Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: A quantitative method for oxidative stress assessment of nanoparticle-treated cells
Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: A quantitative method for oxidative stress assessment of nanoparticle-treated cells
复制标题
DOI：
10.1016/j.tiv.2013.01.016
复制
发表时间：
2013-03-01
影响因子：
3.2
通讯作者：
Gombau, L.
中科院分区：
文献类型：
--
作者：
Aranda, A.;Sequedo, L.;Gombau, L.
关于如何解决纳米材料可能的毒性问题，目前还没有达成共识，因为它们会干扰大多数基于比色和荧光探针的体外筛选试验，例如用于检测氧化物质的二氯二氢荧光素二乙酸酯（DCFH-DA）试验。研究了纳米材料与DCFH-DA的相互作用与其性质和/或测定条件的关系（基于细胞和时间暴露），通过孵育罗丹明（Rhd）标记的25 nm和50 nm二氧化硅（SiO2）、裸的和油酸涂覆的磁铁矿、（Fe 3 O 4）和磁赤铁矿（Fe 2 O3）氧化铁，二氧化钛（TiO 2）和聚（环氧乙烷）-聚（丙交酯/乙交酯）酸（PLGA-PEO）纳米颗粒（NP）与代谢活性大鼠肝细胞4和24小时的时间。数据表明，纳米颗粒摄取与染料荧光发射的淬灭相关。尽管具有掩蔽作用，但当暴露时间比本测试常用的时间更长时，可以在有限的阈值浓度下检测到NP的氧化潜力。然而，在实验条件的变化并没有系统地导致自由基的形成为所有nanomaterials tested.Overall数据表明，尽管淬灭效应的纳米粒子DCFH-DA测定，它可以被认为是一个有用的工具，定量测量的NP诱导的氧化应激的标准化协议的微小修改。(C)2013爱思唯尔有限公司保留所有权利。
No consensus exists on how to address possible toxicity of nanomaterials as they interfere with most in vitro screening tests based on colorimetric and fluorimetric probes such as the dichloro-dihydrofluorescein diacetate (DCFH-DA) assay for detection of oxidative species.In the present research, nanomaterial interaction with DCFH-DA was studied in relation to its nature and/or assay conditions (cell-based and time exposure) by incubating Rhodamine (Rhd)-labeled 25 nm and 50 nm silica (SiO2), naked and oleic acid coated magnetite, (Fe3O4) and maghemite (Fe2O3) iron oxide, titanium dioxide (TiO2) and poly(ethylene oxide)-poly(lactide/glycolide) acid (PLGA-PEO) nanoparticles (NPs) with metabolically active rat hepatocytes for 4 and 24-h periods. Data indicated that nanoparticle uptake correlated with quenching of dye fluorescence emission. In spite of their masking effect, the oxidative potential of NPs could be detected at a limited threshold concentration when exposed for periods of time longer than those frequently used for this test. However, changes in the experimental conditions did not systematically result in free radical formation for all nanomaterials tested.Overall data indicate that despite the quenching effect of nanoparticles on DCFH-DA assay, it can be considered as a useful tool for quantitative measurement of NPs-induced oxidative stress by minor modifications of standardized protocols. (C) 2013 Elsevier Ltd. All rights reserved.