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Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production
Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5120825/ https://www.ncbi.nlm.nih.gov/pubmed/27880838 http://dx.doi.org/10.1371/journal.pone.0166816 |
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author | Mukherjee, Sourav P. Lozano, Neus Kucki, Melanie Del Rio-Castillo, Antonio E. Newman, Leon Vázquez, Ester Kostarelos, Kostas Wick, Peter Fadeel, Bengt |
author_facet | Mukherjee, Sourav P. Lozano, Neus Kucki, Melanie Del Rio-Castillo, Antonio E. Newman, Leon Vázquez, Ester Kostarelos, Kostas Wick, Peter Fadeel, Bengt |
author_sort | Mukherjee, Sourav P. |
collection | PubMed |
description | Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin detection in graphene based material (GBM) samples, including graphene oxide (GO) and few-layered graphene (FLG). Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET) using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay. FLG also triggered TNF-α production in the presence of the LPS inhibitor, pointing to an intrinsic pro-inflammatory effect. Finally, we present guidelines for the preparation of endotoxin-free GO, validated by using the TET. |
format | Online Article Text |
id | pubmed-5120825 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-51208252016-12-15 Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production Mukherjee, Sourav P. Lozano, Neus Kucki, Melanie Del Rio-Castillo, Antonio E. Newman, Leon Vázquez, Ester Kostarelos, Kostas Wick, Peter Fadeel, Bengt PLoS One Research Article Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin detection in graphene based material (GBM) samples, including graphene oxide (GO) and few-layered graphene (FLG). Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET) using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay. FLG also triggered TNF-α production in the presence of the LPS inhibitor, pointing to an intrinsic pro-inflammatory effect. Finally, we present guidelines for the preparation of endotoxin-free GO, validated by using the TET. Public Library of Science 2016-11-23 /pmc/articles/PMC5120825/ /pubmed/27880838 http://dx.doi.org/10.1371/journal.pone.0166816 Text en © 2016 Mukherjee et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Mukherjee, Sourav P. Lozano, Neus Kucki, Melanie Del Rio-Castillo, Antonio E. Newman, Leon Vázquez, Ester Kostarelos, Kostas Wick, Peter Fadeel, Bengt Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title | Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title_full | Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title_fullStr | Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title_full_unstemmed | Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title_short | Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production |
title_sort | detection of endotoxin contamination of graphene based materials using the tnf-α expression test and guidelines for endotoxin-free graphene oxide production |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5120825/ https://www.ncbi.nlm.nih.gov/pubmed/27880838 http://dx.doi.org/10.1371/journal.pone.0166816 |
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