Multifactorial determinants that govern nanoparticle uptake by human endothelial cells under flow

Abstract

Stephen Paul Samuel,1,* Namrata Jain,1,* Frank O’Dowd,2 Toby Paul,2 Dmitry Kashanin,2 Valerie A Gerard,3 Yurii K Gun’ko,3 Adriele Prina-Mello,1,4 Yuri Volkov,1,41Department of Clinical Medicine, Institute of Molecular Medicine, 2Cellix Ltd, Longmile Business Centre, 3School of Chemistry, 4Centre for Research on Adaptive Nanostructures and Nanodevices, Trinity College Dublin, Dublin, Ireland*These authors contributed equally to this workAbstract: Vascular endothelium is a potential target for therapeutic intervention in diverse pathological processes, including inflammation, atherosclerosis, and thrombosis. By virtue of their intravascular topography, endothelial cells are exposed to dynamically changing mechanical forces that are generated by blood flow. In the present study, we investigated the interactions of negatively charged 2.7 nm and 4.7 nm CdTe quantum dots and 50 nm silica particles with cultured endothelial cells under regulated shear stress (SS) conditions. Cultured cells within the engineered microfluidic channels were exposed to nanoparticles under static condition or under low, medium, and high SS rates (0.05, 0.1, and 0.5 Pa, respectively). Vascular inflammation and associated endothelial damage were simulated by treatment with tumor necrosis factor-α (TNF-α) or by compromising the cell membrane with the use of low Triton X-100 concentration. Our results demonstrate that SS is critical for nanoparticle uptake by endothelial cells. Maximal uptake was registered at the SS rate of 0.05 Pa. By contrast, endothelial exposure to mild detergents or TNF-α treatment had no significant effect on nanoparticle uptake. Atomic force microscopy demonstrated the increased formation of actin-based cytoskeletal structures, including stress fibers and membrane ruffles, which have been associated with nanoparticle endocytosis. In conclusion, the combinatorial effects of SS rates, vascular endothelial conditions, and nanoparticle physical and chemical properties must be taken into account for the successful design of nanoparticle–drug conjugates intended for parenteral delivery.Keywords: endothelium, shear stress, quantum dots, membrane ruffling, stress fibers, atomic force microscopy, microfluidics