A novel method in preparation of acellularporcine corneal stroma tissue for lamellar keratoplasty.

Our objective was to develop a novel lamellar cornealbiomaterial for corneal reconstruction.Theporcine acellular corneal stroma discs (ACSDs) were prepared from de-epithelized fresh porcine corneas (DFPCs) by incubation with 100% fresh human serum and additional electrophoresis at 4°C. Such manipulation removed theanterior corneal stromal cells without residual of DNA content and α-Galantigen. Human serum decellularizing activity on porcineanterior corneal stroma cells is through apoptosis, and associated with the presence of α-Gal epitopes in anterior stroma. ACSDs displayed similar optical, biomechanical properties and ultrastructure to DFPCs, and showed good histocompatibility in rabbit corneal stromal pockets and anterior chamber. Rabbit corneallamellar keratoplasty (LKP) using ACSDs showed no rejection and high transparency of cornea at 2 months after surgery. In vivo confocal laser scanning microscopy and immunostaining analysis showed complete re-epithelization and stromal cell in growth of ACSDs without inflammatory cell infiltration, new blood vessel ingrowth and excessive wound healing. In conclusion, this novel decellularization method may be valuable for preparation of xenogenic corneal tissue for clinical application, ACSDs resulted from this method may be served as a matrix equivalent for LKP in corneal xenotransplantation.