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目的探究纳米二氧化钛对小鼠成纤维(Balb/3T3)细胞的毒性作用。方法将处于对数生长期的Balb/3T3细胞暴露于含终浓度分别为0(对照)、1、10、50、100μg/ml纳米二氧化钛颗粒(粒径分别为30、50、100 nm)的DMEM完全培养基中培养24 h。采用CCK-8法测定细胞活性,通过测定细胞培养液上清中LDH的活力来检测细胞膜的完整性,并通过测定细胞内ROS含量来探讨细胞毒性的产生机制。结果与对照组比较,10~100μg/ml不同粒径的纳米二氧化钛染毒组Balb/c3T3细胞的存活率均较低,差异有统计学意义(P<0.05);且随着纳米二氧化钛染毒浓度的升高和纳米二氧化钛粒径的下降,Balb/c3T3细胞的存活率呈下降趋势。与对照组比较,10~100μg/ml不同粒径的纳米二氧化钛染毒组Balb/c3T3细胞培养液上清中LDH的活力和ROS的含量均较高,差异有统计学意义(P<0.05);且随着纳米二氧化钛染毒浓度的升高,Balb/c3T3细胞培养液上清中LDH的活力和ROS的含量均呈上升趋势。结论纳米二氧化钛对小鼠成纤维细胞的毒性作用呈明显的剂量效应和尺寸效应,纳米二氧化钛诱导Balb/c3T3细胞产生大量活性氧(ROS)可能是导致其产生细胞毒性的主要原因。
Objective To investigate the toxic effects of nano-titanium dioxide on Balb / 3T3 cells in mice. Methods Balb / 3T3 cells in logarithmic growth phase were exposed to DMEM containing 0, 1, 10, 50, and 100 μg / ml nanoparticle titania particles (30, 50 and 100 nm, respectively) Complete medium for 24 h. The cell viability was measured by CCK-8 assay. The integrity of cell membrane was measured by measuring the activity of LDH in the supernatant of cell culture medium. The mechanism of cytotoxicity was explored by measuring intracellular ROS content. Results Compared with the control group, the survival rates of Balb / c3T3 cells in 10 ~ 100μg / ml nano-TiO2 with different particle size were lower than those in the control group (P <0.05), and with the concentration of nano-TiO2 Of Balb / c3T3 cells decreased with the decrease of particle size of nano-TiO2. Compared with the control group, the activity of LDH and the content of ROS in Balb / c3T3 cell culture supernatant of 10 ~ 100μg / ml nano - titania exposure group were higher than that of the control group, the difference was statistically significant (P <0.05); And with the concentration of nano-titanium dioxide increased, the activities of LDH and ROS in Balb / c3T3 cell culture supernatant increased. CONCLUSION: The toxic effect of nano-TiO2 on mouse fibroblasts shows obvious dose-effect and size-effect. Nano-TiO2 can induce a large amount of reactive oxygen species (ROS) in Balb / c3T3 cells, which may be the main reason of its cytotoxicity.