Destructive effects on endothelial cells of grafts in cytomegalovirus DNA-positive patients after keratoplasty.

AIM: To investigate corneal graft survival rate and endothelial cell density (ECD) loss after keratoplasty in cytomegalovirus (CMV) positive patients. METHODS: This was a retrospective cohort study. We analyzed the clinical data of patients who underwent viral DNA detection in aqueous humor/corneal tissue collected during keratoplasty from March 2015 to December 2018 at the Peking University Third Hospital, Beijing, China. To further evaluate the effect of CMV on graft survival rate and ECD loss, patients were divided into three groups: 1) CMV DNA positive (CMV+) group; 2) viral DNA negative (virus-) group, comprising virus- group eyes pairwise matched to eyes in the CMV+ group according to ocular comorbidities; 3) control group, comprising virus- group eyes without ocular comorbidities. The follow-up indicators including graft survival rate, ECD, ECD loss, and central corneal thickness (CCT), were analyzed by Tukey honestly significant difference (HSD) test. RESULTS: Each group included 29 cases. The graft survival rate in CMV+ group were lowest among the three groups (P=0.000). No significant difference in donor graft ECD was found among three groups (P=0.54). ECD in the CMV+ group was lower than the virus- group at 12 (P=0.009), and 24mo (P=0.002) after keratoplasties. Furthermore, ECD loss was higher in the CMV+ group than in the virus- group in the middle stage (6-12mo) post-keratoplasty (P=0.017), and significantly higher in the early stage (0-6mo) in the virus- group than in the control group (P=0.000). CONCLUSION: CMV reduces the graft survival rate and exerts persistent detrimental effects on the ECD after keratoplasty. The graft ECD loss associate with CMV infection mainly occurrs in the middle stage (6-12mo postoperatively), while ocular comorbidities mainly affects ECD in the early stage (0-6mo postoperatively).

Fully automated grading system for the evaluation of punctate epithelial erosions using deep neural networks.

PURPOSE: The goal was to develop a fully automated grading system for the evaluation of punctate epithelial erosions (PEEs) using deep neural networks. METHODS: A fully automated system was developed to detect corneal position and grade staining severity given a corneal fluorescein staining image. The fully automated pipeline consists of the following three steps: a corneal segmentation model extracts corneal area; five image patches are cropped from the staining image based on the five subregions of extracted cornea; a staining grading model predicts a score for each image patch from 0 to 3, and automated grading score for the whole cornea is obtained from 0 to 15. Finally, the clinical grading scores annotated by three ophthalmologists were compared with automated grading scores. RESULTS: For corneal segmentation, the segmentation model achieved an intersection over union of 0.937. For punctate staining grading, the grading model achieved a classification accuracy of 76.5% and an area under the receiver operating characteristic curve of 0.940 (95% CI 0.932 to 0.949). For the fully automated pipeline, Pearson's correlation coefficient between the clinical and automated grading scores was 0.908 (p<0.01). Bland-Altman analysis revealed 95% limits of agreement between the clinical and automated grading scores of between -4.125 and 3.720 (concordance correlation coefficient=0.904). The average time required for processing a single stained image during pipeline was 0.58 s. CONCLUSION: A fully automated grading system was developed to evaluate PEEs. The grading results may serve as a reference for ophthalmologists in clinical trials and residency training procedures.

Clinical Manifestations and Long-term Outcomes of Endothelial Keratoplasty in Patients with Proven VZV-related Endothelial Decompensation.

PURPOSE: To report the clinical manifestations, postoperative complications and long-term outcomes of endothelial keratoplasty in VZV-related endothelial decompensation. METHODS: In this retrospective study, thirteen eyes undergoing endothelial keratoplasty (EK) for VZV-related endothelial decompensation were compared with controls for Fuchs endothelial dystrophy or pseudophakic bullous keratopathy. RESULTS: Twelve patients did not have typical dermal pain or blisters. Ten patients had obvious iris abnormalities. Glaucoma was noted in eight patients before surgery. The best spectacle-corrected visual acuity improved from 1.12 ± 0.47 to 0.39 ± 0.43 (p = .002), whereas endothelial cell (EC) loss was 65% ±15% at 12 months that higher than that in the controls (p < .05). Postoperative complications included graft detachment (2/13), recurrence of endotheliitis (3/13), neurotrophic ulcer (1/13) and scleritis (1/13). CONCLUSIONS: The onset of VZV-related endothelial decompensation is generally insidious. Iris segmental atrophy, glaucoma and pigment KPs are highly suspected to be associated with VZV. EK is a reasonable option to rehabilitate vision.

A Fully Automated Segmentation and Morphometric Parameter Estimation System for Assessing Corneal Endothelial Cell Images.

PURPOSE: To develop a fully automated segmentation and morphometric parameter estimation system for assessing corneal endothelial cells from in vivo confocal microscopy images. DESIGN: Artificial intelligence (neural network) study. METHODS: First, a fully automated deep learning system for assessing corneal endothelial cells was developed using the development set (from 99 subjects). Second, 184 images (from 97 subjects) were used to construct the testing set to evaluate the clinical validity and usefulness of the automated segmentation and morphometric system. Third, the automatically calculated endothelial cell density (ECD) values, Topcon's cell density, and manually calculated ECD were compared. RESULTS: After slit lamp examination, 88 healthy subjects, 2 Fuchs endothelial dystrophy patients, and 7 corneal endotheliitis patients were identified among the 97 subjects in the testing set. The automatedly estimated morphometric parameters for the testing set were an average number of 234 cells, an ECD of 2592 cells/mm2, a coefficient of variation in the cell area of 32.14%, and a percentage of hexagonal cells of 54.16%. Pearson's correlation coefficient between the automated ECD and Topcon's cell density and between the manually calculated ECD and Topcon's cell density was 0.932 (P < .01) and 0.818 (P < .01), respectively. The Bland-Altman plot of Topcon's cell density and the automated ECD yielded 95% limits of agreement between 271.94 and -572.46 (concordance correlation coefficient = 0.9). CONCLUSIONS: A fully automated method for segmenting corneal endothelial cells and estimating morphometric parameters using in vivo confocal microscopy images is more efficient and accurate for assessing the normal corneal endothelium.

Loss of endothelial cells in viral DNA-positive grafts after keratoplasty: a 2-year follow-up study.

BACKGROUND: To compare endothelial loss between recipients who received viral DNA-positive grafts and controls 2 years after corneal transplantation. METHODS: We retrospectively analysed the clinical data and endothelial cell density of recipients of viral DNA-positive grafts and age-, sex-, aetiology- and operation-matched controls from April 2017 to July 2019 at the Peking University Third Hospital, Beijing, China. RESULTS: A total of 23/942 (2.44%) donor corneal buttons tested virus-positive by real-time PCR. A total of 27 recipients (except for 2 recipients) of viral DNA-positive grafts and 48 recipients of viral DNA-negative grafts were included in this study. Recipients of viral DNA-positive grafts had a higher endothelial cell (EC) loss rate post-penetrating keratoplasty and post-descemet stripping automated endothelial keratoplasty (p<0.05), but post-deep lamellar keratoplasty, the EC loss rate was similar to that of the controls. Recipients of herpes simplex virus-1-, cytomegalovirus- and varicella-zoster virus-positive grafts all had a higher EC loss rate than the controls during the 12- and 24-month follow-up periods (p<0.05). CONCLUSION: We inferred that viruses might be hidden in corneal grafts and mainly incubate in the corneal endothelium. Viral DNA-positive grafts do not need to be replaced immediately and can be followed up for a long time.

The incidence and influence of the donor corneas positive for herpesviridae DNA in keratoplasty.

PURPOSE: We detected the DNA of herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) in donor corneas and assessed the clinical outcomes of recipients who received virus-positive grafts. METHOD: All donor corneas were analyzed for the presence of HSV-1, HSV-2, VZV, CMV, and EBV by real-time PCR from April 2017 to July 2019. The medical records of the transplant patients who received virus-positive grafts were reviewed. RESULT: Twenty-three (2.44%) donor cornea buttons tested positive for herpesviridae DNA. The positivity rates of HSV-1, CMV, VZV, and EBV were 0.74%, 0.85%, 0.64%, and 0.21%, respectively. CONCLUSION: We suggest that the corneas from donors who had cancer, donors who were inpatients, and donors who had immunodeficiency or who were on immunosuppressive therapy should be tested for herpesviridae DNA before transplantation. Finally, HSV-1 can be transmitted from graft to recipient, but that CMV cannot be transmitted according to our observations. The donor corneas found to be HSV-1-positive have to be discarded and not used for keratoplasty.

Epithelial changes with corneal punctate epitheliopathy in type 2 diabetes mellitus and their correlation with time to healing.

BACKGROUND: To study basal epithelial cell (BEC), sub-basal nerve plexus (SBN) and Langerhans cell (LC) density in patients with type 2 diabetes mellitus (T2DM) with corneal punctate epitheliopathy (CPE) and to assess their association with time to healing of CPE. METHODS: Retrospective study of in vivo confocal microscopy (IVCM) in 160 eyes from 160 patients with T2DM diagnosed with CPE due to a single cause. Key exclusion criteria included multiple-causes for CPE or treatment with autologous serum. A total of 149 eyes from 149 gender- age- and aetiolgy-matched patients with CPE without T2DM comprised the control group. Electronic records were reviewed for demographic features, history of T2DM and aetiology of CPE. Density of BEC, SBN and LC were compared between the two groups. RESULTS: The healing time in days for CPE with different aetiologies in the T2DM and control groups were as follows: dry eye (21.56 ± 2.41; 7.00 ± 2.19; P = 0.001); meibomian gland dysfunction (26.42 ± 6.04; 9.21 ± 2.55; P = 0.001); cataract extraction (38.00 ± 19.62; 25.83 ± 11.49; P = 0.043); drug induced (53.19 ± 18.83; 41.86 ± 23.87; P = 0.018) and exposure (38.25 ± 14.13; 29.00 ± 13.67; P = 0.026). LC density was 38.70 ± 9.65 cells/mm2 in the T2DM group comparedwith 25.53 ± 3.54 cells/mm2 in the controls (P = 0.001). SBN density was 11.76 ± 1.69 mm/mm2 in the T2DM group compared with 20.92 ± 1.43 mm/mm2 in the controls (P = 0.001). BEC density in the T2DM group was 4982 ± 1178 cells/mm2 compared with 5739 ± 394 cells/mm2 in the control group (P = 0.018). Age and duration of T2DM had no relationship with healing time (multiple linear regression, P = 0.618; P = 0.787). The density of LC in the T2DM group showed a negative correlation with SBN density (r = 0.350; R2 = 0.1225; P = 0.034). The density of SBN in the T2DM group showed a positive correlation with BEC density (r = 0.427; R2 = 0.1823; P = 0.008). The density of BEC in the T2DM group showed a negative correlation with healing time (r = 0.931; R2 = 0.8668; P = 0.001). CONCLUSIONS: Utilising IVCM, we have demonstrated increased LC and decreased SBN in patients with T2DM and CPE. Both may be related to lower BEC density and nuclei enhanced reflection. Furthermore, decreased BEC density may lead to delay in cornea epithelium healing in the T2DM group comparedwith controls. An immune-mediated response may play a role in delayed wound closure in patients with T2DM.

Early central and peripheral corneal microstructural changes in type 2 diabetes mellitus patients identified using in vivo confocal microscopy: A case-control study.

The aim of this study was to find early central and peripheral corneal microstructural changes in healthy subjects and type 2 diabetes mellitus (T2DM) patients with/without cornea fluorescein dot staining.This is a prospective case-control study of T2DM patients with/without cornea fluorescein dot staining. Age, sex, duration of diabetes, and serum glycosylated hemoglobin A1c (HbA1c) levels were recorded. Keratograph 5 M (K5 M) and in vivo confocal microscopy were performed on all subjects. The cornea was divided into 5 zones: central, superior, temporal, nasal, and inferior. Basal epithelial cell (BEC) density, the area of BEC, sub-basal nerve plexus (SBN) density, Langerhans cell (LC), and endothelial cell (EC) density were quantitatively analyzed.This study included a total of 87 individuals (28 males and 59 females; mean age, 62.30 ±â€Š9.93 years) with T2DM, without (n = 48; T2DM group 1) and with (n = 39; T2DM group 2) cornea fluorescein staining, as well as 51 age- and sex-matched healthy subjects (18 males and 33 females; mean age, 61.53 ±â€Š10.15 years). Ocular Surface Disease Index scores, Schirmer Ι test, tear meniscus height, the first breakup of tear film occurrence (NIKBUT-first), and the average time of all breakup incidents (NIKBUT-average) values were significantly lower for the T2DM groups than for the healthy group. The corneal sensations of all cornea positions in the T2DM groups were significantly different from the control group. The HbA1c in the T2DM groups showed a negative correlation with central BEC density (R = 0.348, P = .015; R = 0.91, P = .001); there was no correlation of HbA1c with BEC density in the control group. The BEC density, the area of BEC, SBN, and LC density of T2DM group 1 and T2DM group 2 were significantly different compared with the control group in all corneal positions (P < .001). The BEC density of T2DM group 2 was significantly different from T2DM group 1 in the central (P = .044) and inferior (P = .013) zones. The area of BEC in T2DM group 2 was significantly different from T2DM group 1 in inferior zone (P = .014) and other corneal positions showed was no significant difference (P > .05). The SBN density of T2DM group 2 was not significantly different from T2DM group 1 in all corneal positions (P > .05). The LC density of T2DM group 2 was significantly different from T2DM group 1 in the central (P = .006) and inferior (P = .006) zones. Although the LC density in the T2DM groups showed no significant difference in all corneal zones (P > .05), the LC density in the central zone was significantly lower compared with the peripheral zone in the control group (P = .001). The central ECs in the 3 groups were not significantly different (P > .05).LC induced an immune-mediated contribution to corneal nerve damage and may influence the early stages of BEC proliferation and differentiation in T2DM. BEC density was the reliable index for evaluating the early condition of diabetic corneal epitheliopathy. The BEC density of the central and inferior corneal zones was more sensitive.

Repeatability and Reproducibility of Noninvasive Keratograph 5M Measurements in Patients with Dry Eye Disease.

Purpose. To determine the intraexaminer repeatability and interexaminer reproducibility of tear meniscus height (TMH) and noninvasive Keratograph tear breakup time (NIKBUT) measurements obtained with the Keratograph 5M (K5M) in a sample of healthy and dry eye populations. Methods. Forty-two patients with dry eye disease (DED group) and 42 healthy subjects (healthy group) were recruited in this prospective study. In all subjects, each eye received 3 consecutive measurements using the K5M for the TMH and NIKBUTs (NIKBUT-first and NIKBUT-average). And then a different examiner repeated the measurements. The repeatability and reproducibility of measurements were assessed by the coefficient of variation (CV) and intraclass correlation coefficient (ICC). Results. The repeatability and reproducibility of TMH and NIKBUTs were good in both DED and healthy groups (CV% ≤ 26.1% and ICC ≥ 0.75 for all measurements). Patients with DED showed better intraexaminer repeatability for NIKBUTs, but worse for TMH than healthy subjects. Average TMH, NIKBUT-first, and NIKBUT-average were significantly lower in DED group than in healthy group (all P values < 0.05). Conclusions. Measurements of TMH and NIKBUTs obtained with the K5M may provide a simple, noninvasive screening test for dry eye with acceptable repeatability and reproducibility. The NIKBUTs were more reliable, but TMH was less reliable in patients with DED.