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In this manuscript, we elucidated, for the first time, the substructural mechanisms present in our recently developed bioinspired polyurethane-based pancreatic tissue models. Different protein coatings of the model, i.e., collagen and fibronectin were examined. More specifically, analysis took place by combined real-time synchrotron X-ray scattering techniques and confocal laser scanning microscopy, to quantify the structural alteration of uncoated-polyurethane (PU) and protein-coated PU as well as the time-resolved structural reorganisation occurring at the micro-, nano- and lattice length scales during in situ micromechanical testing. We demonstrate that a clear increase of stiffness at the lamellar level following the fibronectin-PU modification, which is linked to the changes in the mechanics of the lamellae and interlamellar cohesion. This multi-level analysis of structural-mechanical relations in this polyurethane-based pancreatic cancer tissue model opens an opportunity in designing mechanically robust cost-effective tissue models not only for fundamental research but also for treatment screening.