Effects of blocking integrin β1 and N-cadherin cellular interactions on mechanical properties of vascular smooth muscle cells.

Experimental measurements of cellular mechanical properties have shown large variability in whole-cell mechanical properties between cells from a single population. This heterogeneity has been observed in many cell populations and with several measurement techniques but the sources are not yet fully understood. Cell mechanical properties are directly related to the composition and organization of the cytoskeleton, which is physically coupled to neighboring cells through adherens junctions and to underlying matrix through focal adhesion complexes. This high level of heterogeneity may be attributed to varying cellular interactions throughout the sample. We tested the effect of cell-cell and cell-matrix interactions on the mechanical properties of vascular smooth muscle cells (VSMCs) in culture by using antibodies to block N-cadherin and integrin β1 interactions. VSMCs were cultured on substrates of varying stiffness with and without tension. Under each of these conditions, cellular mechanical properties were characterized by performing atomic force microscopy (AFM) and cellular structure was analyzed through immunofluorescence imaging. As expected, VSMC mechanical properties were greatly affected by the underlying culture substrate and applied tension. Interestingly, the cell-to-cell variation in mechanical properties within each sample decreased significantly in the antibody conditions. Thus, the cells grown with blocking antibodies were more homogeneous in their mechanical properties on both glass and soft substrates. This suggests that diversified adhesion binding between cells and the ECM is responsible for a significant amount of mechanical heterogeneity that is observed in 2D cell culture studies.