Large Amplitude Oscillatory Shear Rheology of Living Fibroblasts: Path-Dependent Steady States.

Mechanical properties of biological cells play a role in cell locomotion, embryonic tissue formation, and tumor migration among many other processes. Cells exhibit a complex nonlinear response to mechanical cues that is not understood. Cells may stiffen as well as soften, depending on the exact type of stimulus. Here we apply large-amplitude oscillatory shear to a monolayer of separated fibroblast cells suspended between two plates. Although we apply identical steady-state excitations, in response we observe different typical regimes that exhibit cell softening or cell stiffening to varying degrees. This degeneracy of the cell response can be linked to the initial paths that the instrument takes to go from cell rest to steady state. A model of cross-linked, force-bearing filaments submitted to steady-state excitation renders the different observed regimes with minor changes in parameters if the filaments are permitted to self-organize and form different spatially organized structures. We suggest that rather than a complex viscoelastic or plastic response, the different observed regimes reflect the emergence of different steady-state cytoskeletal conformations. A high sensitivity of the cytoskeletal rheology and structure to minor changes in parameters or initial conditions enables a cell to respond to mechanical requirements quickly and in various ways with only minor biochemical intervention. Probing path-dependent rheological changes constitutes a possibly very sensitive assessment of the cell cytoskeleton as a possible tool for medical diagnosis. Our observations show that the memory of subtle differences in earlier deformation paths must be taken into account when deciphering the cell mechanical response to large-amplitude deformations.