Generation of acellular reproductive scaffolds from sow oviduct and uterus

Abstract

The development of biomimetic scaffolds from decellularized porcine reproductive tissues represents a promising approach to addressing challenges in reproductive medicine, with high translational potential. However, most available protocols are based on tissues from prepubertal animals because they are more uniform and are not influenced by estrous-cycle fluctuations. These protocols may not be applicable to adult animals due to increased tissue thickness and higher extracellular matrix (ECM) density. Therefore, this study aimed to optimize a detergent-based decellularization protocol to generate decellularized scaffolds from oviducts and uteri of cyclic sows. To this end, oviductal and uterine tissues from adult females in the early follicular stage were subjected to two, three, or four cycles of detergent exposure. The resulting scaffolds were subsequently characterized by morphometric and histological analysis, DNA quantification, biomechanical testing, and biocompatibility analysis by co-incubation with porcine spermatozoa. Histological results showed acellular scaffolds from the second decellularization cycle. However, DNA quantification showed significantly lower levels after four decellularization cycles. The protocol well preserved ECM components, such as collagen and elastin, while a decrease in glycosaminoglycans was observed independent of the number of cycles. Biomechanical properties were preserved in the oviduct, with no changes observed, whereas uterine decellularization resulted in a decrease in deformation capacity. Biocompatibility assays demonstrated that the generated scaffolds were cytocompatible, largely preserving sperm viability and motility. In conclusion, reproductive tissues of adult animals benefit from increased exposure to detergents, as this ensures greater DNA removal without compromising the integrity of the ECM or the cytocompatibility of the scaffolds.