A corneal endothelial layer which is the bottom layer of cornea perform a role to maintain a clarity of cornea. When the number of corneal endothelial cell decrease due to aging or hereditary condition, it is hard to restore because its mitosis capacity is limited. As fundamental treatment, a corneal transplantation is conducted widely. However, there are lots of transplantation waiting lists through a shortage of donor cornea.
In this study, we optimize a protocol to prepare ultra-thin descemet's membrane (DM) graft using decellularized porcine cornea. Using fresh porcine eyeball, a cornea is excised including sclera and conjunctiva. With biopsy punch, endothelium, DM including stroma are partially incised. Next, the cornea was decellularized in 0.1% SDS for 5 hours. Then, a graft is detached DM including stroma. Again, the graft is decelluarized in 0.1% SDS for 10hour at room temperature. The graft was dried in dry oven at 40 celsius overnight. The induced pluripotent stem cell derived corneal endothelial cells was reseeded. Finally, a thickness of dried acellular DM graft was 57.6±7.92um and recelluarized wet graft was 99.2±5.98um. The clarity of recellularized DM graft was 91.8±2.38%.
Although the future in-vivo trials would be warranted, tissue engineering DM graft using porcine decellularized corneas might be efficient tools to resolve refractory corneal disease.