Abstract

BackgroundNanoparticle (NPs) functionalization has been shown to affect their cellular toxicity. To study this, differently functionalized silver (Ag) and gold (Au) NPs were synthesised, characterised and tested using lung epithelial cell systems.MethodsMonodispersed Ag and Au NPs with a size range of 7 to 10 nm were coated with either sodium citrate or chitosan resulting in surface charges from −50 mV to +70 mV. NP-induced cytotoxicity and oxidative stress were determined using A549 cells, BEAS-2B cells and primary lung epithelial cells (NHBE cells). TEER measurements and immunofluorescence staining of tight junctions were performed to test the growth characteristics of the cells. Cytotoxicity was measured by means of the CellTiter-Blue ® and the lactate dehydrogenase assay and cellular and cell-free reactive oxygen species (ROS) production was measured using the DCFH-DA assay.ResultsDifferent growth characteristics were shown in the three cell types used. A549 cells grew into a confluent mono-layer, BEAS-2B cells grew into a multilayer and NHBE cells did not form a confluent layer. A549 cells were least susceptible towards NPs, irrespective of the NP functionalization. Cytotoxicity in BEAS-2B cells increased when exposed to high positive charged (+65-75 mV) Au NPs. The greatest cytotoxicity was observed in NHBE cells, where both Ag and Au NPs with a charge above +40 mV induced cytotoxicity. ROS production was most prominent in A549 cells where Au NPs (+65-75 mV) induced the highest amount of ROS. In addition, cell-free ROS measurements showed a significant increase in ROS production with an increase in chitosan coating.ConclusionsChitosan functionalization of NPs, with resultant high surface charges plays an important role in NP-toxicity. Au NPs, which have been shown to be inert and often non-cytotoxic, can become toxic upon coating with certain charged molecules. Notably, these effects are dependent on the core material of the particle, the cell type used for testing and the growth characteristics of these cell culture model systems.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-014-0062-4) contains supplementary material, which is available to authorized users.

Highlights

  • Nanoparticle (NPs) functionalization has been shown to affect their cellular toxicity

  • Cell development Understanding the growth characteristics of the cell types used in this study is important in order to fully comprehend the observed responses to NPs insult

  • A different growth pattern was observed for BEAS-2B cells, which were shown to grow on top of each other and formed multilayers which contained functional tight junctions (Figure 1d)

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Summary

Introduction

Nanoparticle (NPs) functionalization has been shown to affect their cellular toxicity. Differently functionalized silver (Ag) and gold (Au) NPs were synthesised, characterised and tested using lung epithelial cell systems. Silver (Ag) and gold (Au) NPs of similar size were synthesised with different coatings to provide 4 classes of surface charge ranging from −50 mV to +70 mV. One class of negatively charged NPs, Schlinkert et al Journal of Nanobiotechnology (2015) 13:1 of special interest, as chitosan-coated NPs are increasingly used in the field of nanobiotechnology. These type of functionalized NPs are very promising drug delivery systems [13,14], due to their low toxicity, high stability and biocompatibility [15]. The inhalation of NPs appears to be a promising method for the delivery of drugs to the lung [19,20]

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