Conjunctival Epithelial Cells Auto-Cultivated In Vivo on Human Amniotic Membrane in Rabbits

ABSTRACT

PURPOSE: The purpose of this study was to investigate the phenotype of conjunctival epithelial cells auto-cultivated in vivo on human amniotic membrane (CIVAMs) in rabbits for ocular surface reconstruction. METHODS: A fornix based-conjunctival flap 8 mm in diameter was made in 12 eyes of rabbits. Amniotic membrane was implanted into the conjunctival defects. Rabbit conjunctival epithelial cells were cultivated in vivo on amniotic membrane for a week. A frozen section was made of the excised specimen. To investigate the phenotype of CIVAMs, Hematoxylin-eosin staining, immunohistochemical staining to anti-MUC5AC (monoclonal antibody to conjunctival goblet cell mucin), and transmission electron microscopy were performed. RESULTS In 11 of 12 eyes, conjunctival epithelial cells grew successfully on amniotic membrane. Light microscopy demonstrated two to three layers of cuboidal epithelial cells and two to three layers of stratified epithelial cells in CIVAMs. CIVAMs exhibited non-goblet epithelial differentiation as determined by immunohistochemistry to anti-MUC5AC. Transmission electron microscopy of CIVAMs showed fine structure similar to that of normal conjunctival epithelial cells. CONCLUSIONS: CIVAMs showed morphological findings similar to normal conjunctival epithelial cells and are expected to accomplish more rapid reconjunctivalization than simple amniotic membrane transplantation. We expect that CIVAMs will be adopted in treating conjunctival burn, symblepharon, conjunctiva-scleral ulcer, and filtering bleb leakage with conjunctival defects.