TY - JOUR
T1 - Cause-and-effect analysis as a tool to improve the reproducibility of nanobioassays
T2 - four case studies
AU - Petersen, Elijah J.
AU - Hirsch, Cordula
AU - Elliott, John T.
AU - Krug, Harald F.
AU - Aengenheister, Leonie
AU - Arif, Ali Talib
AU - Bogni, Alessia
AU - Kinsner-Ovaskainen, Agnieszka
AU - May, Sarah
AU - Walser, Tobias
AU - Wick, Peter
AU - Roesslein, Matthias
N1 - Publisher Copyright:
© 2019 American Chemical Society.
PY - 2020/5/18
Y1 - 2020/5/18
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85073260065&partnerID=8YFLogxK
U2 - 10.1021/acs.chemrestox.9b00165
DO - 10.1021/acs.chemrestox.9b00165
M3 - Article
C2 - 31507156
AN - SCOPUS:85073260065
SN - 0893-228X
VL - 33
SP - 1039
EP - 1054
JO - Chemical Research in Toxicology
JF - Chemical Research in Toxicology
IS - 5
ER -