The hiPSC-derived corneal endothelial progenitor-like cell recovers the rabbit model of corneal endothelial dystrophy.

INTRODUCTION: Corneal endothelial dysfunction results in cornea opacity, damaging sightedness, and affecting quality of life. A corneal transplant is the current effective intervention. Due to the scarcity of donated cornea, such an unmet medical need requires a novel therapeutic modality. OBJECTIVES: Customizing patients' corneal endothelial progenitor cells with proliferative activity and lineage restriction properties shall offer sufficient therapeutic cells for corneal endothelial dystrophy. METHODS: The customized induced human corneal endothelial progenitor-like cell (iHCEPLC) was obtained through cell fate conversions starting from PBMC (peripheral blood mononuclear cell), hiPSC (human induced pluripotent stem cell), and hNCC (human neural crest cell), while it finally reached the iHCEPLC state via a series of induction. Several molecular diagnoses were applied to depict its progenitor state, including RNAseq, FlowCytometer, immunostainings, and rtPCR. Significantly, it can be induced to gain differentiation maturity through contact inhibition. In addition, a BAK-mediated rabbit model of corneal endothelial dystrophy was established in the present study to test the therapeutic effectiveness of the iHCEPLC. RESULTS: After inducing cell fate conversion, the specific HCEC markers were detected by rtPCR and immunostaining in iHCEPLC. Further, RNAseq was applied to distinguish its progenitor-like cell fate from primary human corneal endothelial cells (HECE). FlowCytometry profiled the heterogeneity subpopulation, consistently displaying a subtle difference from primary HCEC. A terminal differentiation can be induced in iHCEPLC, addressing its progenitor-like fate. iHCEPLC can restore the BAK-based rabbit model of corneal endothelial dystrophy. Immunohistochemistry verified that such acuity restoration of the BAK-treated cornea is due to the introduced iHCEPLC, and such therapeutic effectiveness is observed in the long term. CONCLUSION: Here, we demonstrated that customized iHCEPLC has long-term therapeutic efficacy. As a progenitor cell, our iHCEPLC has a restricted cell lineage nature and can proliferate in vitro, supporting sufficient therapeutic candidate cells. Due to the immune-privileged nature of the cornea, our iHCEPLC proves the principle of therapeutical feasibility in both autogenic and allogeneic modalities.

Corneal endothelial cell morphology in children with autosomal recessive Alport syndrome: a longitudinal study.

PURPOSE: To evaluate the corneal endothelial cell morphology in children with autosomal recessive Alport syndrome (ARAS). METHODS: This is a longitudinal, prospective cohort study that evaluated pediatric patients with genetically diagnosed ARAS. Fifty-eight eyes of 29 pediatric patients (12 patients, 17 controls) underwent a full ophthalmic examination. Corneal endothelial cell density (ECD) (cells/mm²), coefficient variation (CV) of cell area (polymegathism), the percentage of hexagonal cells (HEX) (pleomorphism), and central corneal thickness (CCT) were analyzed automatically using a noncontact specular microscopy. RESULTS: The mean ECD was 2904 ± 355.48 cell/mm² in the ARAS group and 3263.20 ± 261.71 cell/mm² in the control group (p = 0.004). In the ARAS group, the mean CV was 46.53 ± 10.43, which was significantly higher than that in controls (p = 0.026). The mean HEX was 48.86 ± 14.71 in the ARAS group and 59.06 ± 10.64 in the control group (p = 0.038). The mean CCT was 565.26 ± 39.77 µm in the ARAS group and 579.66 ± 31.65 µm in the control group (p = 0.282). The comparison of endothelial cell characteristic of the ARAS group with 1-year follow-up is as follows: The mean ECD decreased from 2904 ± 355.48 cell/mm² to 2735 ± 241.58 cell/mm² (p = 0.003). The mean CV increased from 46.53 ± 10.43 to 47.93 ± 10.50 (p = 0.471). The mean HEX decreased from 48.86 ± 14.71 to 48.50 ± 10.06 (p = 0.916). The mean CCT decreased from 565.26 ± 39.77 µm to 542.86 ± 40.39 µm (p = 0.000). CONCLUSION: Measurement of ECD and percentage of hexagonality can also be used as an indicator of the health of the corneal endothelium. In this study, the mean ECD and HEX were significantly lower in ARAS group than in age-matched pediatric controls. Polymegathism, which reflects cellular stress, was statistically significantly higher in ARAS group. The mean ECD and CCT decreased significantly at 1-year follow-up. This study may demostrated that endothelial damages and stress in ARAS patients appear in childhood and show a rapid increase with age.

Corneal endothelial changes seven years after phacoemulsification cataract surgery.

PURPOSE: To evaluate long-term postoperative corneal changes after phacoemulsification cataract surgery. METHODS: Twenty patients who participated in a previous study regarding corneal endothelial changes after phacoemulsification cataract surgery were examined after 7 years. The patients were divided in three groups based on their initial increase in central corneal thickness day one after the surgery: < 5% increase, 6-20% increase and ≥ 20% increase. The primary outcome measures were corneal endothelial cell loss (ECL), endothelial cell count (ECC) and endothelial morphology. RESULTS: After 7 years, a difference in cell loss between the groups was observed, except for groups 1 and 2. Endothelial cell count (ECC) differed significantly between groups 1 and 3 at 3 months. At 7 years, there was no difference in ECC between the three groups. Cell loss was found exclusively in group 1 between 3 months and 7 years. Endothelial cell morphology showed a converging pattern between 3 months and 7 years. CONCLUSION: After phacoemulsification cataract surgery, long-term ECC and morphology appear to converge towards a comparable steady state regardless of initial corneal swelling and endothelial cell loss.

The effect of irrigation and aspiration on the corneal endothelial cell density in patients undergoing Implantable Collamer Lens with a central hole implantation for myopia correction.

PURPOSE: In the conventional technique, viscoelastic agents are typically rinsed away with balanced salt solution (BSS), but it may lead to a series of complications such as viscoelastic residue, anterior chamber instability and intraoperative TICL rotation. The utilization of irrigation and aspiration (I/A) has been shown to be effective in maintaining anterior chamber stability, reducing the incidence of postoperative high intraocular pressure, and minimizing postoperative fundus complications. However, there is a lack of previous studies investigating the impact of I/A on corneal endothelial cells during ICL implantation. The objective of this study was to examine the effect of I/A on corneal endothelial cells in patients undergoing myopia correction through implantation of Implantable Collamer Lens with a central hole (V4c ICL). METHODS: A retrospective selection was made of 344 eyes from 172 patients who underwent V4c ICL implantation and I/A to remove viscoelastic agent from the anterior chamber between 2021 and 2022. The intraocular pressure (IOP) was measured at 1 h, 2 h and 3 h after surgery. Corneal endothelial cell density (ECD), coefficient of variation in cell size (CV), standard deviation of cell area (SD), and percentage of hexagonal cells (HEX) were evaluated at 1 week postoperatively to assess corneal endothelial cells. The first two represent polymegethism or morphological variation, while the third parameter represents the degree of polymorphism of the corneal endothelial cells. Electronic medical records were utilized for data collection purpose. RESULTS: All surgeries proceeded without complications. The IOP was 16.50 ± 3.42 mmHg (range: 11.5-22.3 mmHg) prior to surgery and increased to 21.25 ± 5.61 mmHg (range: 9.5-34.8 mmHg), 19.85 ± 5.18 mmHg (range: 11.4-36.2 mmHg) and finally settled at an average of 18.81 ± 4.57 mmHg (range: 10.1-38.8 mmHg) at the respective time points of 1 h, 2 h and 3 h after surgery. The preoperative ECD was recorded as being approximately 3004 ± 295 cell/mm2, which exhibited a marginal decreased of 1.17% postoperatively, resulting in an average ECD value of 2969 ± 303 cell/mm2 one week after surgery (P = 0.12). Similarly, the preoperative CV was determined as 31.10 ± 3.78%, and it experienced a slight reduction with an average CV value of 30.74 ± 3.77% at week after surgery (P = 0.21). And, the preoperative SD was reported as 104.76 ± 17.26, and it remained virtually unchanged with an average SD value of 104.85 ± 18.75 at one week after surgery (P = 0.95). The preoperative HEX was calculated as 55.38 ± 8.94%, and it remained its stability with an average HEX value of 55.45 ± 8.73% one week after surgery (P = 0.92). CONCLUSION: The utilization of I/A led to a slight decrease in postoperative ECD when compared to conventional surgical techniques. Nevertheless, the reduction in ECD remained within acceptable limits, taking into accout the avervantaged it offered, such as stabilization of the anterior chamber and decreased occurrence of viscoelastic residue after surgery. It is challenging to anticipate the long-term safety of corneal endothelial cells based on current short-term studies. However, this study provides a valuable reference indicating that neither anterior chamber irrigation nor I/A aspiration have an adverse impact on the safety of corneal endothelial cells in the short term. Further research is imperative to enhance our understanding of their effects over an extended period.

U-Net Convolutional Neural Network for Real-Time Prediction of the Number of Cultured Corneal Endothelial Cells for Cellular Therapy.

Corneal endothelial decompensation is treated by the corneal transplantation of donor corneas, but donor shortages and other problems associated with corneal transplantation have prompted investigations into tissue engineering therapies. For clinical use, cells used in tissue engineering must undergo strict quality control to ensure their safety and efficacy. In addition, efficient cell manufacturing processes are needed to make cell therapy a sustainable standard procedure with an acceptable economic burden. In this study, we obtained 3098 phase contrast images of cultured human corneal endothelial cells (HCECs). We labeled the images using semi-supervised learning and then trained a model that predicted the cell centers with a precision of 95.1%, a recall of 92.3%, and an F-value of 93.4%. The cell density calculated by the model showed a very strong correlation with the ground truth (Pearson's correlation coefficient = 0.97, p value = 8.10 × 10-52). The total cell numbers calculated by our model based on phase contrast images were close to the numbers calculated using a hemocytometer through passages 1 to 4. Our findings confirm the feasibility of using artificial intelligence-assisted quality control assessments in the field of regenerative medicine.

Electrospun Nanofiber Membrane for Cultured Corneal Endothelial Cell Transplantation.

The corneal endothelium, comprising densely packed corneal endothelial cells (CECs) adhering to Descemet's membrane (DM), plays a critical role in maintaining corneal transparency by regulating water and ion movement. CECs have limited regenerative capacity within the body, and globally, there is a shortage of donor corneas to replace damaged corneal endothelia. The development of a carrier for cultured CECs may address this worldwide clinical need. In this study we successfully manufactured a gelatin nanofiber membrane (gelNF membrane) using electrospinning, followed by crosslinking with glutaraldehyde (GA). The fabricated gelNF membrane exhibited approximately 80% transparency compared with glass and maintained a thickness of 20 µm. The gelNF membrane demonstrated desirable permeability and degradability for a Descemet's membrane analog. Importantly, CECs cultured on the gelNF membrane at high densities showed no cytotoxic effects, and the expression of key CEC functional biomarkers was verified. To assess the potential of this gelNF membrane as a carrier for cultured CEC transplantation, we used it to conduct Descemet's membrane endothelial keratoplasty (DMEK) on rabbit eyes. The outcomes suggest this gelNF membrane holds promise as a suitable carrier for cultured CEC transplantation, offering advantages in terms of transparency, permeability, and sufficient mechanical properties required for successful transplantation.

Corneal endothelial validation in the eye bank: differences in automated methods and repeatability.

PURPOSE: To evaluate reproducibility of endothelial cell density (ECD) measurements using the Konan Cell Check D in donor corneas by two different ophthalmologists and to compare the two automated cell count methods (center and flex-center) available in the software of this specular microscope. METHODS: ECD values were quantified in 54 donor corneas by two independent investigators using the Cell Check D (Konan Medical USA Inc) with both automated cell count methods. In the center method, at least 30 contiguous cells are marked. For the flex-center method, an area is delineated and only the cells within the designated area are counted. RESULTS: The mean ECD was 2473.81±378.22 cells/mm2. Good ECD intergrader reproducibility for the center (ICC=0.821) and the flex-center method (ICC=0.784) were noted. Poor reliability was observed for coefficient of variation and hexagonality (ICC≤0.265). When both methods for ECD analysis were compared, a moderate correlation for the two independent graders using the two manual (center and flex-center) methods was detected (correlation coefficient of 0.678 and 0.745 for each of the investigators). Comparison between methods yielded significantly higher ECD with the flex-center method (P=0.013). When corneas were divided by ECD, those under 2200 cells/mm2 and those between 2200 and 2700 cells/mm2 also had significantly higher ECD with the flex-center method (P<0.022). CONCLUSIONS: ECD values are reproducible with both methods, although the flex-center method ECDs tend to be higher, particularly in cases of low ECD. Eye banks and surgeons should exercise caution in making decisions based only on small differences in ECD.

Comparisons of the clinical outcomes of Centurion® active fluidics system with a low IOP setting and gravity fluidics system with a normal IOP setting for cataract patients with low corneal endothelial cell density.

Background: During cataract phacoemulsification surgery, the Alcon Centurion with Active Sentry can achieve a more stable anterior chamber, which allows a lower intraocular pressure (IOP) setting than the gravity fluidics system. In this randomized controlled trial, we compared these two systems' damage to the cornea under different IOP settings. Methods: Seventy-eight eyes of 53 patients with corneal endothelial cell density (ECD) of 500∼1500/mm2 were enrolled and randomly divided into the active fluidics system (AFS) group using an Active Sentry handpiece with 30 mmHg IOP setting (40 eyes) and the gravity fluidics system (GFS) group using an Ozil handpiece with 80 cmH2O IOP setting (38 eyes). Intraoperative parameters, visual acuity, corneal edema ratio, central corneal thickness (CCT) changes as well as loss rate of ECD were analyzed. Results: We observed no significant differences in best corrected visual acuity (BCVA), cumulative dissipated energy (CDE), total case time, estimated fluidics usage (EFU) and ophthalmic viscoelastic devices (OVDs) usage between the two groups. The enrolled eyes were further divided into soft nucleus (27 eyes) and hard nucleus (51 eyes) subgroups. And we found less pain complaint during surgeries, lower corneal edema ratio at 1-day and 1-week visit, smaller CCT changes at 1-day visit and lower ECD loss rate at 1-month visit (p < 0.05) in both subgroups of the AFS group than in the GFS group, implying higher intraoperative comfort levels and less corneal damage of the AFS group with a low IOP setting. Conclusion: Owing to a lower IOP setting, Centurion® Vision System with Active Sentry handpiece causes less corneal damage and pain perception during phacoemulsification for patients with low pre-operative ECD. Clinical Trial Registration: https://www.chictr.org.cn, identifier ChiCTR2300077865.

Integrated Analysis of Transcriptome and Proteome of the Human Cornea and Aqueous Humor Reveal Novel Biomarkers for Corneal Endothelial Cell Dysfunction.

Earlier studies have reported that elevated protein levels in the aqueous humor (AH) are associated with corneal endothelial cell dysfunction (CECD), but the details of the underlying mechanism as well as specific biomarkers for CECD remain elusive. In the present study, we aimed to identify protein markers in AH directly associated with changes to corneal endothelial cells (CECs), as AH can be easily obtained for analysis. We carried out an in-depth proteomic analysis of patient-derived AH as well as transcriptomic analysis of CECs from the same patients with bullous keratopathy (BK) resulting from CECD. We first determined differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) from CECs and AH in CECD, respectively. By combining transcriptomic and proteomic analyses, 13 shared upregulated markers and 22 shared downregulated markers were observed between DEGs and DEPs. Among these 35 candidates from biomarker profiling, three upregulated markers were finally verified via data-independent acquisition (DIA) proteomic analysis using additional individual AH samples, namely metallopeptidase inhibitor 1 (TIMP1), Fc fragment of IgG binding protein (FCGBP), and angiopoietin-related protein 7 (ANGPTL7). Furthermore, we confirmed these AH biomarkers for CECD using individual immunoassay validation. Conclusively, our findings may provide valuable insights into the disease process and identify biofluid markers for the assessment of CEC function during BK development.

A cross-sectional study investigating the effects of argon laser retinal photocoagulation on lens clarity and corneal endothelial cells.

Background: The mechanism of argon laser retinal photocoagulation (ALRP) treatment is to apply thermal-induced retinal pigment epithelium damage. Light passes through the anterior optical segments of the eye to reach the retina. Lens densitometry is a noninvasive and quantitative measurement providing information about corneal and lens clarity. Objectives: This study aimed to investigate whether laser light affects lens clarity and corneal endothelial cells. Design: This was a prospective, cross-sectional study. Methods: Lens densitometric (LD) analysis and specular microscopy were performed before, after, and 1 month after ALRP treatment, by an expert ophthalmologist, blinded to the medical status of the patients. LD analysis was performed using a Pentacam HR (Oculus, Wetzlar, Germany) and a Specular Microscope CEM-530 (Nidek, Japan) was used for endothelial cell analysis. Results: The evaluation was made on 81 eyes of 41 patients, with a mean age of 54.46 ± 6.7 years. LD, after ALRP, was significantly more than before ALRP (p < 0.001). There was no statistically significant difference in LD, before ALRP, and 1 month after ALRP (p = 0.262). There was a statistically significant increase in LD after ALRP compared to before ALRP, but it decreased after 1 month. There was an increase in the coefficient of variance (CV) after ALRP compared to before ALRP but it was not statistically significant (p = 0.188). There was no statistically significant difference in CV between before ALRP and 1 month after ALRP (p = 1.000). There was no statistically significant difference in the cell density, the hexagonality percentage between before ALRP, after ALRP, and 1 month after ALRP (p = 0.993, and 0.863, respectively). Conclusion: ALRP may affect the lens densitometry temporarily during the procedure. Thermal damage may be the reason for increased lens densitometry.

Long-term safety and efficacy of a foldable iris-fixated phakic intraocular lens for the correction of myopia.

PURPOSE: To analyze and report the long-term outcomes in terms of efficacy and safety of eyes implanted with the spherical version of a foldable iris-fixated phakic intraocular lens (pIOL) for the correction of myopia. METHODS: Retrospective analysis of the results of 56 eyes of 32 patients (age, 19-45 years) who underwent implantation of the spherical model of the Artiflex pIOL (Ophtec B.V., Groningen, The Netherlands) for the correction of myopia. Visual, refractive, biometric, intraocular pressure (IOP) and corneal endothelial changes were evaluated during a long-term follow-up: 2, 7, 10 and 12 years for more than 50, 30, 20 and 10 eyes, respectively. RESULTS: At 4 weeks postoperatively, a significant reduction of manifest sphere and spherical equivalent (SE), with a significant improvement of uncorrected distance visual acuity were found (all p < 0.001). No significant changes were found during the rest of follow-up in sphere (p ≥ 0.072). The percentage of eyes with SE within ± 1.00 D was over 83% during the whole follow-up. A non-significant trend to IOP increase was observed at 4 weeks postoperatively (p = 0.530), with a significant reduction at 1 year after (p = 0.039) and no significant changes during the rest of follow-up (p = 0.180). There was a significant reduction of anterior chamber depth at 4 weeks after surgery (p < 0.001), with no significant changes during the following 9 years of follow-up (p = 0.118). However, an additional significant decrease of this parameter was observed between 10 and 13 years after surgery (p = 0.027). Mean endothelial cell loss changed from 2.01 ± 4.49% at 4 weeks after surgery to 9.11 ± 2.24% at the end of the follow-up. No complications were reported during the follow-up. CONCLUSIONS: Myopia correction with the Artiflex pIOL is an effective and safe procedure in the long term.

Clinical observation of phacoemulsification under the low perfusion pattern and low negative pressure in patients with low corneal endothelial cell density.

BACKGROUND: To observe the safety and effect of phacoemulsification combined with intraocular lens (IOL) implantation in patients with low corneal endothelial cell density (CD) under the low perfusion pattern with low negative pressure. METHODS: In this retrospective case series study, a total of 16 patients (17 eyes) were studied. They had all been diagnosed with low corneal endothelial (CD) and cataracts in the First Affiliated Hospital of Fujian Medical University from December 2019 to October 2021. They underwent phacoemulsification combined with IOL implantation under the low perfusion pattern with low negative pressure. The variations of corneal endothelial( CD), coefficient of variation (CV) of the cell area, central corneal thickness (CCT), visual acuity, and intraocular pressure before and after the operation were observed and assessed. Then a paired t-test, repeated measures analysis of variance, and Pearson correlation analysis were adopted for data analysis. RESULTS: The mean intraocular pressure of the 17 eyes was 16.88 ± 6.47 mmHg before the operation and 14.41 ± 3.10 mmHg after the operation, showing a statistically significant difference of t = 2.222, and p = 0.041. Before the operation, the mean visual acuity was 0.16 ± 0.09, and after the operation, it was 0.45 ± 0.16, displaying a statistically significant difference of t = -9.917, p < 0.001. Before and after the operation, four of the 17 eyes had no detectable CD. The mean CD of the other 13 eyes at one month after the operation (644.308 ± 106.24 cells/mm2) was lower than that before the operation (709.62 ± 119.19 cells/mm2), and the differences were statistically significant (F = 20.044, p < 0.001). However, no statistically significant differences were found in the mean CV before the operation (31.23 ± 4.21), and at one month after the operation (32.62 ± 3.80; F = 2.130, p = 0.157). Moreover, the mean CCT of 14 eyes at one month after the operation (562.72 ± 27.82 µm) was larger than that before the operation (534.79 ± 24.69 µm). CONCLUSIONS: The low perfusion pattern with low negative pressure is safe and effective for corneal endothelial dysfunction patients complicated with cataracts.

Crossover Randomized Study of Pharmacologic Effects of Ripasudil-Brimonidine Fixed-Dose Combination Versus Ripasudil or Brimonidine.

INTRODUCTION: Multidrug regimens for glaucoma treatment often result in adherence issues due to inconvenience; these issues may be improved with fixed-dose combination drugs. The ophthalmic solution of ripasudil-brimonidine fixed-dose combination (RBFC; K-232) is the first treatment combining a Rho kinase inhibitor and an α2-adrenoceptor agonist, and has demonstrated ability to lower intraocular pressure (IOP) and have various effects on conjunctival hyperemia and corneal endothelial cell morphology. This study evaluates the pharmacologic effects of RBFC treatment versus its separate components-ripasudil or brimonidine. METHODS: This single-center, prospective, randomized, open-label, blinded endpoint study with 3 × 3 crossover design randomly assigned healthy adult men to three groups (1:1:1) to undergo consecutive 8-day administration phases (with drug-free intervals of at least 5 days). Subjects received twice-daily instillation of RBFC → ripasudil → brimonidine (group A), ripasudil → brimonidine → RBFC (group B), or brimonidine → RBFC → ripasudil (group C). Endpoints included change in IOP, severity of conjunctival hyperemia, corneal endothelial cell morphology, pupil diameter, and pharmacokinetics. RESULTS: Eighteen subjects were assigned in total (six to each group). RBFC significantly reduced IOP from baseline at 1 h post-instillation on days 1 and 8 (12.7 vs. 9.1 and 9.0 mmHg, respectively; both P < 0.001), and provided significantly greater IOP reductions than ripasudil or brimonidine at several time points. The most common adverse drug reaction with all three treatments was mild conjunctival hyperemia, which transiently increased in severity with RBFC or ripasudil, peaking at 15 min post-instillation. In post hoc analyses, conjunctival hyperemia scores were lower with RBFC than with ripasudil at several time points. Transient morphologic changes in corneal endothelial cells occurred for up to several hours with RBFC or ripasudil, but not with brimonidine. Pupil diameter did not change with RBFC. CONCLUSION: RBFC significantly reduced IOP compared with each agent alone. A combination of each agent's pharmacologic profile was observed in that of RBFC. TRIAL REGISTRATION: Japan Registry of Clinical Trials; Registration No. jRCT2080225220.

Micropulse Cyclophotocoagulation vs Selective Laser Trabeculoplasty: Effects on Corneal Endothelial Cells and Intraocular Pressure.

Purpose: To evaluate the effects of micropulse transscleral cyclophotocoagulation (mCPC) and selective laser trabeculoplasty (SLT) on endothelial cell density (ECD) and reduction of the intraocular pressure (IOP) in patients with primary-open angle glaucoma (POAG). Patients and Methods: In this observational, retrospective study, 40 eyes with POAG were included. Patients were divided into three groups-group I was treated with SLT (n = 13), group II was treated with mCPC (n = 13), and group III included age-matched patients with medically treated glaucoma (n = 14) (control group). In both treatment groups (SLT and mCPC) preoperative and postoperative findings of best-corrected visual acuity (BCVA), ECD, and IOP were compared to the control group. Results: The mean time interval before and after the treatment was 215 ± 120 days in group I (SLT) and 273 ± 177 days in group II (mCPC). The follow-up for group III (control group) was 209 ± 103 days. In both treatment groups (SLT and mCPC) there was a statistically significant reduction of the IOP postoperatively (group I: 3.5 ± 3.7 mm Hg (p = 0.005) and group II: 4.3 ± 4.1 mm Hg (p = 0.003)). The mean IOP for group III was 12.9 ± 3.7 mm Hg at visit 1 and did not change significantly (p = 0.353) at visit 2. In all three groups, there was no statistically significant change in ECD at the last visit. Conclusion: According to our results, both SLT and mCPC seem to be effective in lowering the IOP, without showing any statistically significant effect on ECD in patients with POAG.However, larger and longer-term studies are necessary to understand the effects of the SLT and mCPC procedures on ECD. How to cite this article: Garip Kuebler A, Priglinger S, Reznicek L. Micropulse Cyclophotocoagulation vs Selective Laser Trabeculoplasty: Effects on Corneal Endothelial Cells and Intraocular Pressure. J Curr Glaucoma Pract 2023;17(1):40-43.

Analysis of corneal topographic and endothelial cell properties in newly diagnosed obstructive sleep apnea patients: A case-control study.

PURPOSE: To investigate the topographic features and endothelial cell changes in the corneas of patients with newly diagnosed obstructive sleep apnea (OSA). BACKGROUND: This cross-sectional case-control study included 98 patients with OSA and 90 healthy controls. OSA was diagnosed on the basis of a polysomnographic sleep examination. Patients with OSA were grouped according to their apnea-hypopnea index (AHI) scores. Corneal topographic measurements were obtained using the Scheimpflug-Placido disk topographer, and keratometry values, anterior chamber parameters, thinnest corneal thickness, symmetry index (SI), keratoconus vertex (KV), and the Baiocchi-Calossi-Versaci index, including keratoconus (KC) screening indexes, were recorded. The corneal endothelium was visualizedby specular microscopy, and the endothelial cell density, coefficient of variability (CV), and percentage of hexagonal cells (HEX) were recorded. RESULTS: In this study, 32 patients had mild, 34 patients had moderate, and 32 patients had severe OSA. The mean back KV (KVb) and back SI values were significantly higher in the OSA group than in the control group (P <0.001, P = 0.035, respectively). As body mass index (BMI) increased, KVb also increased significantly (r = 0.281; P = 0.009). The mean CV and HEX values differed significantly between the OSA and control groups (P = 0.012 and P = 0.002, respectively). In addition, a significant correlation was found between AHI and KVb and CV values (r = 0.315; P <0.001 and r = 0.231; P = 0.011, respectively). CONCLUSIONS: This study revealed abnormal changes in the corneal endothelial morphology and topographic KC screening indexes in relation to OSA severity in patients and controls. Further research is required to determine the effect of BMI on corneal topographic findings in patients with OSA.

Long-Term Surgical Outcomes and Possible Postoperative Complication with Severe Corneal Endothelial Cell Loss After Trabeculectomy for Cytomegalovirus-Associated Anterior Uveitis with Secondary Glaucoma.

PURPOSE: We assess long-term surgical outcomes after an initial trabeculectomy for cytomegalovirus-associated anterior uveitis with secondary glaucoma (CMV-SG). METHODS: We retrospectively reviewed the medical records of 16 eyes of 15 patients with CMV-SG and 157 eyes of 157 patients with primary open-angle glaucoma. The average follow-up period was approximately 3 years. Surgical success was defined as intraocular pressure (IOP) below 18 mmHg and at least 20% lower than baseline. RESULTS: Kaplan-Meier survival analysis revealed that bleb survival rates were not significantly different in the CMV-SG and POAG groups (P = 0.75). Bullous keratopathy occurred in 2 of 16 eyes with CMV-SG postoperatively but did not occur in the POAG group. The corneal endothelial cell density decreased by 34.2 ± 22.7% in the CMV-SG group during an average follow-up period of 2.7 ± 2.0 years. CONCLUSION: Trabeculectomy effectively controlled IOP in CMV-SG, but attention must be paid to corneal endothelial cell loss.

Carteolol triggers senescence via activation of ß-arrestin-ERK-NOX4-ROS pathway in human corneal endothelial cells in vitro.

Carteolol is a commonly-used topical medication for primary open-angle glaucoma. However, long-term and frequent ocular application of carteolol entails its residuals at low concentration in the aqueous humor for a long duration and may exert latent toxicity in the human corneal endothelial cells (HCEnCs). Here, we treated the HCEnCs in vitro with 0.0117% carteolol for 10 days. Thereafter, we removed the cartelolol and normally cultured the cells for 25 days to investigate the chronical toxicity of carteolol and the underlying mechanism. The results exhibited that 0.0117% carteolol induces senescent features in the HCEnCs, such as increased senescence-associated ß-galactosidase positive rates, enlarged relative cell area and upregulated p16INK4A and senescence-associated secretory phenotypes, including IL-1α, TGF-ß1, IL-10, TNF-α, CCL-27, IL-6 and IL-8, as well as decreased Lamin B1 expression and cell viability and proliferation. Thereby, further exploration demonstrated that the carteolol activates ß-arrestin-ERK-NOX4 pathway to increase reactive oxygen species (ROS) production that imposes oxidative stress on energetic metabolism causing a vicious cycle between declining ATP and increasing ROS production and downregulation of NAD+ resulting in metabolic disturbance-mediated senescence of the HCEnCs. The excess ROS also impair DNA to activate the DNA damage response (DDR) pathway of ATM-p53-p21WAF1/CIP1 with diminished poly(ADP-Ribose) polymerase (PARP) 1, a NAD+-dependent enzyme for DNA damage repair, resulting in cell cycle arrest and subsequent DDR-mediated senescence. Taken together, carteolol induces excess ROS to trigger HCEnC senescence via metabolic disturbance and DDR pathway.

Mechanical protective effect of lens anterior capsule disc on corneal endothelial cells during femtosecond laser-assisted cataract surgery in a rabbit model.

PURPOSE: To evaluate the effects of a novel technique using an isolated lens anterior capsule disc (LACD) to protect corneal endothelial cells in rabbit eyes during femtosecond laser-assisted cataract surgery. METHODS: Experimental study. 40 rabbits were divided into endothelium-protected (experimental) and control groups, with 20 rabbits in each group. In the experimental group, after femtosecond laser capsulotomy, the isolated capsule disc was lifted to the corneal endothelium by an ophthalmic viscosurgical device. The endothelium was damaged for 1 min with an ultrasonic probe. The control group underwent the same surgery, except that the disc was removed immediately after capsulorhexis. Corneal endothelioscopy was performed preoperatively and on postoperative days (PODs) 3 and 7 to observe endothelial cell counts (ECC) and endothelial cell loss rate. Central corneal thickness (CCT) was measured before and at PODs 1, 3 and 7. RESULTS: There were 3.59%±1.88% (p < 0.001) and 2.92%±2.14% (p < 0.001) loss of ECC in experimental group at POD3 and POD7, respectively, while those in the control group were 11.62%±7.43% and 10.34%±5.77%, respectively. On POD 1, the difference in central corneal thickness was significant(P = 0.019) between the two groups. At POD 3 and POD 7, CCT was not significantly different (P = 0.597;0.913) between the two groups. CONCLUSIONS: The isolated LACD technique significantly reduced damage to the endothelium caused by ultrasonic energy and protects corneal endothelial cells during phacoemulsification.

Modified gellan gum-based hydrogel with enhanced mechanical properties for application as a cell carrier for cornea endothelial cells.

Recently, the number of people suffering from visual loss due to eye diseases is increasing rapidly around the world. However, due to the severe donor shortage and the immune response, corneal replacement is needed. Gellan gum (GG) is biocompatible and widely used for cell delivery or drug delivery, but its strength is not suitable for the corneal substitute. In this study, a GM hydrogel was prepared by blending methacrylated gellan gum with GG (GM) to give suitable mechanical properties to the corneal tissue. In addition, lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), a crosslinking initiator, was added to the GM hydrogel. After the photo-crosslinking treatment, it was named GM/LAP hydrogel. GM and GM/LAP hydrogels were analyzed for physicochemical properties, mechanical characterization, and transparency tests to confirm their applicability as carriers for corneal endothelial cells (CEnCs). Also, in vitro studies were performed with cell viability tests, cell proliferation tests, cell morphology, cell-matrix remodeling analysis, and gene expression evaluation. The compressive strength of the GM/LAP hydrogel was improved compared to the GM hydrogel. The GM/LAP hydrogel showed excellent cell viability, proliferation, and cornea-specific gene expression than the GM hydrogel. Crosslinking-improved GM/LAP hydrogel can be applied as a promising cell carrier in corneal tissue engineering.

The Biologic Character of Donor Corneal Endothelial Cells Influences Endothelial Cell Density Post Successful Corneal Transplantation.

Purpose: Corneal endothelial cell density (ECD) gradually decreases after corneal transplantation by unknown biologic, biophysical, or immunologic mechanism. Our purpose was to assess the association between donor corneal endothelial cell (CEC) maturity in culture and postoperative endothelial cell loss (ECL) after successful corneal transplantation. Design: Prospective cohort study. Participants: This cohort study was conducted at Baptist Eye Institute, Kyoto, Japan, between October 2014 and October 2016. It included 68 patients with a 36-month follow-up period who had undergone successful Descemet stripping automated endothelial keratoplasty (DSAEK) or penetrating keratoplasty. Methods: Human CECs (HCECs) from remaining peripheral donor corneas were cultured and evaluated for maturity by surface markers (CD166+, CD44-/dull, CD24-, and CD105-) using fluorescence-activated cell sorting. Postoperative ECD was assessed according to the mature-differentiated HCEC contents: high-maturity group: > 70%, middle-maturity group: 10% to 70%, low-maturity group: < 10%. The successful rate of ECD maintained at 1500 cells/mm2 at 36 months postoperative was analyzed using the log-rank test. Main Outcome Measures: Endothelial cell density and ECL at 36 months postoperative. Results: The 68 included patients (mean [standard deviation] age 68.1 [13.6] years, 47.1% women, 52.9% DSAEK). The high, middle, and low-maturity groups included 17, 32, and 19 eyes, respectively. At 36 months postoperative, the mean (standard deviation) ECD significantly decreased to 911 (388) cells/mm2 by 66% in the low-maturity group, compared with 1604 (436) by 40% and 1424 (613) cells/mm2 by 50% in the high and middle-maturity groups (P < 0.001 and P = 0.007, respectively) and the low-maturity group significantly failed to maintain ECD at 1500 cells/mm2 at 36 months postoperative (P < 0.001). Additional ECD analysis for patients who underwent DSAEK alone displayed a significant failure to maintain ECD at 1500 cells/mm2 at 36 months postoperative (P < 0.001). Conclusions: The high content of mature-differentiated HCECs expressed in culture by the donor peripheral cornea was coincident with low ECL, suggesting that a high-maturity CEC content predicts long-term graft survival. Understanding the molecular mechanism for maintaining HCEC maturity could elucidate the mechanism of ECL after corneal transplantation and aid in developing effective interventions. Financial Disclosures: Proprietary or commercial disclosure may be found after the references.