A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression

A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression

Joyce M Njoroge, Jeffrey J Yourick, Mary Ann PrincipatoDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, US Food and Drug Administration, Laurel, MD 20708, USA Background: Nanoparticles exhibit unique physiochemical chara...

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Autores principales: Njoroge JM, Yourick JJ, Principato MA
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2018
Materias:
Silver Nanoparticle
flow cytometry
innate immunity
macrophage
toll receptor
Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/3eec6c3ef3d040d697e5f39308945ad4
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spelling oai:doaj.org-article:3eec6c3ef3d040d697e5f39308945ad42021-12-02T05:06:40ZA flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression1178-2013https://doaj.org/article/3eec6c3ef3d040d697e5f39308945ad42018-12-01T00:00:00Zhttps://www.dovepress.com/a-flow-cytometric-analysis-of-macrophage-nanoparticle-interactions-in--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Joyce M Njoroge, Jeffrey J Yourick, Mary Ann PrincipatoDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, US Food and Drug Administration, Laurel, MD 20708, USA Background: Nanoparticles exhibit unique physiochemical characteristics that provide the basis for their utilization. The diversity of potential and actual applications compels a thorough understanding regarding the consequences of their containment within the cellular environment.Purpose: This paper presents a flow cytometric examination of the biologic effects associated with the internalization of citrate-buffered silver (Ag) nanoparticles (NP) by the murine macrophage cell line, RAW264.7.Materials and methods: Cells were cultured with varying concentrations of citrate-buffered Ag nanoparticle and analyzed for changes in cellular volume, fluorescence emissions, and surface receptor expression.Results: Notable changes in side scatter (SSC) signal occurred following the phagocytosis of citrate-buffered Ag NP representative of the 10 nm, 50 nm, and 100 nm particle size by cultured RAW 264.7 cells. A characteristic associated with the internalization of all the citrated Ag NP sizes tested, was the detection of emitted infra-red and near-infrared wavelength emissions. This characteristic consistently permitted the detection of 10 nm, 50 nm, and 100 nm Ag NP particles internalized within the RAW cells by flow cytometry. A functional distinction between monocyte subsets within the RAW 264.7 cell line was noted as Ag NP are taken up by the F4/80+ subset of cells within the culture. Further, the internalization of Ag NP by the cells resulted in an increased cell surface expression of the Toll-like Receptor (TLR) 3, but not TLR4.Conclusion: Taken together, these results implicate the more mature macrophage in the ingestion of Ag NP; and an influence upon at least one of the Toll receptors present in macrophages following exposure to Ag NP. Further, our flow cytometric approach presents a potentially viable detection method for the identification of occult Ag NP material using an indicator cell line. Keywords: silver nanoparticle, flow cytometry, innate immunity, macrophage, Toll-like receptorNjoroge JMYourick JJPrincipato MADove Medical PressarticleSilver Nanoparticleflow cytometryinnate immunitymacrophagetoll receptorMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 13, Pp 8365-8378 (2018)
institution DOAJ
collection DOAJ
language EN
topic Silver Nanoparticle
flow cytometry
innate immunity
macrophage
toll receptor
Medicine (General)
R5-920
spellingShingle Silver Nanoparticle
flow cytometry
innate immunity
macrophage
toll receptor
Medicine (General)
R5-920
Njoroge JM
Yourick JJ
Principato MA
A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
description Joyce M Njoroge, Jeffrey J Yourick, Mary Ann PrincipatoDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, US Food and Drug Administration, Laurel, MD 20708, USA Background: Nanoparticles exhibit unique physiochemical characteristics that provide the basis for their utilization. The diversity of potential and actual applications compels a thorough understanding regarding the consequences of their containment within the cellular environment.Purpose: This paper presents a flow cytometric examination of the biologic effects associated with the internalization of citrate-buffered silver (Ag) nanoparticles (NP) by the murine macrophage cell line, RAW264.7.Materials and methods: Cells were cultured with varying concentrations of citrate-buffered Ag nanoparticle and analyzed for changes in cellular volume, fluorescence emissions, and surface receptor expression.Results: Notable changes in side scatter (SSC) signal occurred following the phagocytosis of citrate-buffered Ag NP representative of the 10 nm, 50 nm, and 100 nm particle size by cultured RAW 264.7 cells. A characteristic associated with the internalization of all the citrated Ag NP sizes tested, was the detection of emitted infra-red and near-infrared wavelength emissions. This characteristic consistently permitted the detection of 10 nm, 50 nm, and 100 nm Ag NP particles internalized within the RAW cells by flow cytometry. A functional distinction between monocyte subsets within the RAW 264.7 cell line was noted as Ag NP are taken up by the F4/80+ subset of cells within the culture. Further, the internalization of Ag NP by the cells resulted in an increased cell surface expression of the Toll-like Receptor (TLR) 3, but not TLR4.Conclusion: Taken together, these results implicate the more mature macrophage in the ingestion of Ag NP; and an influence upon at least one of the Toll receptors present in macrophages following exposure to Ag NP. Further, our flow cytometric approach presents a potentially viable detection method for the identification of occult Ag NP material using an indicator cell line. Keywords: silver nanoparticle, flow cytometry, innate immunity, macrophage, Toll-like receptor
format article
author Njoroge JM
Yourick JJ
Principato MA
author_facet Njoroge JM
Yourick JJ
Principato MA
author_sort Njoroge JM
title A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
title_short A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
title_full A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
title_fullStr A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
title_full_unstemmed A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression
title_sort flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered toll-like receptor expression
publisher Dove Medical Press
publishDate 2018
url https://doaj.org/article/3eec6c3ef3d040d697e5f39308945ad4
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