Impact of postoperative intraocular pressure elevation on graft endothelial cells in non-preexisting glaucoma eyes undergoing descemet membrane endothelial keratoplasty: a cohort study.

BACKGROUND: Descemet membrane endothelial keratoplasty (DMEK) has become the dominant keratoplasty procedure. However, the impact of high intraocular pressure (IOP) on the DMEK prognosis in patients without preexisting glaucoma remains unknown. METHODS: Non-glaucoma patients who underwent DMEK in Peking University Third Hospital between July 2017 and March 2023 with a follow-up duration longer than six months were included in this cohort study. Eyes were divided into three groups: Group A) normal IOP; Group B) early IOP elevation (IOP ≥ 30 mmHg or increase of more than 10 mmHg from baseline within 3 days); Group C) intermediate-term IOP elevation (IOP > 21 mmHg or increase of more than 10 mmHg from baseline after 14 days postoperatively). The postoperative IOP, endothelial cell density (ECD), central corneal thickness (CCT), best-corrected visual acuity (BCVA) and rate of graft failure were analysed. RESULTS: Forty-seven eyes from forty-seven patients were included. Thirty-seven eyes were bullous keratopathy, and ten were Fuchs endothelial corneal dystrophy. Twenty-five eyes were classified as Group A, six as Group B and sixteen as Group C. The mean peak IOP was 49.00 ± 4.99 mmHg in Group B eyes and 31.89 ± 11.75 mmHg in Group C eyes. The postoperative BCVA significantly differed from that before surgery (P < 0.001). The ECD at 3 months after surgery in eyes with intermediate-term IOP elevation was lower (P = 0.032). Four eyes with intermediate-term IOP elevation developed graft failure (P = 0.001). CONCLUSIONS: Intermediate-term IOP elevation after DMEK may reduce the graft ECD and lead to graft failure within six months after surgery. However, early IOP elevation had no effect on the prognosis. Careful IOP monitoring and intermediate-term IOP management should be conducted for graft protection.

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

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

Corneal endothelial density and morphology in long-term soft contact lens users in Japan: a retrospective cross-sectional study of 17,732 eyes.

PURPOSE: There is a lack of long-term and large-scale studies on the adverse effects of soft contact lenses (SCLs) on the corneal endothelia of Asian populations. Here, we aimed to examine the effects of long-term SCL use on corneal endothelial density and morphology. MATERIALS AND METHODS: This retrospective study involved consecutive patients at the Miyata Eye Hospital (Miyazaki, Japan), who had used SCLs for more than 1 year. Patients with ophthalmological disorders without refractive errors were excluded. The period of SCL use, SCL type, corneal endothelial cell density (ECD), appearance rate of hexagonal cells (HEX), and coefficient of variation of cells (CV) were analyzed. RESULTS: In total, 17,732 eyes of 8866 patients were included in the analysis (age, 26.0 ± 8.8 years). The mean period of SCL use was 6.3 ± 5.4 years. Multivariate regression analysis revealed that ECD and HEX were significantly negatively correlated with the period of SCL use, age, and sex (p < 0.001 for all). The CV was significantly positively correlated with the period of use (p < 0.001), sex (p = 0.002), and age (p < 0.001). CONCLUSIONS: Corneal ECD, HEX, and CV were significantly associated with the period of SCL use in long-term users. It is essential to regularly check the corneal endothelium in patients with a history of long-term SCL use.

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.

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

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

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.

Inhibition of Rho-associated protein kinase activity enhances oxidative phosphorylation to support corneal endothelial cell migration.

Corneal endothelial cell (CEC) dysfunction causes corneal edema and severe visual impairment that require transplantation to restore vision. To address the unmet need of organ shortage, descemetorhexis without endothelial keratoplasty has been specifically employed to treat early stage Fuchs endothelial corneal dystrophy, which is pathophysiologically related to oxidative stress and exhibits centrally located corneal guttae. After stripping off central Descemet's membrane, rho-associated protein kinase (ROCK) inhibitor has been found to facilitate CEC migration, an energy-demanding task, thereby achieving wound closure. However, the correlation between ROCK inhibition and the change in bioenergetic status of CECs remained to be elucidated. Through transcriptomic profiling, we found that the inhibition of ROCK activity by the selective inhibitor, ripasudil or Y27632, promoted enrichment of oxidative phosphorylation (OXPHOS) gene set in bovine CECs (BCECs). Functional analysis revealed that ripasudil, a clinically approved anti-glaucoma agent, enhanced mitochondrial respiration, increased spare respiratory capacity, and induced overexpression of electron transport chain components through upregulation of AMP-activated protein kinase (AMPK) pathway. Accelerated BCEC migration and in vitro wound healing by ripasudil were diminished by OXPHOS and AMPK inhibition, but not by glycolysis inhibition. Correspondingly, lamellipodial protrusion and actin assembly that were augmented by ripasudil became reduced with additional OXPHOS or AMPK inhibition. These results indicate that ROCK inhibition induces metabolic reprogramming toward OXPHOS to support migration of CECs.

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.

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.

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.

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.

CHIR99021 balance TGFß1 induced human corneal endothelial-to-mesenchymal transition to favor corneal endothelial cell proliferation.

Corneal endothelial cells (CECs) play a major role in the maintenance of stromal hydration via the barrier and pump function for clear vision. Adult CECs cannot regenerate after injury. CECs cultured in vitro can undergo mitosis but may undergo corneal endothelial-to-mesenchymal transition (EnMT) and lose their endothelial characteristics. In this study, we examined the effects of CHIR99021 on transforming growth factor beta-1(TGFß1)-induced EnMT in human CECs (hCECs) lines. CHIR99021 kept hCECs in the hexagonal shape and could downregulate the EnMT markers alpha-smooth muscle actin (α-SMA) and fibronectin (FN1), meanwhile maintained the hCECs function markers Na+/K+-ATPase and zonula occludens-1 (ZO-1) at levels comparable to those in the normal control. Interestingly, we found that the combination of CHIR99021 and TGFß1 at appropriate concentrations would significantly promote the proliferation and migration of hCECs. These effects may be related to the inhibition of RhoA or Rac1, as well as the activation of Wnt and Erk pathway, with a calcium homeostasis. Our findings indicate that CHIR99021 inhibit EnMT and that the combination of CHIR99021 and TGFß1 may provide new ideas for corneal endothelial regeneration and wound healing.

The role of Wnt/ß-catenin pathway for skin-derived precursors differentiating into corneal endothelial cell-like cells.

Bullous keratopathy is a serious blinding eye disease requiring corneal endothelial transplantation. However, the lack of cornea donors forced us to search for new sources of functional corneal endothelial cells (CECs). In our previous study, we have successfully differentiated the SKPs into CEC-like cells with unclear mechanism. The Wnt/ß-catenin signaling pathway is essential for maintaining embryonic eye development and formation. This study aimed to clarify the activity of Wnt/ß-catenin pathway in the process of skin-derived precursors (SKPs) differentiating into CEC-like cells. We showed that the expression of active ß-Catenin, p-GSK3ß, P-LRP6 and LRP6 upregulated, indicating the activation of Wnt/ß-catenin pathway during CEC-like cells induction. What's more, when the pathway was inhibited with a specific inhibitor, the process of induction was obviously suppressed. These findings indicates that Wnt/ß-catenin pathway plays an important role in the CEC-like cells induction from SKPs. Our study lays an experimental foundation for providing abundant corneal endothelial cells and promotes CEC-like cells to be clinically applied in cellular replacement therapy or regenerative medicine in the future.

Corneal endothelial cell density and its correlation with birth weight, anthropometric parameters, and ocular biometric parameters in Chinese school children.

BACKGROUND: To describe the distribution of corneal endothelial cell density (ECD), and to explore its correlation with birth weight (BW), anthropometric parameters, and ocular biometric parameters in Chinese school children. METHODS: In the population-based cross-sectional Nanjing Eye Study, children were measured for anthropometric information, for ECD by the noncontact specular microscope and for ocular biometric parameters by the optic low-coherent reflectometer. Data from right eyes were analyzed to illustrate the distribution of ECD and for determining correlated factors with ECD using univariate and multiple linear regression analysis. Comparisons among three different BW groups were performed using a one-way ANOVA analysis followed by the Bonferroni correction for pairwise comparisons. RESULTS: Of 1171 children, the mean (± standard deviation) ECD was 2875.34 ± 195.00 cells/mm2. In the Multiple Linear Regression analysis, BW, gender and central corneal thickness were significantly associated with ECD. The ECD increased by 36.16 cells/mm2 with BW increasing by 1 kg (P = 0.001) and increased by 0.44 cells/mm2 for every additional 1 mm in central corneal thickness (P = 0.01). The ECD of girls was 54.41 cells/mm2 higher than boys (P < 0.001). Children born with low BW presented significantly lower ECD than those born with normal BW (P < 0.05) and high BW (P < 0.05). Age and axial length were not significantly associated with ECD (P = 0.06 and P = 0.21, respectively). CONCLUSIONS: In Chinese school children aged 82 to 94 months, the ECD is positively correlated with BW and central corneal thickness, in which BW is a newly identified associated factor. It is like that gender plays an important role in ECD distribution while girls have relatively greater ECD than boys.

Evaluation of corneal endothelial cell morphology by specular microscopy in patients with acute anterior uveitis.

PURPOSE: To determine the structure and morphology of corneal endothelial cell layer in patients with acute anterior uveitis. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

Overestimation of corneal endothelial cell density by automated method in glaucomatous eyes with impaired corneal endothelial cells.

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 < 0.001 to 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 < 1000 cells/mm2 recorded an ECD-Au < 1000 cells/mm2. Compared with the center method, the automated method had higher and lower median CV and HEX values, respectively (P < 0.001). The between-method differences in both CV and HEX were negatively correlated with ECD-Ce (r = -0.49, P < 0.001 and r = -0.25, P < 0.001, respectively). CONCLUSION: The automated method of the CellCheck software overestimates ECD in eyes with lower ECD values and may overlook risk of corneal decompensation.

Corneal endothelial expansion using human umbilical cord mesenchymal stem cell-derived conditioned medium.

Rabbit corneal endothelial cells are frequently used in pharmacological experiments and are useful for corneal transplant experiments. We performed the present study to analyze the effect of conditioned medium (CM) derived from human umbilical cord mesenchymal stem cells (HUMSCs) on the growth of rabbit corneal endothelial cells (RCECs) and to establish a program for expansion of RCECs in vitro. RCECs were cultured using a CM derived from HUMSCs (HUMSCs-CM) in vitro. The proliferation ability of RCECs cultured in the presence of HUMSCs-CM was evaluated by conducting 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, colony formation, and scratch migration assays. The proliferation ability of RCECs maintained in HUMSCs-CM was significantly enhanced as compared to RCECs cultivated in the control group. Immunofluorescence indicated that zonula occludens-1 (ZO-1) and N-cadherin were located at intercellular junctions. Real-time PCR and western blot analyses demonstrated that the CEC-relative functional markers were expressed in RCECs maintained in HUMSCs-CM. Flow cytometry analyses demonstrated that HUMSCs-CM promoted the G0/G1 entrance to the S phase in RCECs. Our results demonstrated that HUMSCs-CM induced the proliferation of RCECs in vitro and maintained the necessary characteristic phenotypes. The expanded RCECs may provide a promising cell source for experimental research and clinical therapy.

The impact of biomechanics on corneal endothelium tissue engineering.

The integrity of innermost layer of the cornea, the corneal endothelium, is key to sustaining corneal transparency. Therefore, disease or injury causing loss or damage to the corneal endothelial cell population may threaten vision. Transplantation of corneal tissue is the standard treatment used to replace malfunctioning corneal endothelial cells. However, this surgery is dependent upon donor tissue, which is limited in supply. Hence, tissue engineers have attempted to construct alternative transplantable tissues or cell therapies to alleviate this problem. Nevertheless, the intrinsic non-dividing nature of corneal endothelial cells continues to foil scientists in their attempts to yield large numbers of cells in the laboratory for use in such novel therapies. Interestingly, the contribution of the biomechanical properties of the underlying extracellular matrix (ECM) on cell division, tissue development and maintenance has been extensively investigated in other many cell types. However, the impact of biomechanics on corneal endothelial cell behaviour is relatively unexplored. Here, we describe contemporary tissue engineering solutions aimed at circumventing donor tissue scarcity. We review the ECM structure and biomechanical features of corneal endothelial cells. We discuss the alterations of ECM in endothelial disease development and progression and point out the role of ECM in developing a tissue-engineered corneal endothelium. We highlight the main biomechanical cues, including topographical and mechanical features, that impact cellular behaviors. Finally, we discuss the influence of biomechanical cues on cell and tissue development, and how corneal endothelial cells response to individual biomechanical stimuli in tissue engineering, which have implications for designing an engineered endothelium and maintaining cell function.

Profile of central corneal thickness and corneal endothelial cell morpho-density of in healthy Congolese eyes.

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 < 0.001), BMI (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.

Primary cell culture of canine corneal endothelial cells.

OBJECTIVE: To establish a primary cell culture and clarify the characteristics of canine corneal endothelial cells in vitro. PROCEDURES: The eyes were enucleated from dogs that were euthanized for reasons unrelated to this study. Enucleated canine eyes were dissected, and the intact corneas were isolated from the globes. Using enzymes, the corneal endothelial cells were dispersed from the cornea. The obtained canine corneal endothelial cells were cultured in a cell culture dish. Cultured corneal endothelial cells were morphologically evaluated using phase-contrast microscopy. Immunohistochemical analysis of the cultured cells, particularly of the corneal endothelial cell marker, zonula occludens-1 (ZO-1), Na+ /K+ -ATPase, and vimentin, was performed to clarify whether the cultured cells were actually corneal endothelial cells. Furthermore, the post-passage morphology of cultured cells was evaluated. RESULTS: Canine primary cultured corneal endothelial cells showed morphologically small, cobblestone-like structures. The isolated cells had proliferative ability in vitro and demonstrated positive expression of the corneal endothelial cell markers, ZO-1, Na+ /K+ -ATPase, and vimentin. However, repeated passages resulted in larger cell sizes as assessed by phase-contrast microscopy. Repeated passages also resulted in lower cell density. CONCLUSIONS: This study demonstrated the successful culture of canine corneal endothelial cells. This might enhance the understanding of corneal endothelial cell characteristics in dogs.