Multilayer and Biomimetic Polycaprolactone/Uterine ECM Scaffold for Uterine Tissue Engineering and Anti-Adhesion Barrier Applications via 3D-Printing Near-Field Melt Electrowriting

Abstract

Background: Post-operative uterine adhesions remain a major clinical challenge, often leading to infertility and severe pelvic pain. Current anti-adhesion barriers lack the mechanical strength, tissue specificity, or bioactivity required for optimal regeneration of uterine tissue. To address these limitations, we developed a biomimetic, multilayer scaffold that replicates the structural and functional complexity of the myometrial and serosal layers of the uterus by combining a tunable polycaprolactone (PCL) mesh with a bioactive hydrogel derived from decellularized uterine extracellular matrix (dUECM) reinforced with sodium alginate (Alg). Materials and Methods: Composite scaffolds were fabricated using near-field melt electrowriting (MEW) to create precision-engineered PCL meshes with various infill angles (90{degrees}, 60{degrees}, 45{degrees}, and 30{degrees}), followed by impregnation with an alginate-dUECM hydrogel. A comprehensive physicochemical characterization was conducted using Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric Analysis (TGA), and contact angle measurements. Mechanical properties were evaluated in both dry and wet states via uniaxial tensile testing. Swelling and degradation were assessed over 10 days. hTERT-HM cells were used for in vitro evaluation through Live/Dead staining, MTT, SEM, and immunocytochemistry for -SMA and DAPI. Results: The successful incorporation of the hydrogel into the PCL mesh was confirmed by the results obtained from FTIR and TGA analyses; these findings were further supported by contact angle studies, which showed a marked increase in hydrophilicity, resulting in a reduction of the static contact angle from 113 {+/-} 4{degrees} in the pure PCL to 60 {+/-} 12{degrees} in the hydrogel-infiltrated samples in the 30{degrees} group. Mechanical testing showed that the wet composite scaffolds maintained higher stiffness than hydrated native uterine tissue. Cell culture assays confirmed excellent cytocompatibility and proliferation, especially in 30 infill architecture. Immunofluorescence revealed strong -SMA expression and cytoskeletal organization, indicating phenotypic maturation of uterine smooth muscle cells for all groups. Conclusion: Among all groups, the 30 MEW mesh impregnated with Alg-dUECM hydrogel demonstrated the most balanced combination of mechanical resilience, degradation profile, wettability, and cellular compatibility. This structure most closely recapitulates the biological characteristics of uterine outer myometrium tissue, positioning it as a highly promising scaffold for regeneration and biofunctional anti-adhesion barrier applications.