a Department of Biology , Faculty of Sciences, Akdeniz University , Antalya , Turkey.
J Toxicol Environ Health A. 2013;76(16):990-1002. doi: 10.1080/15287394.2013.830584.
In this study a genotoxic evaluation of titanium dioxide (TiO2, 2.3 nm), zirconium oxide (ZrO2, 6 nm), aluminum oxide (Al2O3, 16.7 nm) nanoparticles (NP) and their ionic forms was conducted using human peripheral blood lymphocytes and cultured human embryonic kidney (HEK293) cells by means of a modified alkaline comet assay with/without the formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease III (Endo III) enzymes. Modifications to the comet assay by using lesion-specific endonucleases, such as Endo III and Fpg, detect DNA bases with oxidative damage. Both human peripheral blood lymphocytes and cultured embryonic kidney cells were incubated with TiO2, ZrO2, or Al2O3 NP at concentrations of 1, 10, or 100 μg/ml. Our results showed no significant induction in DNA damage by the comet assay with/without the Endo III and Fpg enzymes at all concentrations of ZrO2 and Al2O3. In the case of TiO2 NP only the highest concentration of 100 μg/ml significantly induced a genotoxic response. Data thus indicate that both ZrO2 and Al2O3 NP were not genotoxic in our system and in the case of TiO2 the lowest-observed-adverse-effect level (LOAEL) for genotoxicity was 100 μg/ml. Evidence indicates that these metallic NP are considered safe in light of the fact that no genotoxicity was noted with ZrO2 and Al2O3 and that the highest TiO2 concentration is not environmentally relevant.
本研究采用改良碱性彗星试验,结合/不结合甲酸嘧啶-DNA 糖基化酶(Fpg)和内切酶 III(Endo III),对二氧化钛(TiO2,2.3nm)、氧化锆(ZrO2,6nm)、氧化铝(Al2O3,16.7nm)纳米颗粒(NP)及其离子形式进行了遗传毒性评价,分别用人外周血淋巴细胞和培养的人胚肾(HEK293)细胞进行实验。通过使用内切酶 III 和 Fpg 等损伤特异性内切酶对彗星试验进行改良,可以检测到具有氧化损伤的 DNA 碱基。将 TiO2、ZrO2 或 Al2O3 NP 分别以 1、10 或 100μg/ml 的浓度孵育人外周血淋巴细胞和培养的胚胎肾细胞。结果显示,在所有浓度的 ZrO2 和 Al2O3 下,彗星试验结合/不结合 Endo III 和 Fpg 酶均未诱导 DNA 损伤。对于 TiO2 NP,仅最高浓度 100μg/ml 显著诱导遗传毒性反应。数据表明,在我们的系统中,ZrO2 和 Al2O3 NP 均无遗传毒性,而对于 TiO2,遗传毒性的最低观察到有害效应水平(LOAEL)为 100μg/ml。有证据表明,鉴于 ZrO2 和 Al2O3 未观察到遗传毒性,并且 TiO2 的最高浓度与环境无关,这些金属 NP 被认为是安全的。