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The Effect of Surface Charges on the Cellular Uptake of Liposomes Investigated by Live Cell Imaging

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Abstract

Purpose

Liposomes have been developed as versatile nanocarriers for various pharmacological agents. The effect of surface charges on the cellular uptake of the liposomes has been studied by various methods using mainly fixed cells with inevitable limitations. Live cell imaging has been proposed as an alternative methods to overcome the limitations of the fixed cell-based analysis. In this study, we aimed to investigate the effects of surface charges on cellular association and internalization of the liposomes using live cell imaging.

Methods

We studied the cellular association and internalization of liposomes with different surface charge using laser scanning confocal microscopy (LSCM) equipped with live cell chamber system. Flow cytometry was also carried out using flow cytometer (FACS) for comparison.

Results

All of the cationic, neutral and anionic liposomes showed time-dependent cellular uptake through specific endocytic pathways. In glioblastoma U87MG cells, the cationic and anionic liposomes were mainly taken up via macropinocytosis, while the neutral liposomes mainly via caveolae-mediated endocytosis. In fibroblast NIH/3T3 cells, all of the three liposomes entered into the cell via clathrin-mediated endocytosis.

Conclusions

This study provides a better understanding on the cellular uptake mechanisms of the liposomes, which could contribute significantly to development of liposome-based drug delivery systems.

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Abbreviations

CD:

Cytochalasin D

CPZ:

Chlorpromazine

DIC:

Differential interference contrast

DLS:

Dynamic light scattering

DMEM:

Dulbecco’s modified eagle’s medium

FACS:

Flow cytometer

GS:

Genistein

HBG:

HEPES buffered glucose (10 mM HEPES, 5% glucose, pH 7.4)

HBS:

HEPES buffered saline (10 mM HEPES, 150 mM NaCl, pH 7.4)

LSCM:

Laser scanning confocal microscopy

PBS:

Phosphate buffered saline

PDI:

Polydispersity index

PEG-PE:

1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N-[Methoxy(Polyethylene glycol)-2000]

POPC:

1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphocholine

POPG:

1-Palmitoyl-2-Oleoyl-sn-Glycero-3-[Phospho-rac-91-glycerol]

PTRF:

Polymerase I and transcript release factor

Rh-PE:

1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl)

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ACKNOWLEDGMENTS AND DISCLOSURES

This research was supported by the Basic Science Research Program (NRF2014R1A1A2053373) and the Pioneer Research Center Program (NRF2014M3C1A3054153) of the National Research Foundation of Korea, funded by the Ministry of Education, Science and Technology.

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    Authors and Affiliations

    1. College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University, 191 Hambakmoe-ro, Yeonsu-gu, Incheon, South Korea, 21936

      Ji Hee Kang, Woo Young Jang & Young Tag Ko

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    1. Ji Hee Kang
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    2. Woo Young Jang
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    3. Young Tag Ko
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    Correspondence to Young Tag Ko.

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    Ji Hee Kang and Woo Young Jang contributed equally to this work.

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    Kang, J.H., Jang, W.Y. & Ko, Y.T. The Effect of Surface Charges on the Cellular Uptake of Liposomes Investigated by Live Cell Imaging. Pharm Res 34, 704–717 (2017). https://doi.org/10.1007/s11095-017-2097-3

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