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Deciphering the mechanisms of cellular uptake of engineered nanoparticles by accurate evaluation of internalization using imaging flow cytometry.

hal.structure.identifierUnité de Biologie Fonctionnelle et Adaptative [BFA (UMR_8251 / U1133)]
dc.contributor.authorVRANIC, Sandra
hal.structure.identifierInstitut Jacques Monod [IJM (UMR_7592)]
dc.contributor.authorBOGGETTO, Nicole
hal.structure.identifierInstitut Jacques Monod [IJM (UMR_7592)]
dc.contributor.authorCONTREMOULINS, Vincent
hal.structure.identifierInstitut de Chimie de la Matière Condensée de Bordeaux [ICMCB]
dc.contributor.authorMORNET, Stéphane
hal.structure.identifierInstitut de Chimie de la Matière Condensée de Bordeaux [ICMCB]
dc.contributor.authorREINHARDT, Nora
hal.structure.identifierUnité de Biologie Fonctionnelle et Adaptative [BFA (UMR_8251 / U1133)]
dc.contributor.authorMARANO, Francelyne
hal.structure.identifierUnité de Biologie Fonctionnelle et Adaptative [BFA (UMR_8251 / U1133)]
dc.contributor.authorBAEZA-SQUIBAN, Armelle
hal.structure.identifierUnité de Biologie Fonctionnelle et Adaptative [BFA (UMR_8251 / U1133)]
dc.contributor.authorBOLAND, Sonja
dc.date.issued2013
dc.identifier.issn1743-8977
dc.description.abstractEnBACKGROUND: The uptake of nanoparticles (NPs) by cells remains to be better characterized in order to understand the mechanisms of potential NP toxicity as well as for a reliable risk assessment. Real NP uptake is still difficult to evaluate because of the adsorption of NPs on the cellular surface.RESULTS: Here we used two approaches to distinguish adsorbed fluorescently labeled NPs from the internalized ones. The extracellular fluorescence was either quenched by Trypan Blue or the uptake was analyzed using imaging flow cytometry. We used this novel technique to define the inside of the cell to accurately study the uptake of fluorescently labeled (SiO2) and even non fluorescent but light diffracting NPs (TiO2). Time course, dose-dependence as well as the influence of surface charges on the uptake were shown in the pulmonary epithelial cell line NCI-H292. By setting up an integrative approach combining these flow cytometric analyses with confocal microscopy we deciphered the endocytic pathway involved in SiO2 NP uptake. Functional studies using energy depletion, pharmacological inhibitors, siRNA-clathrin heavy chain induced gene silencing and colocalization of NPs with proteins specific for different endocytic vesicles allowed us to determine macropinocytosis as the internalization pathway for SiO2 NPs in NCI-H292 cells.CONCLUSION: The integrative approach we propose here using the innovative imaging flow cytometry combined with confocal microscopy could be used to identify the physico-chemical characteristics of NPs involved in their uptake in view to redesign safe NPs.
dc.language.isoen
dc.publisherBioMed Central
dc.subject.enLung epithelial cells
dc.subject.enImageStreamX
dc.subject.enEndocytosis
dc.subject.enClathrin
dc.subject.enMacropinocytosis
dc.subject.enTiO2
dc.subject.enSiO2
dc.title.enDeciphering the mechanisms of cellular uptake of engineered nanoparticles by accurate evaluation of internalization using imaging flow cytometry.
dc.typeArticle de revue
dc.identifier.doi10.1186/1743-8977-10-2
dc.subject.halSciences du Vivant [q-bio]/Toxicologie
dc.subject.halSciences du Vivant [q-bio]/Biologie cellulaire/Interactions cellulaires [q-bio.CB]
bordeaux.journalParticle and Fibre Toxicology
bordeaux.page2
bordeaux.volume10
bordeaux.peerReviewedoui
hal.identifierhal-00813978
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-00813978v1
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