Comprehensive assessment of corneal microstructural changes following V4c implantable collamer lens surgery using in vivo confocal microscopy.

BACKGROUND: Implantable Collamer Lense (ICL) presents a viable alternative to conventional refractive surgeries, but their impact on corneal microstructure remains unclear. By employing in vivo confocal microscopy (IVCM), we examined changes in stromal and endothelial cells following the insertion of V4c ICLs, with the goal of enhancing post-surgical care and outcomes. METHODS: In this longitudinal investigation, we conducted detailed preoperative assessments on 103 eyes from 53 participants. Follow-up evaluations were carried out after surgery at set intervals: one day, one week, one month, three months, six months, and twelve months. We used IVCM to analyze changes in stromal and endothelial cells. To assess differences between pre- and post-surgery variables and to investigate correlations with age, axial length (AL), and spherical equivalent refraction (SER), we applied a repeated measures mixed-effects model, with statistical significance set at P < 0.05. RESULTS: No vision-threatening complications were reported post-surgery. Significant reductions in stromal cell density (SCD) were observed postoperatively, with anterior and mid- SCD reaching their lowest values at 3 months and posterior SCD at 1 month, remaining below baseline at 12 months. endothelial cell density (ECD) and percentage of hexagonal cells (PHC) decreased initially, recovering by 12 months. Conversely, endothelial cellular area (ECA) and coefficient of variation of cell size (CoV) increased postoperatively, with the most significant change at 1 week. Endothelial deposits were detected in 49 of 101 eyes on postoperative day 1, half of them were absorbed within 3 months post-surgery. Changes in posterior SCD were negatively related to AL, while AL, SER, lens thickness showed associated with endothelium changes. CONCLUSION: Our findings elucidate the corneal microstructural changes following V4c ICL implantation, particularly the significant early reductions in stromal and endothelial cell densities. We recommend careful management of viscoelastics during surgery to minimize endothelial deposits that may harm the endothelium. Enhanced early postoperative monitoring and these surgical adjustments can lead to improved surgical and post-surgical care, ultimately supporting better patient recovery.

Hepatocyte Growth Factor Modulates Corneal Endothelial Wound Healing In Vitro.

In this study, we assessed the impact of hepatocyte growth factor (HGF) on corneal endothelial cells (CECs), finding that HGF concentrations of 100-250 ng/mL significantly increased CEC proliferation by 30%, migration by 32% and improved survival under oxidative stress by 28% compared to untreated controls (p < 0.05). The primary objective was to identify non-fibrotic pharmacological strategies to enhance corneal endothelial regeneration, addressing a critical need in conditions like Fuchs' endothelial dystrophy (FED), where donor tissue is scarce. To confirm the endothelial nature of the cultured CECs, Na+/K+-ATPase immunohistochemistry was performed. Proliferation rates were determined through BrdU incorporation assays, while cell migration was assessed via scratch assays. Cell viability was evaluated under normal and oxidative stress conditions using WST-1 assays. To ensure that HGF treatment did not trigger epithelial-mesenchymal transition, which could lead to undesirable fibrotic changes, α-SMA staining was conducted. These comprehensive methodologies provided robust data on the effects of HGF, confirming its potential as a therapeutic agent for corneal endothelial repair without inducing harmful EMT, as indicated by the absence of α-SMA expression. These findings suggest that HGF holds therapeutic promise for enhancing corneal endothelial repair, warranting further investigation in in vivo models to confirm its clinical applicability.

Rapid detection of guttae area using aniline blue staining in Fuchs endothelial corneal dystrophy mouse model.

Fuchs endothelial corneal dystrophy (FECD) is a leading cause of corneal endothelial degeneration resulting in impaired visual acuity. Excessive deposition of extracellular matrix (guttae) on Descemet's membrane (DM) is the hallmark of FECD. We sought to detect the guttae area rapidly using aniline blue (AB) staining in FECD mouse model. FECD mouse model was established via ultraviolet A (UVA) exposure. Masson's trichrome staining was utilized to stain the corneal sections. AB staining was utilized to stain both whole cornea tissues and stripped Descemet's membrane-endothelium complex (DMEC) flat mounts, while immunofluorescence staining of collagen I was employed to stain guttae areas. In Masson's trichrome staining, corneal collagen fibrils were stained blue with AB. The DMEC flat mounts were stained into relative dark blue areas and relative light blue areas using 2% AB staining. The areas of dark blue could almost overlap with collagen I-positive areas, and have an acellular centre and a moderately distinct boundary line with the surrounding corneal endothelial cells. In conclusion, AB staining is a rapid and effective method for the evaluation of the guttae areas in the FECD mouse model.

MitoQ relieves mitochondrial dysfunction in UVA and cigarette smoke-induced Fuchs endothelial corneal dystrophy.

Fuchs endothelial corneal dystrophy (FECD), a degenerative corneal condition, is characterized by the droplet-like accumulation of the extracellular matrix, known as guttae and progressive loss of corneal endothelial cells ultimately leading to visual distortion and glare. FECD can be influenced by environmental stressors and genetic conditions. However, the role of mitochondrial dysfunction for advancing FECD pathogenesis is not yet fully studied. Therefore, in the present study we sought to determine whether a combination of environmental stressors (ultraviolet-A (UVA) light and cigarette smoke condensate (CSC)) can induce mitochondrial dysfunction leading to FECD. We also investigated if MitoQ, a water-soluble antioxidant, can target mitochondrial dysfunction induced by UVA and CSC in human corneal endothelial cells mitigating FECD pathogenesis. We modeled the FECD by increasing exogenous oxidative stress with CSC (0.2%), UVA (25J/cm2) and a combination of UVA + CSC and performed a temporal analysis of their cellular and mitochondrial effects on HCEnC-21T immortalized cells in vitro before and after MitoQ (0.05 µM) treatment. Interestingly, we observed that a combination of UVA + CSC exposure increased mitochondrial ROS and fragmentation leading to a lower mitochondrial membrane potential and increased levels of cytochrome c release leading to apoptosis and cell death. MitoQ intervention successfully mitigated these effects and restored cell viability. The UVA + CSC model could be used to study stress induced mitochondrial dysfunction. Additionally, MitoQ can serve as a viable antioxidant in attenuating mitochondrial dysfunction, underscoring its potential as a molecular-focused treatment approach to combat FECD pathogenesis.

Neuropeptide alpha-Melanocyte stimulating hormone preserves corneal endothelial morphology in a murine model of Fuchs dystrophy.

Fuchs endothelial corneal dystrophy is a heterogenous disease with multifactorial etiology, and genetic, epigenetic, and exogenous factors contributing to its pathogenesis. DNA damage plays a significant role, with ultraviolet-A (UV-A) emerging as a key contributing factor. We investigate the potential application of neuropeptide α-melanocyte stimulating hormone (α-MSH) in mitigating oxidative stress induced endothelial damage. First, we examined the effects of α-MSH on a cultured human corneal endothelial cell line (HCEnC-21T) exposed to hydrogen peroxide (H2O2) induced oxidative DNA damage. We performed immunofluorescence and flow cytometry to assess DNA damage and cell death in the cultured cells. Additionally, we used an established mouse model that utilizes ultraviolet light to induce corneal endothelial cell damage resulting in decreased CEnC number, increased cell size variability, and decreased percentage of hexagonal cells. This endothelial decompensation leads to an increase in corneal thickness. Following UV-A exposure, the mice were systemically treated with α-MSH, either immediately after exposure (early treatment) or beginning two weeks post-exposure (delayed treatment). To evaluate treatment efficacy, we analyzed CEnC density and morphology using in vivo confocal microscopy, and central corneal thickness using anterior segment optical coherence tomography. Our findings demonstrated that α-MSH treatment effectively protects HCEnC-21T from free-radical induced oxidative DNA damage and subsequent cell death. In vivo, α-MSH treatment, mitigated the loss of CEnC density, deterioration of cell morphology and suppression of the resultant corneal swelling. These results underline the potential application of α-MSH as a therapeutic agent for mitigating corneal endothelial damage.

Correlation between corneal endothelial layer features and age-related macular degeneration severity.

PURPOSE: This study aimed to investigate the relationship between corneal endothelial layer features and the severity of age-related macular degeneration (AMD). METHODS: The study included 119 patients, with 47 females and 72 males. Patients were categorized into four groups based on the AREDS grading system: no AMD (group 1), mild AMD (group 2), moderate AMD (group 3), and advanced AMD (group 4). Only the right eye of patients with both eyes suitable for the study was included. Corneal endothelial cell density (CD), coefficient of variation (CoV), hexagonal cell ratio (HEX), and central corneal thickness (CCT) were measured using specular microscopy (Konan Medical Inc., Nishinomiya, Japan). RESULTS: Group 1 had 40 patients, group 2 had 27 patients, and groups 3 and 4 had 26 patients each. Significant differences were observed between the mean endothelial CD, CoV, and HEX values among the groups, while no significant difference was found in CCT values (p = 0.049, p = 0.002, p = 0.004, and p = 0.883, respectively). A mild negative correlation was observed between AMD severity and CD and HEX values, while a mild positive correlation was found between AMD severity and CoV. CONCLUSION: Increasing severity of AMD may negatively impact corneal endothelial layer values.

Safety and prognosis of phacoemulsification using active sentry and active fluidics with different IOP settings - a randomized, controlled study.

PURPOSE: To explore the impact of different intraoperative intraocular pressure (IOP) settings on the safety and prognosis in phacoemulsification. METHODS: Age related cataract patients who met the inclusion criteria and underwent phacoemulsification by using active sentry handpiece and active fluidics system. According to different intraoperative IOP settings during surgery, they were randomly divided into two groups: the 20mmHg group and the 60mmHg group. The best corrected visual acuity (BCVA), cumulative dissipated energy (CDE), total U/S time, active surge mitigation (ASM), estimated fluid usage (EFU) as well as the changes in corneal thickness (CT), corneal epithelial layer thickness (CELT) and endothelial cell density (ECD) were collected. The post-operative follow-up was only 1 day. RESULTS: A total of 110 cases (110 eyes) were included in the study. There were 55 eyes in each group. There was no statistically significant difference in postoperative BCVA (p = 0.839). The CDE, total U/S time and EFU during surgery were (5.22 ± 3.31), (30.60 ± 15.06), (45.07 ± 12.68) and (4.70 ± 2.83), (27.39 ± 13.75), (42.38 ± 11.93) in the 20mmHg group and 60mmHg group (p = 0.381, 0.246, 0.254). The ASM during surgery in the 20mmHg group and 60mmHg group were (0.95 ± 2.77) and (7.24 ± 6.34), respectively. The 20mmHg group showed a significant decrease in ASM (p < 0.001). There was no statistically significant difference in the changes in CT, CELT and ECD before and after surgery between the two groups (p = 0.913, 0.825, 0.624). Both groups did not experience any intraoperative complications, such as posterior capsule rupture. CONCLUSION: A lower IOP setting of 20 mmHg can significantly reduce the occurrence of intraoperative surges during phacoemulsification. And there was no increase in rate of complications. TRIAL REGISTRATION: The trial registration number is ChiCTR2100050240. The registered date is August 24th, 2021.

Comparison of Pacman cutting technique and trisection cutting technique for hydrophobic acrylic intraocular lens removal.

The objective of this retrospective cohort study is to compare the Pacman cutting technique with the trisection cutting technique for hydrophobic acrylic intraocular lens extraction. A total of 29 eyes of 29 patients requiring IOL removal were selected and enrolled in this study. The patients in group 1 (13 cases) underwent the conventional trisection cut technique from May 2022 to Dec. 2022. The patients in group 2 (16 cases) underwent the modified partial IOL cutting technique (Pacman cutting technique) from Jan. 2023 to June 2023. Corneal endothelial cells (CECs) count, slit-lamp examination, and intraocular pressure (IOP) measurement were conducted at all preoperative and postoperative visits. Surgical time, intraoperative and postoperative complications were recorded. The mean surgical time was 3.48 ± 1.54 minutes (range: 2 to 7 minutes) for the trisection cut technique group and 2.52 ± 1.18 minutes (range: 2 to 5 minutes) for the Pacman cut technique group. There was a statistically significant difference between the two groups in terms of surgical duration (P < .05). There was no difference in the mean preoperative CEC counts (group 1: 1924 ± 462, group 2: 1876 ± 516, P > 0.05). However, three months after the operation, the count of CECs decreased by 213 ± 72 (11%) in group 1 and 142 ± 57 (8%) in group 2. The decrease in CECs in group 1 was greater than that in group 2, and the difference was statistically significant (P < .05). There were no serious complications affecting vision in either group during the follow-up period. This study showed that the Pacman cutting technique is more effective and safer than the trisection cutting technique for hydrophobic acrylic IOL removal.

Automatic Determination of Endothelial Cell Density From Donor Cornea Endothelial Cell Images.

Purpose: To determine endothelial cell density (ECD) from real-world donor cornea endothelial cell (EC) images using a self-supervised deep learning segmentation model. Methods: Two eye banks (Eversight, VisionGift) provided 15,138 single, unique EC images from 8169 donors along with their demographics, tissue characteristics, and ECD. This dataset was utilized for self-supervised training and deep learning inference. The Cornea Image Analysis Reading Center (CIARC) provided a second dataset of 174 donor EC images based on image and tissue quality. These images were used to train a supervised deep learning cell border segmentation model. Evaluation between manual and automated determination of ECD was restricted to the 1939 test EC images with at least 100 cells counted by both methods. Results: The ECD measurements from both methods were in excellent agreement with rc of 0.77 (95% confidence interval [CI], 0.75-0.79; P < 0.001) and bias of 123 cells/mm2 (95% CI, 114-131; P < 0.001); 81% of the automated ECD values were within 10% of the manual ECD values. When the analysis was further restricted to the cropped image, the rc was 0.88 (95% CI, 0.87-0.89; P < 0.001), bias was 46 cells/mm2 (95% CI, 39-53; P < 0.001), and 93% of the automated ECD values were within 10% of the manual ECD values. Conclusions: Deep learning analysis provides accurate ECDs of donor images, potentially reducing analysis time and training requirements. Translational Relevance: The approach of this study, a robust methodology for automatically evaluating donor cornea EC images, could expand the quantitative determination of endothelial health beyond ECD.

Outcomes of retropupillary iris claw lens implantation in patients with intraocular lens dislocation and low (less than 1000 cells/mm2) corneal endothelial cell density.

BACKGROUND: Posterior chamber intraocular lens (IOL) dislocation is a common complication of cataract surgery. Dislocated IOLs often require surgical intervention due to the potentially severe risks of leaving this condition untreated. If a patient with extremely low corneal endothelial cell density (ECD) presents with IOL dislocation, the surgeon faces a crucial dilemma of choosing the most optimal surgical treatment option. We sought to investigate the efficacy and safety of retropupillary iris claw intraocular lens (R-IOL) implantation in patients with IOL dislocation and extremely low (< 1000 cells/mm2) ECD. METHODS: We retrospectively reviewed the medical records of nine patients (all men) whose pre-operative ECD was < 1000 cells/mm2 and who underwent R-IOL implantation due to intraocular subluxation or total dislocation into the vitreous cavity between 2014 and 2020. We evaluated corneal endothelial function and visual outcomes after surgery. RESULTS: Nine patients were included in this study. The mean age at diagnosis was 64.89 ± 7.15 years (range 57-76 years), and the follow-up duration was 37.93 ± 23.72 months (range 18.07-89.07 months). No patients developed bullous keratopathy during follow-up. Compared to the initial ECD, corneal thickness (CT), coefficient variation of cell area (CV) and percentage of hexagonal cells (HEX), there was no statistically significant decrease in the ECD, CV, and HEX at last follow-up (P = 0.944, 0.778, 0.445, 0.443). There was significant improvement in the mean uncorrected distance visual acuity (UDVA) at the last follow-up (average 0.13 logMAR, 20/27 Snellen) compared to the pre-operative mean UDVA (average 1.09 logMAR, 20/250 Snellen) (P < 0.01). CONCLUSIONS: R-IOL implantation did not result in a statistically significant decline in corneal endothelial function in patients with preoperatively low ECD, and it significantly improved the mean UDVA postoperatively. R-IOL implantation appears to be a safe and effective treatment modality for intraocular lens dislocation in patients with low ECD (< 1000 cells/mm²); however, long-term follow-up studies are warranted to corroborate these findings.

The progress and future of corneal endothelial transplantation.

Endothelial transplantation has recently been accepted worldwide, in the long history of corneal transplantation. The introduction of endothelial keratoplasty (Descemet stripping automated endothelial keratoplasty and Descemet membrane endothelial keratoplasty) has enabled us to expand the surgical indications owing to the low incidence of rejection and quick recovery of visual function. New technologies have been developed to ensure stable postoperative outcomes with a shorter learning curve, such as transplantation using cultured human endothelial cells and induced pluripotent stem cells (iPS) or new devices such as artificial endothelium. This review discusses the history and characteristics of corneal transplantation alongside new treatment options that may offer hope for patients with endothelial disease in the future.

Guttae Morphology After Cultured Corneal Endothelial Cell Transplant in Fuchs Endothelial Corneal Dystrophy.

Importance: Whether guttae in Fuchs endothelial corneal dystrophy (FECD) can be removed by polishing without Descemet stripping and whether postoperative maintenance of reduced guttae can be achieved through cultured corneal endothelial cell (CEC) transplant therapy are critical issues to be addressed. Objective: To investigate the decrease of guttae through polishing degenerated CECs and abnormal extracellular matrix (ECM) without Descemet stripping and to observe the behavior of guttae following cultured CEC transplant. Design, Setting, and Participants: This case series prospective observational study was conducted in a hospital outpatient clinic setting. Between December 2013 and January 2019, 22 eyes with corneal endothelial failure caused by FECD received cultured CEC transplant therapy at Kyoto Prefectural University Hospital. Of these, 15 eyes were consistently monitored at the same central corneal area during the preoperative phase, as well as in the early (within 1 year) and late (after 3 years) postoperative phases. The images from these phases were categorized into 3 groups: typical guttae, atypical guttae, and no guttae. Exposures: Cultured CEC transplant therapy. Main Outcomes: Proportion of guttae in the observable area was measured, comparing the early and late postoperative phases for each group. Results: The mean age of the patients at the time of surgery was 69 years (range, 49-79 years). All 15 eyes exhibited the presence of confluent guttae preoperatively (100%). Among these, 3 of 15 eyes belonged to male patients. The early postoperative phase of guttae morphologies was classified into 3 groups: 5 eyes with typical guttae, 7 with atypical guttae, and 3 with no guttae. The decrease in the number of these guttae was achieved by surgical procedures. The median percentage of guttae in the typical guttae, atypical guttae, and no guttae groups was 41.8%, 44.4%, and 16.2%, respectively, in the early phase, and 42.2%, 38.2%, and 18.8%, respectively, in the late phase. Conclusions and Relevance: The findings demonstrate that in some cases of FECD, guttae can be removed by scraping and polishing abnormal ECM and degenerated CECs, while preserving the Descemet membrane. Furthermore, cultured CEC transplant resulted in no increase in guttae for up to 3 years, providing insights into surgically eliminating guttae.

Corneal parameters in diabetics versus non-diabetics and correlation with various blood sugar parameters.

Aim: To compare corneal parameters in diabetics versus age-group-matched non-diabetics; also, to correlate these parameters with the duration of diabetes, glycated haemoglobin (HbA1c) levels, and severity levels of diabetic retinopathy (DR). Materials and methods: A comparative study was conducted at a tertiary eye-care center from January 2020 to December 2020. Two-hundred patients (400 eyes) with type-2 diabetes (100) and age-sex-matched non-diabetics (100) were included. Corneal morphological parameters like central corneal thickness (CCT), endothelial cell density (ECD), coefficient of variance (CoV), hexagonality (6A), and average cell area were recorded by specular microscopy. These parameters were correlated with the duration of diabetes, severity of disease based upon fasting blood glucose levels, HbA1c, and grade of DR. Mean and standard deviation were calculated, and regular distribution of continuous data was tested using independent sample t-test and ANOVA. Results: Mean ECD (2447.32 ± 269.89/mm2), 6A (45.03 ± 6.71%), and IOP (15.47 ± 2.02 mmHg) changed in diabetic cases and were significantly low in diabetics, whereas, mean average cell area (413 ± 50.19 mm2), standard deviation (167.05 ± 77.91), CCT (525.81 ± 36.69) and CoV (39.84 ± 15.59%), were significantly high in diabetics. Mean CCT had insignificant variation. Subgroup analysis within diabetics showed a statistically significant reduction of ECD, cell count, and 6A with increased duration of diabetes, poor glycaemic control, and raised HbA1c. Discussion: The corneal endothelial analysis is vital in daily clinical practice and provides valuable evidence concerning the viability of corneal endothelium in various intraocular surgeries. Uncontrolled DM harms the cornea with 70% of diabetics resulting in complications like keratopathy. The study highlighted that the increased duration of diabetes raised HbA1c, and poor glycemic control negatively affected corneal morphology. Our study showed a definite reduction in ECD and 6A in diabetics compared to non-diabetics. Conclusion: A definite reduction in the corneal endothelial counts, cell density, and hexagonality was found in type-2 diabetics compared to non-diabetics. Abbreviations: DM = Diabetes Mellitus, CCT = central corneal thickness, ECC = endothelial cell counts, ECD = endothelial cell density, CoV = coefficient of variance, 6A = hexagonality, DR = Diabetic retinopathy, SD = Standard of deviation, IOP = Intraocular pressure.

Corneal endothelial density loss in patients after gonioscopy­assisted transluminal trabeculotomy.

PURPOSE: To compare short-term changes in corneal endothelial cells after gonioscopy-assisted transluminal trabeculotomy(GATT). METHODS: This retrospective comparative study included 138 patients(138 eyes), and 98 of these patients underwent GATT procedure and 40 underwent SLT procedure as a control group. Changes in the corneal endothelium in patients who underwent GATT and SLT were analyzed retrospectively. Endothelial changes in the central cornea were examined using specular microscopy before and 6 months after the GATT and SLT procedure. Intraocular pressure(IOP), number of glaucoma medications, and side effects were evaluated at visits before and after two methods. RESULTS: One hundred and thirty-eight eyes of 138 patients with a mean age of 62.9±12.7 years in the SLT group and 62.5±11.8 years in the GATT group were included in this study. Pre-procedure mean ± SD IOP was 27.7±3.6 mmHg and 27.4±5.3 mmHg (p=0.173) 2.8±0.5 and 2.9±0.8 (p=0.204) glaucoma drugs are in the SLT and GATT group, respectively. The mean corneal endothelial cell density (CECD) in the SLT group was 2433.1±581.4 cells/mm2 before the procedure and 2435.1±585 cells/mm2 6 months after the procedure, a change of 0.1±0.6% which was not statistically significant (p>0.967).The mean CECD at baseline in the GATT group was 2443.4±508.2 cells/mm2 and decreased to 2290.2±527.7 cells/mm2 6 months after this procedure, representing a cell loss of 6,2±9,1% (p<0.001). CONCLUSION: GATT caused more CECD damage than SLT at the sixth month after the procedure. Considering the loss of CECD in candidates for GATT, sufficient number of endothelial cells in the central cornea is recommended.

Comparison of rebubbling rate between preloaded endothelium-in and preloaded no-touch endothelium-out Descemet membrane endothelial keratoplasty transplantation.

BACKGROUND: To compare the difference in rebubbling rates between patients undergoing Descemet membrane endothelial keratoplasty (DMEK) with endothelium-in using a standard IOL cartridge and those with endothelium-out DMEK utilizing a no-touch technique with borosilicate glass cartridge transplantation. METHODS: This retrospective study included all eyes that underwent preloaded endothelium-in or endothelium-out DMEK transplantation from June 2019 to December 2023 at the Hanusch Hospital, Vienna, Austria. All DMEKs were harvested, prepared and preloaded at the European Eye Bank of Venice, Italy. DMEK surgeries were done by one experienced surgeon and the procedure was completed by air tamponade of the anterior chamber. RESULTS: Overall, 32 eyes each of 31 endothelium-out patients and of 29 endothelium-in patients were included. 32 preloaded endothelium-in procedures were followed by 32 preloaded endothelium-out procedures. Rebubbling rate for endothelium-in was 15/32 (47%) and for endothelium-out was 7/25 (28%) (p = 0.035, Pearson's chi-squared test). Donor age was the most important variable for rebubbling in a random forest algorithm model (ROC: 0.69). CONCLUSIONS: Rebubbling rate in endothelium-out DMEK was less than two-thirds compared to endothelium-in DMEK favoring no-touch endothelium-out DMEK as the preferred technique of DMEK transplantation.

Enhanced Migration of Fuchs Corneal Endothelial Cells by Rho Kinase Inhibition: A Novel Ex Vivo Descemet's Stripping Only Model.

Descemet's Stripping Only (DSO) is a surgical technique that utilizes the peripheral corneal endothelial cell (CEnC) migration for wound closure. Ripasudil, a Rho-associated protein kinase (ROCK) inhibitor, has shown potential in DSO treatment; however, its mechanism in promoting CEnC migration remains unclear. We observed that ripasudil-treated immortalized normal and Fuchs endothelial corneal dystrophy (FECD) cells exhibited significantly enhanced migration and wound healing, particularly effective in FECD cells. Ripasudil upregulated mRNA expression of Snail Family Transcriptional Repressor (SNAI1/2) and Vimentin (VIM) while decreasing Cadherin (CDH1), indicating endothelial-to-mesenchymal transition (EMT) activation. Ripasudil activated Rac1, driving the actin-related protein complex (ARPC2) to the leading edge, facilitating enhanced migration. Ex vivo studies on cadaveric and FECD Descemet's membrane (DM) showed increased migration and proliferation of CEnCs after ripasudil treatment. An ex vivo DSO model demonstrated enhanced migration from the DM to the stroma with ripasudil. Coating small incision lenticule extraction (SMILE) tissues with an FNC coating mix and treating the cells in conjunction with ripasudil further improved migration and resulted in a monolayer formation, as detected by the ZO-1 junctional marker, thereby leading to the reduction in EMT. In conclusion, ripasudil effectively enhanced cellular migration, particularly in a novel ex vivo DSO model, when the stromal microenvironment was modulated. This suggests ripasudil as a promising adjuvant for DSO treatment, highlighting its potential clinical significance.

Analysis after posterior chamber phakic intraocular lens implantation: 17- to 19-year follow-up study.

PURPOSE: To evaluate the clinical outcomes of the Visian implantable collamer lens (ICL) in eyes with ≥17 years of follow-up. SETTING: Single-center. DESIGN: Retrospective observational study. METHODS: 70 eyes from 38 patients who underwent surgery between 2002 and 2006 with V4 ICL implantation were analyzed. Preintervention data included corrected distance visual acuity (CDVA), refraction, and intraocular pressure (IOP). At the 19-year mark, CDVA, refraction, IOP, endothelial count, and vault were measured using Tomey Casia 2 anterior pole tomography, along with recording long-term events. RESULTS: The operated patients had a spherical correction between -4 diopters (D) and -21 D and a cylinder up to 7.5 D. The mean CDVA (decimal) at the 17-year check-up was 0.89 ± 0.18, with a spherical equivalent of -1.05 ± 1.36 D. IOP has remained stable because mean measurements before surgery and currently were 15.16 ± 2.54 mm Hg and 16.19 ± 3.29 mm Hg, respectively. The endothelial cell count showed values of 2191 ± 386 cells/mm2, with a maximum of 2804 cells and a minimum of 1125 cells. Finally, the vault obtained was 348.53 ± 234.58 mm. Over the years, 2 eyes developed angle-closure glaucoma and 9 eyes were operated on for the development of anterior subcapsular cataracts. CONCLUSIONS: Visian ICL implantation demonstrated long-term visual and refractive stability, indicating it as a low-risk procedure. The ICL V4 model yielded satisfactory results for myopia and astigmatism correction after 17 years.

Comparison of automated corneal endothelial cell analysis in healthy and postoperative eyes with phakic intraocular lens: a cross-sectional study and literature review.

PURPOSE: Measurement of corneal endothelial cells is critical for postoperative evaluation of phakic intraocular lens (pIOL) surgery. However, inter-instrument differences in corneal endothelial cell density (ECD) after pIOL implantation have not yet been reported. This study aimed to compare automated corneal endothelial cell analysis between CellChek-20 (Konan Medical, Hyogo, Japan) and EM-4000 (Tomey, Nagoya, Japan) in healthy and postoperative eyes with pIOL. METHODS: We retrospectively analyzed 154 healthy and 236 postoperative eyes after pIOL surgery. Endothelial cell measurements were performed using CellChek-20 and EM-4000 with autofocusing and automated image analysis. ECD, percentage of hexagonal cells (HEX), coefficient of variation in cell size (CoV), and central corneal thickness (CCT) were compared between the two devices. RESULTS: The ECDs of the two devices were highly correlated in both healthy (Spearman's correlation coefficient [r] = 0.805; p < 0.001) and postoperative (r = 0.901; p < 0.001) groups. ECD from CellChek-20 was higher than EM-4000 in both healthy (mean difference = 228.9 cells/mm2; p < 0.001) and postoperative (mean difference = 115.6 cells/mm2; p < 0.001) groups. The CCT values also showed a strong correlation in healthy eyes (r = 0.974; p < 0.001) and in postoperative eyes (r = 0.936; p < 0.001); however, significant inter-instrument differences were observed. HEX and CV showed significant differences and relatively weak correlations (r < 0.7) between the two devices in both healthy and postoperative groups. CONCLUSION: The ECD values between the two instruments were correlated, but that of the CellChek-20 was significantly higher than that of the EM-4000 in both healthy and postoperative eyes after pIOL surgery. Most previous studies have also shown that the Konan software overestimated the ECD compared to other products in automatic measurement mode. The possibility of measurement bias should be considered when replacing equipment used for corneal endothelial cell measurements.

Comparison of corneal endothelial cell density reduction between primary open-angle glaucoma and pseudo-exfoliation glaucoma patients at 3 years after Ex-Press® surgery.

PURPOSE: We compared corneal endothelial cell (CED) loss after Ex-Press (EXP) surgery between patients with primary open-angle glaucoma (POAG) and pseudo-exfoliation glaucoma (PEX). PATIENTS AND METHODS: This was a single-facility retrospective study. We included glaucoma patients who had undergone EXP surgery and were followed up > 3 years. We measured the CED before and after (at 12, 24, and 36 months) EXP surgery by noncontact specular microscopy and compared the means of the CED values and CED survival ratios after EXP surgery by paired t-test. RESULTS: We included 119 eyes that underwent EXP surgery, including 60 POAG eyes and 59 PEX eyes. In the POAG group, the mean CED decreased from 2389 ± 321 at baseline to 2230 ± 424 cells/mm2 after 3 years. In the PEX group, the mean CED decreased from 2111 ± 510 at baseline to 1845 ± 628 cells/mm2 after 3 years. At the 3-year follow-up, the CED survival ratio was 93.3 ± 12.5% in the POAG group and significantly lower, at 85.0 ± 19.5%, in the PEX group (p = 0.0064). Two cases in the PEX group developed bullous keratopathy. CONCLUSIONS: EXP surgery decreased the corneal endothelial cell populations in PEX patients faster than POAG patients.

Clinical outcome analysis of two approaches to trypan blue dyeing for DMEK.

To evaluate the clinical implications of the different trypan blue dyeing techniques used during liquid bubble (LBT) and manual peel (MPT) DMEK lenticule preparation techniques. This study retrospectively compared the degree to which endothelial cells are preserved using selective Descemet Membrane (DM) staining (LBT) versus bath-staining (MPT) when performed by a single surgeon, sourced from a single eye bank. Endothelial cell density measured after the 3-month follow-up was 1805 and 1916 cells/mm2 respectively, differing significantly (p = 0.012). A double-scroll graft formation was found and maintained until implantation in 94% of preparations with bath staining and 50% of preparations using selective DM staining. Preoperative visual acuity was comparable between preparation techniques at 0.4 logMAR as well as postoperatively, at an average of 0.1 logMAR. Reducing chemical stress on the endothelium by avoiding any contact with trypan blue allows for a significantly higher degree of cell preservation. However, achieving the often-desired double-scroll graft formation was possible less frequently. It remains unclear which factors define the differences graft scrolling behavior observed between LBT and MPT.