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Cytotoxicity, Cellular Uptake And Apoptotic Responses In Human Coronary Artery Endothelial Cells Exposed To Ultrasmall Superparamagnetic Iron Oxide Nanoparticles

Teresa Palacios-Hernandez, Daysi M Diaz-Diestra, A. Nguyen, Shelby A. Skoog, Bhaskara Vijaya Chikkaveeraiah, X. Tang, Y. Wu, P. Petrochenko, Eric M. Sussman, P. Goering
Published 2020 · Chemistry, Medicine

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Ultrasmall superparamagnetic iron oxide nanoparticles (USPION) possess reactive surfaces, are metabolized and exhibit unique magnetic properties. These properties are desirable for designing novel theranostic biomedical products; however, toxicity mechanisms of USPION are not completely elucidated. The goal of this study was to investigate cell interactions (uptake and cytotoxicity) of USPION using human coronary artery endothelial cells as a vascular cell model. Polyvinylpirrolidone‐coated USPION were characterized: average diameter 17 nm (transmission electron microscopy [TEM]), average hydrodynamic diameter 44 nm (dynamic light scattering) and zeta potential −38.75 mV. Cells were exposed to 0 (control), 25, 50, 100 or 200 μg/mL USPION. Concentration‐ and time‐dependent cytotoxicity were observed after 3‐6 hours through 24 hours of exposure using Alamar Blue and Real‐Time Cell Electronic Sensing assays. Cell uptake was evaluated by imaging using live‐dead confocal microscopy, actin and nuclear fluorescent staining, and TEM. Phase‐contrast, confocal microscopy, and TEM imaging showed significant USPION internalization as early as 3 hours after exposure to 25 μg/mL. TEM imaging demonstrated particle internalization in secondary lysosomes with perinuclear localization. Three orthogonal assays were conducted to assess apoptosis. TUNEL staining demonstrated a marked increase in fragmented DNA, a response pathognomonic of apoptosis, after a 4‐hour exposure. Cells subjected to agarose gel electrophoresis exhibited degraded DNA 3 hours after exposure. Caspase‐3/7 activity increased after a 3‐hour exposure. USPION uptake resulted in cytotoxicity involving apoptosis and these results contribute to further mechanistic understanding of the USPION toxicity in vitro in cardiovascular endothelial cells.
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