Single cell analysis of short-term dry eye induced changes in cornea immune cell populations.

Background: Dry eye causes corneal inflammation, epitheliopathy and sensorineural changes. This study evaluates the hypothesis that dry eye alters the percentages and transcriptional profiles of immune cell populations in the cornea. Methods: Desiccating stress (DS) induced dry eye was created by pharmacologic suppression of tear secretion and exposure to drafty low humidity environment. Expression profiling of corneal immune cells was performed by single-cell RNA sequencing (scRNA-seq). Cell differentiation trajectories and cell fate were modeled through RNA velocity analysis. Confocal microscopy was used to immunodetect corneal immune cells. Irritation response to topical neurostimulants was assessed. Results: Twelve corneal immune cell populations based on their transcriptional profiles were identified at baseline and consist of monocytes, resident (rMP) and MMP12/13 high macrophages, dendritic cells (cDC2), neutrophils, mast cells, pre T/B cells, and innate (γDT, ILC2, NK) and conventional T and B lymphocytes. T cells and resident macrophages (rMP) were the largest populations in the normal cornea comprising 18.6 and 18.2 percent, respectively. rMP increased to 55.2% of cells after 5 days of DS. Significant changes in expression of 1,365 genes (adj p < 0.0001) were noted in rMP with increases in cytokines and chemokines (Tnf, Cxcl1, Ccl12, Il1rn), inflammatory markers (Vcam, Adam17, Junb), the TAM receptor (Mertk), and decreases in complement and MHCII genes. A differentiation trajectory from monocytes to terminal state rMP was found. Phagocytosis, C-type lectin receptor signaling, NF-kappa B signaling and Toll-like receptor signaling were among the pathways with enhanced activity in these cells. The percentage of MRC1+ rMPs increased in the cornea and they were observed in the basal epithelium adjacent to epithelial nerve plexus. Concentration of the chemokine CXCL1 increased in the cornea and it heightened irritation/pain responses to topically applied hypertonic saline. Conclusion: These findings indicate that DS recruits monocytes that differentiate to macrophages with increased expression of inflammation associated genes. The proximity of these macrophages to cornea nerves and their expression of neurosensitizers suggests they contribute to the corneal sensorineural changes in dry eye.

Robust and accurate corneal interfaces segmentation in 2D and 3D OCT images.

Segmentation of corneal layer interfaces in optical coherence tomography (OCT) images is important for diagnostic and surgical purposes, while manual segmentation is a time-consuming and tedious process. This paper presents a novel technique for the automatic segmentation of corneal layer interfaces using customized initial layer estimation and a gradient-based segmentation method. The proposed method was also extended to three-dimensional OCT images. Validation was performed on two corneal datasets, one with 37 B-scan images of healthy human eyes and the other with a 3D volume scan of a porcine eye. The approach showed robustness in extracting different layer boundaries in the low-SNR region with lower computational cost but higher accuracy compared to existing techniques. It achieved segmentation errors below 2.1 pixels for both the anterior and posterior layer boundaries in terms of mean unsigned surface positioning error for the first dataset and 2.6 pixels (5.2 µm) for segmenting all three layers that can be resolved in the second dataset. On average, it takes 0.7 and 0.4 seconds to process a cross-sectional B-scan image for datasets one and two, respectively. Our comparative study also showed that it outperforms state-of-the-art methods for quantifying layer interfaces in terms of accuracy and time efficiency.

Evaluation of the results of contact lens assisted corneal cross-linking treatment in keratoconus patients with thin corneas.

PURPOSE: We aimed to compare the efficacy and safety of accelerated contact lens-assisted cross-linking (CA-CXL) with Lotrafilcon B and Comfilcon A lenses in keratoconus (KC) patients with thin corneas. STUDY DESIGN: Retrospective, single-center study. MATERIALS AND METHODS: We retrospectively included 51 eyes of 39 KC patients with corneal thickness <400µm after epithelial scraping (Epi-off), who underwent accelerated CA-CXL treatment with Lotrafilcon B (n=20) and Comfilcon A (n=31). Uncorrected and corrected distance visual acuity (UDVA and CDVA), manifest refraction values, corneal topographic data and endothelial cell density were recorded at preoperative and postoperative 1st, 3rd and 6th month controls. RESULTS: CDVA in the Comfilcon A group was higher than CDVA before surgery at 6 months postoperatively (p<0.001). When the two lenses were compared, CDVA was found to be significantly higher in the Lotrafilcon B group in the preoperative, postoperative 1st month and 3rd month values, but there was no significant difference between the postoperative 6th month values (p=0.028, p=0.018, p=0.044, p=0.181, respectively). The maximum keratometry (Kmax) value at the 6th month after surgery in the Comfilcon A group was significantly lower than in the Lotrafilcon B group (p=0,009). There was no significant difference between the endothelial cell density values between the groups (p=0.623, p=0.609, p=0.794, p=0.458, respectively). There was no significant difference between the progression, regression, and stability rates of the two groups (p=0.714). CONCLUSIONS: Accelerated CA-CXL with Lotrafilcon B and Comfilcon A silicone hydrogel lenses is a safe and effective method to stop progression in patients with thin corneas.

Modified sutureless and glue-free method versus conventional sutures for conjunctival autograft fixation in primary pterygium surgery: a systematic review and meta-analysis.

BACKGROUND: Pterygium is a common ocular surface disorder that requires surgical intervention for treatment. Conjunctival autografts are preferred over simple excision due to lower recurrence rates. This systematic review and meta-analysis compared the modified sutureless glue-free (MSGF) method with conventional sutures (CS) for conjunctival autograft fixation in primary pterygium surgery. METHODS: A comprehensive search was conducted in MEDLINE, Embase, CENTRAL, Google Scholar and ClinicalTrials.gov for randomised controlled trials (RCTs) comparing MSGF and CS conjunctival autografts. Outcome measures included operation time, recurrence and postoperative complications. Standardised mean difference (SMD) and risk ratio (RR) were used for continuous and dichotomous outcomes, respectively. RESULTS: 11 RCTs involving 833 participants were included. The analysis revealed that MSGF had a significantly shorter operation time compared with CS (SMD -3.704, 95% CI -5.122 to -2.287, p<0.001). CS was associated with a higher risk of foreign body sensation (RR 0.22, 95% CI 0.06 to 0.74, p=0.01). MSGF was associated with a higher risk of graft dehiscence (RR 9.01, 95% CI 2.74 to 29.68, p=0.000) and graft retraction (RR 2.37, 95% CI 1.17 to 4.77, p=0.02). No significant differences were found in recurrence, graft haemorrhage, granuloma, Dellen and conjunctival oedema. CONCLUSION: Using the MSGF technique in conjunctival autograft fixation for pterygium surgery reduces operation time by relying solely on the patient's blood for fixation. However, it increases the risk of graft dehiscence and retraction. However, CS is linked to a higher likelihood of experiencing foreign body sensations. Understanding the learning curve and surgeon familiarity with novel techniques is crucial for optimising patient care and surgical outcomes, while individualised decision-making is necessary considering the advantages and disadvantages of each approach. Further research is warranted to minimise complications and optimise surgical outcomes.

Femtosecond laser-assisted astigmatic keratotomy versus toric IOL implantation for correcting astigmatism in cataract patients: a systematic review and meta-analysis with trial sequential analysis.

AIMS: To compare the refractive and visual outcomes of femtosecond laser-assisted astigmatic keratotomy (FSAK) and toric intraocular lens (IOL) implantation for correcting astigmatism in cataract patients. METHODS: Studies were retrieved from the Ovid-Medline, EMBASE, Cochrane Central Register of Controlled Trials and Scopus which compared FSAK and toric IOL for astigmatism correction in cataract patients. Outcome measures included postoperative refractive cylinder, correction index, uncorrected distance visual acuity (UDVA), the proportion of patients achieving a residual refractive cylinder of 1.00 dioptre or less, target-induced astigmatism (TIA) and surgically induced astigmatism (SIA). The trial sequential analysis (TSA) was used to collect firm evidence supporting our conclusion. RESULTS: 9 studies encompassing 590 participants were analysed. The meta-analysis revealed that toric IOLs could result in less postoperative refractive cylinder and provide better UDVA compared with FSAK. The TSA disclosed strong evidence of lower postoperative refractive cylinder in the toric IOL group compared with that of the FSAK group. FSAK showed a smaller correction index and lower mean TIA and SIA compared with toric IOLs. CONCLUSIONS: For cataract patients, both FSAK and toric IOLs are effective methods for correcting astigmatism. However, toric IOLs offer less postoperative astigmatism and result in better postoperative UDVA compared with FSAK. In vector analysis of astigmatism, toric IOLs can also produce higher TIA and SIA. Additionally, neither method is associated with severe untreatable complications. Therefore, the conclusion is that toric IOLs are the preferred choice for astigmatism correction in cataract patients and FSAK serves as a viable alternative when toric IOLs are contraindicated.

Validation of the Reichert® Tono-Vera® Vet rebound tonometer in normal ex vivo canine eyes.

OBJECTIVE: To determine the accuracy of the Reichert® Tono-Vera® Vet rebound tonometer for canine intraocular pressure (IOP) measurement. ANIMALS STUDIED: Five normal canine ex vivo globes. PROCEDURES: The anterior chambers of five freshly enucleated normal canine eyes were cannulated and connected to a reservoir of Plasma-Lyte A and a manometer. Starting at a manometric IOP of 5 mmHg, the pressure was progressively increased to 80 mmHg by raising the reservoir. Triplicate IOP measurements were taken with the Tono-Vera® Vet from the central cornea using the dog setting and compared to the manometric pressure by linear regression analysis and Bland-Altman plots. RESULTS: There was a strong positive linear regression trend when comparing central corneal Tono-Vera® Vet IOPs to manometric pressures (r2 = .99) with solid agreement between the two methods. Compared to manometric IOPs, the Tono-Vera® Vet underestimated IOPs at higher pressures ≥70 mmHg. CONCLUSIONS: Measurement of IOPs from the central cornea with the Tono-Vera® Vet provided accurate results over a large range in normal canine globes compared to direct manometry. The mild to moderate underestimation of IOPs at high pressures was not considered clinically relevant.

Ex vivo, in vivo and in silico studies of corneal biomechanics: a systematic review.

Healthy cornea guarantees the refractive power of the eye and the protection of the inner components, but injury, trauma or pathology may impair the tissue shape and/or structural organization and therefore its material properties, compromising its functionality in the ocular visual process. It turns out that biomechanical research assumes an essential role in analysing the morphology and biomechanical response of the cornea, preventing pathology occurrence, and improving/optimising treatments. In this review, ex vivo, in vivo and in silico methods for the corneal mechanical characterization are reported. Experimental techniques are distinct in testing mode (e.g., tensile, inflation tests), samples' species (human or animal), shape and condition (e.g., healthy, treated), preservation methods, setup and test protocol (e.g., preconditioning, strain rate). The meaningful results reported in the pertinent literature are discussed, analysing differences, key features and weaknesses of the methodologies adopted. In addition, numerical techniques based on the finite element method are reported, incorporating the essential steps for the development of corneal models, such as geometry, material characterization and boundary conditions, and their application in the research field to extend the experimental results by including further relevant aspects and in the clinical field for diagnostic procedure, treatment and planning surgery. This review aims to analyse the state-of-art of the bioengineering techniques developed over the years to study the corneal biomechanics, highlighting their potentiality to improve diagnosis, treatment and healing process of the corneal tissue, and, at the same, pointing out the current limits in the experimental equipment and numerical tools that are not able to fully characterize in vivo corneal tissues non-invasively and discourage the use of finite element models in daily clinical practice for surgical planning.

Design of a new 3D printed all-in-one magnetic smartphone adapter for fundus and anterior segment imaging.

PURPOSE: To describe and validate a 3D-printed adapter tool which could be used with either a slit lamp or a condensing lens, interchangeable between devices through magnetic fastening, in order to provide physicians a quick, easy and effective method of obtaining clinical photos. MATERIALS AND METHODS: Three specialists, with at least 4-year experience in ophthalmology, gave a rate of image quality obtained by our device and the diagnostic confidence grade. The 3 specialists conducted each 13 or 14 examinations with the smartphone and magnetic adapter. At the end of evaluation, they rated with the Likert scale the ease of use of the device in obtaining clinical images of the anterior segment and ocular fundus respectively. RESULTS: Data of quality perception and confidence demonstrated high values not dissimilar to the "de visu" eye examination. Moreover the instrument we designed turned out to be very user friendly. CONCLUSION: Our adapter coupled with a modern smartphone was able to obtain 4k images and videos of anterior segment, central and peripheral fundus, in an easy and inexpensive way.

Correlation Between the Cost and Safety of Corneal Graft Types.

Background Corneal diseases are the fourth most common cause of blindness worldwide. In the majority of these diseases, vision reduction is reversible and can be restored to a large extent by replacing the cornea through specific surgery and, in particular, transplantation. In Greece, due to a lack of organized eye banks as well as donors, the grafts intended for corneal transplantation usually come from eye banks abroad. This study focuses on the dynamics of cost versus value in the decision-making process for the procurement of corneal grafts, ultimately investigating the safety that the procured grafts provide to patients. Methodology A total of 267 patients with severe vision problems who underwent 301 corneal and amniotic membrane transplants from years 2020 to 2023 at the Transplant Unit of the Athens General Hospital "Georgios Gennimatas" were included in this study. All patients who were deemed appropriate to undergo corneal transplant operations, the diagnosis that led to the specific surgery, and other relevant data were recorded and evaluated. Results There was no significant difference in the ratio between males and females (51.3% male and 48.7% female). The mean age of the patients was 66.5 years (SD = 13.7 years). Graft problems were faced by 13.9% of the patients, with the amniotic membrane by 1.5% (in the total number of surgical operations) and both eyes by 4.5% of patients. The majority of the patients had undergone only one surgery (88.8%). Reoperation was needed in 14% of the cases, and 7.6% of the cases were surgeries that occurred due to graft rejection or non-functioning grafts from surgeries performed at another hospital or clinic. In the majority of surgeries (60.8%), a Descemet's stripping automated endothelial keratoplasty graft was used. The mean cost was 3,167 euro (SD = 960.3 euro). Furthermore, in 35.8% of the surgeries, the graft was preserved with amphotericin. Conclusions The present study draws useful conclusions about the effectiveness of surgical interventions through the correlation of cost and safety of the grafts that are approved and finally used in corneal transplants, as well as the submission of proposals to improve the procedures and lead to patient benefits.

An untargeted metabolomics approach to study changes of the medium during human cornea culture.

INTRODUCTION: Two main approaches (organ culture and hypothermia) for the preservation and storage of human donor corneas are globally adopted for corneal preservation before the transplant. Hypothermia is a hypothermic storage which slows down cellular metabolism while organ culture, a corneal culture performed at 28-37 °C, maintains an active corneal metabolism. Researchers, till now, have just studied the impact of organ culture on human cornea after manipulating and disrupting tissues. OBJECTIVES: The aim of the current work was to optimize an analytical procedure which can be useful for discovering biomarkers capable of predicting tissue health status. For the first time, this research proposed a preliminary metabolomics study on medium for organ culture without manipulating and disrupting the valuable human tissues which could be still used for transplantation. METHODS: In particular, the present research proposed a method for investigating changes in the medium, over a storage period of 20 days, in presence and absence of a human donor cornea. An untargeted metabolomics approach using UHPLC-QTOF was developed to deeply investigate the differences on metabolites and metabolic pathways and the influence of the presence of the cornea inside the medium. RESULTS: Differences in the expression of some compounds emerged from this preliminary metabolomics approach, in particular in medium maintained for 10 and 20 days in presence but also in the absence of cornea. A total of 173 metabolites have been annotated and 36 pathways were enriched by pathway analysis. CONCLUSION: The results revealed a valuable untargeted metabolomics approach which can be applied in organ culture metabolomics.

Comparative Analysis of Corneal Higher-Order Aberrations after Laser-Assisted In Situ Keratomileusis, Photorefractive Keratectomy, and Small Incision Lenticule Extraction with Correlations to Change in Myopic Q-Value and Spherical Equivalent with and without Astigmatism.

Background: This retrospective chart review compared the higher-order aberrations (HOAs) among photorefractive keratectomy (PRK), laser-assisted in situ keratomileusis (LASIK), and small incision lenticule extraction (SMILE) alongside changes in spherical equivalent (SEQ) and corneal shape (Q-value). Methods: Analyzing 371 myopic eyes, including 154 LASIK, 173 PRK, and 44 SMILE cases, Pentacam imaging was utilized pre-operatively and at one-year post-operative visits. Results: All procedures resulted in 100% of patients achieving an uncorrected distance visual acuity (UDVA) of 20/40 or better, with 87% of LASIK and PRK, and 91% of SMILE patients having 20/20 or better. Significant increases in HOAs were observed across all procedures (p < 0.05), correlating positively with SEQ and Q-value changes (LASIK (0.686, p < 0.05), followed by PRK (0.4503, p < 0.05), and SMILE (0.386, p < 0.05)). Vertical coma and spherical aberration (SA) were the primary factors for heightened aberration magnitude among the procedures (p < 0.05), with the largest contribution in SMILE, which is likely attributed to the centration at the corneal apex. Notably, PRK showed insignificant changes in vertical coma (-0.197 µm ± 0.0168 to -0.192 µm ± 0.0198, p = 0.78), with an increase in oblique trefoil (p < 0.05). Conclusions: These findings underscore differences in HOAs among PRK, LASIK, and SMILE, helping to guide clinicians.

Aquaporins in the Cornea.

The cornea is an avascular, transparent tissue that allows light to enter the visual system. Accurate vision requires proper maintenance of the cornea's integrity and structure. Due to its exposure to the external environment, the cornea is prone to injury and must undergo proper wound healing to restore vision. Aquaporins (AQPs) are a family of water channels important for passive water transport and, in some family members, the transport of other small molecules; AQPs are expressed in all layers of the cornea. Although their functions as water channels are well established, the direct function of AQPs in the cornea is still being determined and is the focus of this review. AQPs, primarily AQP1, AQP3, and AQP5, have been found to play an important role in maintaining water homeostasis, the corneal structure in relation to proper hydration, and stress responses, as well as wound healing in all layers of the cornea. Due to their many functions in the cornea, the identification of drug targets that modulate the expression of AQPs in the cornea could be beneficial to promote corneal wound healing and restore proper function of this tissue crucial for vision.

Corneal endothelial cell density and associated factors among adults at a regional referral hospital in Uganda: a cross-sectional study.

BACKGROUND: To assess the prevalence of low corneal endothelial cell density and correlates of corneal endothelial cell density among adults attending Mbarara University and Referral Hospital Eye Centre in Uganda. METHODS: In this hospital-based cross-sectional study, participants 18 years and older, were enrolled. We obtained informed consent, and basic demographic data. We also conducted visual acuity, a detailed slit lamp examination, intra-ocular pressure, corneal diameter, tear-film break-up time, keratometry, A-scan, and pachymetry on all participants. A confocal microscope Heidelberg HRT3 was used to examine the central cornea and to obtain the mean cell density (cells/mm2). To calculate the proportion of low endothelial cell density, descriptive statistics were used, whereas correlates of endothelial cell density were assessed, using linear regression analyses. RESULTS: We evaluated a total of 798 eyes of 404 participants aged between 18 and 90 years (males = 187, females = 217). The average endothelial cell density was 2763.6 cells/mm2, and there was a decrease in endothelial cell density with increasing age, irrespective of gender. There was no significant difference in endothelial cell density between males and females. Increasing age (adjusted coefficient - 10.1, p < 0.001), history of smoking (adjusted coefficient - 439.6, p = 0.004), history of ocular surgery (adjusted coefficient - 168.0, p = 0.023), having dry eye (adjusted coefficient - 136.0, p = 0.051), and having arcus senilis (adjusted coefficient - 132.0, p = 0.08), were correlated with lower endothelial cell density. However, increasing corneal diameter (adjusted coefficient 134.0, p = 0.006), increasing central corneal thickness (adjusted coefficient 1.2, p = 0.058), and increasing axial length (adjusted coefficient 65.8, p = 0.026), were correlated with higher endothelial cell density. We found five eyes (0.63%) from different participants with a low endothelial cell density (< 1000cells/mm2). CONCLUSION: Our study established baseline normal ranges of ECD in a predominantly black African population, and found that low ECD is rare in our population. The elderly, smokers, and those with past ocular surgery are the most vulnerable. The low prevalence could be due to a lack of reference values for the black African population.

U-NTCA: nnUNet and nested transformer with channel attention for corneal cell segmentation.

Background: Automatic segmentation of corneal stromal cells can assist ophthalmologists to detect abnormal morphology in confocal microscopy images, thereby assessing the virus infection or conical mutation of corneas, and avoiding irreversible pathological damage. However, the corneal stromal cells often suffer from uneven illumination and disordered vascular occlusion, resulting in inaccurate segmentation. Methods: In response to these challenges, this study proposes a novel approach: a nnUNet and nested Transformer-based network integrated with dual high-order channel attention, named U-NTCA. Unlike nnUNet, this architecture allows for the recursive transmission of crucial contextual features and direct interaction of features across layers to improve the accuracy of cell recognition in low-quality regions. The proposed methodology involves multiple steps. Firstly, three underlying features with the same channel number are sent into an attention channel named gnConv to facilitate higher-order interaction of local context. Secondly, we leverage different layers in U-Net to integrate Transformer nested with gnConv, and concatenate multiple Transformers to transmit multi-scale features in a bottom-up manner. We encode the downsampling features, corresponding upsampling features, and low-level feature information transmitted from lower layers to model potential correlations between features of varying sizes and resolutions. These multi-scale features play a pivotal role in refining the position information and morphological details of the current layer through recursive transmission. Results: Experimental results on a clinical dataset including 136 images show that the proposed method achieves competitive performance with a Dice score of 82.72% and an AUC (Area Under Curve) of 90.92%, which are higher than the performance of nnUNet. Conclusion: The experimental results indicate that our model provides a cost-effective and high-precision segmentation solution for corneal stromal cells, particularly in challenging image scenarios.

Increased Anti-Inflammatory Therapeutic Potential and Progenitor Marker Expression of Corneal Mesenchymal Stem Cells Cultured in an Optimized Propagation Medium.

There is a huge unmet need for new treatment modalities for ocular surface inflammatory disorders (OSIDs) such as dry eye disease and meibomian gland dysfunction. Mesenchymal stem cell therapies may hold the answer due to their potent immunomodulatory properties, low immunogenicity, and ability to modulate both the innate and adaptive immune response. MSC-like cells that can be isolated from the corneal stroma (C-MSCs) offer a potential new treatment strategy; however, an optimized culture medium needs to be developed to produce the ideal phenotype for use in a cell therapy to treat OSIDs. The effects of in vitro expansion of human C-MSC in a medium of M199 containing fetal bovine serum (FBS) was compared to a stem cell medium (SCM) containing knockout serum replacement (KSR) with basic fibroblast growth factor (bFGF) and human leukemia inhibitory factor (LIF), investigating viability, protein, and gene expression. Isolating populations expressing CD34 or using siRNA knockdown of CD34 were investigated. Finally, the potential of C-MSC as a cell therapy was assessed using co-culture with an in vitro corneal epithelial cell injury model and the angiogenic effects of C-MSC conditioned medium were evaluated with blood and lymph endothelial cells. Both media supported proliferation of C-MSC, with SCM increasing expression of CD34, ABCG2, PAX6, NANOG, REX1, SOX2, and THY1, supported by increased associated protein expression. Isolating cell populations expressing CD34 protein made little difference to gene expression, however, knockdown of the CD34 gene led to decreased expression of progenitor genes. C-MSC increased viability of injured corneal epithelial cells whilst decreasing levels of cytotoxicity and interleukins-6 and -8. No pro-angiogenic effect of C-MSC was seen. Culture medium can significantly influence C-MSC phenotype and culture in SCM produced a cell phenotype more suitable for further consideration as an anti-inflammatory cell therapy. C-MSC show considerable potential for development as therapies for OSIDs, acting through anti-inflammatory action.

Prevalence and associated factors of corneal opacity among adults in Kolladiba town, Northwest Ethiopia: a cross-sectional study.

OBJECTIVE: This study aimed to assess the prevalence and associated factors of corneal opacity among adults in Kolladiba town, Northwest Ethiopia. METHODS AND ANALYSIS: A community-based cross-sectional study was conducted using a systematic random sampling technique. A total of 846 adult individuals were recruited for the study. Ethical approval was obtained from the University of Gondar School of Medicine Ethical Review Committee. A standardised, semistructured questionnaire plus an ocular examination were used to collect the data. The data were entered into Epi Info V.7 and cleaned and analysed using SPSS V.26. Binary and multivariable logistic regression analyses were performed to select candidate variables and identify statistically significant factors. Variables with a p value of less than 0.05 according to the multivariable logistic regression analysis were considered to be statistically significant. RESULTS AND CONCLUSION: The prevalence of corneal opacity among the study participants was 27.2% (95% CI 24.4% to 30.4%). In this study, age 49-60 years (adjusted OR (AOR): 1.90; 95% CI 1.03 to 3.32), age ≥61 years (AOR=2.12; 95% CI 1.17 to 3.87), inability to read and write (AOR=2.65; 95% CI 1.68 to 4.16), middle-income level (AOR=2.12; 95% CI 1.30 to 3.47) and poor income level (AOR=4.96; 95% CI 3.04 to 8.09) were factors that were significantly associated with corneal opacity.In this study, the prevalence of corneal opacity was considerably high. Being poor and unable to read and write were the primary factors significantly associated with corneal opacity. Hence, concerned stakeholders should strive to reverse the effects of corneal opacity on the quality of life of the study and causal studies should be considered in the future.

[Impact of the ratio between graft and host corneal size on immune rejection, re-bubbling rate and postoperative endothelial cell loss in 457 eyes after Descemet membrane endothelial keratoplasty (DMEK)]. / Einfluss des Verhältnisses von Transplantatgröße zu Hornhautgröße auf Immunreaktion, Re-Bubbling-Rate und postoperativen Endothelzellverlust bei 457 Augen nach Descemet-Membrane-Endothelial-Keratoplastik (DMEK).

BACKGROUND: The aim of this study was to assess the impact of the ratio between the graft and host corneal size (RGH) on postoperative complications, such as immune reactions, re-bubbling rate and endothelial cell loss (ECL) after Descemet membrane endothelial keratoplasty (DMEK). PATIENTS AND METHODS: Retrospectively, 457 patient eyes were included which had undergone surgery between 2016 and 2019 in the Department of Ophthalmology, Saarland University Medical Center in Homburg/Saar using DMEK or triple DMEK, diagnosed as Fuchs' endothelial dystrophy (n = 431), pseudophakic bullous keratopathy (n = 9) and others (n = 17). The follow-up period extended until the end of 2020. Main outcome measures included immune reaction (IR), re-bubbling rate and the postoperative endothelial cell loss (ECL) at 6 weeks, 6 months and 12 months and whether these measures depended on the RGH. RESULTS: The RGH in this study ranged from 0.35 to 0.62 (0.46 ± 0.04). There were 33 (7.2%) postoperative IRs (DMEK n = 25; triple DMEK n = 8). The average RGH without IR (0.46 ± 0.04) was significantly (p = 0.038) smaller than in the group with IR (0.47 ± 0.05). Re-bubbling was necessary in 159 of 457 (34.8%) patient eyes. The RGH in patient eyes with re-bubbling (0.47 ± 0.04) was significantly (p = 0.014) higher than that in eyes without re-bubbling (0.45 ± 0.04). The mean preoperative endothelial cell count (ECD) was 2603 ± 251 cells/mm2 (min: 2161, max: 3500 cells/mm2). It was shown that a larger RGH had no positive influence on endothelial cell loss (r = 0.001; p = 0.974). CONCLUSION: Our results suggest that a larger graft diameter compared to host corneal size is associated with an increased rate of immune reactions and a higher re-bubbling rate after DMEK. Otherwise, a larger RGH had no positive influence on endothelial cell loss after DMEK. Accordingly, the graft size for DMEK should not be unnecessarily large, especially in eyes with Fuchs' endothelial dystrophy.

Elucidating the mechanism of corneal epithelial cell repair: unraveling the impact of growth factors.

The repair mechanism for corneal epithelial cell injuries encompasses migration, proliferation, and differentiation of corneal epithelial cells, and extracellular matrix remodeling of the stromal structural integrity. Furthermore, it involves the consequential impact of corneal limbal stem cells (LSCs). In recent years, as our comprehension of the mediating mechanisms underlying corneal epithelial injury repair has advanced, it has become increasingly apparent that growth factors play a pivotal role in this intricate process. These growth factors actively contribute to the restoration of corneal epithelial injuries by orchestrating responses and facilitating specific interactions at targeted sites. This article systematically summarizes the role of growth factors in corneal epithelial cell injury repair by searching relevant literature in recent years, and explores the limitations of current literature search, providing a certain scientific basis for subsequent basic research and clinical applications.

Evidence for persistent UV-induced DNA damage and altered DNA damage response in xeroderma pigmentosa patient corneas.

Xeroderma pigmentosum (XP) is a rare genetic disorder characterized by injury to the ocular surface due to exposure to ultraviolet (UV) radiation. UV-induced damage in the cells leads to the formation of cyclobutane pyrimidine dimers (CPDs) and 6-4 pyrimidine-pyrimidone photoproducts that are repaired by the NER (Nucleotide Excision Repair) pathway. Mutations in the genes coding for NER proteins, as reported in XP patients, would lead to sub-optimal damage repair resulting in clinical signs varying from photo-keratitis to cancerous lesions on the ocular surface. Here, we aimed to provide evidence for the accumulation of DNA damage and activation of DNA repair pathway proteins in the corneal cells of patients with XP. Corneal buttons of patients who underwent penetrating keratoplasty were stained to quantify DNA damage and the presence of activated DNA damage response proteins (DDR) using specific antibodies. Positive staining for pH2A.X and thymidine dimers confirmed the presence of DNA damage in the corneal cells. Positive cells were found in both control corneas and XP samples however, unlike normal tissues, positive cells were found in all cell layers of XP samples indicating that these cells were sensitive to very low levels of UV. pH2A.X-positive cells were significantly more in XP corneas(p<0.05) indicating the presence of double strand breaks in these tissues. A positive expression of phosphorylated-forms of DDR proteins was noted in XP corneas (unlike controls) such as ataxia telangiectasia mutated/Rad-3 related proteins (ATM/ATR), breast cancer-1 and checkpoint kinases-1 and -2. Nuclear localization of XPA was noted in XP samples which co-localized (calculated using Pearson's correlation) with pATM (0.9±0.007) and pATR (0.6±0.053). The increased presence of these in the nucleus confirms that unresolved DNA damage was accumulating in these cells thereby leading to prolonged activation of the damage response proteins. An increase in pp53 and TUNEL positive cells in the XP corneas indicated cell death likely driven by the p53 pathway. For comparison, cultured normal corneal epithelial cells were exposed to UV-radiation and stained for DDR proteins at 3, 6 and 24 hours after irradiation to quantify the time taken by cells with intact DDR pathway to repair damage. These cells, when exposed to UV showed nuclear translocation of DDR proteins at 3 and 6 hours which reduced significantly by 24 hours confirming that the damaged DNA was being actively repaired leading to cell survival. The persistent presence of the DDR proteins in XP corneas indicates that damage is being actively recognized and DNA replication is stalled, thereby causing accumulation of damaged DNA leading to cell death, which would explain the cancer incidence and cell loss reported in these patients.

Exploring clinical variability in gelsolin amyloidosis: Brazilian family case study with confocal microscopy.

INTRODUCTION: Genetic mutations or inflammatory, degenerative, or neoplastic conditions can trigger amyloidosis. Hereditary gelsolin amyloidosis is a genetic disorder primarily marked by amyloid fibrils composed of misfolded gelsolin fragments. CASE REPORT: We present three sisters with AGel amyloidosis, illustrating its clinical diversity. Patient 1, a 51-year-old, had bilateral ptosis, ocular discomfort, and dry eye syndrome due to cranial nerve involvement. Patient 2, a 53-year-old, experienced progressive bilateral visual impairment. Patient 3, a 50-year-old, exhibited right eye ectropion. Genetic analysis, with the identical mutation, heterozygous c.640G > A (p.Asp214Asn) mutation, confirmed AGel amyloidosis diagnoses, with common findings including lattice corneal amyloidosis, reduced corneal sensitivity, and recurrent corneal erosions. Neurological manifestations included ataxia and peripheral neuropathy, with skin abnormalities observed in patient 1. Ocular involvement severity and distribution varied among patients. DISCUSSION: Common ocular and neurological manifestations validated AGel amyloidosis diagnoses, reinforcing its hereditary basis. Neurological symptoms highlighted the disorder's impact on various organ systems, while skin abnormalities contributed to ocular discomfort. Variable ocular involvement emphasized the disorder's heterogeneity. These patients emphasize hereditary gelsolin amyloidosis's clinical diversity and suggest potential environmental influences on disease expression. Genetic confirmation and confocal microscopy findings reaffirm the genetic basis while raising questions about assessing systemic disease severity, necessitating further investigation in larger cohorts. Ophthalmologists' specialized care is crucial for managing ocular symptoms, given the absence of a universal cure.