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PURPOSE To investigate the safety and efficacy of cultured human corneal endothelial cell (hCEC)-injection therapy with mature differentiated (mature) cell subpopulations (SPs) for corneal endothelial failure (CEF). DESIGN Comparative, interventional case series. METHODS This study involved 18 eyes with CEF that underwent cultured hCEC-injection therapy, categorized into two groups: 1) 11 eyes administered a relatively lower proportion (0.1 to 76.3%) of mature cell SPs [Group 1 (Gr1)], and 2) 7 eyes administered a relatively higher proportion (>90%) of mature cell SPs [Group 2 (Gr2)]. From 1-week to 3-years postoperative, corneal endothelial cell (CEC) density (CECD), central corneal thickness (CCT), and best-corrected visual acuity (BCVA) were recorded, and the CEC parameter's 'spring constant' was calculated. The proportion of mature SPs was evaluated by fluorescence-activated cell sorting analysis based on cell-surface markers. RESULTS At 3-years postoperative, corneal restoration with improved BCVA was attained in 10 of the 11 Gr1 eyes and all Gr2 eyes, the median CECD in Gr2 (3,083 cells/mm2; range, 2,182-4,417 cells/mm2) was higher than that in Gr1 (1,349 cells/mm2; range, 746-2,104 cells/mm2) (P < 0.001), and the spring constant verified the superiority of the mature cultured hCECs. From 24-weeks through 3-years postoperative, the median percentage of CECD decrease was 3.2% in Gr2 and 23.6% in Gr1 (P < 0.005). CCT recovery was prompt and constant in Gr2, while diverse in Gr1. No adverse events were observed. CONCLUSION Our findings showed that mature cell SPs for hCEC-injection therapy provide rapid recovery of CCT, better CECD, and low CECD attrition over 3 years post surgery.

Cell therapies are emerging as a unique class of clinical therapeutics in medicine. In 2015, Holoclar (ex vivo expanded autologous human corneal epithelial cells containing stem cells) gained the regulatory approval for treating limbal stem cell deficiency after chemical eye burn. This has set a precedent in ophthalmology and in medicine, reinforcing the therapeutic promise of cell therapy. However, to generalize and commercialize cell therapies on a global scale, stringent translational and regulatory requirements need to be fulfilled at both local and international levels. Over the past decade, the Singapore group has taken significant steps in developing human corneal endothelial cell (HCEnC) therapy for treating corneal endothelial diseases, which are currently the leading indication for corneal transplantation in many countries. Successful development of HCEnC therapy may serve as a novel solution to the current global shortage of donor corneas. Based on the experience in Singapore, this review aims to provide a global perspective on the translational and regulatory challenges for bench-to-bedside translation of cell therapy. Specifically, we discussed about the characterization of the critical quality attributes (CQA), the challenges that can affect the CQA, and the variations in the regulatory framework embedded within different regions, including Singapore, Europe, and the United States. Impact statement Functional corneal endothelium is critical to normal vision. Corneal endothelial disease-secondary to trauma, surgery, or pathology-represents an important cause of visual impairment and blindness in both developed and developing countries. Currently, corneal transplantation serves as the current gold standard for treating visually significant corneal endothelial diseases, although limited by the shortage of donor corneas. Over the past decade, human corneal endothelial cell therapy has emerged as a promising treatment option for treating corneal endothelial diseases. To allow widespread application of this therapy, significant regulatory challenges will need to be systematically overcome.

To determine the structure and morphology of corneal endothelial cell layer in patients with acute anterior uveitis. Thirty-four eyes of 34 acute anterior uveitis patients and 34 eyes of 34 healthy subjects were included. Mean cell density, coefficient of variation, maximum cell area, minimum cell area, average cell area and hexagonality ratio values were evaluated by non-contact specular microscopy. Parameters recorded in both groups were compared. The mean maximum cell area was 1054,44 ± 251,14 µm2, minimum cell area was 152.29 ± 53.65 µm2 and average cell area was 386.91 ± 41.73 µm2 in acute anterior uveitis group and the mean maximum cell area was 1057.65 ± 261.23 µm2, minimum cell area was 147.26 ± 20.45 µm2 and average cell area was 383.53 ± 43.12 µm2 in the control group. There were no significant differences between the two groups in terms of maximum, minimum and average cell area (respectively, p = 0.080, p = 0.72, p = 0.62, p = 0.67). The mean cell density was 2607.74 ± 277.63 cells/ µm2 in acute anterior uveitis group and 2669.35 ± 265.22 cells/µm2 in the control group. (p = 0.358). In acute anterior uveitis group the mean coefficient of variation was 31.68 ± 8.16, hexagonality ratio was 63.85 ± 11.14 and mean central corneal thickness was 571.47 ± 55.99 µm; in control group the mean coefficient of variation was 25.29 ± 3.00, mean hexagonality ratio was 72.6 ± 4.80% and mean central corneal thickness was 534.82 ± 33.84 µm. Statistically significant differences were seen between the two groups (respectively, P = 0,00, P = 0,00, P = 0.02). The mean central corneal thickness and coefficient of variation values were found higher, and the hexagonality ratio was found lower in acute anterior uveitis group. Our findings suggest that intraocular inflammation in anterior chamber negatively affects the endothelial function in patients with acute anterior uveitis.

PURPOSE To investigate the corneal endothelium damage in Graves ophthalmopathy (GO) and its role as a promising quantitative index to evaluate GO activity. DESIGN Cross-sectional study. METHODS This study included 128 eyes of 64 patients with GO. All subjects underwent ophthalmologic examinations, including proptosis, tear break-up time (BUT), corneal fluorescein staining, and Schirmer test. Corneal endothelium was measured by noncontact specular microscope and ocular biometric parameters were measured by IOLMaster 700. Each eye was assigned a specific clinical activity score (CAS), then grouped as active (CAS ≥3 points) or inactive (CAS

Background: Mitomycin C is an antimetabolite and affects corneal endothelial cells as measured by noncontact specular microscopy after trabeculectomy with mitomycin-C (MMC). Objective: To evaluate the corneal endothelial cells changes after trabeculectomy with MMC. Patients and methods: Patients were categorized into 2 groups, group A(25 eyes) were managed by trabeculectomy with mitomycin-C injection, group B (25 eyes) were managed by trabeculectomy with mitomycin-C sponge-applied technique which was carried out on patients attending at Ophthalmology Department of Al-Azhar University Hospitals from March 2019 to February 2021. Follow-up by specular microscopy preoperatively and 1 month and 6 months postoperatively to evaluate corneal endothelial cells density and morphological changes. Results: There was a significant reduction in the endothelial cell density and percentage of hexagonal cells and increased variation in cell size in the two studied groups after 1 month and 6 months compared with preoperative values. With no differences between the two studied groups. Conclusion: Trabeculectomy with both MMC injection and sponge applied caused a significant loss of the corneal endothelial cells intraoperatively or at the early postoperative period. Progressive cell losses due to MMC were not a major concern.

AIM: To investigate the safety and efficacy of using a one-step viscoelastic agent technique for posterior chamber phakic implantable collamer lens with a central hole (ICL V4c) implantation for myopia correction. METHODS: The one-step viscoelastic agent technique for ICL V4c implantation was used in 100 eyes of 52 patients. Refractive outcomes, intraocular pressure (IOP), corneal endothelial cell, and corneal densitometry values were evaluated at 1d, 1wk, 1 and 3mo postoperatively. RESULTS: All the surgeries were uneventful. No corrected distance visual acuity was lost after 3mo. IOP was 16.12±3.18 mm Hg before surgery, and 14.74±3.08 mm Hg at 1d and 14.50±2.56 mm Hg at 3mo after surgery (P0.05). The corneal densitometry values of the central 2 mm and 2 to 6 mm areas showed similar regularities. After surgery, the values significantly increased at 1d, then decreased to the preoperative values at 1wk, and then continued to decrease at 3mo (P

Corneal endothelial cells (CECs) facilitate the function of maintaining the transparency of the cornea. Damage or dysfunction of CECs can lead to blindness, and the primary treatment is corneal transplantation. However, the shortage of cornea donors is a significant problem worldwide. Thus, cultured CEC therapy has been proposed and found to be a promising approach to overcome the lack of tissue supply. Unfortunately, CECs in humans rarely proliferate in vivo and, therefore, can be extremely challenging to culture in vitro. Several promising cell isolation and culture techniques have been proposed. Multiple factors affecting the success of cell expansion including donor characteristics, preservation and isolation methods, plating density, media preparation, transdifferentiation and biomarkers have been evaluated. However, there is no consensus on standard technique for CEC culture. This review aimed to determine the challenges and investigate potential options that would facilitate the standardization of CEC culture for research and therapeutic application.

Purpose: To investigate the effects of horizontal and oblique strabismus surgery on the corneal endothelial cell layer. Methods: Eighty-three eyes of 56 patients were prospectively evaluated. Corneal endothelial cell layer analysis was performed with a specular microscopy device (EM-3000; Tomey Corporation) preoperatively and 1 day, 2 weeks, and 2 months postoperatively. Endothelial cell density, coefficient of variation, average cell area, standard deviation (SD) of the endothelial cell area, hexagonality, and central corneal thickness (CCT) were evaluated. Results: Endothelial cell density, average cell area, and SD of the endothelial cell area did not show a statistically significant change postoperatively. In the one and two horizontal muscle surgery groups, the hexagonality measurements 2 weeks postoperatively were significantly lower compared to preoperative values. In the one horizontal muscle surgery group and when all patients were examined, CCT 1 day postoperatively was significantly higher than 2 weeks postoperatively. No significant change was observed postoperatively in any of the endothelial parameters in the oblique surgery group. Spherical equivalent was positively correlated with endothelial cell density and CCT, and it was negatively correlated with the average cell area and age. Age was positively correlated with the average cell area and the SD of the endothelial cell area, negatively correlated with endothelial cell density and hexagonality, and was not correlated with the coefficient of variation or CCT. Conclusions: There is a temporary effect of horizontal strabismus surgery on hexagonality and corneal thickness. A definite decrease in cellular hexagonality occurs as the number of endothelial cells decrease and cell areas increase with age. [ J Pediatr Ophthalmol Strabimus . 2021;58(4):254–260.]

In 2013, a clinical trial was initiated to investigate cell therapy for the treatment of corneal endothelial decompensation. Cultivating human corneal endothelial cells (CECs) while maintaining their functional phenotype is challenging; therefore, establishment of a confirmed protocol is pivotal for obtaining approval from regulatory authorities for use of cellular therapy products. In this study, we evaluated organ culture (OC) as a storage method for donor corneas used as a raw material for establishing CEC cultures. OC allows storage of corneal tissue for conventional corneal transplantation at 31-37 °C for up to 5 weeks, whereas storage at 4 °C is limited to 2 weeks. We investigated 20 pairs of corneas: one cornea of each pair was stored in OC and the other in cold storage for one week before CEC culture. In 15/20 cases, the CECs assumed a hexagonal sheet-like monolayer structure and expressed endothelial function-related markers. CECs were also obtained from OC corneas that had been stored for 1 (n = 19) and 2 (n = 7) months. As a further test, CECs were cultivated from 5 OC corneas that had been transported from France to Japan. In all cases, these corneas, even after international transport, generated CECs that formed hexagonal monolayers with clinically applicable and sufficiently high cell densities. In conclusion, the CEC cultures required for endothelial cell therapy can be obtained from OC corneas without changing the standard storage operating procedures of the eye banks.

Objective: To compare the effect of Hydroxypropyl methylcellulose versus Sodium Hyaluronate on corneal endothelial cell count in patients undergoing cataract surgery by phacoemulsification. Design: It was a randomized clinical trial. Study Settings: This study was conducted at the Eye Unit III, Mayo Hospital / KEMU, Lahore over 12 months period from June 2014 to May 2015. Hypothesis: There is a significant difference in protective effect on corneal endothelial cell loss with dispersive (Hydroxypropyl methylcellulose) viscoelastic agent as compared to cohesive (sodium hyaluronate) viscoelastic agent used during phacoemulsification with intraocular lens for the treatment of cataract. Material and Methods: Total 80 patients from both the genders aged between 40-70 years with cataract undergoing phacoemulsification surgery were involved in this study. Two equal treatment groups were made by random allocation. Patients in Group-A (n=40) received Hydroxypropyl methylcellulose as viscoelastic agent while patients in Group-B (n=40) received treatment with Sodium Hyaluronate. Surgery was carried out by phacoemulsification technique and follow-up was done at first week, 1st month and 3rd month post-operatively. Data was collected in terms of endothelial cell count by noncontact specular microscope. Results: Patients age was in the range of 40 to 70 years with a mean of 57.26 ± 8.00 years. There were 31 (38.75%) females and 49 (61.25%) male patients with a female to male ratio of 1.6:1. Both the groups had no statistically significant difference in the mean corneal cell count before surgery (2839.67±368.71 cells/mm2 vs. 2801.36 ± 332.52 cells/mm2; p-value= 06.27). Post-operative mean corneal cell count at 1 week, 1 month and 3 months was significantly higher in patients in Group-A compared to Group-B. Similarly, significant difference was noted across various subgroups of both the groups on the basis of patient’s age and gender. Conclusion: This study concluded that there is less corneal endothelial cell loss with dispersive Hydroxypropyl methylcellulose viscoelastic agent as compared to cohesive sodium hyaluronate viscoelastic agent used during phacoemulsification surgery with intraocular lens implant for the treatment of cataract. Keywords: Cataract, phacoemulsification, viscoelastic agents, corneal, endothelial cell.

BACKGROUND The main attributes of cornea which makes it optically important is its ability to maintain its dehydrated state. This corneal dehydration can be achieved and maintained by various factors among which corneal endothelial cell count and morphology play a significant role. Corneal attributes considered in this study i.e. the corneal endothelial cell count and central corneal thickness are extremely variable, and changes are seen even with smaller changes in anterior segment or overall structural changes in eyes. We wanted to compare central corneal thickness in emmetropia and axial myopia and compare corneal endothelial cell count in emmetropia and axial myopia. METHODS The sample size was decided taking into consideration various prevalence studies. The patients were consecutively recruited for the study considering the inclusion and exclusion criteria. The subjects were divided into two groups emmetropia and axial myopia and were differentiated on the basis of the axial lengths. Specular microscopy was performed for the subjects and corneal endothelial cell count, morphology and central corneal thickness were measured, and comparative study was performed. RESULTS This study of 80 eyes shows us that the difference between mean the corneal endothelial cell count between emmetrope (2812.80 cells / mm2) and axial myopes (2653 cells / mm2) is statistically significant where p was < 0.05 while the mean central corneal thickness measurements didn’t show statistically significant change between emmetrope (490.05 microns) and axial myope. (489.37 microns). CONCLUSIONS Axial length has an indirect correlation with the corneal endothelial cell count which is statistically significant. While significant correlation between the central corneal thickness and axial length cannot be established in this study. KEY WORDS Emmetropia, Axial Myopia, Corneal Endothelial Cell Count, Central Corneal Thickness, Comparison

Endothelial keratoplasty (EK) is increasingly used to treat corneal endothelial disease. However, issues remain with EK including a non-zero rejection rate, shortage of donor tissue, and technical challenges for Descemet’s membrane endothelial keratoplasty. Technologies that might solve these issues include small-molecule drugs, surgical innovations, cell therapy, tissue engineering, and gene therapy. Our purpose is to inform ophthalmologists, eye bankers, and eye health professionals of technologies that could impact practice in the near future. Recent advances are targeted at solving known issues with EK. Although none is yet routine, some are in clinical trials. Technologies that are closest to an ideal solution are those facing the highest regulatory and commercial hurdles. In the future, there will multiple options for the treatment of corneal endothelial cell failure. Which of these differing treatments will be used will depend on their efficacy as well as commercial, regulatory, and patient factors.

To summarize the recent advances of clinical and preclinical studies for corneal endothelial tissue bioengineering. The challenges facing the generation of a clinical applicable corneal endothelial graft can be broadly classified into cell source selection, culture medium optimization, scaffold establishment, and the following three- and four-dimensional (4D) corneal construction. Based on the current advances in primary human corneal endothelial cell (HCEC) culture and good manufacturing practice (GMP)–compliant medium development, the first clinical trial of bioengineered HCEC injection therapy has been conducted with encouraging results. Other significant findings include the in vivo experiments of the stem cell–derived HCEC, the development of serum-, xeno-, and additive-free media, and the construction of 4D scaffold. It can be anticipated that tissue engineering–based therapy toward corneal endothelial diseases will replace the current keratoplasty method as a promising treatment option in the near future.

AIM: To compare the difference in corneal endothelial cell density and morphology after phacoemulsification using ringer lactate (RL) and balanced salt solution (BSS) irrigating solutions. METHODS: The prospective randomized controlled trial study was conducted between February 2017 and April 2017 in Dr. YAP Eye Hospital, Yogyakarta, Indonesia. There were a total of 52 subjects (52 eyes) who were senile cataract patients further grouped into two, 26 patients undergoing the phacoemulsification procedure using RL irrigating solution and the other 26 patients with BSS irrigating solution, both conducted by one operator. On the 1, 7, and 28d post operative, an evaluation was done to measure the density and corneal endothelial cell morphology, as well as the variable of inflammation in the two groups. RESULTS: Fifty-two eyes had undergone phacoemulsification with posterior intraocular lens implantation. Both groups were evaluated for the endothelial cell reduction and corneal endothelial cell morphology change, along with postoperative inflammation. On the 28d postoperative, endothelial cell reduction in the BSS group (173.96 cell/mm2, 8.12%) was lower than the RL group (253.20 cell/mm2, 10.25%), percentage of corneal endothelial cell variation coefficient increase in the BSS group (2.92%, 8.36%) was lower compared to the RL group (3.42%, 9.96%), decrease of hexagonal cells of corneal endothelium cells presentation percentage in the BSS group (4.30%, 8.17%) was lower compared to the RL group (4.84%, 8.97%), and the percentage increase of central corneal thickness in the BSS group (4.69 μm, 0.89%) was almost equal to the RL group (4.53 μm, 0.90%). All of the results regarding difference in density and corneal cell endothelium morphology between the two groups did not reveal any statistically significant difference (P>0.05). Inflammatory variable in the two groups were even. CONCLUSION: BSS and RL were equal in their capability of maintaining endothelial cell loss and endothelial cell morphologic change in senile cataract patients after phacoemulsification.

Background: Corneal endothelial tissues are susceptible to mechanical trauma from ultrasound energy during phacoemulsification. Several studies have reported various results of phacoemulsification effect on corneal endothelial cells between diabetic and non-diabetic patients. Purpose: To compare endothelial density, hexagonality, Coefficient of Variance (CV) and Central Corneal Thickness (CCT) changes between diabetic and non-diabetic patients at four weeks after phacoemulsification. Methods: Specular microscopy examinations prior to phacoemulsification and at four-weeks after phacoemulsification were performed on diabetic and non-diabetic groups with cataract. Later, patients in the diabetic group were divided based on their HbA1c level. The changes in endothelial density, the percentage of hexagonality, CV and CCT were evaluated. Results: At a four-weeks follow up, there were no statistical differences in endothelial cells density, CV and CCT changes between diabetic and non-diabetic groups. Mean (SD) of hexagonality percentage at four weeks of follow-up was lower (p-value=0.001) in diabetic group [(14.6) 41.7%] compared to non-diabetic group [(14.5) 50.1%]. There were no differences in endothelial cells density, hexagonality, CV and CCT values between the diabetic group with HbA1c level Conclusion: There were no statistically significant differences in the endothelial loss, reduction of CV and CCT changes between the diabetic and non-diabetic group at four-weeks follow up after phacoemulsification. Diabetic group showed greater hexagonality decrease compared to non-diabetic group at four weeks after phacoemulsification. Different HbA1c levels did not affect the changes in endothelial density, the percentage of hexagonality, CV and CCT after phacoemulsification.

Purpose: To compare corneal endothelium parameters taken by two common noncontact specular microscopes in healthy subjects. Methods: Healthy participants visiting the outpatient eye clinic at the Rabin Medical Center, Petah Tikva, Israel, were recruited prospectively. All participants underwent three consecutive corneal endothelial cell photographs with both the Konan-Noncon Robo SP-6000 and the Tomey EM-3000 specular microscopes. Endothelial cell density (ECD) was evaluated using the manual center technique in both machines. Bland-Altman graphs were used to assess the agreement between the devices, and intraclass correlation coefficient (ICC) served to assess intraobserver variability for each device. Results: Recruited were 49 healthy subjects with a mean age of 48.9 +/- 15.6 years, 49 right eyes were included. The mean ECD was comparable between the Tomey EM-3000 and the Konan-Noncon Robo SP-6000 (2,713.2 +/- 242.4 vs. 2,700.8 +/- 300.5 cells/mm(2), respectively, P=0.47) with a mean difference of 12.4 cells/mm(2) (0.67%), a mean ECD absolute difference of 93.3 cells/mm(2), and low 95% limits of agreement of -222.0 to +246.9 cells/mm(2). A folded empirical distribution function curve showed that all differences fell within 525.4 cells/mm(2), centered around a median of 13.3 cells/mm(2). Intraclass correlation coefficient was high for both the Konan-Noncon Robo SP-6000 (0.93, 95% confidence interval [CI]: 0.89-0.95) and the Tomey EM-3000 (0.88, 95% CI: 0.82-0.93). Conclusions: The difference in endothelial cell measurements between the Konan SP-6000 and the Tomey EM-3000 specular microscopes through the center and the L-count analyzing techniques, respectively, is clinically small and not statistically significant. Nevertheless, caution should be taken when used interchangeably because ECD difference between the two machines can be as high as 525.4 cells/mm(2).

Background/AimsTo investigate the time-dependent change of corneal endothelial cell (CEC) morphology and density (CECD) in patients with glaucoma post instillation of rho-associated protein kinase inhibitor ripasudil (Rip) 0.4% eye drops.MethodsThis observational study involved 163 eyes of 163 patients with glaucoma in whom CEC morphological change was evaluated by CECD calculated via non-contact specular microscopy (NCSM) before and at 1 or 3 months post-Rip instillation. The change of CECD was plotted along with elapsed time from last instillation of Rip. The patients were divided into the following three groups based on the elapsed time post-Rip instillation: Early Group (ResultsMorphological changes in the CECs appeared within 1 hour and recovered to normal within 6 hours post instillation. In the Early, Middle and Late Group, the median rate of CECD change as calculated by the NCSM automated software was −5.68%, −4.95% and −0.07%, respectively. The CEC images showed the same morphological changes with observational study in all four cases.ConclusionDue to transient morphological changes, the NCSM software produced misleading data for determining CECD within 1 hour post-Rip instillation, yet revealed that CEC morphology gradually recovered to normal within 6 hours.

To evaluate the changes in the corneal endothelial cell parameters and macular thickness after intraocular application of epinephrine $$\frac{1}{{10,000}}$$ and epinephrine $$\frac{1}{{100,000}}$$ . In this study, 210 eyes from 210 patients with age-related cataracts who underwent uncomplicated surgery were included. For all patients, specular microscopy of the corneal endothelium and macular OCT were performed before surgery and 3 months after the surgery. Patients were divided randomly into three groups: without drug (control group), epinephrine $$\frac{1}{{10,000}}$$ , and epinephrine $$\frac{1}{{100,000}}$$ . Three months after the surgery, specular microscopy of the cornea and macular OCT measurements were performed. Measurements were compared between the three groups. Postoperative measurements were also compared with those measurements obtained before surgery. All the three groups showed a statistically significant decrease in the endothelial cell density after surgery; the reduction in endothelial cell density in the epinephrine $$\frac{1}{{10,000}}$$ group was significantly more than those of the other two groups (P value

Purpose To determine whether intracameral moxifloxacin 500 μg is noninferior to 250 μg for central endothelial cell loss (ECL) after phacoemulsification. Setting Aravind Eye Care System. Design Prospective masked randomized study. Methods Eyes with bilateral nuclear cataracts, central endothelial cell density (ECD) of more than 2000 cells/mm, and ECD not differing between eyes by more than 200 cells/mm underwent phacoemulsification at least 14 days apart. Intraoperatively, the first eye was randomized to receive either a 500 or 250 μg dose of moxifloxacin intracamerally and received the other dose for the second-eye surgery. Postoperative course was monitored at 1 day, 1 week, 1 month, and 3 months. Preoperative and 30-day and 90-day postoperative central ECD was determined by a reading center for a masked analysis of ECL at 3 months postoperatively. Results Fifty eyes of 25 patients (aged 48 to 69 years) underwent uneventful surgery and had normal postoperative courses. The point estimate (PE) and 95% CI for the mean difference in % ECL between the 500 μg and 250 μg doses at 3 months postoperatively was 0.8% (-5.8%, 7.4%). Upon identifying and removing 2 outliers, noninferiority was proven with a mean difference of the PE, -2.2% (CI, -6.5%, 2.1%). Conclusions Clinical and corneal endothelial cell were comparable in this study population for the 250 μg and 500 μg doses of intracameral moxifloxacin. Both doses were well tolerated clinically, supporting the use of the higher dose for improved antimicrobial coverage for the prevention of postoperative endophthalmitis.

Purpose: Corneal endothelial cells (CECs) serve as a barrier and foothold for the corneal stroma to maintain the function and transparency of the cornea. Loss of CECs during aging or disease states leads to blindness, and cell replacement therapy using either donated or artificially differentiated CECs remains the only curative approach.Methods: Human induced pluripotent stem cells (hiPSCs) that were cultured in chemically defined medium were induced with dual-SMAD inhibition to differentiate into neural crest cells (NCCs). A small-molecule library was screened to differentiate the NCCs into corneal endothelial-like cells. The characteristics of these cells were identified with real-time PCR and immunofluorescence. Western blotting was applied to detect the signaling pathways and key factors regulated by the small molecules.Results: We developed an effective protocol to differentiate hiPSCs into CECs with defined small molecules. The hiPSC-CECs were characterized by ZO-1, AQP1, Vimentin and Na+/K+-ATPase. Based on our small-molecule screen, we identified a small-molecule combination, A769662 and AT13148, that enabled the most efficient production of CECs. The combination of A769662 and AT13148 upregulated the PKA/AKT signaling pathway, FOXO1 and PITX2 to promote the conversion of NCCs to CECs.Conclusion: We established an efficient small molecule-based method to differentiate hiPSCs into corneal endothelial-like cells, which might facilitate drug discovery and the development of cell-based therapies for corneal diseases.

PURPOSE To determine the limits of normal interocular symmetry and the repeatability of central corneal thickness (CCT) and endothelial cell morphology and density. METHODS Two hundred and seventy-eight subjects (age: 10.9-80.7 years) with healthy eyes were studied. CCT as well as endothelial cell morphology and density were measured by non-contact specular microscopy. Three measurements were obtained per eye per participant. Upper and lower tolerance limits of normal interocular symmetry were defined as the 2.5th and 97.5th percentiles of the interocular differences in CCT, endothelial cell density (CECD), coefficient of variation (CoV), hexagonality, and cell size. Repeatability was determined using the intraclass coefficient correlation (ICC), the coefficient of variation (CV), and the test-retest variability standard deviation (TRTSD). RESULTS The mean interocular differences were -1.05 ± 17.5 µm (p = .32) for CCT and 0.5 ± 90.8 cells/mm2 (p = .76) for CECD. The 2.5th and 97.5th percentiles of the interocular differences were -13.9 and 15.4 µm for CCT and -177.4 cells/mm2 to 182.0 cells/mm2 for CECD. Only interocular difference in CoV correlated weakly with age (r = -0.22, P

To evaluate corneal endothelium damage with silicone oil (SO) presence in the anterior chamber after pars plana vitrectomy. We investigated the medical records of consecutive 54 eyes of 53 patients undergoing SO removal after pars plana vitrectomy with SO tamponade at Saitama Medical Center, Jichi Medical University, Japan. We recorded SO tamponade retention period, anterior chamber SO with gonioscope, area of SO attachment to the corneal endothelium before SO removal surgery, and the lens status. We then retrospectively investigated the correlation between SO presence in the anterior chamber and the decrease rate of corneal endothelial cell (CEC) density during SO tamponade. The average decrease rate of CEC density was 7.6 (0–38.1) %. The correlation between SO tamponade retention period and decrease rate of CEC density was high (p = 0.0001). However, there was no correlation between anterior chamber SO under gonioscope, SO attaching area, and lens status with the decrease rate of CEC density (p = 0.11, p = 0.93, p = 0.16). No correlation was observed between CEC loss and the existence of anterior chamber SO, although CEC decrease rate was relatively high after a long SO tamponade period. These findings suggest that SO presence in the anterior chamber may not directly injure CEC.

Objective To compare corneal endothelial cell changes after femtosecond laser-assisted cataract surgery (FLACS) in diabetic and nondiabetic patients. Methods This retrospective study included patients with cataract who underwent FLACS performed by a single surgeon between August 2018 and November 2020. Changes in corneal endothelial cell density (ECD), hexagonality, coefficient of variation in cell size (CV), and central corneal thickness (CCT) at baseline and at the 1-month and 3-month postoperative follow-ups were investigated by dividing the patients into diabetic and nondiabetic groups. Results We included 75 patients (60% men, mean age: 57.7±11.4 years, range: 27-80 years) in the analysis: 31 diabetic patients (64% men, mean age: 58.7±11.9 years, range: 27-79 years) and 44 nondiabetic patients (56% men, mean age: 57.1±11.3 years, range: 34-80 years). No differences were observed between the two groups as regards preoperative and intraoperative parameters, the mean postoperative ECD, hexagonality, and CV. At 1 month postoperatively, the CCT was significantly greater in the diabetic group (P=0.034); however, at 3 months, there was no significant difference between the two groups (P=0.927). Conclusions Changes in postoperative corneal endothelial cells were comparable between patients with and without diabetes after FLACS. Femtosecond laser-assisted cataract surgery seems to cause less damage to the corneal endothelium in diabetic patients because it uses less phacoemulsification energy.

Purpose To determine between-method differences in corneal endothelial cell parameters using center and automated methods of non-contact specular microscopy (CellCheck software of Konan, Inc.) in glaucomatous eyes. Methods We analyzed the central corneal endothelial cell density (ECD) of 245 glaucomatous eyes using center (ECD-Ce) and automated methods (ECD-Au). Based on the ECD-Ce, we allocated subjects to Groups 1 to 10 (at 250 cells/mm2 intervals) and evaluated the ECD, coefficient of variation in cell area (CV), and percentage of hexagonal cells (HEX). Results There was a close correlation (r = 0.91) between the ECD values measured using both methods. However, ECD-Au were significantly higher than those measured by the center method when ECD-Ce was less than 2500 (in Groups 1 to 8; P P = 0.006). The regression equation of (ECD-Au—ECD-Ce) = 1028–0.397*ECD-Ce shows greater deviation in eyes with lower ECD, and this difference became 0 when ECD -Ce was 2593 cells/mm2. None of the 44 subjects with an ECD-Ce of 2 recorded an ECD-Au 2. Compared with the center method, the automated method had higher and lower median CV and HEX values, respectively (P r = −0.49, P r = −0.25, P Conclusion The automated method of the CellCheck software overestimates ECD in eyes with lower ECD values and may overlook risk of corneal decompensation.

We performed an investigation of the temporal impact on corneal endothelial cell (CEC) shape after ophthalmic instillation of one drop of the ROCK inhibitor, ripasudil. Subjects comprised 32 healthy adult volunteers, each of whom instilled ripasudil in the right eye. CEC shape [defined as CEC density (CECD), coefficient of variation (CV), and hexagonal cell rate (6A)], central corneal thickness (CCT), intraocular pressure (IOP), and conjunctival hyperemia were evaluated before instillation (baseline) and at 15 min, 1 h, 2 h, and 4–6 h after instillation by non-contact specular microscopy and slit-lamp microscopy. The incidence of pseudo gutta was also assessed. The CECD (cells/mm2) significantly decreased after instillation as follows: 2661 cells/mm2 at baseline, 2578 cells/mm2 at 15 min, 2527 cells/mm2 at 1 h, 2575 cells/mm2 at 2 h, and 2613 cells/mm2 at 4–6 h. After instillation, CV significantly increased and 6A significantly decreased. Pseudo gutta appeared in 18 subjects (56.3%) after instillation. There was no difference in CCT before and after instillation. IOP significantly decreased after instillation. The most severe conjunctival hyperemia was observed at 15 min after instillation, but had nearly disappeared within 2 h. Pseudo gutta appeared in more than half of the subjects, and the CEC shape changed upon instillation of one drop of ripasudil; however, it recovered at 4–6 h after instillation. Therefore, these changes would not greatly impact clinical treatment. Similar to the presence of conjunctival hyperemia, changes in CEC shape occur at early stages after instillation of ripasudil.

Background: Mitomycin-C was suggested for application during surface ablation procedures to reduce post-operative haze formation. It can effectively reduce haze formation and hence, improve the predictability of visual outcomes following refractive surgery. Despite these advantages, MMC can po-tentially damage all three main corneal cell types including epithelial (differentiated epithelium and limbal cells), stromal (keratocytes), and endothelial cells.Aim of Study: To investigate the croneal endothelial cell changes after Photorefractive Keratectomy (PRK) with and without Mitomycin-C 0.02% for low to moderate myopia.Patient and Methods: This prospective, randomized, comparative study included 60 eyes of 30 patients who were chosen from Ophthalmology Clinic at Tanta University Hos-pital and Shorouk Laser Vision Correction Centre for 6 month from December 2017 to May 2018 and scheduled for Photo-refractive Keratectomy (PRK) using193nm wavelength exci-mer laser system Allegretto EX500, (Wave Light Technologie., USA). These patients were randomly divided into two groups. Group (A) were scheduled for (PRK) with adjunctive mito-mycin-C 0.02% and patients of Group (B) were scheduled for (PRK) without mitomycin-C. Informed consents were obtained from all study participants.Results: In Group A the mean Endothelial Cell Density (ECD) was 2891.61±272.41 and at three months after surgery was 28 13.04±276.55cells/mm2 (p=0.289), pre-operative mean of Coefficient of Variation (CV) was 34.87±4.37 and post-operatively was 34.70±4.06 (p=0.877), the pre-operative mean of Central Corneal Thickness (CCT) was 523.00±26.14 m and 467.36±40.61 m post-operatively (p=0.001), the mean of hexagonality pre-operatively was 64.26±8.64% and 59.96± 8.15% post-operatively (p=0.059) which was statistically insignificant. In Group B the mean Endothelial Cell Density was (ECD) 2874.50±314.48 and at three months after surgery was 2777.23±302.44cells/mm2 (p=0.227), pre-operative mean of Coefficient of Variation (CV) was 33.49±4.98 and post-operatively 33.55±4.10 (p=0.962), pre-operative mean of central corneal thickness was 538.03±26.03 and 484.50±20.55 post-operatively (p=0.001), the mean of hexagonality pre-operatively was 63.00±7.74 and 61.43±7.29% post-operatively (p=0.423).Conclusions: There's no significant effect on endothelium in patients underwent to PRK in low to moderate myopia with or without mitomycin-C. We can't judge the same results for patients with high myopia because the ablation depth will be more and theoretically the diffusion of mitomycin-C to en-dothelium would affect its function or count.

The prospective case series aimed to examine the agreement between the use of a slit-scanning contact specular microscope and a noncontact specular microscope in corneal endothelial cell (CEC) analysis and to evaluate the differences between the central and peripheral regions in normal corneas.After confirming normal corneal endothelium with slit-lamp microscopy, CEC images of 56 eyes of 56 cataractous patients were analyzed in the central and 4 peripheral regions using a slit-scanning contact specular microscope. A noncontact specular microscope was used for the analysis in the central region. The endothelial cell density (ECD), the percentage of hexagonal shape cells (HEX), and the coefficient of variation (CV) in the central region were compared. Differences between central and peripheral CECs were also evaluated.The mean ECD was 2778 cell/mm and was not different from the results using the noncontact specular microscope (2736 cell/mm, P = 0.051). There was a significant correlation (P0.001, R = 0.72). The analysis of HEX resulted in larger values with the slit-scanning contact microscope (53.13% vs. 48.89%, P0.001), whereas there was no difference in the CV (38.48 vs. 38.04, P = 0.56). On comparing the central and peripheral regions, there was no significant difference in the ECD, whereas significant differences were found in the superior region in the HEX and CV (P0.001) and in the nasal region in CV (P = 0.023).The analysis of ECD with the use of the slit-scanning contact specular microscope did not differ from the noncontact specular microscope, and the results demonstrated no difference between the central and peripheral ECD.

To assess corneal endothelial cell (CEC) quantity and quality in eye-bank prepared lamellar grafts for Descemet Membrane Endothelial Keratoplasty (DMEK) from organ cultured donor corneas that have not undergone de-swelling by media supplementation with dextran. Prior to graft preparation, corneas that had not undergone de-swelling (n = 30) were placed into fresh storage medium without dextran (KMI). Corneas in the control group (n = 30) were placed in dehydration medium containing 5% dextran (KMII). Subtotal stripping of Descemet’s membrane (DM) was performed manually. Following graft preparation, 10 corneas of each group were cultured further in their respective medium for 24 h, 72 h, or 120 h, respectively. Before and after DM stripping, as well as at the end of culture, CEC numbers were obtained. At the end of culture, CEC morphology was graded using a scoring system and CEC metabolism was assessed by detection of adenosine triphosphate. At 24 h after DM stripping, mean CEC counts (in cells/mm2) were 2204 in corneas stored in KMII, and 2391 in corneas stored in KMI (p = 0.003). This corresponds to a mean relative CEC loss of 12.4% with dextran versus 9.7% without dextran (p = 0.04). At 72 h, CEC counts were 1946 in KMII, and 2289 in KMI (p = 0.004). This corresponds to CEC loss of 23% with dextran versus 14% without dextran (p = 0.009). At 120 h, CEC counts were 2047 in KMII, and 2230 in KMI (p = 0.14). This corresponds to CEC loss of 22.7% with dextran versus 17.2% without dextran (p = 0.14). Also, at 120 h after DM stripping, 6/10 corneas fell below a threshold of 2000 cells/mm2 if stored in medium containing dextran, versus 1/9 corneas if stored without dextran (p = 0.003). Morphological assessment of CEC quality revealed equal scores for cell polymorphism (median = 1), granulation (median = 0) and segmentation (median = 1) in all groups. Lower ATP/protein ratios were observed in corneas stored in medium without dextran at 24 h (p

To determine how the Rho kinase inhibitor, ripasudil, affects metabolic function and cell viability in donor human corneal endothelial cells (HCECs).Endothelial cell-Descemet membrane (EDM) tissues were treated with 10 μM ripasudil and assayed for mitochondrial and glycolytic activity using extracellular flux analysis and then compared to untreated controls. In addition, EDM tissues with a 24-h ripasudil treatment and control tissues were exposed to 1 μM staurosporine to induce apoptosis and then analyzed for cell viability using apoptosis and necrosis assays.Mitochondrial respiration metrics, specifically maximal respiration (P = 0.758) and spare respiratory capacity (P = 0.777), did not differ among the 1-h ripasudil treatment, 24-h treatment, and untreated tissues. Glycolytic activity assays showed an increase in glycolytic capacity at 1 h compared to the 24-h exposure group (P = 0.049) and controls (P = 0.009). Following exposure to staurosporine, the percentage of apoptotic HCECs was lower (P = 0.009) in ripasudil-treated tissues (2.473%, standard error of the mean [SEM] 0.477%) compared to untreated controls (3.349%, SEM 0.566%). In contrast, the percentage of necrotic HCECs decreased but did not differ statistically (P = 0.158) between ripasudil-treated (3.789%, SEM 0.487%) and untreated (4.567%, SEM 0.571%) tissues.Exposures to ripasudil did not result in any detectable reduction in metabolic function for HCECs in an ex vivo donor tissue model, and an increase in glycolytic activity at the 1-h time point was detected. In addition, HCECs treated with ripasudil gained a protective effect against induced apoptosis, suggesting that ripasudil may help improve the integrity of the corneal endothelium.

Purpose The purpose of this study was to evaluate the features of and relationships among corneal diameter, central corneal thickness (CCT), endothelial cell density (ECD), and corneal refractive properties in cases of microcornea (MC) with normal axial length. Methods We included 22 patients (35 eyes) with MC and cataracts and 41 patients (48 eyes) with isolated cataracts as age-matched controls. They were divided into patients with MC younger than 18 years (MC-child), patients with MC 18 years or older (MC-adult), children with congenital cataracts (CCs), and adults with senile cataracts (SCs). Corneal diameter, axial and anterior chamber length, and keratometry were measured; central corneal endothelial cell imaging was performed. Results The mean horizontal corneal diameter was 7.71 ± 1.51 and 8.78 ± 0.52 mm in MC-child and MC-adult groups, respectively, and 11.89 ± 0.59 and 11.52 ± 2.42 mm in child and adult controls, respectively. The mean CCT was 641.26 ± 63.37 (MC-child) and 617.38 ± 45.40 mm (MC-adult), and 554.92 ± 34.64 (CC) and 551.58 ± 28.47 mm (SC). The mean ECD was 2898.47 ± 443.90 (MC-child) and 2825.81 ± 484.65 cells/mm2 (MC-adult), and 3155.13 ± 372.67 (CC) and 2749.33 ± 399.63 cells/mm2 (SC). The average keratometry was 44.22 ± 3.14 D (MC-child) and 43.86 ± 2.59 D (MC-adult), and 44.19 ± 1.44 D (CC) and 43.94 ± 1.34 D (SC). Conclusions Patients with MC and normal axial length possess specific parameters, including significantly smaller corneal diameter and thicker CCT than the patients in the control groups. There were no significant differences in ECD and average keratometry. These parameters should be taken into account in the follow-up and treatment.

Background Contact lenses can cause complications in 2/3 of the total population, but in most cases, they are mild and easy to manage. However, most of the studies are short-term. The aim of the study was to analyze the publications on the long-term effects of contact lens wear to understand the current research landscape through the analysis of dating networks as well as determining the different research areas and the most cited publications. Methods The Web of Science database was used to perform the publication search, looking for the terms “Long-term” AND “contact lens wear” within the period between 1977 and January 2021. The CitNetExplorer and CiteSpace software were then used to conduct the publication analysis. To obtain the graphics, the VOSviewer software was used. Results A total of 721 publications were found with 2999 citations generated within the network. The year 2010 was singled out as a “key year”, considering the number of publications that emerged in that year and increase in adaptations of contact lens worldwide. The most widely cited publication was “Effects of long-term extended contact lens wear on the human cornea”, an article by Holden et al., which was published in 1985. By using the clustering function, we were able to establish three clusters that encompassed the different research areas within this field: morphological changes, the loss of corneal sensitivity, tear film characteristics and longitudinal changes in ocular physiology. Conclusions The main research topic in this field focuses on the morphological changes that take place in the corneal endothelial cell layer.

Purpose To determine the pachymetric and corneal endothelial cell morphometric features and their relationship to ocular and systemic factors in healthy Congolese subjects. Methods Non-contact specular microscopy was used in 278 healthy eyes (278 subjects) to measure central corneal thickness (CCT), corneal endothelial cell density (CECD) along with cell size, coefficient of variation (CV) in cell size, and hexagonality (HEX). The lower and upper reference limits and average values for each parameter were determined. Correlation and association of average values with anthropo-demographic and clinical variables were assessed. Results The mean age was 38.9 ± 17.2 years (10.9–80.7 years). Average values were 504.2 ± 30.7 μm (CCT), 2907.1 ± 290.9 cells/mm2 (CECD), 348.5 ± 38.4 μm2 (cell size), 32.9 ± 3.6% (CV), and 51.8 ± 7.2% (HEX). CCT was 504.9 ± 33.6 μm in men and 503.6 ± 28.3 μm in women (p = .73); values for CECD were 2917.1 ± 253.5 cells/mm2 and 2899.2 ± 317.8 cells/mm2 (p = 0.61), respectively. Lower and upper reference limits were 449.6 μm and 566.0 μm for CCT, and 2165.3 cells/mm2 and 3414.4 cells/mm2 for CECD, respectively. CCT correlated with body mass index (BMI), (r = − 0.12, P = 0.04). CECD decreased with age (r = − 0.49, P r = − 0.20, P = 0.001), intraocular pressure (r = − 0.13, P = 0.029) and ocular perfusion pressure (r = − 0.28, P = 0.028). CECD decayed by 8.3 cells/mm2 or 0.30% per year of age and CCT decreased by 0.72 μm per kg/m2. Conclusions Mean central cornea was thinner, CECD higher, and references limits lower than reported in other African populations. The CCT and CECD normative values reported herein will be useful for both clinical and research purposes in this population.

PRCIS We reduced the tube length through small limbal incision in eyes underwent Ahmed glaucoma valve implantation and found rate of corneal endothelial cell density was markedly reduced from -12.2%/year to -3.5%/year after this simple procedure. PURPOSE To evaluate the effect of postoperative tube tip trimming (TT) after Ahmed glaucoma valve (AGV) implantation on corneal endothelial cell (CEC) loss. METHODS In this retrospective cohort study, a total of 75 eyes from 67 patients (41 eyes in the AGV implantation only group (AGVG: mean follow up, 42.2±13.8▒mo) and 34 eyes in the TT after AGV implantation group (TAGVG: mean follow up, 64.9±18.5▒mo) were analyzed. TT was performed through a small limbal incision at a mean of 41 months after AGV implantation. Both groups were followed up for at least 1 year and underwent at least two specular microscopic (SM) examinations after AGV implantation and/or TT. RESULTS In the AGVG, CEC density was reduced from 2247.0±527.3 to 1665.7±643.3▒cells/mm2 (P

To compare the benefit of femtosecond laser-assisted cataract surgery (FLACS) versus phacoemulsification (PE) and 2 fragmentation patterns in managing dense cataracts.Randomized controlled trial.Patients with nuclear opacity (NO) grade5 (Lens Opacities Classification System III) were enrolled at the Singapore National Eye Centre. Patients who were unsuitable for FLACS, whose corneal endothelial cell count (ECC) was1,500 cells/mmNinety-three patients were randomized to PE (48), FLACSg (22), and FLACS16 (23). Majority were Chinese (87; 93.5%). Mean age was 74.3 ± 8.8 years of age. Cataracts were mostly graded as NO 5-6 (49; 61.3%). EPT among treatment arms was not different (P = .097, one-way ANOVA) but was significantly higher for NO6 than NO6 (P.001, general linear model). ECC loss was significantly less in FLACSg than in PE (P = .018, Bonferroni correction). Mean 1-month postoperative LogMAR BCVA (0.23 ± 0.20) was significantly better than preoperative BCVA (1.02 ± 0.85; P.001, paired t test) but not different between PE and FLACS.FLACSg but not FLACS16 significantly lowered the mean ECC loss during phacoemulsification in dense cataracts.

Funded by the NIHR Health Technology Assessment Panel (project reference number HTA 13/04/46) and sponsored by University College London (UCL). Supported by the NIHR Biomedical Research Centre (A. C. Day) based at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology. Partly supported by the NIHR Biomedical Research Centre, (C. Bunce) based at Guy's and St Thomas' NHS Foundation Trust and King's College London. Purpose : To report the 1-year outcomes of a randomized trial comparing femtosecond laser-assisted cataract surgery (FLACS) and phacoemulsification cataract surgery (PCS). Setting : Moorfields Eye Hospital, New Cross Hospital, and Sussex Eye Hospital, United Kingdom. Design : Multicenter, randomized controlled noninferiority trial. Methods : Patients undergoing cataract surgery were randomized to FLACS or PCS. Postoperative assessments were masked. Outcomes included uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), complications, corneal endothelial cell count, and patient-reported outcomes measures. Results : The study enrolled 785 participants. A total of 311 of 392 (79%) participants were allocated to FLACS and 292 of 393 (74%) participants were allocated to PCS attended follow-up at 1 year. Mean UDVA was 0.14 (SD = 0.22) for FLACS and 0.17 (0.25) for PCS with difference of -0.03 logarithm of the minimum angle of resolution (logMAR) (95%, -0.06 to 0.01, P = .17). Mean CDVA was 0.003 (0.18) for FLACS and 0.03 (0.23) for PCS with difference of -0.03 logMAR (95% CI, -0.06 to 0.01, P = .11); 75% of both FLACS (230/307) and PCS (218/290) cases were within +/- 0.5 diopters (D) refractive target, and 292 (95%) of 307 eyes of FLACS and 279 (96%) of 290 eyes of PCS groups were within +/- 1.0 D. There were no significant differences between arms for all other outcomes with the exception of binocular CDVA mean difference -0.02 (-0.05 to 0.002) logMAR (P = .036) favoring FLACS. Mean cost difference was £167.62 per patient greater for FLACS (95% iterations between -£14.12 and £341.67). Conclusions : PCS is not inferior to FLACS regarding vision, patient-reported health, and safety outcomes after 1-year follow-up. A difference was found for binocular CDVA, which, although statistically significant, was not clinically important. FLACS was not cost-effective. Publisher PDF

Purpose: To investigate the reduction in corneal endothelial cell density associated with gonioscopy-assisted transluminal trabeculotomy (GATT) in a short-term follow-up period. Methods: A retrospective analysis of the medical charts of patients with open-angle glaucoma who underwent gonioscopy-assisted transluminal trabeculotomy isolated or combined with phacoemulsification (phaco-gonioscopy-assisted transluminal trabeculotomy) was conducted. Patients who underwent phacoemulsification alone were included as controls. The endothelial cell density data (assessed using a specular microscope) before and at the first month after operation were collected and then compared. Results: Sixty-two eyes previously treated with gonioscopy-assisted transluminal trabeculotomy (gonioscopy-assisted transluminal trabeculotomy, n=39 eyes; phaco-gonioscopy-assisted transluminal trabeculotomy, n=23 eyes) fulfilled the inclusion criteria. The mean age of the study patients was 61.3 ± 18.4 years in the stand-alone gonioscopyassisted transluminal trabeculotomy group and 60.4 ± 11.9 in phaco-gonioscopy-assisted transluminal trabeculotomy group. Men comprised 66.6% of the patients in the isolated gonioscopyassisted transluminal trabeculotomy group and 56.5% of those in the phaco-gonioscopy-assisted transluminal trabeculotomy group. The mean visual field defects (mean deviation index) were -13.9 ± 9.2 and -10.3 ± 7.7 dB in the isolated gonioscopy-assisted and phaco-gonioscopy-assisted transluminal trabeculotomy groups, respectively. The patients in the former group presented a mean endothelial cell density reduction of 28.8 cells/mm² (1.31%; p=0.467). In the latter group, the mean endothelial cell density loss was 89.4 cells/mm² (4.36%; p=0.028). The control eyes (23 patients) presented a mean endothelial cell density change of 114.1 ± 159.8 cells/mm² (4.41%; p=0.505). The endothelial cell density reduction in the phaco-gonioscopy-assisted transluminal trabeculotomy group was not significantly different from that in the controls (p=0.81). Conclusions: Gonioscopy-assisted transluminal trabeculotomy appears to be a safe procedure for the corneal endothelial cell layer when performed either isolated or combined with cataract extraction in a short-term follow-up period. RESUMO Objetivo: Investigar a redução na densidade celular endotelial corneana associada à trabeculotomia transluminal assistida por gonioscopia (GATT) em curto prazo. Métodos: Análise retrospectiva de prontuários médicos de pacientes com glaucoma de ângulo aberto que foram submetidos à trabeculotomia transluminal assistida por gonioscopia isolada ou combinada com facoemulsificação. Pacientes que foram submetidos à facoemulsificação isolada foram incluídos como controles. Dados da densidade celular endotelial corneana (avaliada através de microscópio especular) pré-operatória e ao primeiro mês pós-operatório foram coletados e comparados. Resultados: Sessenta e dois olhos que foram submetidos à trabeculotomia transluminal assistida por gonioscopia (trabeculotomia transluminal assistida por gonioscopia=39 olhos; faco com trabeculotomia transluminal assistida por gonioscopia=23 olhos) passaram pelos critérios de inclusão. A idade média dos pacientes estudados era 61,3 ± 18,4 anos no grupo trabeculotomia transluminal assistida por gonioscopia isolada e 60,4 ± 11,9 anos no grupo faco com trabeculotomia transluminal assistida por gonioscopia. Homens eram 66,6% do grupo trabeculotomia transluminal assistida por gonioscopia isolada e 56,5% do grupo faco com trabeculotomia transluminal assistida por gonioscopia. O defeito perimétrico médio (Mean Deviation) era -13,9 ± 9,2 dB e -10,3 ± 7,7 dB nos grupos trabeculotomia transluminal assistida por gonioscopia isolada e faco com trabeculotomia transluminal assistida por gonioscopia respectivamente. O grupo que fora submetido à trabeculotomia transluminal assistida por gonioscopia isolada apresentou redução média da densidade celular endotelial corneana de 28,8 células/mm² (1,31%; p=0,467). No grupo faco com trabeculotomia transluminal assistida por gonioscopia, a redução média da densidade celular endotelial corneana foi de 89,4 células/mm² (4,36%; p=0,028). Olhos controle (23 olhos) apresentaram redução média da densidade celular endotelial corneana de 114,1 ± 159,8 células/mm² (4,41%; p=0,505). A redução na densidade celular endotelial corneana no grupo faco com trabeculotomia transluminal assistida por gonioscopia não foi significativamente diferente do grupo controle (p=0,81). Conclusões: A trabeculotomia transluminal assistida por gonioscopia parece ser segura para a camada endotelial corneana em um curto prazo quando realizada de forma isolada ou combinada com cirurgia de catarata.

Purpose We sought to assess the correlation of corneal endothelial cell (CEC) density to alterations of collagen composition of Descemet membrane (DM) in advanced Fuchs endothelial corneal dystrophy (FECD) and to image such changes by slit-lamp biomicroscopy in vivo. Design Prospective, observational consecutive case series. Methods Fifty eyes (50 subjects) with advanced FECD were enrolled. After slit-lamp biomicroscopy and corneal Scheimpflug imaging, the Descemet endothelium complex (DEC) was retrieved during DM endothelial keratoplasty (DMEK) surgery. The expression of collagens I, III, and IV (COL I, COL III, and COL IV) and corresponding CEC density were analyzed by immunofluorescence flat mount-staining. Presence, diameter and surface area of collagen expression, and CEC density served as the main outcome measures. Results Immunofluorescence staining revealed central coherent collagen positive areas (mean surface area = 10 mm2 ± 6 mm2) corresponding to a fibrillar layer burying the guttae of DM in 84% (42/50) of DECs. CEC density overlying the fibrillar layer compared with the periphery was significantly reduced (−54.8%, P Conclusion A fibrillar layer with a clear geographic pattern marks areas of pronounced loss of CEC density in advanced FECD eyes and may be imaged by slit-lamp biomicroscopy in vivo.

Purpose To assess how trypan blue staining affects Descemet membrane endothelial keratoplasty (DMEK) graft visibility and corneal endothelial cell (CEC) mitochondrial respiration. Methods DMEK grafts (n = 20) were stained with trypan blue 0.06% for 1, 3, 5, or 10 minutes. Each graft was injected into an artificial anterior chamber. Surgery was simulated with tapping and sweeping motions on the corneal surface and injections of balanced salt solution (BSS). Graft visibility was assessed at 5, 10, 20, and 30 minutes. Effects of trypan blue on mitochondrial respiration were assessed using primary CECs cultured from donor corneas (n = 43). Treatment wells exposed to trypan blue 0.06% (1, 5, or 30 minutes) and donor-matched control wells to methylene blue 1% (1 minute) or BSS (1, 5, or 30 minutes) were assayed for key respiration parameters. Results After 5 minutes of surgical manipulation, grafts stained for 5 minutes were significantly more visible than grafts stained for 1 or 3 minutes; there was no added benefit of staining for 10 minutes. After 10 minutes of surgical manipulation, grafts stained for 3 minutes were more visible than grafts stained for 1 minute, without additional benefits of staining ≥5 minutes. No visibility differences were observed after ≥20 minutes of surgical manipulation. CEC mitochondrial respiration did not change significantly following trypan blue exposure for all intervals tested compared to BSS. Conclusions Staining DMEK grafts with trypan blue for 3 to 5 minutes optimizes visibility during surgical manipulation without mitochondrial impairment. Corneal surgeons learning DMEK will benefit from optimizing this critical step.

Objective: The aim of the present study was to assess the anterior segment parameters of patients with chronic obstructive pulmonary disease (COPD) by corneal specular microscopy and biometry and to compare the results with healthy controls. Materials and Methods: Our prospective study consisted of 122 eyes of 61 patients with COPD and 110 eyes of 55 patients without any systemic diseases. A comprehensive medical history was obtained, and corrected visual acuity, bio microscopic and fundus examination, intraocular pressure (IOP) measurement with applanation tonometry, and non-contact specular microscopy were performed in all participants. Results: The endothelial cell density was lower in the COPD group than in the control group (p

Panpan Li,1,2 Yujian Zhang,1 Lihua Kang,1 Yu Guan,1 Jian Wu,1 Huaijin Guan1 1Department of Ophthalmology, Affiliated Hospital of Nantong University, Nantong, China; 2Department of Ophthalmology, The First People’s Hospital of Nantong, Nantong, China Purpose: To compare corneal variations in patients undergoing one-handed and two-handed coaxial phacoemulsification. Setting: Eye Institute, Affiliated Hospital of Nantong University, Nantong, China. Design: Prospective consecutive nonrandomized comparative cohort study. Methods: Patients with cataracts were subject to one-handed (one-handed group) or two-handed coaxial phacoemulsification (two-handed group). Intraoperative phaco parameters and postoperative outcomes, such as visual acuity, surgically induced astigmatism (SIA), corneal volume, central corneal thickness, and corneal endothelial cell counts/size were compared. Results: No significant differences in the intraoperative phaco parameters were noted between the 2 groups. At postoperative week 1, visual outcomes were significantly improved in the one-handed compared with the two-handed group (all P

Background: The corneal complications of a successful vitreoretinal surgery are not the main interest of the surgeon operating on a difficult retinal detachment. However, the surgeon should pay attention to avoid corneal decompensation. Aim: To evaluate corneal endothelial cell changes in patients undergoing pars plana vitrectomy with and without silicon oil (SO) injection.Subjects and Methods: This prospective observational study includes 54 eyes divided into two equal groups: silicone-filled group (group one) and saline-filled group (group two). Endothelial cell densities (ECD), coefficient of variation (CV), central corneal thickness (CCT), and percentage of hexagonal cells at the corneal center were measured preoperatively and at the first week, the first month, and the third month postoperatively and compared between the two groups. All data were collected and statistically analyzed using SPSS program. Results: Three months after PPV, mean ECD in group one and in group two was highly significant (p

Background Complete nuclear disassembly of superhard cataracts cannot always be achieved by phaco chop, which is considered one of the best techniques for dealing with hard cataracts. We present a phaco chop-progressive cracking technique to divide superhard cataracts completely. Case presentation We presented a case of cataract with over Grade V nucleus sclerosis and very low density of corneal endothelial cell (812 cells/mm2). By performing the cataract surgery with our phaco chop-progressive cracking technique, the corneal endothelial cells were well protected and the patient’s visual acuity was markedly improved from finger counting at 40 cm to 20/200 the day after surgery without obvious corneal edema. Conclusions Although an initial learning curve was needed, this phaco chop-progressive cracking technique could be of particular benefit to the superhard cataract, especially in patients with low density of corneal endothelial cells.

The main treatment available for restoration of the corneal endothelium is keratoplasty and DMEK provides faster visual recovery and better postoperative visual acuity when compared to DSAEK. However, the technical challenges related to this technique and the steep technical learning curve seem to prevent the overcoming of DSAEK in favor of DMEK. Furthermore, the outcome of lamellar keratoplasty techniques is influenced by problems related to corneal grafting tissue availability, management, and quality. On the other hand, improvements in the field of cell engineering have opened the way for the use of stem cells-derived corneal endothelial cells with regenerative intent. In this overview, latest findings in endothelial cell engineering are reported, and perspectives of clinical application of mesenchymal stem cells for corneal endothelial replacement and regeneration are evaluated.

Purpose: To measure corneal endothelial cell (EC) quality and quantity following Descemet membrane (DM) stripping of human donor corneas and continued storage in organ culture medium containing dextran.Methods: DM stripping was performed in 30 organ cultured, corneoscleral discs. Corneas were divided into 3 groups of 10 corneas each. Baseline mean EC density (cells/mm2) was 2,372 (SD ± 259) in group 1, 2,540 (SD ± 266) in group 2, and 2,665 (SD ± 263) in group 3. Following subtotal DM stripping, culture was continued at 31°C for 24 hours (group 1), 72 hours (group 2), and 120 hours (group 3), respectively. EC density was measured before stripping and at the end of culture. At the end of culture, corneal EC morphology was graded using a scoring system and EC viability was measured by detection of adenosine triphosphate.Results: At the end of culture, mean EC density was 2,159 (SD ± 293) in group 1, 1,946 (SD ± 182) in group 2, and 2,047 (SD ± 225) in group 3. This constitutes an EC loss of 9,1% (SD...

Corneal endothelial cell abnormalities may be associated with a number of corneal and systemic diseases. Damage to the endothelial cells can significantly affect corneal transparency by altering hydration of the corneal stroma, which can lead to irreversible endothelial cell pathology requiring corneal transplantation. To date, quantitative analysis of endothelial cell abnormalities has been manually performed by ophthalmologists using time consuming and highly subjective semi-automatic tools, which require an operator interaction. We developed and applied a fully-automated and real-time system, termed the Corneal Endothelium Analysis System (CEAS) for the segmentation and computation of endothelial cells in images of the human cornea obtained by in vivo corneal confocal microscopy.First, a Fast Fourier Transform (FFT) Band-pass filter is applied to reduce noise and enhance the image quality to make the cells more visible. Secondly, endothelial cell boundaries are detected using watershed transformations and Voronoi tessellations to accurately quantify the morphological parameters of the human corneal endothelial cells. The performance of the automated segmentation system was tested against manually traced ground-truth images based on a database consisting of 40 corneal confocal endothelial cell images in terms of segmentation accuracy and obtained clinical features. In addition, the robustness and efficiency of the proposed CEAS system were compared with manually obtained cell densities using a separate database of 40 images from controls (n = 11), obese subjects (n = 16) and patients with diabetes (n = 13).The Pearson correlation coefficient between automated and manual endothelial cell densities is 0.9 (p 0.0001) and a Bland-Altman plot shows that 95% of the data are between the 2SD agreement lines.We demonstrate the effectiveness and robustness of the CEAS system, and the possibility of utilizing it in a real world clinical setting to enable rapid diagnosis and for patient follow-up, with an execution time of only 6 seconds per image.