期刊
SMALL
卷 6, 期 15, 页码 1669-1678出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/smll.201000528
关键词
cells; coatings; gold; nanoparticles; stereology
类别
资金
- Animal Free Research Foundation
- Doerenkamp-Zbinden Foundation
- Deutsche Forschungsgemeinschaft (DFG) [SPP 1313]
- Swiss National Science Foundation [3100A0, 118420]
- Helmholtz Association
This study addresses the cellular uptake and intracellular trafficking of 15-nm gold nanoparticles (NPs), either plain (Le., stabilized with citrate) or coated with polyethylene glycol (PEG), exposed to human alveolar epithelial cells (A549) at the air liquid interface for 1, 4, and 24h. Quantitative analysis by stereology on transmission electron microscopy images reveals a significant, nonrandom intracellular distribution for both NP types. No particles are observed in the nucleus, mitochondria, endoplasmic reticulum, or golgi. The cytosol is not a preferred cellular compartment for both NP types, although significantly more PEG-coated than citrate-stabilized NPs are present there The preferred particle localizations are vesicles of different sizes (<150, 150-1000, >1000 nm). This is observed for both NP types and indicates a predominant uptake by endocytosis. Subsequent inhibition of caveolin- and clathrin-mediated endocytosis by methyl-beta-cyclodextrin (M beta CD) results in a significant reduction of intracellular NPs. The inhibition, however, is more pronounced for PEG-coated than citrate-. stabilized NPs. The latter are mostly found in larger vesicles; therefore, they are potentially taken up by macropinocytosis. which is not inhibited by M beta CD). With prolonged exposure tunes, both NPs are preferentially localized in larger-sized intracellular vesicles such as lysosomes, thus indicating intracellular particle trafficking. This quantitative evaluation reveals that NP surface coatings modulate endocytotic uptake pathways and cellular NP trafficking. Other nonendocytotic entry mechanisms are found to be involved as well, as indicated by localization of a minority of PEG-coated NPs in the cytosol.
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