A versatile mesoindentation system to evaluate the micromechanical properties of soft, hydrated substrates on a cellular scale

It has become increasingly important to study mechanical properties of substrates on the cellular scale since cells sense and respond to changes in the microenvironment in which they are grown. To study the effects of mechanical substrate properties on the cellular scale, an existing microindentation system has been modified to perform indentation tests on highly hydrated polymeric substrates and tissues. The highly sensitive, modified indentation system, labeled as a mesoindenter, is versatile and can be customized to perform a variety of tests useful for studying tissue mechanics, stress relaxation in polymers, and interfacial adhesion phenomena. To validate the efficacy and accuracy of the system, soft, hydrated hydrogels made from agarose (1-5 wt %), poly(2-hydroxyethyl methacrylate) (p(HEMA)) (60-90% water), and unfixed, saline-perfused rat spinal cord tissue were tested. The results demonstrate that moduli vary with water content and are in line with previously published studies. We also demonstrate that the modulus of hydrogels is sensitive to the preload applied, with modulus increasing with preload. Stress relaxation indentation testing of p(HEMA) showed relaxation behavior that can be modeled using a heredity integral and standard linear model. The mesoindenter is versatile, capable of scanning and testing immersed samples, and easily customized to ascertain mechanical properties of substrates ranging from the kPa to GPa range.