Abstract
Purpose: :
Evaluation of single vs. double amniotic membrane technique for the transplantation of limbal epithelial cells.
Methods: :
A limbal stem cell deficiency (LSCD) in the eye of normal New Zealand white rabbits was created by performing a 360º limbal peritomy and limbalectomy. Intact human amniotic membranes (AM), with (n=9) or without (n=6) cultured limbal epithelial cells (LEC), were secured to the cornea. In the single membrane technique, the LEC-AM graft was sutured with the cells facing the cornea (n=6). In the double membrane technique, the LEC-AM graft was sutured with the cells (basement membrane) up, and a second HAM was secured to cover the first with the basement membrane side face down (n=3). Post operatively, the degree of corneal neovascularization and fibrosis was monitored in all rabbits, and scored on a scale of 1-4 depending upon the number of quadrants affected. The degree of corneal vascularization in each animal was measured at completion of the study. Animals were euthanized at 4 weeks (AMs only) or 6 weeks (AM-LEC). Results were compared using Student's t-test.
Results: :
Mean values of neovascularization/corneal fibrosis, respectively, were: AM-only controls, 4.0/4.0; double membrane LEC-AM, 4.0/4.0; and single membrane LEC-AM, 2.7/3.0. The distance of neovascularization at the 12:00 position was statistically larger in eyes that received AM-only grafts (3.8± 0.5mm) relative to those receiving LEC-AM grafts using either the single (1.4±1.2mm) or double membrane (2.3±0.6mm) technique.
Conclusions: :
Single membrane transplant technique is equally effective as double membrane technique in functionally reconstructing the corneal epithelium in a LSCD model.
Keywords: cornea: epithelium • keratitis