This article presents a very detailed study on the mechanical characterization of a highly nonlinear material, the immature equine zona pellucida (ZP) membrane. The ZP is modeled as a visco-hyperelastic soft matter. The Arruda-Boyce constitutive equation and the two-term Prony series are identified as the most suitable models for describing the hyperelastic and viscous components, respectively, of the ZP's mechanical response. Material properties are identified via inverse analysis based on nonlinear optimization which fits nanoindentation curves recorded at different rates. The suitability of the proposed approach is fully demonstrated by the very good agreement between AFM data and numerically reconstructed force-indentation curves. A critical comparison of mechanical behavior of two immature ZP membranes (i.e., equine and porcine ZPs) is also carried out considering the information on the structure of these materials available from electron microscopy investigations documented in the literature.
Keywords: atomic force microscopy; equine zona pellucida; inverse analysis; nanoscale mechanical characterization; visco-hyperelasticity.