F1 - Investigating Regional Differences in Porcine Uterine dECM for Targeted Regenerative Therapies

Introduction: : Decellularized extracellular matrix (dECM) biomaterials have consistently shown success across various applications. Some of these dECM include small intestinal submucosa (SIS), urinary bladder matrix (UBM), and vocal fold lamina propria (VFLP). A common characteristic among these dECMs is their derivation from specialized outer layers of complex organs, suggesting a correlation between their native physiological roles relative to location and their regenerative properties. The present study focuses on investigating the regional heterogeneity of porcine uterine dECM. Our primary objective is to identify and characterize the tissue–and region-specific regenerative potential within different layers of the uterus, thereby developing novel and targeted therapeutic approaches for uterine-specific applications.

Materials and

Methods: : Uterine tissue will be harvested from healthy pigs to explore the hypothesized regional differences in dECM properties. The collected tissue will undergo anatomical division into two distinct regions: the body and the top horn. Subsequently, each of these regions will be further dissected to separate the endometrial (inner) and myometrial (outer) layers. Following this dissection, optimized decellularization protocols will be used to generate unique regional dECM hydrogels. The efficacy of decellularization will be quantitatively assessed using the QuantiFluor dsDNA System, with a target of achieving a greater than 95% reduction in double-stranded DNA content relative to native tissue. The resulting dECM powder will be enzymatically digested with pepsin and self-assembled into hydrogels through pH and salt adjustments. Rheological testing will be conducted on the dECM-hydrogels to measure stiffness and viscoelasticity and to determine their anisotropic or isotropic characteristics. In vitro assays will be used to evaluate cell adhesion, proliferation rates, and overall cell health and viability when cultured on distinct dECM hydrogels derived from varying uterine regions.

Results, Conclusions, and Discussions:: This study aims to clarify how inherent regional differences within the uterus affect the regenerative potential of dECM derived from these areas. This study compares dECM derived from the decidual endometrial matrix versus that of the uterine musculature. The findings will offer important insights that will guide the design of specialized biomaterials for repairing and regenerating uterine tissue. Additionally, this research has wider implications for regenerative medicine.

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