Effect of Storage Conditions on Rheological Properties of Decellularized Extracellular Matrix‐Based Injectable Hydrogels

ABSTRACT Decellularized extracellular matrix‐based scaffolds have gained significant attention for tissue regeneration applications. However, there is a limited understanding of how storage conditions influence the rheological and biological properties of decellularized matrix‐based injectable formulations. This study investigates the effects of different storage conditions (4°C, −80°C, and freeze‐dried) on the rheological properties of a decellularized porcine peripheral nerve‐derived extracellular matrix hydrogel. Our results demonstrated that, compared to storage at 4°C, formulations stored at −80°C and freeze‐dried maintained comparable gelation kinetics (complete gelation within 11 min), mechanical properties (storage modulus around 300–350 Pa), and shear‐thinning behavior across multiple time‐points (Days 0, 3, 7, and 14 for −80°C), similar to freshly prepared formulations at Day 0. Solution stored at 4°C demonstrated a decrease in mechanical stiffness of hydrogel with time, however their gelation kinetics remained similar to Day 0. The solutions stored at −80°C and resuspended freeze‐dried solutions maintained their complex viscosity, while 4°C stored solutions had relatively low complex viscosity as compared to freshly digested solutions. Overall, our findings suggest that −80°C storage and freeze‐drying are suitable approaches for preserving decellularized porcine peripheral nerve‐derived extracellular matrix formulations. Given the translational potential of these injectable biomaterials, understanding the influence of storage conditions on rheological performance is critical for developing appropriate storage and shipping strategies.