TGFB2-mediated regeneration of pelvic ligament equivalents using a stem cell-fibroblast-decellularized membrane composite

Yingying Dong, Y Zhang, Yinan Duan, Zhijun Xia

Pelvic organ prolapse (POP) is a prevalent condition characterized by weakened pelvic floor tissues, significantly impacting women’s quality of life. Current treatments, including synthetic meshes and native tissue repair, face challenges of high recurrence rates and complications. This study developed a novel tissue-engineered strategy utilizing a decellularized human amniotic membrane (HAAM) scaffold seeded with autologous adipose-derived mesenchymal stem cells (ADSCs) and vaginal wall fibroblasts (HVFs) to construct a bioactive pelvic ligament equivalent. We first validated the successful preparation of HAAM with preserved extracellular matrix integrity and characterized the phenotypic markers of ADSCs and HVFs. In a rat abdominal wall defect model, the HAAM+ADSCs+HVFs composite demonstrated superior tissue regeneration and integration, reduced fibrosis, and effective modulation of the host immune microenvironment —evidenced by enhanced repair and decreased infiltration of pro-inflammatory cells —compared to all control groups (including HAAM alone and single-cell groups). Transcriptomic analysis revealed that the composite treatment promoted extracellular matrix organization and collagen synthesis while suppressing matrix degradation (MMP2/MMP9) and inflammatory pathways. Most importantly, mechanistic studies identified TGFB2 as the key paracrine mediator through which ADSCs exert their effects: TGFB2 inhibition abolished ADSCs-induced improvements in HVFs proliferation, migration, and anti-apoptosis, while its overexpression replicated these benefits. Western blot confirmed that TGFB2 coordinately upregulates COL1A1/COL3A1 and downregulates MMP2/MMP9 to restore ECM homeostasis. These comprehensive findings demonstrate that the HAAM-based ADSCs-HVFs composite promotes functional tissue regeneration via TGFB2-mediated mechanisms, offering a safe, effective, and regenerative therapeutic strategy with strong translational potential for POP.