Abstract
Developing methods that evaluate the cellular uptake of magnetic nanoparticles (MNPs) and nanotoxicity effects at single-cellular level are needed. In this study, magnetophoresis combining fluorescence based cytotoxicity assay was proposed to assess the viability and the single-cellular MNPs uptake simultaneously. Malignant cells (SKHep-1, HepG2, HeLa) were incubated with 10 nm anionic iron oxide nanoparticles. Prussian blue stain was performed to visualize the distribution of magnetic nanoparticles. MTT and fluorescence based assay analyzed the cytotoxicity effects of the bulk cell population and single cell, respectively. DAPI/PI stained was applied to evaluate death mechanism. The number of intracellular MNPs was found to be strongly correlated with the cell death. Significant differences between cellular MNP uptake in living and dead cells were observed. The method could be useful for future study of the nanotoxicity induced by MNPs.
Highlights
Nanoparticles (NPs) have raised a broad interest in the field of material science or medicine [1], [2], [3], [4]
The main process involved in magnetic NPs (MNPs) internalization is endocytosis [14], which is subjected to cell-to-cell variations [15]; nanotoxicity effects induced by MNPs should be analyzed more accurately on individual cells instead of masking by the average value of bulk measurements
Prussian blue staining To verify the intracellular localizations of MNPs, the cells treated with MNPs were washed with phosphate-buffered saline (PBS) for three times and fixed in 4% paraformaldehyde
Summary
Nanoparticles (NPs) have raised a broad interest in the field of material science or medicine [1], [2], [3], [4]. Cells must be labeled with large amounts of magnetic NPs (MNPs) in order to be manipulated by the above techniques, but may suffer from nanotoxicity. Methods that assess the cellular uptake of MNPs and nanotoxicity effects at single-cell level are more reasonable than those for bulk-cell assays. Most of the current methods evaluate the internalized iron oxide MNP from a population of cells rather than a single cell. Those techniques including Prussian blue staining [16], T2 relaxometry [17], UV/VIS spectrometry [18], [19], and atomic absorption spectroscopy (AAS) [20], [21] analyze the cells that are non-viable. Later in 2008, Jing et al showed that magnetophoresis could analyze cells that remain active and reveal the differences of uptake capacities between individual cells [23]
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