Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Mukherjee, Sourav P., Lozano, Neus, Kucki, Melanie, Del Rio-Castillo, Antonio E., Newman, Leon, Vázquez, Ester, Kostarelos, Kostas, Wick, Peter, Fadeel, Bengt
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
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
_version_ 1782469306303381504
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
work_keys_str_mv AT mukherjeesouravp detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT lozanoneus detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT kuckimelanie detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT delriocastilloantonioe detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT newmanleon detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT vazquezester detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT kostareloskostas detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT wickpeter detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction
AT fadeelbengt detectionofendotoxincontaminationofgraphenebasedmaterialsusingthetnfaexpressiontestandguidelinesforendotoxinfreegrapheneoxideproduction