Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages.

Kuhn, Dagmar A; Vanhecke, Dimitri; Michen, Benjamin; Blank, Fabian; Gehr, Peter; Petri-Fink, Alke; Rothen-Rutishauser, Barbara (2014). Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages. Beilstein journal of nanotechnology, 5, pp. 1625-1636. Beilstein-Institut zur Förderung der Chemischen Wissenschaften 10.3762/bjnano.5.174

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Precise knowledge regarding cellular uptake of nanoparticles is of great importance for future biomedical applications. Four different endocytotic uptake mechanisms, that is, phagocytosis, macropinocytosis, clathrin- and caveolin-mediated endocytosis, were investigated using a mouse macrophage (J774A.1) and a human alveolar epithelial type II cell line (A549). In order to deduce the involved pathway in nanoparticle uptake, selected inhibitors specific for one of the endocytotic pathways were optimized regarding concentration and incubation time in combination with fluorescently tagged marker proteins. Qualitative immunolocalization showed that J774A.1 cells highly expressed the lipid raft-related protein flotillin-1 and clathrin heavy chain, however, no caveolin-1. A549 cells expressed clathrin heavy chain and caveolin-1, but no flotillin-1 uptake-related proteins. Our data revealed an impeded uptake of 40 nm polystyrene nanoparticles by J774A.1 macrophages when actin polymerization and clathrin-coated pit formation was blocked. From this result, it is suggested that macropinocytosis and phagocytosis, as well as clathrin-mediated endocytosis, play a crucial role. The uptake of 40 nm nanoparticles in alveolar epithelial A549 cells was inhibited after depletion of cholesterol in the plasma membrane (preventing caveolin-mediated endocytosis) and inhibition of clathrin-coated vesicles (preventing clathrin-mediated endocytosis). Our data showed that a combination of several distinguishable endocytotic uptake mechanisms are involved in the uptake of 40 nm polystyrene nanoparticles in both the macrophage and epithelial cell line.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Gastro-intestinal, Liver and Lung Disorders (DMLL) > Clinic of Pneumology
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50 > Forschungsgruppe Pneumologie (Erwachsene)

UniBE Contributor:

Blank, Fabian; Gehr, Peter and Rothen-Rutishauser, Barbara

Subjects:

600 Technology > 610 Medicine & health

ISSN:

2190-4286

Publisher:

Beilstein-Institut zur Förderung der Chemischen Wissenschaften

Language:

English

Submitter:

Rahel Holderegger

Date Deposited:

10 Mar 2015 12:57

Last Modified:

10 Mar 2015 12:57

Publisher DOI:

10.3762/bjnano.5.174

PubMed ID:

25383275

Uncontrolled Keywords:

cell lines, endocytosis, inhibition, nanoparticles, uptake proteins

BORIS DOI:

10.7892/boris.64402

URI:

https://boris.unibe.ch/id/eprint/64402

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