Adhesion Complex in Cultivated Limbal Epithelium on Amniotic Membrane after Transplantation into Chemical Burn Model

ABSTRACT

PURPOSE: To investigate adhesion complex formation in cultivated human limbal epithelium after transplantation into the chemical burn model. METHODS: human limbal epithelial cells were cultured on amniotic membrane that had not undergone dispase treatment. Laminin V was evaluated using immunohistochemistry. The adhesion complex was examined by electron microscopy. Cultured epithelium was transplanted into limbal deficient rabbits induced by chemical burn and mechanical limbal removal. The transplanted rabbits and the controls with mechanical wounding were sacrificed at 1, 2, 3, and 4 weeks. The adhesion complex was examined by electron microscopy. RESULTS: Linear staining was observed against laminin V at 4-week culture but matured adhesion complex was not found. Graft failure developed in 3 rabbits (25%) after transplantation. Morphologically identifiable hemidesmosomes appeared at 1 week and matured adhesion complex with continuous basement membrane was found at 3 weeks. The mean numbers of hemidesmosomes/2.25 micro meter were 2.3 +/- 0.9, 2.5 +/- 0.5, 5.2 +/- 1.0, and 4.0 +/- 0.9 at 1, 2, 3, and 4 weeks, respectively. The adhesion assembly nearly recovered to the level of that in the human cornea (3.7 +/- 60.11) at 3 weeks. CONCLUSIONS: Adhesion complex of cultivated limbal epithelium did not developed in vitro, but the assembly was almost completed at 3 weeks after transplantation in vivo.