶ 20,1 ∶ 40,1 ∶ 80 with complete DMEM after filtration. The diluted solutions were considered as experimental groups and complete DMEM without specimen immersed was used as negative control. All the groups were co-cultured with human periodontal ligament fibroblasts for 24 hours.Then the morphology and amount of cells were observed under the microscope. The cell proliferation rates were detected by Cell Counting Kit-8. Besides,the cell apoptosis was measured by Annexin V-FITC and PI staining using a flow cytometry-based method. The data was statistically analyzed by one-way ANOVA at a significance level of P0.05. Results The cell proliferation rates of two resin cements under chemical curing mode were significantly lower than that under dual curing mode at any concentration（ P0.05）. Besides,the cell proliferation rates of Biscem under chemical curing mode were significantly lower than the value of Rely X U200 under chemical curing mode（ P0.05）. Nevertheless,under dual curing mode,no significant difference was found between Biscem and Rely X U200（ P0.05）. Two resin cements both showed that the apoptosis rates under chemical curing mode were significantly higher than the value under dual curing mode（ P0.05）. And the apoptosis rates of Biscem were significantly higher than Rely X U200 regardless of curing mode（ P0.05）. Conclusions The cell proliferation rate and apoptosis rate of human periodontal ligament fibroblasts can be influenced by the type of self-adhesive resin cements and their curing mode.