Phenotypic investigation of human eyes with transplanted autologous cultivated oral mucosal epithelial sheets for severe ocular surface diseases

Ophthalmology. 2007 Jun;114(6):1080-8. doi: 10.1016/j.ophtha.2006.09.034. Epub 2007 Feb 1.

Abstract

Purpose: To determine the epithelial lineage of origin of surgically removed grafts after autologous cultivated oral mucosal epithelial transplantation (COMET).

Design: Retrospective comparative case series.

Participants: We studied 6 eyes from 5 patients with total corneal stem cell destruction; 3 eyes were from patients with Stevens-Johnson syndrome and 3 eyes had sustained chemical injury.

Methods: Autologous cultivated oral mucosal epithelial sheets on human amniotic membrane (AM) were transplanted onto the ocular surface. Regrafting (2 eyes) or penetrating keratoplasty (4 eyes) was performed after the initial transplantation procedure for further visual rehabilitation.

Main outcome measures: The excised grafts were subjected to clinical evaluation and to light, scanning, and transmission electron microscopic (EM) study and to immunohistochemical analysis.

Results: In clinically failed grafts, EM and immunohistochemical analysis disclosed only small areas where the original cultivated oral epithelial cells persisted. Neighboring conjunctival epithelial cells had apparently invaded a large portion of the corneal surface (keratin 3[-], Muc5ac[+]); there were many blood vessels and inflammatory cells. In clinically successful grafts, transplanted cultivated oral epithelial cells survived and had adapted well to the host corneal tissues (keratin 3[+], Muc5ac[-]); there was no infiltration by inflammatory cells, nor was there dissolution of the AM substrate.

Conclusions: We posit that the process of graft opacification after COMET is responsible for the loss of transplanted cultivated oral epithelial cells and that this is followed by conjunctival cell invasion onto the corneal surface. We confirmed that in clinically successfully grafted eyes, autologous cultivated oral epithelial cells survived on the corneal surface and maintained ocular surface integrity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Biomarkers / metabolism
  • Cell Lineage
  • Cell Survival
  • Cell Transplantation*
  • Cells, Cultured
  • Corneal Diseases / surgery*
  • Epithelial Cells / metabolism
  • Epithelial Cells / transplantation*
  • Epithelial Cells / ultrastructure
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Keratin-3 / metabolism
  • Keratoplasty, Penetrating
  • Male
  • Middle Aged
  • Mouth Mucosa / cytology*
  • Mucin 5AC
  • Mucins / metabolism
  • Phenotype
  • Retrospective Studies
  • Stem Cells / pathology
  • Transplantation, Autologous

Substances

  • Biomarkers
  • KRT3 protein, human
  • Keratin-3
  • MUC5AC protein, human
  • Mucin 5AC
  • Mucins