Depth profiling via nanoindentation for characterisation of the elastic modulus and hydraulic properties of thin hydrogel layers.

The accurate determination of the mechanical properties of hydrogels is of fundamental importance for a range of applications, including in assessing the effect of stiffness on cell behaviour. This is a particular issue when using thin hydrogel layers adherent to stiff substrate supports, as the apparent stiffness can be significantly influenced by the constraint of the underlying impermeable substrate, leading to inaccurate measurements of the elastic modulus and permeability of thin hydrogel layers. This study used depth profiling nanoindentation and a poroelastic model for spherical indentation to identify the elastic moduli and hydraulic conductivity of thin polyacrylamide (PAAm) hydrogel layers (∼27 μm-782 μm thick) on impermeable substrates. The apparent stiffness of thin PAAm layers increased with indentation depth and was significantly greater than those of thicker hydrogels, which showed no influence of indentation depth. The hydraulic conductivity decreased as the geometrical confinement of hydrogels increased, indicating that the fluid became more constrained within the confinement areas. The impact of geometrical confinement on the apparent modulus and hydraulic conductivity of thin PAAm hydrogel layers was then established, and their elastic moduli and intrinsic permeability were determined in relation to this effect. This study offers valuable insights into the mechanical characterisation of thin PAAm hydrogel layers used for the fundamental study of cell mechanobiology.