Mechanical interactions between a cell and its environment regulate migration, contractility, gene expression, and cell fate. We integrated micropatterned substrates to engineer adhesive area and a hydrodynamic assay to analyze fibroblast adhesion strengthening on fibronectin. Independently of cell spreading, integrin binding and focal adhesion assembly resulted in rapid sevenfold increases in adhesion strength to steady-state levels. Adhesive area strongly modulated adhesion strength, integrin binding, and vinculin and talin recruitment, exhibiting linear increases for small areas. However, above a threshold area, adhesion strength and focal adhesion assembly reached a saturation limit, whereas integrin binding transitioned from a uniform distribution to discrete complexes. Adhesion strength exhibited exponential increases with bound integrin numbers as well as vinculin and talin recruitment, and the relationship between adhesion strength and these biochemical events was accurately described by a simple mechanical model. Furthermore, adhesion strength was regulated by the position of an adhesive patch, comprised of bound integrins and cytoskeletal elements, which generated a constant 200-nN adhesive force. Unexpectedly, focal adhesion assembly, in particular vinculin recruitment, contributed only 30% of the adhesion strength. This work elucidates the roles of adhesive complex size and position in the generation of cell-extracellular matrix forces.

A macroscopic compression test utilizing a simple custom-built instrument was employed to measure polydimethylsiloxane (PDMS) elastic modulus. PDMS samples with varying crosslinking density were prepared with the elastomer base to the curing agent ratio ranging from 5 : 1 to 33 : 1. The PDMS network elastic modulus varied linearly with the amount of crosslinker, ranging from 0.57 MPa to 3.7 MPa for the samples tested. PDMS elastic modulus in MPa can be expressed as 20 MPa/PDMS base to curing agent ratio. This article describes a simple method for measuring elastic properties of soft polymeric materials.

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