Fabrication of three-dimensional (3-D) corneal transplantable model from silk fibroin/gelatin-based biomaterial

Abstract

Canine corneal blindness is a common cause of vision loss and visual impairment worldwide. While the demand of canine corneal graft is continuously elevated, the clinical useable grafts are limited to acellular biomaterials and allograft. This study aimed to generate the tissue-engineered canine cornea in the part of corneal epithelium and underlying stroma based on cLESCs (canine limbal epithelial stem cells) seeded silk fibroin and gelatin (SF/G) film and cCSSCs (canine corneal stromal stem cells) seeded SF/G scaffold respectively. Both cell types were successfully isolated by collagenase I. SF/G films and scaffolds served as the prospective substrates for cLESCs and cCSSCs by promoting cell adhesion, cell viability and cell proliferation. Furthermore, gene expression levels were compared among cells seeded tissue culture plate (TCP) and cells seeded their own scaffolds. Here, the results revealed the upregulation of P63 and Abcg2 of cLESCs as well as Kera, Lum, Aldh3a1 and Aqp1 which are the marker of cells differentiated into keratocytes and extracellular matrix production like a native cornea. Moreover, immunohistochemistry was illustrated the positive staining of P63, lumican, Aldh3a1 and collagen I. The results manifested the feasible platform to construct tissue-engineered canine cornea as the functional transplantable corneal grafts and provided the profitable acknowledgement in the area of canine corneal stem cells to develop stem cell-based therapy in the future.