Autologous Limbal Transplantation With Or Without Amniotic Membrane Transplantation For Ocular Surface Reconstruction In Rabbit With Total Limbal Stem Cell Deficiency

Objective: To investigate the efficacy of autologous limbal transplantation (ALT) with or without amniotic membrane transplantation (AMT) for ocular surface reconstruction in rabbit with unilateral total limbal stem cell deficiency.Method: The model of unilateral total limbal stem cell deficiency was created in one eye of each of the 24 rabbits by excising all the conjunctival tissue within 3 mm of the limbus and performing a superficial keratectomy of the entire corneal surface, including the limbal epithelial cells. After preparation of the model, in all cases, the fibrovascular membrane over corneal surface, including the 3-mm zone of adjacent conjunctival tissue, was carefully excised. The cases wastreated with one quadrant, two quadrants, three quadrants limbal grafts removed from the contralateral healthy eye, with or without AMT. The control group was not treated with ALT or AMT. At the end of the surgery, the eyelids were closed using a temporary tarsorrhaphy. After the tarsorrhaphy suture was removed on the fifth postoperative day, the corneal epithelialization was observed. Corneal vascularization was documented with external photographs at postoperative days 30 and the corneal surface conjunctivalization was evaluated by impression cytologic examination.Results: Finished models showed severe corneal neovascularization and severe opacity. Corneal impression cytology examination indicated conjunctival epithelium. After the tarsorrhaphy suture was removed on the fifth postoperative day, the corneal epithelialization was partly complete, from which the limbal area was graft by autologous limbal transplantation. The time which full corneal epithelialization was accomplished follow as: the group of one quadrant ALT in 27.63 ±1.53 days; the group of two quadrants ALT in 20.67 ±3.21 days; the group of three quadrants ALT in 17.67 ±1.53 days; the group of AMT with one quadrant ALT in 21.00 ±1.00 days; the group of AMT with two quadrants ALT in 13.67 ±1.53 days; the group of AMT with three quadrants ALT in 13.33 ±1.53 days; control group in 32.67±2.08 days.Corneal vascularization was documented at postoperative days 30: in the group of one quadrant ALT and of AMT with one quadrant ALT, neovascular invaded into the two quadrants cornea, from which the limbal area was not graft by autologous limbal transplantation, and corneal impression cytology examination indicated conjunctival epithelium at the vascularization area; in the group of two quadrants ALT, of three quadrants ALT, of AMT with two quadrants ALT and AMT with three quadrants ALT, no neovascular invaded into the cornea; in the control group, neovascular invaded into the total corneal surface, and corneal impression cytology examination indicated conjunctival epithelium.Conclusions: One quadrant ALT can partly reconstruct the normal corneal surface, two quadrants or more ALT can totally reconstruct the normal corneal surface. The concurrent AMT can accelerate corneal epithelialization at a certain extent, but it does not add benefits in this rabbit model at last.