Development Of A Biomimetic 3D Ovarian Scaffold Using Decellularized Extracellular Matrix And Mechanically Tuned Hydrogels

Artificial ovarian scaffolds represent a promising therapeutic strategy for preserving reproductive health in patients. However, current in vitro approaches are limited by inadequate biomimicry of the native tissue microenvironment, leading to poor development of in vitro ovarian models. In this study, we developed region-specific hydrogel scaffolds incorporating solubilized decellularized ovarian extracellular matrix (dECM) with mechanically tuned properties to enhance the functionality of engineered 3D ovarian models. Ovine ovarian dECM was isolated by mechanical and chemical decellularization methods and subsequently solubilized and incorporated in varying concentrations in homogenous alginate (0.5%) and a composite mixture of 1% gelatin with 0.5% alginate (1:1). The synthesized hydrogels were characterized for rheological properties, including Youngs modulus, pore size, and viscosity, and cytocompatibility assays were conducted using Chinese hamster ovary (CHO) cells. The study demonstrated that both 0.5% alginate and the composite gelatin-alginate hydrogels successfully replicated the mechanical properties of native human ovarian cortical and medullary tissue, with Youngs modulus of 0.84 {+/-} 0.16 kPa, pore size (60-150 nm), and toughness of 0.4Pa, respectively. Zonal hydrogel scaffolds incorporating ovarian dECM demonstrated significantly enhanced cell viability compared to hydrogels supplemented with dECM. The study emphasises the critical role of integrating both mechanical and biochemical attributes while developing functional artificial ovarian constructs for transplantation and regenerative medicine applications. This work contributes to advancing strategies for creating physiologically relevant in vitro models of ovarian tissue.