Cytoskeletal remodeling induced by substrate rigidity regulates rheological behaviors in endothelial cells.

Altered microenvrionmental mechanical cues induce cytoskeletal remodeling in cells and have a profound impact on their functions as well as rheological properties. This article is aimed to characterize the viscoelastic behavior of endothelial cells, cultivated on variably compliant substrates. Synthetic tunable poly(dimethylsyloxane) substrates, with elastic moduli ranging from 1.5 MPa to 3 kPa, were used to trigger cytoskeletal remodeling of endothelial cells, verified by morphological analysis and actin fluorescent labeling. Elasticity and stress relaxation tests were conducted using an AFM, resulting in a wide range of data. To account for this heterogeneity, fuzzy c-means clustering algorithm was applied to partition elastic data into biologically meaningful groups, representative of different regions in cells. Nanocharacterization of biomechanical properties, along with cytoskeletal studies, proved a significant correlation between substrate flexibility and viscoelasticity of the cells. Regardless of the viscoelastic model applied, increasing substrate rigidity was related to an overall increase in cell stiffness and apparent viscosity (2.95 ± 1.56 kPa and 921.45 ± 102.46 Pa.s for the stiff substrate; 2.17 ± 1.30 kPa and 557.37 ± 494.11 Pa.s for the intermediate substrate), associated with an organized actin cytoskeleton. Conversely, cells on soft substrate were more deformable (1.84 ± 1.3 kPa) and less viscous (327.13 ± 124.25 Pa.s), exhibiting an increased actin disorganization. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 71-80, 2019.