We tested the hypothesis that mechanical tension in the cytoskeleton (CSK) is a major determinant of cell deformability. To confirm that tension was present in adherent endothelial cells, we either cut or detached them from their basal surface by a microneedle. After cutting or detachment, the cells rapidly retracted. This retraction was prevented, however, if the CSK actin lattice was disrupted by cytochalasin D (Cyto D). These results confirmed that there was preexisting CSK tension in these cells and that the actin lattice was a primary stress-bearing component of the CSK. Second, to determine the extent to which that preexisting CSK tension could alter cell deformability, we developed a stretchable cell culture membrane system to impose a rapid mechanical distension (and presumably a rapid increase in CSK tension) on adherent endothelial cells. Altered cell deformability was quantitated as the shear stiffness measured by magnetic twisting cytometry. When membrane strain increased 2.5 or 5%, the cell stiffness increased 15 and 30%, respectively. Disruption of actin lattice with Cyto D abolished this stretch-induced increase in stiffness, demonstrating that the increased stiffness depended on the integrity of the actin CSK. Permeabilizing the cells with saponin and washing away ATP and Ca2+ did not inhibit the stretch-induced stiffening of the cell. These results suggest that the stretch-induced stiffening was primarily due to the direct mechanical changes in the forces distending the CSK but not to ATP- or Ca2+-dependent processes. Taken together, these results suggest preexisting CSK tension is a major determinant of cell deformability in adherent endothelial cells.
Adherent cells sense their mechanical environment, which, in turn, regulates their functions. During the past decade, a growing body of evidence has indicated that a deformable, solid-state intracellular structure known as the cytoskeleton (CSK) plays a major role in transmitting and distributing mechanical stresses within the cell as well as in their conversion into a chemical response. Therefore in order to understand mechanical regulation and control of cellular functions, one needs to understand mechanisms that determine how the CSK changes its shape and mechanics in response to stress. In this survey, we examined commonly used structurally based models of the CSK. In particular, we focused on two classes of these models: open-cell foam networks and stress-supported structures. We identified the underlying mechanisms that determine deformability of those models and compare model predictions with data previously obtained from mechanical tests on cultured living adherent cells at steady state. We concluded that stress-supported structures appear more suitable for describing cell deformability because this class of structures can explain the central role that the cytoskeletal contractile prestress plays in cellular mechanics.
A tensegrity structure composed of six slender struts interconnected with 24 linearly elastic cables is used as a model of cell deformability. Struts are allowed to buckle under compression and their post-buckling behavior is determined from an energy formulation of the classical pin-ended Euler column. At the reference state, the cables carry initial tension balanced by forces exerted by struts. The structure is stretched uniaxially and the stretching force versus axial extension relationships are obtained for different initial cable tensions by considering equilibrium at the joints. Structural stiffness is calculated as the ratio of stretching force to axial extension. Predicted dependences of structural stiffness on initial cable tension and on stretching force are consistent with behaviors observed in living cells. These predictions are both qualitatively and quantitatively superior to those obtained previously from the model in which the struts are viewed as rigid.
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