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ARTICLE

Geometric Confinement Influences Cellular Mechanical Properties I -- Adhesion Area Dependence

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∗ Department of MechanicalEngineering, MIT. Present address: Biochemistry and Molecular Biophysics Option, Caltech.
† Department of Chemistry and Chemical Biology, Harvard University. Present address: National Center for NanoScienceand Technology,China.
‡ Statistics and ManagementScience, MIT.
§ Department of Chemistry and Chemical Biology, Harvard University
¶ Department of Biological Engineering and Department of Mechanical Engineering, MIT.

Molecular & Cellular Biomechanics 2007, 4(2), 87-104. https://doi.org/10.3970/mcb.2007.004.087

Abstract

Interactions between the cell and the extracellular matrix regulate a variety of cellular properties and functions, including cellular rheology. In the present study of cellular adhesion, area was controlled by confining NIH 3T3 fibroblast cells to circular micropatterned islands of defined size. The shear moduli of cells adhering to islands of well defined geometry, as measured by magnetic microrheometry, was found to have a significantly lower variance than those of cells allowed to spread on unpatterned surfaces. We observe that the area of cellular adhesion influences shear modulus. Rheological measurements further indicate that cellular shear modulus is a biphasic function of cellular adhesion area with stiffness decreasing to a minimum value for intermediate areas of adhesion, and then increasing for cells on larger patterns.\nobreakspace {} We propose a simple hypothesis: that the area of adhesion affects cellular rheological properties by regulating the structure of the actin cytoskeleton.\nobreakspace {} To test this hypothesis, we quantified the volume fraction of polymerized actin in the cytosol by staining with fluorescent phalloidin and imaging using quantitative 3D microscopy. The polymerized actin volume fraction exhibited a similar biphasic dependence on adhesion area. Within the limits of our simplifying hypothesis, our experimental results permit an evaluation of the ability of established, micro-mechanical models to predict the cellular shear modulus based on polymerized actin volume fraction. We investigated the ``tensegrity'', ``cellular-solids'', and ``biopolymer physics'' models that have, respectively, a linear, quadratic, and 5/2 dependence on polymerized actin volume fraction. All three models predict that a biphasic trend in polymerized actin volume fraction as a function of adhesion area will result in a biphasic behavior in shear modulus. Our data favors a higher-order dependence on polymerized actin volume fraction. Increasingly better experimental agreement is observed for the tensegrity, the cellular solids, and the biopolymer models respectively. Alternatively if we postulate the existence of a critical actin volume fraction below which the shear modulus vanishes, the experimental data can be equivalently described by a model with an almost linear dependence on polymerized actin volume fraction; this observation supports a tensegrity model with a critical actin volume fraction.

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