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The toxicity outcome of silica nanoparticles (Ludox®) is influenced by testing techniques and treatment modalities

机译:二氧化硅纳米颗粒(Ludox®)的毒性结果受测试技术和治疗方式的影响

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摘要

We analyzed the influence of the kind of cytotoxicity test and its application modality in defining the level of hazard of the in vitro exposures to nanostructures. We assessed the cytotoxicity induced by two different Ludox® silica nanoparticles (NPs), AS30 and SM30, on three human cell lines, CCD-34Lu, A549, and HT-1080. Dynamic light scattering measurements showed particle agglomeration when NPs are diluted in culture medium supplemented with fetal calf serum. We examined the impact of such particle aggregation on the cytotoxicity by exposing the cells to NPs under different treatment modalities: short incubation (2 h) in serum-free medium or long incubation (24–72 h) in serum-containing medium. Under this last modality, NP suspensions tended to form aggregates and were toxic at concentrations five- to tenfold higher than in serum-free medium. The results of cell survival varied considerably when the long-term clonogenic assay was performed to validate the data of the short-term MTS assay. Indeed, the half maximum effective concentrations (EC50) in all the three cell lines were four- to fivefold lower when calculated from the data of clonogenic assay than of MTS. Moreover, the mechanisms of NP toxicity were cell-type-specific, showing that CCD-34Lu are prone to the induction of plasma membrane damages and HT-1080 are prone to DNA double-strand break and apoptosis induction. Taken together, our results demonstrate that the choice of testing strategy and treatment conditions plays an important role in assessing the in vitro toxicity of NPs.>Figure 
机译:我们分析了细胞毒性试验的种类及其应用方式对确定体外暴露于纳米结构的危害程度的影响。我们评估了两种不同的Ludox®二氧化硅纳米颗粒(NPs)AS30和SM30对三种人类细胞系CCD-34Lu,A549和HT-1080诱导的细胞毒性。动态光散射测量显示,当在添加了胎牛血清的培养基中稀释NP时,颗粒会团聚。我们通过在不同处理方式下将细胞暴露于NP来检查此类颗粒聚集对细胞毒性的影响:在无血清培养基中短暂孵育(2小时)或在含血清培养基中长期孵育(24–72小时)。在这种最后的方式下,NP悬浮液倾向于形成聚集体,并且在比无血清培养基中高五到十倍的浓度下具有毒性。当进行长期克隆形成测定以验证短期MTS测定的数据时,细胞存活的结果差异很大。实际上,从克隆形成试验的数据计算得出,所有三种细胞系的半数最大有效浓度(EC50)比MTS低四到五倍。此外,NP毒性的机制是特定于细胞类型的,表明CCD-34Lu易于诱导质膜损伤,而HT-1080则易于DNA双链断裂和凋亡诱导。两者合计,我们的结果表明,选择测试策略和治疗条件在评估NP的体外毒性中起着重要作用。<!-fig ft0-> <!-fig @ position =“ anchor” mode =文章f4-> <!-fig mode =“ anchored” f5-> > Figure <!-fig / graphic | fig / alternatives / graphic mode =“ anchored” m1-> <!-标题a7->

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