Substrate stiffness is emerging as an effective tool for the regulation of cell behaviours such as locomotion, proliferation and differentiation. In order to explore the potential application of this biophysical tool, material platforms displaying lateral and continuously graded stiffness are advantageous since they allow the systematic exploration of adherent cell response to substrate stiffness and the tuning of the material to elicit the desired cell behaviour. Here, we demonstrate a simple approach towards the fabrication of polydimethylsiloxane (PDMS) stiffness gradients (with an indentation modulus of 190 kPa-3.1 MPa across a 12 mm distance) by means of a temperature gradient during curing. We then apply these stiffness gradients to the screening of osteogenic differentiation in rat mesenchymal stem cells (rMSCs). Our proof-of-principle results show that mineralization of rMSCs is strongly dependent on the PDMS substrate stiffness, but is also influenced by the display of extracellular matrix proteins preadsorbed on the gradients. This screening capability holds tremendous potential for the design of improved implant materials and tissue engineering scaffolds.
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