20 nm nanoparticles trigger calcium influx to endothelial cells <i>via</i> a TRPV4 channel

While increased intracellular calcium (Ca²⁺) has been identified as a key effect of nanoparticles on endothelial cells, the mechanism has not been fully elucidated or examined under shear stress. Here, we show the effect of several types of 20 nm particles on Ca²⁺ in the presence of shear stress in human umbilical vein endothelial cells (HUVECs), human coronary artery endothelial cells (HCAECs), and human cardiac microvascular endothelial cells (HMVEC-Cs). Intracellular Ca²⁺ levels increased by nearly three-fold in these cell types upon exposure to 100 μg mL^-1 20 nm Au particles, which was not seen in response to larger or smaller particles. An antagonist to the calcium channel - transient receptor potential vanilloid-type 4 (TRPV4) - drastically reduced the amount of calcium by 9.3-fold in HUVECs exposed to 0.6 Pa shear stress and 100 μg mL^-1 20 nm gold particles, a trend upheld in both HCAECs and HMVEC-Cs. Cell alignment in the direction of fluid flow is a well-known phenomenon in endothelial cells, and interestingly, cells in the presence of 20 nm particles with fluid flow had a higher alignment index than cells in the fluid flow alone. When compared with previous works, these results indicated that 20 nm particles may be inducing endothelial permeability by activating the TRPV4 channel <i>in vitro</i>. The potential of nanoparticle delivery technologies hinges on an improved understanding of this effect toward improved delivery with limited toxicity.