AbstractSide chain functionalization of gelatin with tyrosine-derived moieties, desaminotyrosine (DAT) or desaminotyrosyl tyrosine (DATT), has been reported to lead to physical networks stabilized by aromatic interactions and hydrogen bonds, while the inherent ability of gelatin chains to organize in helices is suppressed. Here, the treatment of DAT and DATT gelatin films at defined temperatures (drying at 5 °C, freeze-drying, and freeze-thawing) were explored for the potential to additionally stabilize the hydrogels by increasing the content of helical domains as additional physical netpoints. The influence of the drying procedures on the hydrogel properties such as network morphology and mechanical properties were analyzed by WAXS, swelling, and rheological measurements. The triple helix content had a stabilizing effect on gelatin-based hydrogels at temperatures below the helix-to-coil transition. However, this effect was less pronounced at physiological conditions above the transition temperature, resulting in rapid dissolution of the physical gelatin networks.