Adhesion complex in cultivated limbal epithelium on amniotic membrane after in vivo transplantation

Abstract

PURPOSE: To investigate adhesion complex formation in cultivated human limbal epithelium after transplantation into the limbal deficient model. METHODS: Cultivated epithelium on amniotic membrane was transplanted into limbal deficient rabbits. The transplanted rabbits and the controls were sacrificed at 1, 2, 3, and 4 weeks. The adhesion complex was examined by electron microscopy and immunohistochemistry. RESULTS: Morphologically identifiable hemidesmosomes appeared at 1 week, and matured adhesion complex was found at 3 weeks. Collagen VII was partly stained after transplantation. The mean numbers of hemidesmosomes/2.25 microm 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, and all they were smaller than those in the control, respectively (p < 0.05). It reached 137.4% of the density of hemidesmosomes in human cornea at 3 weeks. The average depths of anchoring fibril were 0.10 +/- 0.03, 0.27 +/- 0.06, 0.45 +/- 0.06, and 0.46 +/- 0.12 microm at 1, 2, 3, and 4 weeks, reaching 75.0% of that in the human cornea after 3 weeks, although they were shallower than that of the control, respectively (p < 0.05). CONCLUSIONS: Assembly of adhesion complex in cultivated epithelium transplanted in limbal deficient rabbit might recover to the level of that in the human after 3 weeks, although it was delayed compared with that in normal wound healing of the rabbit.