Polystyrene nanoparticle trafficking across MDCK-II

Fazlollahi, Farnoosh; Angelow, Susanne; Yacobi, Nazanin R.; Marchelletta, Ronald R.; Yu, Alan; Borok, Zea; Kim, Kwang‐Jin; Crandall, Edward D.

doi:10.1016/j.nano.2011.01.008

Cited by 59 publications

(50 citation statements)

References 33 publications

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“…Although it is possible to image live cells, it is difficult to gain detailed information about subcellular structures due to the limited spatial resolution. Nanoparticles which have been labeled by fluorescent dyes have been extensively studied and Confocal Laser Scanning Microscopy (CLSM) is an invaluable tool for the visualization of nanoparticles for in vitro studies [19][20][21][22] . By the use of these fluorescent based techniques, fluorescent emmission of nanoparticles and the subcellular location of nanoparticles have been determined.…”

Section: Intracellular Trafficking Of Nanoparticles Ros Production Amentioning

confidence: 99%

Determination of nanoparticle localisation within subcellular organelles in vitro using Raman spectroscopy

et al. 2015

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Ease of sample preparation, narrow spectral bandwidth and minimal influence from water are features of Raman spectroscopy which make it a powerful, label-free way to study a wide range of biological structures and phenomena. In this context, given the concerns over their toxicology arising from their increased production and use, evaluation of nanoparticle uptake and localisation in biological systems and determination of the mechanisms of subcellular interaction and trafficking can provide long-term solutions for nanotoxicology, and potential strategies for nanomedicine. In this study, Raman spectroscopy is explored to monitor the sequential trafficking of nanoparticles through subcellular organelles in-vitro and to establish the spectroscopic signatures of those organelles. A549 human lung carcinoma cells were exposed to 40 nm carboxylate-modified and fluorescently-labelled polystyrene nanoparticles for 4, 12 and 24hrs. Raman spectroscopy was applied to nanoparticle exposed cells to determine the localisation within cellular compartments. Confocal laser scanning fluorescence microscopy (CLSM) with different organelle staining kits confirmed the localisation of the nanoparticles in organelles at the chosen exposure periods and colocalization was quantified using ImageJ with the JACoP colocalisation plugin. To confirm nanoparticle localisation and elucidate the spectroscopic signatures of the different subcellular organelles, a combination of K-means clustering (KMCA) and Principal components analysis (PCA) was applied to the Raman spectroscopic maps. The study showed the applicability of the techniques for elucidation of the localisation of polystyrene nanoparticles within the cell as well as determination of their local environment, differentiating the spectral signatures of intracellular compartments such as endosomes, lysosomes and endoplasmic reticulum, in a completely label free manner.

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Section: Intracellular Trafficking Of Nanoparticles Ros Production Amentioning

confidence: 99%

Determination of nanoparticle localisation within subcellular organelles in vitro using Raman spectroscopy

et al. 2015

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“…We think this is a reasonable result because cationic nanoparticles have been reported to strongly interact with the negatively charged region of the cell membrane surface via electrostatic forces. 36,38) In addition, we observed that the cellular associated amount of magnetic liposomes at 37°C was much higher than that at 4°C, and the highest quantitative change between 37 and 4°C was observed with magnetic cationic liposomes (Fig. 3).…”

Section: Discussionmentioning

confidence: 79%

Influence of Physicochemical Properties and PEG Modification of Magnetic Liposomes on Their Interaction with Intestinal Epithelial Caco-2 Cells

Kono

Jinzai

Kotera

et al. 2017

Biological & Pharmaceutical Bulletin

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The present study aimed to investigate the effect of particle size (100, 500 nm), surface charge (cationic, neutral and anionic) and polyethylene glycol (PEG) modification of magnetic liposomes on their interaction with the human intestinal epithelial cell line, Caco-2. The cellular associated amount of all the magnetic liposomes was significantly increased by the presence of a magnetic field. The highest association and internalization into Caco-2 cells was observed with magnetic cationic liposomes. Moreover, small magnetic liposomes were more efficiently associated and taken up into the cells, than large ones. In contrast, PEG modification significantly attenuated the enhancing effect of the magnetic field on the cellular association of magnetic liposomes. We also found that magnetic cationic liposomes had the highest retention properties to Caco-2 cells. Moreover, the retention of large magnetic liposomes to the cells was much longer than that of small ones. In addition, magnetic cationic and neutral liposomes had relatively high stability in Caco-2 cells, whereas magnetic anionic liposomes rapidly degraded. These results indicate that the physicochemical properties and PEG modification of magnetic liposomes greatly influences their intestinal epithelial transport.

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“…We have reported that translocation of polystyrene nanoparticles across RAECM and Madin Darby canine kidney type II cell monolayers (MDCK-II) was not decreased in the presence of 10-200 µM methyl-β-cyclodextrin. 30,32 Clathrin is composed of three light and three heavy chains that form a triskelion. Assembly of the triskelion leads to formation of a net-like basket (clathrin-coated pit) at the cell plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

“…To explore if quantum dot translocation across RAECM involves endocytotic mechanisms, methyl-β-cyclodextrin, chlorpromazine, and dynasore were used to disrupt lipid raft-mediated endocytosis, 30,32,36,37 clathrin-mediated endocytosis, 30,38 and dynamin-dependent endocytosis (including clathrin-mediated and caveolin-mediated endocytosis), 30,32,39 respectively. Briefly, RAECM were pretreated for 30 minutes with methyl-β-cyclodextrin (200 µM, Sigma), chlorpromazine (28 µM, Sigma), or dynasore (80 µM, Sigma) in both apical and basolateral fluids.…”

Section: Effects Of Endocytosis Inhibitorsmentioning

confidence: 99%

“…[24][25][26][27][28][29] It is currently thought that both the size and surface properties of nanoparticles affect the cytotoxicity and intracellular fate(s) of nanoparticles. [30][31][32][33] In this regard, we recently reported the trafficking properties of polystyrene nanoparticles with variable sizes and surface charges across RAECM. 30,31 In brief, trafficking rates of polystyrene nanoparticles across RAECM are dependent on both surface charge density and size, in that we found an approximately 20-40 times greater flux for positively (versus negatively) charged polystyrene nanoparticles of similar size and that larger polystyrene nanoparticles (about 100 nm) cross RAECM about 3-4 times slower than smaller polystyrene nanoparticles (20 nm) of similar surface charge.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Translocation of PEGylated quantum dots across rat alveolar epithelial cell monolayers

Fazlollahi¹,

Sipos²,

Kim³

et al. 2011

IJN

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Background: In this study, primary rat alveolar epithelial cell monolayers (RAECM) were used to investigate transalveolar epithelial quantum dot trafficking rates and underlying transport mechanisms. Methods: Trafficking rates of quantum dots (PEGylated CdSe/ZnS, core size 5.3 nm, hydrodynamic size 25 nm) in the apical-to-basolateral direction across RAECM were determined. Changes in bioelectric properties (ie, transmonolayer resistance and equivalent active ion transport rate) of RAECM in the presence or absence of quantum dots were measured. Involvement of endocytic pathways in quantum dot trafficking across RAECM was assessed using specific inhibitors (eg, methyl-β-cyclodextrin, chlorpromazine, and dynasore for caveolin-, clathrin-, and dynamin-mediated endocytosis, respectively). The effects of lowering tight junctional resistance on quantum dot trafficking were determined by depleting Ca 2+ in apical and basolateral bathing fluids of RAECM using 2 mM EGTA. Effects of temperature on quantum dot trafficking were studied by lowering temperature from 37°C to 4°C. Results: Apical exposure of RAECM to quantum dots did not elicit changes in transmonolayer resistance or ion transport rate for up to 24 hours; quantum dot trafficking rates were not surface charge-dependent; methyl-β-cyclodextrin, chlorpromazine, and dynasore did not decrease quantum dot trafficking rates; lowering of temperature decreased transmonolayer resistance by approximately 90% with a concomitant increase in quantum dot trafficking by about 80%; and 24 hours of treatment of RAECM with EGTA decreased transmonolayer resistance by about 95%, with increased quantum dot trafficking of up to approximately 130%. Conclusion: These data indicate that quantum dots do not injure RAECM and that quantum dot trafficking does not appear to take place via endocytic pathways involving caveolin, clathrin, or dynamin. We conclude that quantum dot translocation across RAECM takes place via both transcellular and paracellular pathways and, based on comparison with our prior studies, interactions of nanoparticles with RAECM are strongly dependent on nanoparticle composition and surface properties.

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Polystyrene nanoparticle trafficking across MDCK-II

Cited by 59 publications

References 33 publications

Determination of nanoparticle localisation within subcellular organelles in vitro using Raman spectroscopy

Determination of nanoparticle localisation within subcellular organelles in vitro using Raman spectroscopy

Influence of Physicochemical Properties and PEG Modification of Magnetic Liposomes on Their Interaction with Intestinal Epithelial Caco-2 Cells

Translocation of PEGylated quantum dots across rat alveolar epithelial cell monolayers

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