Investigation on neural and ocular surface regeneration after cultivated mucosal epithelial sheet transplantation using amniotic membrane

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Cultivated mucosal epithelial transplantation using corneal epithelial stem cells and oral mucosal epithelium dramatically improves the clinical outcome of severe ocular surface reconstruction. However, the biological mechanisms behind the cell proliferation and differentiation still remain unclear. In this study, the in vivo condition of the regenerated tissue structure composed of transplanted epithelium was investigated using in vivo laser confocal microscopy. The surviving oral mucosal epithelium showed a well-differentiated, stratified epithelial structure similar to corneal epithelium. The high density and large number of basal cells indicated the active potential for cell proliferation to maintain the ocular surface. No neural regeneration was observed in the epithelial cell layer and sub-amniotic membrane region even after an extended period of time. This evidence is consistent with the low amount of ocular surface sensation.Oral mucosal stem- and progenitor cells expressed p75 that exhibited a high affinity with neurotrophin, indicating that surviving oral mucosal epithelium may be regulated by nerve growth factor (NGF) and neurotrophin 3 and 4. Differentiated oral mucosal epithelium on the ocular surface does not express keratin 1 and 4, keratins that are specific for keratinization, but does express keratin 3 and 13. Surviving oral mucosal epithelium does not express keratin 12 that is cornea specific or MUCSAC that is specific for goblet cells. This study concluded that surviving oral mucosal epithelium on the ocular surface is able to retain the oral mucosal epithelial lineage without transdifferentiation into ocular surface epithelium.