Extracellular matrix properties of porcine reproductive scaffolds depend on storage temperature

Abstract

Decellularized extracellular matrix (dECM) scaffolds show great promise in the field of reproductive medicine for the treatment of reproductive tract pathologies, the engineering of artificial organs to restore fertility, or the improvement of assisted reproduction techniques. Although protocols for decellularizing oviducts and uteri to obtain dECM scaffolds are well established, the optimal storage conditions required to maintain the structural and biochemical integrity of these scaffolds remain poorly defined. In this study, we evaluated the effects of one-month storage at three different temperatures (4 °C, −20 °C, and −80 °C) on the stability of dECM scaffolds from porcine oviducts and uteri. Although storage did not alter the macroscopic structure, microscopic analyses showed that storage at −80 °C resulted in a reduction in glycosaminoglycans in the oviduct, while storage at 4 °C altered the histological pattern of uterine scaffolds. Proteomic analysis revealed that freezing conditions (−20 °C and −80 °C) better preserved matrix proteins associated with key functions, such as regeneration and cell adhesion. Frozen storage was also associated with greater biomechanical strength, reduced bacterial growth, and improved biocompatibility of the scaffolds. Together, these results demonstrate that storage conditions directly influence the quality of dECM scaffolds, highlighting that freezing temperatures are more suitable for short-term storage.