Development of Decellularized Cornea by Organic Acid Treatment for Corneal Regeneration

Abstract

Because of the lack of donor corneas, an important area of research is the development of bioengineered corneal scaffolds to treat corneal blindness. Decellularized cornea has become a prominent area of research to satisfy the clinical demand. However, the limitation of its application is that a suitable decellularization procedure has not been developed. Organic acids are naturally occurring constituents in animal tissues and plants, and could be safely neutralized into harmless salts. In this study, we developed decellularized porcine corneal (dPC) scaffolds that were prepared by organic acid treatment. Cell removal and intact extracellular matrix preservation were evidenced by histological and biochemical quantitative analysis, and the dPC scaffolds showed porous parallel lamellar microstructure and excellent biomechanical properties. In vitro cell culture demonstrated that the dPC scaffolds had good biocompatibility, and the porous microstructure provided an ideal space for the growth of stroma keratocytes. Moreover, in vivo implantation revealed ideal reepithelialization, stromal recellularization, and complete transparency during the full follow-up period. Thus, dPC scaffolds that were prepared by organic acid treatment could be a promising biological material for use in corneal transplantation.

Impact Statement

This study successfully developed decellularized corneal scaffolds that were prepared by organic acid, which safely exist in animal tissues and plants. The results showed the highly efficient removal of cell debris from porcine corneas, and excellent preservation of optical properties, extracellular matrix (ECM) architecture, and biomolecules. In addition, decellularized corneal scaffolds revealed excellent biocompatibility and recellularization potential in vitro. In an animal model, the transplanted corneas were completely epithelialized, clear, showed no signs of immune response, and effectively supported stromal keratocytes growth. Hence, this could be a promising scaffold material for corneal tissue engineering applications.