Endothelial cell aging detection by means of atomic force spectroscopy

Abstract

Endothelial cell aging is related to changes not only in cell phenotype, such as luminal changes, intimal and medial thickening, and increased vascular stiffness, but encompasses different cell responses to various substances including drugs or nanomaterials. In the present work, time‐ and dose‐dependent elasticity changes evoked by silver nanoparticles in endothelial cells in early (below 15) passages were analyzed. Silver nanoparticle concentrations of 3, 3.6, and 16 μg/mL were selected for elasticity measurements for long incubation (24 hours) and of 1 and 3 μg/mL for monitoring dynamic elasticity changes of 1‐, 3‐, and 6‐hour incubations. Surprisingly, a significant reduction in the cells elasticity modulus at lower number of passages exposed to silver nanoparticles used at 3 μg/mL for 24 hours was demonstrated. These results are in contrast to those obtained for endothelial cells in late (33‐43) passages that may result from cellular aging in response to nanosilver. Furthermore, for short incubation times (1 and 3 hours), SNP‐induced significant increase in the cell elasticity modulus was detected. In current work, we also attempted to answer the question whether the changes in cell elasticity were induced by the silver nanoparticles stabilized with polyvinyl pyrrolidone or by stabilizer itself. Elasticity measurements were supplemented by observations made with transmission electron microscopy and scanning electron microscopy, which confirmed the presence of silver nanoparticles inside the cells and on the cell membrane. Additionally, activation of reactive oxygen species was detected for cells exposed to SNPs for 1 and 3 hours, which was accompanied by increased cell elasticity modulus suggesting a possible mechanism of observed phenomenon.

Different Approaches for Silver Nanoparticle Sterilization for Administration to Cell Culture

“…The toxicity of AgNPs depends on the cell line and cell culture conditions, including whether the tests were conducted in a complete cellular medium with fetal bovine serum. In our previous studies [11][12][13], we conducted tests on EA.hy926 cells exposed to AgNPs in a serum-free cell culture medium, and we observed a decrease in cell viability to about 60% for a concentration of 10 μg/mL [12]. In this study, the tests were conducted in a fetal-bovine-serumsupplemented medium, and the cell viability was above 80%.…”

“…The incubation of cells with AgNPs was conducted in a cellular medium in a physiological environment. Due to the 24-h cell exposure to AgNPs, silver agglomerates were observed on the membrane surface and in the cellular interior, which was published in our previous works [11][12][13]. The agglomeration processes of AgNPs may be influenced by other factors in the cellular medium (e.g., antibiotics, L-glutamine, and glucose), through which we maintain static cell culture under physiological conditions.…”

“…AgNPs can trigger many undesirable processes in cells, including elevated levels of reactive oxygen species, polymerization of actin fibers, cell stiffening, and even the initiation of processes inducing apoptosis [11]. The effect of AgNPs on EA.hy926 cells differs between low and high passages [12,13].…”

Charting the course of blood flow: vessel-on-a-chip technologies in thrombosis studies

“…Costa et al demonstrated that stenotic vessel-on-a-chip channels could easily trigger thrombosis after 15 min of whole blood perfusion compared with healthy vessel-on-a-chip geometries [55] . Furthermore, Zhang et al established a microvasculature-on-a-post chip to explore stenosis-induced thrombosis and endothelial behavior and found that the thrombus-like post structure could disrupt endothelial polarization and trigger microvascular thrombosis [73] [Figure 3B]. These studies suggest that there may be a positive correlation between vascular narrowing and thrombus formation, and that stenosis may also affect the integrity and stability of blood vessels, thus triggering thrombosis.…”