Phenotypic investigation of human eyes with transplanted autologous cultivated oral mucosal epithelial sheets for severe ocular surface diseases.
OBJECTIVE
To determine the epithelial lineage of origin of surgically removed grafts after autologous cultivated oral mucosal epithelial transplantation (COMET).


METHODS
Retrospective comparative case series.


METHODS
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.


METHODS
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.
Nakamura
Takahiro
T
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Graduate School of Medicine, Kyoto, Japan. tnakamur@ophth.kpu-m.ac.jp
Inatomi
Tsutomu
T
Cooper
Leanne J
LJ
Rigby
Helen
H
Fullwood
Nigel J
NJ
Kinoshita
Shigeru
S
D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000378:metabolism; D019070:Cell Lineage; D002470:Cell Survival; D017690:Cell Transplantation; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000601:surgery*; D004847:Epithelial Cells / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D053558:Keratin-3 / Q000378:metabolism; D015948:Keratoplasty, Penetrating; D008297:Male; D008875:Middle Aged; D009061:Mouth Mucosa / Q000166:cytology*; D055271:Mucin 5AC; D009077:Mucins / Q000378:metabolism; D010641:Phenotype; D012189:Retrospective Studies; D013234:Stem Cells / Q000473:pathology; D014182:Transplantation, Autologous
D015415:Biomarkers; C479360:KRT3 protein, human; D053558:Keratin-3; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins