Quantifying size-dependent interactions between fluorescently labeled polystyrene nanoparticles and mammalian cells

Juan A Varela,Mariana G Bexiga,Kenneth A Dawson,Jeremy C Simpson,Christoffer Åberg

doi:10.1186/1477-3155-10-39

Juan A Varela, Mariana G Bexiga + Show 3 more

Open Access

https://doi.org/10.1186/1477-3155-10-39

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Abstract

BackgroundNanoparticles (NPs) are currently used in a wide variety of fields such as technology, medicine and industry. Due to the novelty of these applications and to ensure their success, a precise characterization of the interactions between NPs and cells is essential.FindingsThe current study explores the uptake of polystyrene NPs by 1321N1 human astrocytoma and A549 human lung carcinoma cell lines. In this work we show for the first time a comparison of the uptake rates of fluorescently labeled carboxylated polystyrene (PS) NPs of different sizes (20, 40 and 100 nm) in two different cell types, keeping the number of NPs per unit volume constant for all sizes. We propose a reliable methodology to control the dose of fluorescently labeled NPs, by counting individual NPs using automated particle detection from 3D confocal microscopy images. The possibility of detecting individual NPs also allowed us to calculate the size of each nanoparticle and compare the fluorescence of single NPs across different sizes, thereby providing a robust platform for normalization of NP internalization experiments as measured by flow cytometry.ConclusionsOur findings show that 40 nm NPs are internalized faster than 20 nm or 100 nm particles in both cell lines studied, suggesting that there is a privileged size gap in which the internalization of NPs is higher.

Highlights

Nanoparticles (NPs) are currently used in a wide variety of fields such as technology, medicine and industry
Our findings show that 40 nm NPs are internalized faster than 20 nm or 100 nm particles in both cell lines studied, suggesting that there is a privileged size gap in which the internalization of NPs is higher
The hydrodynamic size of the NPs in phosphate buffered saline (PBS) was measured by dynamic light scattering (DLS), revealing that they were well dispersed in all cases (Table 1), as all measurements presented a low polydispersity index

Summary

Conclusions

Our correlation between single particle tracking (SPT) analysis of NPs in solution and flow cytometry experiments measuring NP internalization proved to be a robust method to investigate bio-nano interactions. NP uptake rates were shown to differ between the two cell lines under study (Additional file 1: Figure S4) with the uptake rates being NP-size-dependent for each cell line. For both cell lines under study 40 nm NPs were internalized faster than 20 nm or 100 nm NPs, consistent with previous literature on gold nanoparticles [24]. Authors’ contributions JAV conceived, designed, performed and analyzed experiments and wrote the manuscript.

Background

Results and discussion