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 μ m 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 μ m 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.

KEYWORDS:

• anchoring fibril
• amniotic membrane
• basement membrane
• hemidesmosome
• limbal epithelium