Ocular Surface Reconstruction Using Cultivated Mucosal Epithelial Stem Cells

Abstract

To investigate the possibility of using cultivated oral epithelial cells in ocular surface reconstruction.

An ocular surface injury was created in adult albino rabbits by a lamellar keratectomy. Oral mucosal biopsy specimens taken from both adult albino rabbits and human volunteers were cultivated for 2–3 weeks on a denuded amniotic membrane (AM) carrier. The cultivated epithelium was examined by histologic and immunohistochemical analysis. At 3–4 weeks after the ocular surface injury, the rabbit conjunctivalized corneal surfaces were surgically reconstructed by transplanting both the rabbit and human cultivated oral epithelial cells on the AM carrier.

Both the rabbit and human cultivated oral epithelial sheets had 5 or 6 layers of stratified, well-differentiated cells. Histologic examination revealed that the cultivated epithelial cells were similar in appearance to those of in vivo normal corneal epithelium. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin 3 in the cultivated oral epithelial cells. Corneas that were grafted with rabbit and human cultivated oral epithelial cells on an AM carrier were clear and were epithelialized 10 and 2 days after surgery, respectively.

We have generated confluent cultures of both rabbit and human oral epithelial cells on AM expanded ex vivo from biopsy-derived oral mucosal tissues. We have successfully carried out xeno- and autologous transplantation of these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. We believe that xeno- and autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction.