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Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials
Engineered nanomaterials have been found to induce oxidative stress. Cellular oxidative stress, in turn, can result in the induction of antioxidant and detoxification enzymes which are controlled by the nuclear erythroid 2-related factor 2 (NRF2) transcription factor. Here, we present the results of...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9511406/ https://www.ncbi.nlm.nih.gov/pubmed/36171865 http://dx.doi.org/10.3389/ftox.2022.974429 |
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author | Martin, Sebastin de Haan, Laura Miro Estruch, Ignacio Eder, Kai Moritz Marzi, Anne Schnekenburger, Jürgen Blosi, Magda Costa, Anna Antonello, Giulia Bergamaschi, Enrico Riganti, Chiara Beal, David Carrière, Marie Taché, Olivier Hutchison, Gary Malone, Eva Young, Lesley Campagnolo, Luisa La Civita, Fabio Pietroiusti, Antonio Devineau, Stéphanie Baeza, Armelle Boland, Sonja Zong, Cai Ichihara, Gaku Fadeel, Bengt Bouwmeester, Hans |
author_facet | Martin, Sebastin de Haan, Laura Miro Estruch, Ignacio Eder, Kai Moritz Marzi, Anne Schnekenburger, Jürgen Blosi, Magda Costa, Anna Antonello, Giulia Bergamaschi, Enrico Riganti, Chiara Beal, David Carrière, Marie Taché, Olivier Hutchison, Gary Malone, Eva Young, Lesley Campagnolo, Luisa La Civita, Fabio Pietroiusti, Antonio Devineau, Stéphanie Baeza, Armelle Boland, Sonja Zong, Cai Ichihara, Gaku Fadeel, Bengt Bouwmeester, Hans |
author_sort | Martin, Sebastin |
collection | PubMed |
description | Engineered nanomaterials have been found to induce oxidative stress. Cellular oxidative stress, in turn, can result in the induction of antioxidant and detoxification enzymes which are controlled by the nuclear erythroid 2-related factor 2 (NRF2) transcription factor. Here, we present the results of a pre-validation study which was conducted within the frame of BIORIMA (“biomaterial risk management”) an EU-funded research and innovation project. For this we used an NRF2 specific chemically activated luciferase expression reporter gene assay derived from the human U2OS osteosarcoma cell line to screen for the induction of the NRF2 mediated gene expression following exposure to biomedically relevant nanobiomaterials. Specifically, we investigated Fe(3)O(4)-PEG-PLGA nanomaterials while Ag and TiO(2) “benchmark” nanomaterials from the Joint Research Center were used as reference materials. The viability of the cells was determined by using the Alamar blue assay. We performed an interlaboratory study involving seven different laboratories to assess the applicability of the NRF2 reporter gene assay for the screening of nanobiomaterials. The latter work was preceded by online tutorials to ensure that the procedures were harmonized across the different participating laboratories. Fe(3)O(4)-PEG-PLGA nanomaterials were found to induce very limited NRF2 mediated gene expression, whereas exposure to Ag nanomaterials induced NRF2 mediated gene expression. TiO(2) nanomaterials did not induce NRF2 mediated gene expression. The variability in the results obtained by the participating laboratories was small with mean intra-laboratory standard deviation of 0.16 and mean inter laboratory standard deviation of 0.28 across all NRF2 reporter gene assay results. We conclude that the NRF2 reporter gene assay is a suitable assay for the screening of nanobiomaterial-induced oxidative stress responses. |
format | Online Article Text |
id | pubmed-9511406 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95114062022-09-27 Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials Martin, Sebastin de Haan, Laura Miro Estruch, Ignacio Eder, Kai Moritz Marzi, Anne Schnekenburger, Jürgen Blosi, Magda Costa, Anna Antonello, Giulia Bergamaschi, Enrico Riganti, Chiara Beal, David Carrière, Marie Taché, Olivier Hutchison, Gary Malone, Eva Young, Lesley Campagnolo, Luisa La Civita, Fabio Pietroiusti, Antonio Devineau, Stéphanie Baeza, Armelle Boland, Sonja Zong, Cai Ichihara, Gaku Fadeel, Bengt Bouwmeester, Hans Front Toxicol Toxicology Engineered nanomaterials have been found to induce oxidative stress. Cellular oxidative stress, in turn, can result in the induction of antioxidant and detoxification enzymes which are controlled by the nuclear erythroid 2-related factor 2 (NRF2) transcription factor. Here, we present the results of a pre-validation study which was conducted within the frame of BIORIMA (“biomaterial risk management”) an EU-funded research and innovation project. For this we used an NRF2 specific chemically activated luciferase expression reporter gene assay derived from the human U2OS osteosarcoma cell line to screen for the induction of the NRF2 mediated gene expression following exposure to biomedically relevant nanobiomaterials. Specifically, we investigated Fe(3)O(4)-PEG-PLGA nanomaterials while Ag and TiO(2) “benchmark” nanomaterials from the Joint Research Center were used as reference materials. The viability of the cells was determined by using the Alamar blue assay. We performed an interlaboratory study involving seven different laboratories to assess the applicability of the NRF2 reporter gene assay for the screening of nanobiomaterials. The latter work was preceded by online tutorials to ensure that the procedures were harmonized across the different participating laboratories. Fe(3)O(4)-PEG-PLGA nanomaterials were found to induce very limited NRF2 mediated gene expression, whereas exposure to Ag nanomaterials induced NRF2 mediated gene expression. TiO(2) nanomaterials did not induce NRF2 mediated gene expression. The variability in the results obtained by the participating laboratories was small with mean intra-laboratory standard deviation of 0.16 and mean inter laboratory standard deviation of 0.28 across all NRF2 reporter gene assay results. We conclude that the NRF2 reporter gene assay is a suitable assay for the screening of nanobiomaterial-induced oxidative stress responses. Frontiers Media S.A. 2022-09-05 /pmc/articles/PMC9511406/ /pubmed/36171865 http://dx.doi.org/10.3389/ftox.2022.974429 Text en Copyright © 2022 Martin, de Haan, Miro Estruch, Eder, Marzi, Schnekenburger, Blosi, Costa, Antonello, Bergamaschi, Riganti, Beal, Carrière, Taché, Hutchison, Malone, Young, Campagnolo, La Civita, Pietroiusti, Devineau, Baeza, Boland, Zong, Ichihara, Fadeel and Bouwmeester. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Toxicology Martin, Sebastin de Haan, Laura Miro Estruch, Ignacio Eder, Kai Moritz Marzi, Anne Schnekenburger, Jürgen Blosi, Magda Costa, Anna Antonello, Giulia Bergamaschi, Enrico Riganti, Chiara Beal, David Carrière, Marie Taché, Olivier Hutchison, Gary Malone, Eva Young, Lesley Campagnolo, Luisa La Civita, Fabio Pietroiusti, Antonio Devineau, Stéphanie Baeza, Armelle Boland, Sonja Zong, Cai Ichihara, Gaku Fadeel, Bengt Bouwmeester, Hans Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title | Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title_full | Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title_fullStr | Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title_full_unstemmed | Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title_short | Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
title_sort | pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials |
topic | Toxicology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9511406/ https://www.ncbi.nlm.nih.gov/pubmed/36171865 http://dx.doi.org/10.3389/ftox.2022.974429 |
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