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Cause-and-Effect Analysis as a Tool To Improve the Reproducibility of Nanobioassays: Four Case Studies.
Chemical Research in Toxicology ( IF 3.7 ) Pub Date : 2019-10-03 , DOI: 10.1021/acs.chemrestox.9b00165
Elijah J Petersen 1 , Cordula Hirsch 2 , John T Elliott 1 , Harald F Krug 3 , Leonie Aengenheister 2 , Ali Talib Arif 4, 5 , Alessia Bogni 6 , Agnieszka Kinsner-Ovaskainen 6 , Sarah May 2 , Tobias Walser 7 , Peter Wick 2 , Matthias Roesslein 2
Affiliation  

One of the challenges in using in vitro data to understand the potential risks of engineered nanomaterials (ENMs) is that results often differ or are even contradictory among studies. While it is recognized that numerous factors can influence results produced by nanobioassays, there has not yet been a consistently used conceptual framework to identify key sources of variability in these assays. In this paper, we use cause-and-effect analysis to systematically describe sources of variability in four key in vitro nanobioassays: the 2',7'-dichlorofluorescein assay, an enzyme-linked immunosorbent assay for measuring interleukin-8, a flow cytometry assay (Annexin V/propidium iodide), and the Comet assay. These assays measure end points that can occur in cells impacted by ENMs through oxidative stress, a principle mechanism for ENM toxicity. The results from this analysis identify control measurements to test for potential artifacts or biases that could occur during conduct of these assays with ENMs. Cause-and-effect analysis also reveals additional measurements that could be performed either in preliminary experiments or each time the assay is run to increase confidence in the assay results and their reproducibility within and among laboratories. The approach applied here with these four assays can be used to support the development of a broad range of nanobioassays.

中文翻译:

因果分析作为提高纳米生物测定可重复性的工具:四个案例研究。

使用体外数据来了解工程纳米材料(ENM)的潜在风险的挑战之一是研究之间的结果常常不同甚至矛盾。尽管人们认识到有许多因素会影响纳米生物测定法产生的结果,但尚未有一致使用的概念框架来确定这些测定法中变异的关键来源。在本文中,我们使用因果分析系统地描述了四个关键的体外纳米生物测定中的变异性来源:2',7'-二氯荧光素测定,用于测量白介素-8的酶联免疫吸附测定,流式细胞术分析法(Annexin V /碘化丙啶)和彗星分析法。这些测定法可测量ENM通过氧化应激(ENM毒性的一种主要机理)而受到ENM影响的细胞中可能发生的终点。此分析的结果确定了对照测量值,以测试在使用ENM进行这些测定过程中可能发生的潜在伪像或偏差。因果分析还揭示了可以在初步实验中或每次运行测定时都可以执行的其他测量,以增加对测定结果及其在实验室内部和实验室之间的可重复性的信心。这四种检测方法在此处应用的方法可用于支持广泛的纳米生物检测方法的开发。因果分析还揭示了可以在初步实验中或每次运行测定时都可以执行的其他测量,以增加对测定结果及其在实验室内部和实验室之间的可重复性的信心。这四种检测方法在此处应用的方法可用于支持广泛的纳米生物检测方法的开发。因果分析还揭示了可以在初步实验中或每次运行测定时都可以执行的其他测量,以增加对测定结果及其在实验室内部和实验室之间的可重复性的信心。这四种检测方法在此处应用的方法可用于支持广泛的纳米生物检测方法的开发。
更新日期:2019-09-11
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