Abstract
BackgroundNanoparticles have been used for biomedical applications, including drug delivery, diagnosis, and imaging based on their unique properties derived from small size and large surface-to-volume ratio. However, concerns regarding unexpected toxicity due to the localization of nanoparticles in the cells are growing. Herein, we quantified the number of cell-internalized nanoparticles and monitored their cellular localization, which are critical factors for biomedical applications of nanoparticles.MethodsThis study investigates the intracellular trafficking of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate dye [MNPs@SiO2(RITC)] in various live single cells, such as HEK293, NIH3T3, and RAW 264.7 cells, using site-specific direct stochastic optical reconstruction microscopy (dSTORM). The time-dependent subdiffraction-limit spatial resolution of the dSTORM method allowed intracellular site-specific quantification and tracking of MNPs@SiO2(RITC).ResultsThe MNPs@SiO2(RITC) were observed to be highly internalized in RAW 264.7 cells, compared to the HEK293 and NIH3T3 cells undergoing single-particle analysis. In addition, MNPs@SiO2(RITC) were internalized within the nuclei of RAW 264.7 and HEK293 cells but were not detected in the nuclei of NIH3T3 cells. Moreover, because of the treatment of the MNPs@SiO2(RITC), more micronuclei were detected in RAW 264.7 cells than in other cells.ConclusionThe sensitive and quantitative evaluations of MNPs@SiO2(RITC) at specific sites in three different cells using a combination of dSTORM, transcriptomics, and molecular biology were performed. These findings highlight the quantitative differences in the uptake efficiency of MNPs@SiO2(RITC) and ultra-sensitivity, varying according to the cell types as ascertained by subdiffraction-limit super-resolution microscopy.Graphical
Highlights
Nanoparticles have been used for biomedical applications, including drug delivery, diagnosis, and imaging based on their unique properties derived from small size and large surface-to-volume ratio
Our research focused on the internalization of magnetic nanoparticles (MNPs)@SiO2(RITC) in macrophage cell lines, a comparison was made with HEK293 and NIH3T3 cell lines to form a better understanding of the sensitivity of internalization
The resulting super-resolution images obtained with direct stochastic optical reconstruction microscopy (dSTORM) enabled individual MNPs@SiO2(RITC) in the aggregated region to be distinguished and provided improved resolution over the results obtained from conventional wide-field total internal reflection fluorescence (TIRF) images
Summary
Nanoparticles have been used for biomedical applications, including drug delivery, diagnosis, and imaging based on their unique properties derived from small size and large surface-to-volume ratio. We quantified the number of cell-internalized nanoparticles and monitored their cellular localization, which are critical factors for biomedical applications of nanoparticles. Engineered magnetic nanoparticles (MNPs), because of their inert chemical composition and ease of modification, serve as important nanomedicine probes for the potent delivery of genes and drugs to cells [1]. MNPs and MNPs coated with biocompatible compounds are widely studied for uses in drug delivery, as contrast agents in magnetic resonance imaging (MRI), and in cell labeling [1,2,3]. The MNPs@SiO2(RITC) have novel applications as drug and gene nanocarriers, MRI contrast agents, and in antibody engineering. Precise quantitative estimation of cellular NP uptake and the consequent internalization of NPs at the intracellular sites is needed to support the use of MNPs for highprofile medical applications. The quantitative estimation of such drug carriers at intracellular sites is essential for determining the pharmacokinetics, that is, the molecules delivered by native carriers and their respective cellular effects
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