Aligned vascular endothelial growth factor-functionalized gelatin/polycaprolactone/heparin nanofibrous scaffolds with cyclic stretching for mesothelial tissue regeneration.

Mesothelial tissue regeneration requires scaffolds that integrate structural, biochemical, and mechanical cues to mimic the native microenvironment. In this work, random and aligned gelatin/polycaprolactone/heparin nanofibrous scaffolds (rGPH and aGPH) were fabricated by electrospinning. The aligned scaffold (aGPH) provided topographical guidance for mesothelial cell orientation and cytoskeletal organization. To enhance bioactivity, vascular endothelial growth factor (VEGF) was immobilized via heparin binding. This enables the combination of topographical cues with tailored biochemical signaling. Mesothelial cells cultured on aGPH/VEGF exhibited improved proliferation, migration, and epithelial marker expression, forming a mature mesothelial cell layer with preserved phenotype. Under cyclic stretching during dynamic culture of the cell/scaffold construct to reflect physiological mechanical stimulation, cells demonstrated enhanced proliferation, alignment, and cytoskeletal remodeling. The synergistic effects of fiber orientation, VEGF bioactivity, and dynamic culture established a biomimetic environment that maintained mesothelial identity and function. In vivo implantation of cell-seeded aGPH/VEGF scaffolds pre-cultured in vitro under cyclic stretching repaired rat peritoneum, regenerating a neo-mesothelium structurally comparable to native tissue. These findings establish a platform for mesothelial tissue engineering and underscore the value of integrating topographical, biochemical, and biophysical modulation for serosal membrane regeneration.