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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| Format: | Article |
| Language: | English |
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Dove Medical Press
2018-12-01
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| Series: | International Journal of Nanomedicine |
| Subjects: | |
| Online Access: | https://www.dovepress.com/a-flow-cytometric-analysis-of-macrophage-nanoparticle-interactions-in--peer-reviewed-article-IJN |
| _version_ | 1818993355602788352 |
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| author | Njoroge JM Yourick JJ Principato MA |
| author_facet | Njoroge JM Yourick JJ Principato MA |
| author_sort | Njoroge JM |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-20T20:40:44Z |
| format | Article |
| id | doaj.art-3eec6c3ef3d040d697e5f39308945ad4 |
| institution | Directory Open Access Journal |
| issn | 1178-2013 |
| language | English |
| last_indexed | 2024-12-20T20:40:44Z |
| publishDate | 2018-12-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | International Journal of Nanomedicine |
| spelling | doaj.art-3eec6c3ef3d040d697e5f39308945ad42022-12-21T19:27:08ZengDove Medical PressInternational Journal of Nanomedicine1178-20132018-12-01Volume 138365837842999A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expressionNjoroge JMYourick JJPrincipato MAJoyce 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 receptorhttps://www.dovepress.com/a-flow-cytometric-analysis-of-macrophage-nanoparticle-interactions-in--peer-reviewed-article-IJNSilver Nanoparticleflow cytometryinnate immunitymacrophagetoll receptor |
| spellingShingle | Njoroge JM Yourick JJ Principato MA A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression International Journal of Nanomedicine Silver Nanoparticle flow cytometry innate immunity macrophage toll receptor |
| title | 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_short | A flow cytometric analysis of macrophage–nanoparticle interactions in vitro: induction of altered Toll-like receptor expression |
| title_sort | flow cytometric analysis of macrophage ndash nanoparticle interactions in vitro induction of altered toll like receptor expression |
| topic | Silver Nanoparticle flow cytometry innate immunity macrophage toll receptor |
| url | https://www.dovepress.com/a-flow-cytometric-analysis-of-macrophage-nanoparticle-interactions-in--peer-reviewed-article-IJN |
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