Abstract
Congenital deficiency or extensive cervical injury leads to female genital tract obstruction and cervical laxity, resulting in infertility or miscarriage. Current clinical approaches could be conducted to restore the continuity of cervix. However, full-thickness and large-scale cervical regeneration with complete structural and functional restoration have not been reported. In this study, we fabricated a double-layered collagen membrane (CM) scaffold based on decellularized extracellular matrix. Each layer was adapted to support the growth of epithelial cells and stromal cells, respectively. Further, the thickness and folded "sandwich" structure were tailored to match the cervical structure. When transplanted into rabbit full-thickness and total ectocervix excision models, only CM scaffold groups enabled the regeneration of neo-ectocervix tissue, including epithelium, stroma and muscular layers after 3 and 6 months. The neo-ectocervix regenerated by CM scaffolds exhibited significantly higher expression of secretory glands and estrogen receptors, more secretion of neutral and acidic mucins, showing functional maturity of regenerated epithelium. Notably, CM scaffolds supported the regeneration of stroma and muscular layers. The mechanical strength of neo ectocervix was comparable to that of normal ectocervix. CM scaffolds demonstrate good biocompatibility, support different cell growth, and enhance superior regeneration of epithelium, stromal and muscular tissue, confirming its construction capacity as new strategies for addressing cervical deficiency and damage.