Abstract
The aim of this study is to assess the effect of PAMAM dendrimers of second, fourth, and seventh generations on human umbilical vein endothelial cells. Primary endothelial cells were exposed to PAMAM dendrimers for 24 h, using concentrations reducing cellular viability to the levels of 90, 75, and 50%. We assumed, that changes in mechanical properties reflect toxicity of PAMAM dendrimers. The mechanical properties were investigated using atomic force spectroscopy (AFS) technique with the use of two approaches for measuring cell elasticity: global, where the tests were performed using a micrometer-hemispherical probe, and local, where a nanometer-sized probe was used. For the sharp probe, a reduction in the elasticity modulus was observed in comparison to untreated control cells, that is related to the depolymerization of the cytoskeleton and the processes leading to cell apoptosis. In the case of the hemispherical probe, cell softening was also observed in comparison to control cells, but with increasing PAMAM concentrations, the modulus of elasticity increases. It is related to the sensing of numerous intracellular vesicles with the use of this probe, e.g. endosomal and empty plasmalemmal which can also alter cell elasticity. The presence of external and intracellular vesicles was confirmed by scanning and transmission electron microscopy. The relationship between the elasticity of HUVEC cells exposed to PAMAM dendrimers of selected generations and their toxic effects was presented herein for the first time. In the transmission electron microscopy images of the cells exposed to PAMAM dendrimers, we have also observed distinctive vesicles with regular multilayer arranged structure.
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