Abstract

Recent investigations showed that the dimensionality of the environment in which living cells are cultured - flat 2D culture wells versus 3D biopolymer networks - has a strong effect on cell morphology, metabolism and migration. The reason for these differences are unclear. What is lacking is a fundamental understanding of the mechanical and morphological properties of 3D matrices at varying length scales.We probe the local microrheology of a series of reconstituted collagen gels with different concentrations (1.2 - 2.4 mg/ml) by applying a calibrated force on embedded magnetic particles (O4.5μm) using magnetic tweezers. The resulting strain field within the matrix is visualized by tracking the positions of polystyrene spheres (O1μm) embedded in the collagen gels. This strain field is compared to expectations from continuum theory. In addition, the local microrheology is compared to bulk rheological properties measured in a cone-plate rheometer. At low forces and strains below 3%, local and bulk rheological properties agree closely, and the strain field follows that of a continuum linear elastic material. At higher strains, marked non-linear strain stiffening occurs, showing an increase in modulus of nearly 20-fold until the material eventually yields. Because of the non-uniform shear conditions around magnetic beads in the local microrheology experiments, the non-linear stiffening appeared to be less pronounced, but the strain field spread much farther out than expected from continuum theory. These data suggest that the strain stiffening behavior of collagen gels, together with the well-documented ability of cells to sense the stiffness of their surroundings, could account for the differences in cell behavior seen in 2D versus 3D culture conditions.

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