Phenotypic Investigation of Human Eyes with Transplanted Autologous Cultivated Oral Mucosal Epithelial Sheets for Severe Ocular Surface Diseases
Received 14 June 2006; accepted 28 September 2006. published online 01 February 2007.
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.
1Department of Ophthalmology, Kyoto Prefectural University of Medicine, Graduate School of Medicine, Kyoto, Japan.
2Research Center for Regenerative Medicine, Doshisha University, Kyoto, Japan.
3Biomedical Sciences, Lancaster University, Lancaster, United Kingdom.
Correspondence to Takahiro Nakamura MD, PhD, Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto 602-0841 Japan.
Manuscript no. 2006-648.
Supported in part by grants-in-aid for scientific research from the Ministry of Health, Labour and Welfare, Tokyo, Japan (no. H16–Saisei-007), and Ministry of Education, Culture, Sports, Science and Technology, Tokyo, Japan (Kobe Translational Research Cluster); a research grant from the Kyoto Foundation for the Promotion of Medical Science, Kyoto, Japan; and the Intramural Research Fund of Kyoto Prefectural University of Medicine.
The authors have no financial or propriety interest in the products mentioned in the article.
ABSTRACT