Descemet Membrane Endothelial Keratoplasty for Endothelial Failure in Primary Angle Closure Suspects.

PURPOSE: To analyze the feasibility and outcome of Descemet membrane endothelial keratoplasty (DMEK) for treatment of endothelial failure in primary angle closure suspect (PACS) eyes. METHODS: Retrospective, single-center case series of eyes treated by DMEK for endothelial failure caused by PACS. Main study parameters were success rate of DMEK, best-corrected visual acuity, anterior chamber depth, central corneal thickness, and endothelial cell density. Mean follow-up time was 16 ± 13 months. RESULTS: Ten eyes of 9 patients receiving DMEK for the treatment of corneal endothelial failure because of PACS were included. Except for 2 eyes that had undergone cataract surgery, none of the eyes had previous ocular surgery. DMEK combined with cataract surgery was performed in 5 eyes, DMEK alone with second-step cataract surgery in 2 eyes. The eyes with corneal edema after cataract surgery received DMEK only. DMEK surgery was successful in nine out of 10 eyes, 1 patient required repeat DMEK because of primary graft failure. In the group of phakic eyes, mean preoperative internal anterior chamber depth was 1.74 ± 0.18 mm. In eyes with corneal edema, central corneal thickness was 849 ± 205 µm before DMEK surgery, and 517 ± 24 µm at the final postoperative visit (P = 0.002). CONCLUSIONS: DMEK is a feasible option in eyes with endothelial failure because of primary angle closure. In case of advanced corneal edema, a second-step procedure (first DMEK, second cataract surgery) is a possible approach if visibility of the lens is too poor for simultaneous cataract surgery.

Dysregulation of the TCF4 Isoform in Corneal Endothelial Cells of Patients With Fuchs Endothelial Corneal Dystrophy.

Purpose: This study evaluated the dysregulation of TCF4 isoforms and differential exon usage (DEU) in corneal endothelial cells (CECs) of Fuchs endothelial corneal dystrophy (FECD) with or without trinucleotide repeat (TNR) expansion in the intron region of the TCF4 gene. Methods: Three RNA-Seq datasets of CECs (our own and two other previously published datasets) derived from non-FECD control and FECD subjects were analyzed to identify TCF4 isoforms and DEU events dysregulated in FECD by comparing control subjects to those with FECD with TNR expansion and FECD without TNR expansion. Results: Our RNA-Seq data demonstrated upregulation of three TCF4 isoforms and downregulation of two isoforms in FECD without TNR expansion compared to the controls. In FECD with TNR expansion, one isoform was upregulated and one isoform was downregulated compared to the control. Additional analysis using two other datasets identified that the TCF4-277 isoform was upregulated in common in all three datasets in FECD with TNR expansion, whereas no isoform was dysregulated in FECD without TNR expansion. DEU analysis showed that one exon (E174) upstream of the TNR, which only encompassed TCF4-277, was upregulated in common in all three datasets, whereas eight exons downstream of the TNR were downregulated in common in all three datasets in FECD with TNR expansion. Conclusions: This study identified TCF4-277 as a dysregulated isoform in FECD with TNR expansion, suggesting a potential contribution of TCF4-277 to FECD pathophysiology.

Differences in Guttae Ultramorphology in Relation to Visual Function in Fuchs Endothelial Corneal Dystrophy.

PURPOSE: The purpose of this study was to investigate the differences in guttae ultramorphology and their relation to visual function in eyes with Fuchs endothelial corneal dystrophy (FECD). METHODS: Thirty FECD eyes without ocular comorbidities were included. Visual functional parameters (best-corrected visual acuity with high-contrast and low-contrast letters and contrast sensitivity/LogCS) and corneal morphology measured with Scheimpflug tomography (Pentacam) were assessed. The surgically removed Descemet membranes were examined by light and transmission electron microscopy. RESULTS: Preoperative mean best-corrected visual acuity (logarithm of the minimum angle of resolution) was 0.52 ± 0.18, LogCS 0.96 ± 0.21 and central corneal thickness 640 ± 55 µm. All eyes had signs of subclinical corneal edema in Scheimpflug tomography; clinically visible corneal edema was present in 40% of eyes. Histological findings included a posterior fibrillar zone (PFZ) in 10 specimens (33%) and abnormal collagen depositions in Descemet membranes in 14 specimens (47%). Guttae buried within the PFZ were present only in eyes with clinically visible edema (n = 4, 13%). There was no difference in visual function results and tomography parameters between eyes with and without PFZ or between protruding guttae and guttae embedded in a PFZ, respectively. CONCLUSIONS: Guttae morphology and density were not correlated with visual functional parameters. Guttae buried in a PFZ occurred only in eyes with clinically manifest edema, and thereby, they are an ultramorphological sign for advanced FECD. Subclinical edema was present in all eyes and might be more relevant for quality of vision than guttae ultramorphology.

Characterisation of ectasia after penetrating keratoplasty in keratoconus eyes using anterior segment optical coherence tomography.

BACKGROUND/AIMS: Ectasia of the cornea can occur decades after penetrating keratoplasty (PK), especially in keratoconus eyes. The purpose of this study was to characterise ectasia after PK by morphological findings in anterior segment optical coherence tomography (AS-OCT). METHODS: In this retrospective, single-centre case series, 50 eyes of 32 patients with a history of PK at an average of 25±10 years earlier were included. The eyes were classified either as ectatic (n=35) or as non-ectatic (n=15). The main parameters included central corneal thickness (CCT), lowest corneal thickness at the interface (LCTI), anterior chamber depth, graft-host interface angle at the thinnest point and host cornea-iris angle. Furthermore, steep and flat keratometry readings obtained by AS-OCT (CASIA-2, Tomey) and Scheimpflug tomography (Pentacam, Oculus) were assessed. OCT findings were correlated with clinical grading of ectasia. RESULTS: There was a highly significant difference in LCTI, graft-host interface angle and anterior chamber depth (in pseudophakic eyes) between the groups. The ratio calculated by the quotient of LCTI divided by CCT was significantly lower in ectatic than non-ectatic eyes (p<0.001). In eyes with an LCTI/CCT ratio of ≤0.7, the OR for the occurrence of a clinical detectable ectasia was 2.4 (CI 1.5 to 3.7). Steep keratometry values were significantly higher in ectatic eyes. CONCLUSION: AS-OCT is a helpful tool to recognise and quantify ectasia in post-PK eyes objectively.

RNA-Seq-based transcriptome analysis of corneal endothelial cells derived from patients with Fuchs endothelial corneal dystrophy.

Fuchs endothelial corneal dystrophy (FECD) is the most common inherited corneal disease. Fibrillar focal excrescences called guttae and corneal edema due to corneal endothelial cell death result in progressive vision loss. Multiple genetic variants have been reported, but the pathogenesis of FECD is not fully understood. In this study, we used RNA-Seq to analyze differential gene expression in the corneal endothelium obtained from patients with FECD. Differential expression analysis of transcriptomic profiles revealed that expression of 2366 genes (1092 upregulated and 1274 downregulated genes) was significantly altered in the corneal endothelium of patients with FECD compared to healthy subjects. Gene ontology analysis demonstrated an enrichment of genes involved in extracellular matrix (ECM) organization, response to oxidative stress, and apoptotic signaling. Several pathway analyses consistently indicated the dysregulation of ECM-associated pathways. Our differential gene expression findings support the previously proposed underlying mechanisms, including oxidative stress and apoptosis of endothelial cells, as well as the phenotypic clinical FECD hallmark of ECM deposits. Further investigation focusing on differentially expressed genes related to these pathways might be beneficial for elucidating mechanisms and developing novel therapies.

Management of Late Descemet's Membrane Detachment After Penetrating Keratoplasty in Keratoconus.

PURPOSE: The purpose of this study was to describe the feasibility of Descemet membrane endothelial keratoplasty (DMEK) as a treatment modality for spontaneous detachment of DM (DMD) decades after penetrating keratoplasty (PK) for keratoconus. METHODS: We describe the clinical characteristics and therapeutic surgical approach in 6 eyes of 5 patients with DMD. Clinical images, anterior segment optical coherence tomography scans, and histological findings are presented. RESULTS: Mean age of patients at time of diagnosis was 60 years (range 56-66 years). Mean interval between PK and occurrence of DM detachment was 36 years (range 29-45 years). In 4 of 6 eyes, air injections into the anterior chamber were initially attempted to reattach DM to the stroma but without long-lasting effect. Two eyes underwent repeat PK because of pronounced ectasia after long-standing DMD and stromal scars. DMEK was performed successfully in 4 eyes leading to an increase in visual acuity and a reduction in central corneal thickness. Electron microscopy showed abnormal vacuolar inclusions and collagenous material in the posterior nonbanded layer and a separation of the anterior banded layer from the posterior nonbanded layer. CONCLUSIONS: This case series provides evidence that DMEK is a viable option in eyes with spontaneous DM detachment after PK. Visual outcome is limited by the persisting high astigmatism in the ectatic cornea. Illustrated by a small series of patients, the results of DMEK in this condition are presented and new findings about the pathophysiology are given.

Descemet membrane endothelial keratoplasty: analysis of clinical outcomes of patients with 8-10 years follow-up.

PURPOSE: This study aimed to evaluate the clinical outcomes up to 10 years after Descemet membrane endothelial keratoplasty (DMEK). METHODS: In this retrospective, consecutive, single-center case series the medical files of eyes which have received DMEK between 2009 and 2012 for the treatment of endothelial dysfunction was evaluated regarding follow-up time and clinical outcomes. Annual examinations of best-corrected visual acuity (BCVA), endothelial cell density (ECD), central corneal thickness (CCT) of 66 eyes which fulfilled the criterion of a minimum of 8 years follow-up were analyzed. RESULTS: BCVA improved from 0.55 ± 0.37 logMAR (n = 54) to 0.15 ± 0.11 (n = 47) in eyes without ocular comorbidities one year after DMEK (p < 0.001), and remained stable up to 10 years after DMEK. Mean ECD decreased to 744 ± 207 cells/mm2 (n = 39) after 9 years, and to 729 ± 167 cells/mm2 (n = 21) after 10 years, respectively. CCT decreased from 650 ± 67 µm before DMEK to 525 ± 40 µm (n = 56) after 1 year, increasing slowly to 563 ± 40 µm (n = 39) after 9 years, and to 570 ± 42 µm (n = 21) after 10 years, respectively. Graft failure occurred in 4 of 66 eyes after year 8. These 4 eyes required repeat DMEK after 101-127 months. CONCLUSION: This study shows the long-term outcomes in a small subset of DMEK grafts. Visual acuity remained stable in spite of slowly increasing corneal thickness and diminishing endothelial cell density during the 10-year period after DMEK.

Potential Functional Restoration of Corneal Endothelial Cells in Fuchs Endothelial Corneal Dystrophy by ROCK Inhibitor (Ripasudil).

PURPOSE: Rho-associated kinase (ROCK) inhibitors have been successfully used as a rescue strategy in eyes that failed to clear after descemetorhexis without endothelial graft for treatment of Fuchs endothelial corneal dystrophy (FECD). The functional mechanisms by which ROCK inhibitors modulate corneal endothelial cell regeneration in FECD patients have, however, not been clarified. Here, we analyzed the effect of the ROCK inhibitor ripasudil on corneal endothelial cells of FECD patients and normal donors using ex vivo tissue and in vitro cellular models. DESIGN: Experimental study: laboratory investigation. METHODS: This institutional study used endothelial cell-Descemet membrane lamellae from FECD patients (n = 450) undergoing Descemet membrane endothelial keratoplasty (FECD ex vivo model), normal research-grade donor corneas (n = 30) after scraping off central endothelial cells (ex vivo wound healing model), normal donor corneas (n = 20) without endothelial injury, and immortalized cell lines (n = 3) generated from FECD patients (FECD in vitro model). Descemet membrane lamellae were dissected into halves and incubated for 24-72 hours in storage medium with or without a single dose of 30 µM ripasudil. The effects of ripasudil on expression of genes and proteins related to endothelial cell proliferation, migration, functionality, and endothelial-to-mesenchymal transition were analyzed and complemented by functional assays on FECD cell lines. RESULTS: A single dose of ripasudil induced significant upregulation of genes and proteins related to cell cycle progression, cell-matrix adhesion and migration, as well as endothelial barrier and pump function up to 72 hours, whereas classical markers of endothelial-to-mesenchymal transition were downregulated in both FECD and normal specimens compared to unstimulated controls ex vivo. In addition to stimulation of proliferation and migration, ripasudil-induced changes in expression of functional signature genes could be also verified in FECD cell lines in vitro. CONCLUSIONS: These data support the concept that inhibition of ROCK signaling represents a potent tool in regenerative therapies in FECD patients through reactivation of cell proliferation and migration as well as restoration of endothelial pump and barrier function without inducing adverse phenotypic changes.

Influence of corneal guttae and nuclear cataract on contrast sensitivity.

AIMS: To evaluate the contrast sensitivity in patients with nuclear cataract and corneal guttae compared to patients with nuclear cataract without guttae. METHODS: In this retrospective, single-centre case series, 50 eyes of 50 patients fulfilling the inclusion criteria were enrolled. Patients with corneal guttae and nuclear cataract (n=25, study group) underwent triple Descemet membrane endothelial keratoplasty (DMEK). Patients with nuclear cataract and healthy corneas underwent cataract surgery (n=25, control group). Inclusion criteria were preoperative best-corrected visual acuity ≥20/40, no corneal oedema and similar lens opacity (nuclear opalescence 2.0-2.9). Outcome measures included MARS letter and OPTEC 6500P contrast sensitivity test, corneal volume, central corneal thickness and anterior and posterior corneal densitometry. RESULTS: Preoperative MARS letter and OPTEC 6500P contrast sensitivity was significantly worse in the study group (MARS: p<0.001; OPTEC 6500P: p<0.007 at low spatial frequencies in daylight with and without glare and nightlight without glare). After surgery, there was no significant difference in MARS letter contrast sensitivity between groups (p=0.225). OPTEC 6500P contrast sensitivity remained significantly lower in the study group in daylight and nightlight with and without glare at most spatial frequencies (p<0.01) postoperatively. Preoperative and postoperative corneal volume, central corneal thickness and anterior corneal densitometry were equal in both groups (p>0.05). Posterior densitometry was significantly higher in the study group than in the control group preoperatively (p<0.001) but turned into equal values postoperatively (p=0.07). CONCLUSIONS: Corneal guttae cause an additional significant decrease in contrast sensitivity in eyes with nuclear cataract. This is in favour of performing a triple DMEK even in eyes with a visual acuity of ≥20/40.

Refractive Outcomes After Descemet Membrane Endothelial Keratoplasty + Cataract/Intraocular Lens Triple Procedure: A Fellow Eye Comparison.

PURPOSE: Descemet membrane endothelial keratoplasty is often combined with phacoemulsification and intraocular lens implantation (DMEK + cataract/IOL triple procedure) in phakic patients. This procedure results in a refractive shift that is difficult to predict. The aim of this study was to evaluate the hypothesis that the refractive shift in the second eye follows the shift in the first eye. METHODS: In this retrospective, single-center, consecutive case series, the refractive outcomes of 254 eyes of 127 patients who underwent DMEK + cataract/IOL triple procedure in both eyes for Fuchs endothelial corneal dystrophy have been analyzed. Main outcome measures were spherical equivalent outcome (shift calculations), best spectacle-corrected visual acuity, central corneal thickness, and posterior simulated keratometry. RESULTS: The mean best spectacle-corrected visual acuity before surgery was 0.51 ± 0.24 and increased to 0.19 ± 0.15 (logMAR) after surgery (P < 0.001). After surgery, a mean hyperopic shift of 0.98 ± 0.89 D was observed. The refractive shift was 1.03 ± 0.93 D and 0.92 ± 1.02 D, in the first and second eyes, respectively (P = 0.435). In a paired analysis, the mean difference of the refractive shift between the first and second eyes was 0.49 ± 0.43 D. CONCLUSIONS: In our fellow eye comparison, the refractive shift after DMEK + cataract/IOL triple procedure in the second eye was comparable with the shift in the first eye. As a consequence, the refractive outcome of the first eye might serve as a reference for optimizing the refractive target in the second eye. Further studies investigating the influence of corneal parameters on refractive shift are needed for a more predictable lens power selection.

SARS-CoV-2: Impact on, Risk Assessment and Countermeasures in German Eye Banks.

INTRODUCTION: Since the beginning of the COVID-19 pandemic there has been some debate regarding the risk of transmission through tissue transplantation and tissue banking processes. AIM OF THE STUDY: To analyze the changes that SARS-CoV-2 has caused regarding the harvesting of corneal donor tissue and eye bank activities in Germany. METHODS: A questionnaire was provided to 26 eye banks in Germany, consisting of questions about adaptations made in the screening of potential donors and the harvesting of corneal tissue following the pandemic spread of SARS-CoV-2. RESULTS: Eighteen eye banks actively reduced recruitment of donors and two banks ceased all activity. Additional diagnostic screening was performed in eight banks, using conjunctival swabs and/or nasopharyngeal swabs. In six eye banks, additional protective measures, such as FFP2 masks and/or facial shields, were implemented. Overall, a mean reduction in the number of obtained donor tissues of 17% was observed. DISCUSSION: Conjunctival and/or nasopharyngeal swabs of donors have been implemented by a minority. Reasons for not performing additional tests may be moderate sensitivity and lack of validation for postmortem use of RT-PCR testing. Also, the hazard of SARS-CoV-2 entering the corneal donor pool with subsequent transmission might be perceived as theoretical. Face shields provide a sufficient barrier against splash and splatter contamination but may be insufficient against aerosols. Additional face masks would provide support against aerosols, but it remains debatable if corneal harvesting can be considered an aerosol-producing procedure. In the future we expect to see changes in current guidelines because of a surge in scientific activities to improve our understanding of the risks involved with cornea donation in the COVID-19 pandemic, and because current practice may reduce the availability of donor corneas due to new exclusion criteria while the demand remains unchanged.

Association of rs613872 and Trinucleotide Repeat Expansion in the TCF4 Gene of German Patients With Fuchs Endothelial Corneal Dystrophy.

PURPOSE: To investigate single nucleotide polymorphisms (SNPs) and trinucleotide repeat (TNR) expansion in the transcription factor 4 (TCF4) gene in a large cohort of German patients with Fuchs endothelial corneal dystrophy (FECD). METHODS: Genomic DNA was obtained from 398 patients with FECD and from 58 non-FECD controls. Thirty-seven previously reported SNPs were evaluated by genotyping. The 398 FECD samples were analyzed for TNR expansions by short tandem repeat assays and Southern blotting. The possible associations between the TNR length and clinical parameters (age, sex, visual acuity, and central corneal thickness) were analyzed in 132 patients. RESULTS: The SNPs in COL8A2, TCF8, LOXHD1, and AGBL1 showed no heterogeneity in 36 cases, although SLCA411 showed 3 nonsense mutations. SNPs were detected for TCF4 (rs613872, rs2123392, rs17089887, rs1452787, and rs1348047), but only rs613872 showed a significant association with FECD (P = 9.93 × 10). Overall, 315/398 (79%) patients harbored TNR lengths >50, whereas no non-FECD controls harbored TNR lengths >50. The TCF4 SNP rs613872 genotype was TT: 39 (67%), TG: 18 (31%), and GG: 1 (2%) in non-FECD controls; TT: 39 (47%), TG: 38 (46%), and GG: 6 (7%) in FECD cases harboring TNR <50; and TT: 23 (8%), TG: 224 (79%), and GG: 38 (13%) in FECD cases harboring TNR >50 (P = 2.93 × 10). No significant association was detected between the TNR length and clinical parameters. CONCLUSIONS: Our large German cohort demonstrated a significant association between the risk allele G in rs613872 and FECD, irrespective of TNR expansion, although this risk allele was more frequent in FECD cases with TNR expansion than without.

Effect of Trinucleotide Repeat Expansion on the Expression of TCF4 mRNA in Fuchs' Endothelial Corneal Dystrophy.

Purpose: CTG trinucleotide repeat (TNR) expansion is frequently found in transcription factor 4 (TCF4) in Fuchs' endothelial corneal dystrophy (FECD), though the effect of TNR expansion on FECD pathophysiology remains unclear. The purpose of this study was to evaluate the effect of TNR expansion on TCF4 expression in corneal endothelium of patients with FECD. Methods: Peripheral blood DNA and Descemet membrane with corneal endothelium were obtained from 203 German patients with FECD. The CTG TNR repeat length in TCF4 was determined by short tandem repeat (STR) assays and Southern blotting using genomic DNA. Genotyping of rs613872 in TCF4 was performed by PCR. TCF4 mRNA levels in corneal endothelium were evaluated by quantitative PCR using three different probes. Control corneal endothelial samples were obtained from 35 non-FECD subjects. Results: The STR assay and Southern blotting showed that 162 of the 203 patients with FECD (80%) harbored CTG trinucleotide repeat lengths larger than 50. Quantitative PCR using all three probes demonstrated that TCF4 mRNA is significantly upregulated in the corneal endothelium of patients with FECD, regardless of the presence of TNR expansion. However, the length of the TNR tended to show a positive correlation with TCF4 expression level. No correlation was shown between the genotype of TCF4 SNP, rs613872, and the level of TCF4 expression. Conclusions: Our findings showed that TCF4 mRNA is upregulated in the corneal endothelium of patients with FECD. Further studies on the effects of TCF4 upregulation on corneal endothelial cell function will aid in understanding the pathophysiology of FECD.

Can we predict the refractive outcome after triple Descemet membrane endothelial keratoplasty?

PURPOSE:: To analyze and correlate corneal parameters with refractive shift after Descemet membrane endothelial keratoplasty combined with cataract surgery (triple Descemet membrane endothelial keratoplasty). METHODS:: This single-center retrospective observational case series included 152 eyes of 152 consecutive patients undergoing triple Descemet membrane endothelial keratoplasty in the first eye for Fuchs endothelial corneal dystrophy. Patients were examined preoperatively, as well as at 3, 6, and 12 months after surgery. The main outcome measures were: refractive shift (predicted refractive outcome based on intraocular lens calculation compared to actual postoperative refractive outcome), central corneal thickness, corneal volume, anterior and posterior corneal curvature, and corneal densitometry. These parameters were analyzed and correlated with the refractive shift after surgery. RESULTS:: After 3 months from surgery, a mean refractive shift of +1.12 ± 1.10 D was observed and remained stable until the last follow-up at 12 months (+1.24 ± 1.07 D). Correlation analysis showed a weak but significant positive correlation between refractive shift and preoperative posterior curvature (rho = 0.314; p = 0.002) or preoperative posterior densitometry (rho = 0.227; p = 0.008). No correlation was found between refractive shift and preoperative central corneal thickness, corneal volume, anterior curvature, or anterior/mid-cornea densitometry. CONCLUSION:: Changes of the posterior cornea may have an influence on the refractive shift. Patients with flatter posterior corneal curvature or higher posterior corneal density seem to exhibit a higher hyperopic shift. The weak correlations indicate a poor predictive value of any preoperative parameter used in our study.

Fungal infection after endothelial keratoplasty: association with hypothermic corneal storage.

PURPOSE: To compare the incidence of fungal infection after endothelial keratoplasty (EK) when donor tissue had been stored in hypothermic medium or organ culture. METHODS: We describe the clinical features of 10 cases of fungal infection (keratitis or endophthalmitis) following EK identified at three European centres. Case definition was the culture of fungus or a positive PCR from the host cornea or anterior chamber after EK. A survey of the incidence of infection after EK was conducted by the European Eye Bank Association. The main outcome measure was the number of cases in which donor tissue had been stored in hypothermic medium compared with organ culture. RESULTS: The 10 cases occurred between 2014 and 2017. All donor corneas had been stored in hypothermic medium sourced from three US eye banks. Three pairs of mate corneas caused infections in six recipients. Candida spp were identified from nine cases, with one isolate of Purpureocillium lilacinum. Data on 16 862 corneas supplied for EK were available from 16 European eye banks for the 5-year period from 2012. There were 17 reported cases of infection, of which 15 (88%) were fungal infections and 14 (82%) were Candida spp. Fungal infection was reported from 3 of 14 476 (0.02%) corneas supplied in organ culture compared with 12 of 2386 (0.50%) corneas supplied in hypothermic medium (p<0.0001). The incidence of infection after hypothermic storage was similar for material sourced from Europe (0.52%) or the USA (0.61%). CONCLUSIONS: Infection after EK is strongly associated with Candida spp. The possible explanations for the higher incidence of infection when tissue is stored in hypothermic medium are discussed.

Fungal Interface Keratitis After Descemet Membrane Endothelial Keratoplasty.

PURPOSE: To evaluate the incidence, clinical course, and management of fungal interface keratitis (IK) after Descemet membrane endothelial keratoplasty (DMEK). METHODS: This is a single-center retrospective observational case series of 3950 eyes undergoing DMEK. Six eyes with fungal IK were detected and analyzed. Analysis included graft storage condition, incidence of fungal IK, identification of the pathogenic agent, topical/systemic and surgical treatment regimen, and best-corrected visual acuity. RESULTS: Fungal IK after DMEK occurred in 6 of 3950 cases (0.15%). Corneal grafts were either stored in Optisol-GS (n = 4) or in organ culture (n = 2). In all cases, Candida species were isolated (Candida tropicalis, Candida albicans, Candida orthopsilosis, and Candida guilliermondii). Four eyes developed fungal IK during the early postoperative period (3-5 d) and 2 eyes later at 16 to 42 days after surgery. All patients received topical and systemic antifungal treatment and intracameral application of antifungal agents. In the case of an early infection, graft removal was performed in 3 of 4 patients. Late infections were eradicated without graft exchange. Recurrence of fungal infection was observed in 1 case after early IK and in both cases after late IK. Final visual acuity ranged from 20/200 to 20/20. CONCLUSIONS: Fungal IK is a rare complication after DMEK. Based on our experience, we believe that treatment of early fungal IK with aggressive presentation should include both immediate graft exchange and intracameral application of voriconazole and amphotericin, in addition to topical and systemic antifungal treatment. Graft exchange seems not to be mandatory in late infections.

Air Versus Sulfur Hexafluoride Gas Tamponade in Descemet Membrane Endothelial Keratoplasty: A Fellow Eye Comparison.

PURPOSE: To perform a fellow eye comparison of outcomes and complications when using air or sulfur hexafluoride (SF6) gas as a tamponade in Descemet membrane endothelial keratoplasty (DMEK). METHODS: One hundred thirty-six eyes of 68 consecutive patients who underwent uneventful DMEK in both eyes for Fuchs endothelial corneal dystrophy were included in this retrospective study. Inclusion criteria were air tamponade (80% of the anterior chamber volume) in the first eye and 20% SF6 gas tamponade (80% of the anterior chamber volume) in the second eye; and same donor tissue culture condition in both eyes. All eyes received laser iridotomy on the day before DMEK. Main outcome measures included preoperative and postoperative best-corrected visual acuity, endothelial cell density, corneal volume, rebubbling rate, and rate of postoperative pupillary block caused by the air/gas bubble. RESULTS: Thirteen of 68 eyes (19.1%) with an air tamponade needed rebubbling compared with 4 of 68 eyes (5.9%) with an SF6 gas tamponade (P = 0.036). Postoperative pupillary block necessitating partial release of air/gas occurred in 1 eye (1.5%) with an air tamponade and 3 eyes (4.4%) with an SF6 gas tamponade (P = 0.301). There were no significant differences in preoperative and postoperative best-corrected visual acuity, endothelial cell density, and corneal volume within 3-month follow-up. CONCLUSIONS: Our results confirm the previously reported better graft adhesion when using an SF6 gas tamponade in DMEK without increased endothelial cell toxicity. The rate of pupillary block in eyes with an SF6 gas tamponade was comparable to that with an air tamponade. As a consequence, we recommend using SF6 gas as the tamponade in DMEK.

"Bubble-in-the-Roll" Technique Using the Endoject DMEK Injector: Influence of the Air Bubble on Endothelial Cell Loss.

PURPOSE: To evaluate the impact of the air bubble on endothelial cell loss using the "bubble-in-the-roll" technique during Descemet membrane endothelial keratoplasty (DMEK). METHODS: Twenty DMEK grafts not suitable for transplantation were manually prepared from organ-cultured corneoscleral discs and injected into culture media using the Endoject DMEK injector (Medicel AG, Wolfhalden, Switzerland). Based on the injection method, the grafts were divided into 2 groups: In group A (n = 10), a small air bubble was placed inside the graft roll while it was in the injector. In group B (n = 10), the grafts were injected without an air bubble inside the graft roll. Main outcome measures included endothelial cell density (ECD) after graft stripping and graft injection. RESULTS: There were no statistically significant differences between groups A and B in donor age, storage duration, and donor ECD. ECD decreased from 1929 ± 145 cells/mm to 1796 ± 303 cells/mm after graft stripping in group A and from 1801 ± 226 cells/mm to 1709 ± 290 cells/mm in group B. ECD after graft injection further decreased to 1683 ± 291 cells/mm in group A and to 1651 ± 292 cells/mm in group B. Endothelial cell loss after graft stripping and graft injection was not statistically significant between groups A and B (P = 0.29 and P = 1, respectively). CONCLUSIONS: The bubble-in-the-roll technique for injection and unfolding of the graft is a safe method for graft delivery into the anterior chamber guaranteeing orientation of the graft without harming the endothelium.

Sustained Activation of the Unfolded Protein Response Induces Cell Death in Fuchs' Endothelial Corneal Dystrophy.

Purpose: The unfolded protein response (UPR) is believed to play a role in the pathogenesis of Fuchs' endothelial corneal dystrophy (FECD). The purpose of this study was to investigate whether unfolded proteins accumulate in the corneal endothelium in FECD and if they are involved in triggering cell death. Methods: Descemet's membranes with corneal endothelial cells (CECs) were obtained during keratoplasty, and expression of aggresomes, type 1 collagen, fibronectin, and agrin was evaluated. Endoplasmic reticulum (ER) stress of immortalized human CECs from non-FECD subjects and from FECD patients (iHCEC and iFECD, respectively) were evaluated. The effect of MG132-mediated aggresome formation on the UPR and intrinsic pathway and the effect of mitochondrial damage on UPR were also examined. The effect of CHOP knockdown on the ER stress-mediated intrinsic pathway was also evaluated. Results: Aggresome formation was higher in iFECD than in iHCEC and was colocalized with type 1 collagen, fibronectin, and agrin. GRP78, phosphorylated IRE1, PERK, and CHOP showed higher activation in iFECD than in iHCEC. MG132-mediated aggresome formation upregulated ER stress sensors, the mitochondrial membrane potential drop, cytochrome c release to the cytoplasm, and activation of caspase-9 and -3. By contrast, staurosporine-mediated mitochondrial damage did not induce ER stress. Knockdown of CHOP attenuated the ER stress-induced cleavage of caspase-9, which is caused by intrinsic pathway activation. Conclusions: Excessive synthesis of extracellular matrix proteins induced unfolded protein accumulation in FECD. Prolonged ER stress-mediated cell death, occurring via the intrinsic apoptotic signaling pathway, therefore might be associated with the pathogenesis of FECD.

Activation of TGF-ß signaling induces cell death via the unfolded protein response in Fuchs endothelial corneal dystrophy.

Fuchs endothelial corneal dystrophy (FECD) is a slowly progressive bilateral disease of corneal endothelium in which accumulation of extracellular matrix (ECM) and loss of corneal endothelial cells (CECs) are phenotypic features. The corneal endothelium maintains corneal transparency by regulating water hydration; consequently, corneal endothelial dysfunction causes serious vision loss. The only therapy for corneal haziness due to corneal endothelial diseases, including FECD, is corneal transplantation using donor corneas, and no pharmaceutical treatment is available. We provide evidence that the expression levels of transforming growth factor-ß (TGF-ß) isoforms and TGF-ß receptors are high in the corneal endothelium of patients with FECD. A cell model based on patients with FECD shows that TGF-ß signaling induced a chronic overload of ECM proteins to the endoplasmic reticulum (ER), thereby enhancing the formation of unfolded protein and triggering the intrinsic apoptotic pathway through the unfolded protein response (UPR). We propose that inhibition of TGF-ß signaling may represent a novel therapeutic target that suppresses cell loss as well as the accumulation of ECM in FECD.