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Cytotoxicity, intracellular localization and exocytosis of citrate capped and PEG functionalized gold nanoparticles in human hepatocyte and kidney cells

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Abstract

Surface-modified gold nanoparticles (AuNPs) are nanomaterials that hold promise in drug delivery applications. In this study, the cytotoxicity, uptake, intracellular localization, and the exocytosis of citrate-stabilized (Cit-AuNP) and polyethylene glycol (PEG)-modified gold nanoparticles with the carboxyl (COOH) terminal functional group were assessed in human embryonic kidney (HEK 293) and the human caucasian hepatocytes carcinoma (Hep G2) cell systems, representing two major accumulation sites for AuNPs. The zeta (ζ)-potential measurements confirmed the negative surface charge of the AuNPs in water and in cell growth medium. The transmission electron microscopy confirmed the size and morphology of the AuNPs. Both types of AuNPs were shown to induce cytotoxic effects in cells. The Hep G2 cells were more sensitive cell type, with the COOH-PEG-AuNPs inducing the highest toxicity at higher concentrations. Dark field microscopy and TEM images revealed that the AuNPs were internalized in cells, mostly as agglomerates. TEM micrographs further revealed that the AuNPs were confined as agglomerates inside vesicle-like compartments, likely to be endosomal and lysosomal structures as well as in the cytosol, mostly as individual particles. The AuNPs were shown to remain in cellular compartments for up to 3 weeks, but thereafter, clearance of the gold nanoparticles from the cells by exocytosis was evident. The results presented in this study may therefore give an indication on the fate of AuNPs on long-term exposure to cells and may also assist in safety evaluation of AuNPs.

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Acknowledgments

This work is supported by the South African Department of Science and Technology (DST) and the National Research Foundation (NRF). The authors wish to thank Prof Jim Phillips from the Pathology Department at the National Institute for Occupational Health (NIOH) for the technical advice in preparing samples for TEM for assistance with acquiring the TEM images. We also thank the Immunology and Microbiology Department of the NIOH for sterility testing of the AuNPs.

Authors’ contributions

AuNPs were synthesized by FKK and AS. Characterization of the AuNPs was done by FKK and NT. xCELLigence, CytoViva, and TEM work was performed by NT. NT, MV, RT, and MG were involved in the inception and planning of the project and in the preparation of the manuscript. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. School of Pathology, Department of Molecular Medicine and Haematology, University of the Witwatersrand, P/Bag 3, Wits, 2050, South Africa

    Nonhlanhla Tlotleng, Melissa A. Vetten & Mary Gulumian

  2. Toxicology and Biochemistry Section, National Institute for Occupational Health, 25 Hospital Street, Constitutional Hill, Johannesburg, 2000, South Africa

    Nonhlanhla Tlotleng, Melissa A. Vetten & Mary Gulumian

  3. DST/Mintek Nanotechnology Innovation Centre Advanced Materials Division, Mintek, Private Bag X3015, Randburg, 2125, South Africa

    Frankline K. Keter, Amanda Skepu & Robert Tshikhudo

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  1. Nonhlanhla Tlotleng
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  2. Melissa A. Vetten
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  3. Frankline K. Keter
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  4. Amanda Skepu
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Correspondence to Mary Gulumian.

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Tlotleng, N., Vetten, M.A., Keter, F.K. et al. Cytotoxicity, intracellular localization and exocytosis of citrate capped and PEG functionalized gold nanoparticles in human hepatocyte and kidney cells. Cell Biol Toxicol 32, 305–321 (2016). https://doi.org/10.1007/s10565-016-9336-y

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  • DOI: https://doi.org/10.1007/s10565-016-9336-y

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