A cell-based screening assay to identify pharmaceutical compounds that enhance the regenerative quality of corneal repair.

The goal of this study was to develop and validate a simple but quantitative cell-based assay to identify compounds that might be used pharmaceutically to give tissue repair a more regenerative character. The cornea was used as the model, and some specific aspects of repair in this organ were incorporated into assay design. A quantitative cell-based assay was developed based on transcriptional promoter activity of fibrotic marker genes ACT2A and TGFB2. Immortalized corneal stromal cells (HTK) or corneal epithelial cells (HCLE) were tested and compared to primary corneal stromal cells. Cells were transiently transfected with constructs containing the firefly luciferase reporter gene driven by transcriptional promoters for the selected fibrotic marker genes. A selected panel of seven chemical test compounds was used, containing three known fibrosis inhibitors: lovastatin (LOV), tyrphostin AG 1296 (6,7-dimethoxy-3-phenylquinoxaline) and SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole), and four potential fibrosis inhibitors: 5-iodotubercidin (4-amino-5-iodo-7-(ß-D-ribofuranosyl)-pyrrolo(2,3-d)pyrimidine), anisomycin, DRB (5,6-dichloro-1-ß-D-ribofuranosyl-benzimidazole) and latrunculin B. Transfected cells were treated with TGFB2 in the presence or absence of one of the test compounds. To validate the assay, compounds were tested for their direct effects on gene expression in the immortalized cell lines and primary human corneal keratocytes using RT-PCR and immunohistochemistry. Three "hits" were validated LOV, SB203580 and anisomycin. This assay, which can be applied in a high throughput format to screen large libraries of uncharacterized compounds, or known compounds that might be repurposed, offers a valuable tool for identifying new treatments to address a major unmet medical need. Anisomycin has not previously been characterized as antifibrotic, thus, this is a novel finding of the study.

Fixed dilated pupil (urrets-zavalia syndrome) and anterior subcapsular cataract formation after descemet stripping endothelial keratoplasty.

PURPOSE: To report a fixed dilated pupil and anterior subcapsular cataract formation following Descemet stripping endothelial keratoplasty (DSAEK). METHODS: A 47-year-old man with Fuchs endothelial dystrophy underwent DSAEK of the right eye. The preoperative slit lamp examination was normal. Mydriatic drops were used at the end of procedure. RESULTS: Postoperatively, the pupil remained fixed and dilated with no right direct or consensual response to light, or constriction during accommodation throughout a 1-year follow-up period. Slit lamp examination after air resorption revealed a transient anterior subcapsular cataract and posterior synechiae. Based on these findings, a diagnosis of Urrets-Zavalia syndrome was made. CONCLUSION: To our knowledge, this is the first report of Urrets-Zavalia syndrome after DSAEK.

Descemet stripping automated endothelial keratoplasty in a child with descemet membrane breaks after forceps delivery.

PURPOSE: To report the use of Descemet stripping automated endothelial keratoplasty (DSAEK) and to treat visual loss in Descemet membrane (DM) breaks after forceps delivery. METHODS: An 8-year-old boy presented with unilateral DM ruptures secondary to forceps delivery. Central, vertically oriented DM breaks significantly decreased visual acuity. Preoperative visual acuity was 20/80. A DSAEK was performed, the DM and endothelium were stripped from the recipient, and an 8.5-mm-diameter donor button consisting of posterior stroma and healthy endothelium was folded and implanted through a 5-mm incision. An air bubble was used to press the donor tissue against the recipient cornea, allowing it to attach without sutures. RESULTS: Graft dislocation in the early postoperative period required surgical repositioning of the donor lenticule, which led to excellent anatomic and functional outcomes. There was a significant and rapid improvement of the best-corrected visual acuity, which was 20/50 at the first month and 20/32 at the seventh month postoperatively. CONCLUSIONS: To our knowledge, this is the first report of a DSAEK in a child with DM tears after forceps delivery. Selective replacement of posterior corneal layers with DSAEK might be a promising alternative to penetrating keratoplasty to successfully treat endothelial or DM lesions during childhood.

Late-onset hyaloideocapsular block syndrome.

PURPOSE: To report an unusual cause of decreased vision in an aphakic patient. DESIGN: Interventional case report. PARTICIPANT: One patient. INTERVENTION: An 80-year-old woman presented with a myopic shift and decreased visual acuity in the left eye 16 years after extracapsular cataract extraction (ECCE) without intraocular lens (IOL) implantation. Slit-lamp examination after pupil dilation revealed a milky fluid accumulation between the posterior lens capsule and the anterior hyaloid membrane. A neodymium:yttrium-aluminium-garnet (Nd:YAG) laser hyaloidotomy was performed on the anterior hyaloid membrane. MAIN OUTCOME MEASURES: Refraction and best-corrected visual acuity. RESULTS: The Nd:YAG laser hyaloidotomy allowed the trapped fluid to flow into the vitreous cavity. The myopic shift disappeared immediately and visual acuity improved. CONCLUSIONS: To our knowledge, this is the first report of a late-onset hyaloideocapsular block syndrome after ECCE without IOL implantation.