A rabbit anterior cornea replacement derived from acellular porcine cornea matrix, epithelial cells and keratocytes

Abstract

The aim of this study was to construct a rabbit anterior cornea replacement with an acellular porcine cornea matrix (APCM) as a scaffold. The scaffold was prepared from fresh porcine corneas which were treated with 0.5% (wt./vol.) sodium dodecyl sulfate (SDS) solution and stirred for 24 h in a 4 °C refrigeration chamber. The complete removal of corneal cells was confirmed by H&E and DAPI staining. The stroma structure and mechanical properties were well preserved. The extracts had no cytotoxicity to rabbit corneal keratocytes, epithelial and endothelial cells as determined by MTT assay. Moreover, there was no sign that an immune reaction occurred in or around the transplanted disks within 6 months of animal implantation. To construct a rabbit anterior cornea replacement, keratocytes were injected into APCM and cultured for 7 days in a dynamic culturing system, followed by culturing corneal epithelial cells on the stroma construct surface for another 7 days. The phenotype of the construct was similar to normal rabbit corneas, with high expression of cytokeratin 3 in the epithelial cell layer and expression of vimentin in the stromal cells. These results suggested that the APCM developed by using SDS might be a suitable scaffold for cornea tissue engineering.