Autologous glueless simple oral mucosal transplantation for the repair of limbal stem cell deficiency ocular surface in a rabbit model

This study aims to investigate the efficacy of glueless simple oral mucosal transplantation to prevent the development of the limbal stem cell deficiency in rabbit model. Rabbit limbal stem cell deficiency (LSCD) models were constructed with alcohol and mechanical scraping. The oral mucosa and limbal tissue were harvested and cut into small pieces. Microincisions were made on the corneal limbus and the precut pieces were inserted in. Animals were divided to three groups: glueless simple oral mucosal epithelium transplantation (GSOMT), glueless simple limbal epithelium transplantation (GSLET) and LSCD group. Corneal epithelium defects, opacity and neovascularization were observed after surgery by slit-lamp microscopy. Neovascularization and epithelialization were scored. HE staining and immunohistochemistry of CK12, CK13, p63 were performed after observation. In the LSCD group, corneal neovascularization and opacification were present and epithelial defects were still observed by 4 months. In the GSOMT group, the cornea was clear and epithelialization was complete by 15 days; the fluorescein staining scores and neovascularization were significantly lower in GSOMT group comparing with LSCD group. There were no significant differences between GSOMT group and GSLET group. Histological study showed that the transplanted oral mucosal graft blend into the recipient corneal tissue well after surgery and the morphology or phenotype of the corneal epithelium surrounding the graft were similar to that of normal corneal epithelium. The glueless simple oral mucosal transplantation successfully prevented the development of the limbal stem cell deficiency in rabbit model. This method avoids the limited graft sources and the allogeneic rejection problem especially in binocular LSCDs. GSOMT could become a new surgery method for treating LSCD and this basic research lays a foundation for next clinical promotion and application.