Effects of corneal epithelial superficial keratectomy in patients with focal limbal stem cell disease.

PURPOSE: Irregular corneal epithelium in limbal stem cell disease can cause visual acuity to deteriorate substantially when it reaches the pupil. In this case series, we assessed the effectiveness of simple corneal epithelial superficial keratectomy in improving visual acuity in patients with irregular corneal epithelium in focal limbal stem cell disease covering the visual axis. OBSERVATIONS: We performed simple corneal epithelial superficial keratectomy in four patients (five eyes) with irregular corneal epithelium covering the visual axis. The main outcome measures were best-corrected visual acuity, slit lamp findings with fluorescein staining, anterior segment optical coherence tomography and histopathology. In all five eyes, slit lamp findings showed uneven fluorescein staining in a spiral pattern, with impaired corneal epithelial smoothness and visual disturbance. We removed the irregular epithelium in all five eyes. Visual acuity in all the eyes was improved immediately after surgery, and good visual acuity and stable epithelium were maintained for the duration of the observation periods. Hematoxylin and eosin staining showed, normal squamous and columnar epithelial cells. Goblet cells were not detected. CONCLUSIONSAND IMPORTANCE: Corneal epithelial superficial keratectomy can lead to a pathological diagnosis by examining the removed epithelial tissues, and result in excellent therapeutic outcomes in focal limbal stem cell disease reaching the pupil.

Generation of mouse model of TGFBI-R124C corneal dystrophy using CRISPR/Cas9-mediated homology-directed repair.

Mutations in transforming growth factor-beta-induced (TGFBI) gene cause clinically distinct types of corneal dystrophies. To delineate the mechanisms driving these dystrophies, we focused on the R124C mutation in TGFBI that causes lattice corneal dystrophy type1 (LCD1) and generated novel transgenic mice harbouring a single amino acid substitution of arginine 124 with cysteine in TGFBI via ssODN-mediated base-pair substitution using CRISPR/Cas9 technology. Eighty percent of homozygous and 9.1% of heterozygous TGFBI-R124C mice developed a corneal opacity at 40 weeks of age. Hematoxylin and eosin and Masson trichrome staining showed eosinophilic deposits in subepithelial corneal stroma that stained negative for Congo-red. Although amyloid deposition was not observed in TGFBI-R124C mice, irregular amorphous deposits were clearly observed via transmission electron microscopy near the basement membrane. Interestingly, we found that the corneal deposition of TGFBI protein (TGFBIp) was significantly increased in homozygous TGFBI-R124C mice, suggesting a pathogenic role for the mutant protein accumulation. Furthermore, as observed in the LCD1 patients, corneal epithelial wound healing was significantly delayed in TGFBI-R124C mice. In conclusion, our novel mouse model of TGFBI-R124C corneal dystrophy reproduces features of the human disease. This mouse model will help delineate the pathogenic mechanisms of human corneal dystrophy.

Simple oral mucosal epithelial transplantation in a rabbit model.

This study investigated a rabbit model of autologous simple oral mucosal epithelium transplantation (SOMET) for limbal stem cell deficiency (LSCD). LSCD was created in the SOMET group and the Control group. In the SOMET group, oral mucosa harvested from the buccal region was treated with dispase, cut into small pieces, and placed on the exposed corneal stroma without using graft sutures, amniotic membrane, and/or glue. A soft contact lens was positioned and tarsorrhaphy was performed in both groups. Postoperative corneal neovascularization and fluorescein staining scores were evaluated by slit lamp microscopy in both groups. At 2 weeks postoperatively, eyes were excised and subjected to immunohistochemical staining for CK3, CK13, CK15, and p63. In the SOMET group, transplantation of oral mucosa led to complete recovery of LSCD, as indicated by low neovascularization scores, low fluorescein staining scores, and detection of stratified K3/K13-positive cells on the stroma at 2 weeks after surgery. In contrast, corneal epithelial defects persisted in the Control group at 2 weeks. SOMET achieved re-epithelialization of the corneal surface in this rabbit LSCD model. It is a simple technique that does not require culture and could be a promising option for ocular surface reconstruction in bilateral LSCD.