Prevalence of Herpes Simplex and Varicella-Zoster Virus DNA in Corneal Grafts Is Higher than Expected.

PURPOSE: (1) To determine the prevalence of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), varicella-zoster virus (VZV), and cytomegalovirus (CMV) DNA in donor corneas; (2) To evaluate the clinical outcome of the grafts with viral DNA and to compare donors with and without viral DNA. METHODS: We analyzed data from all donors and recipients who underwent penetrating keratoplasty (PK) or Descemet membrane endothelial keratoplasty (DMEK) between September 2022 and March 2023. Donor corneoscleral rims and excised recipients' corneal buttons were tested for the presence of HSV-1, HSV-2, VZV, and CMV DNA by polymerase chain reaction (PCR). The results were known 2-3 days after the surgery. We closely followed up on patients whose grafts tested positive for viral DNA. We compared the medical histories of donors with and without viral DNA. RESULTS: We included 85 corneas from 67 donors. Seven (8.2%) donor corneas tested positive for HSV-1 (n = 3) or VZV (n = 4) DNA. We did not detect any HSV-2 or CMV DNA. In the postoperative follow-up of patients with positive PCR, a graft failure was observed in one and infections in two eyes. Re-operation was necessary in three of these cases (42.9%). Patients without herpes DNA in the donor cornea needed reoperation in 7.7% of the cases. Cultural duration, the cause of the donor's death, and the death-to-explantation interval did not differ significantly between donors with and without viral DNA. Additionally, 3 of the 7 (42.9%) donors with positive PCR were in a septic status at the time of death, compared to 21 of the 78 (26.9%) donors with negative PCR (p = 0.52). CONCLUSIONS: The prevalence of herpes DNA in the donor corneas was 8.2% and thus higher than previously reported. We did not notice any evidence for a donor-to-host transmission, but a higher rate of postoperative complications in recipients of the grafts with viral DNA. The donors with and without herpetic DNA did not differ significantly regarding systemic diagnoses or cultural conditions, but sepsis was more frequent in the group with viral DNA.

In situ donor keratometry in deceased patients as a novel screening technique for eye banking.

PURPOSE: To investigate the potential role of keratometry on whole globes in situ of deceased patients by assessing its repeatability and comparing it with sterile donor tomography after excision and preservation in organ culture. METHODS: A sequence of 5 measurements was taken from 40 eyes in situ of deceased patients < 24 h after death using the portable Retinomax K-plus 3 (Bon, Tokyo, Japan). Keratometry of whole globes in situ, from which sclerocorneal discs were taken for organ culture, was compared to those obtained after measuring these sclerocorneal disks through their cell culture flask in medium I after 5 ± 4 days using the anterior segment optical coherence tomograph Casia 2 (Tomey Corp., Nagoya, Japan), and to 964 different donor corneas in medium II. RESULTS: Cronbach's alpha of the in situ keratometry was 0.891 and 0.942 for the steepest and flattest corneal power (P). The steepest (44.5D) and flattest (41.1D) P as well as the astigmatism (3.4D) of in situ corneas remained unchanged after preserving sclerocorneal discs in medium I (respectively 44.7D [p = 0.09]; 41.4D [p = 0.17]; 3.3D [p = 0.09]). The comparison of the in situ values with the 964 measured different donor corneas in medium II showed significantly (p < 0.001) higher P at the steep (45.4D) and flat (43.9D) meridian and smaller astigmatism (1.4D) for sterile donor tomography. CONCLUSIONS: Measuring deceased patients' eyes in situ with the portable Retinomax K-plus 3 represents a feasible and reliably repeatable screening method in the eye bank. In comparison to donor tomography in medium I, it measures a similar power and astigmatism.

Factors affecting the epithelial integrity of human donor corneas.

Purpose: To evaluate various factors affecting the integrity of human donor corneal epithelium. Methods: Donor corneal buttons were evaluated for epithelial defect (ED) and exposure. The slit-lamp photographs were taken on day 01, and the data such as age and gender of the donor, cause of death, refrigeration of cadavers, death-to-preservation time (DPT), experience of technician, and distance from site of collection to eye bank were collected. Results: A total of 100 consecutive corneal buttons belonging to 56 donors were evaluated. The median age of donors was 50 years. Males constituted 45 (80.4%). The mean DPT was 9.7 ± 5.3 hours. After death, 34 donors (60.7%) were refrigerated before the collection/retrieval. Most of the corneas (80%) were recovered by technicians having an experience of 0-5 years. Thirty-one donors (55.3%) were located at 1-50 km from the eye bank. The mean area of exposure was 15 ± 4.3 mm2. The mean area of ED was 28.7 ± 5.9 mm2. ED was significantly associated with refrigeration of cadavers and longer DPT. On multivariate analysis, only DPT was found to be significantly associated (P = 0.006; odds ratio [OR] = 1.54 ± 0.24) with the presence of ED. After transplantation, only two corneas had persistent epithelial defects and were treated successfully using various interventions. Conclusion: Integrity of donor corneal epithelium is mainly influenced by the refrigeration of cadavers and DPT.

Donor related corneal graft infection: a review of literature and preventive strategies.

PURPOSE: Donor-related infections are a serious threat to patient safety after corneal transplantation. We provide a concise review of literature from the last decade on donor-related graft infections, sources of contamination and means to reduce the contamination of donor tissue and preservation media. METHODS: We reviewed 50 papers from year 2005 to 2021 related to donor-related graft infections. We included 14 studies related to the risk factors associated with post-keratoplasty infection and preventive methods. RESULTS: Incidence of post-keratoplasty infections has been reported to be approximately 0.2%-0.77% for endophthalmitis and 6.5%-10.5% for microbial keratitis. We analyzed six important studies regarding the risk factors related to donor contamination. It was observed that younger donor age, increased death to retrieval time, warming cycles and increased eye bank processing time and positive corneo-scleral rim cultures were important risk factors for donor-related infections post keratoplasty. Eye banks have adapted newer protocols over the time period for prevention of donor-related contamination. Recommended preventive strategies were published in about eight important studies over the past decade. In addition to meticulous donor screening, rapid warming cycles, double contact with povidone iodine during retrieval and addition of antifungals like amphotericin B, Voriconazole and cycloheximide have been suggested over the last decade although their use is still in debate with regard to the efficacy, toxicity and cost-effectiveness. CONCLUSION: The last decade has witnessed a relative rise of fungal infections and multidrug resistant bacterial infections post-keratoplasty. Eye bank prepared corneas for lamellar surgeries are at increased risk for donor contamination due to increased exposure to the higher temperatures during their processing. Addition of antifungals and broad spectrum antibiotics to the hypothermic preservation media needs to be considered in the new era of increasing trends of lamellar keratoplasty.

Deep Anterior Lamellar Keratoplasty Over Penetrating Keratoplasty to Mitigate Positive Vitreous Pressure Complications.

This report describes deep anterior lamellar keratoplasty over penetrating keratoplasty (DALK-over-PKP) as an alternative technique to mitigate complications related to positive vitreous pressure (PVP) during PKP. We accomplished this by repairing the punctured cornea and performing a modified DALK where a full-thickness donor graft is placed over the host Descemet membrane, which is then removed after partial suturing of the graft. This mitigates the driving force behind the PVP by maintaining a closed-anterior chamber.

Intraoperative optical coherence tomography-guided donor corneal tissue assessment and preparation.

Purpose: To evaluate the role of intraoperative optical coherence tomography (i-OCT) in donor grading, selection, and preparation during different types of keratoplasty. Methods: Seventy-one consecutive donor corneas collected over 6 months, after clinical grading, were observed by an experienced corneal surgeon under an i-OCT equipped microscope. The donor preparation (manual/automated) for different types of keratoplasty procedures was also undertaken under i-OCT. Results: The mean central corneal thickness of optical and nonoptical grade tissues was 533 ± 19 and 662 ± 52 µm, respectively. The i-OCT-based grading matched with clinical grading in 98.5% cases. Irregular thickness, anterior stromal hyperreflectivity, and previous scars were appreciated in 1.4, 1.4, and 7.04% donors, respectively. During Descemet stripping automated endothelial keratoplasty, i-OCT facilitated selection of appropriate microkeratome head for automated donor preparation in all cases, besides allowing manual dissection of partially dissected lenticule, identification of site of inadvertent perforation, and eccentric trephination in one case each. During Descemet membrane endothelial keratoplasty, i-OCT-based assessment of preexisting scar (five cases) guided careful tissue selection (2/5) and preparation. During predescemetic endothelial keratoplasty, precise needle advancement allowed successful type-1 bubble formation in all cases. All manually punched donors demonstrated an extra endothelial ledge, while those with automated preparation showed tapering donor margins. Conclusion: i-OCT might serve as a useful imaging tool for objective assessment of donor characteristics. The modality may complement clinical evaluation for donor grading, selection, and preparation.

Qualitative and Quantitative Evaluation of Donor Corneal Tissue by Slit Lamp and Specular Microscopy.

Background In India, donor eye collection and promotion of eye banking are insufficient to meet the needs. By adequately evaluating donor corneas, eye banks can maximize the number of viable corneas for transplantation. This study evaluated donor corneal tissue based on age, lens status, and cause of death by their morphology and endothelial cell count via slit lamp and specular microscopy. Methods We conducted a prospective observational study of all eye bank donor corneas indicated for eye donation at a tertiary hospital and research center in Western Maharashtra between September 2019 to December 2021. We evaluated the corneoscleral discs by slit-lamp microscopy specular microscopy. We analyzed donor corneas quantitatively and qualitatively and graded them accordingly. We also collected blood samples for serological testing and the donor's behavioral and family medical histories. Results We collected 94 eyes from 47 donors; the mean age of the donor population was 48.2 years, and most donors were aged 41 to 80 years. Thirty-one donors (65.96%) were male, and 16 were female (34.04%. For preservation, we used Cornisol (Aurolab, Madurai, India) in 36 cases (77%) and McCarey-Kaufman medium in 11 cases (23%). We found a mean endothelial cell density (ECD) of 2214.40/mm2, with hexagonality of 53.05%, and a coefficient of variation of 38.01. Further, we observed that ECD and hexagonality of cells in phakic donors were significantly greater than that of pseudophakic (PP) donors. Moreover, ECD and hexagonality significantly decreased in donors with the chronic disease compared to those who had a sudden, unexpected death. Conclusion Corneal grafts from younger donors, phakic donors, and donors who experienced an acute cause of death were qualitatively and quantitatively significantly better than those of older donors, PP donors, and donors who experienced sudden, unexpected death. Therefore, eye bank specular examination can improve tissue utilization and transplantation success. Therefore, we strongly recommend that eye bank personnel evaluate their donor tissue with a specular microscope to enhance the quality of eye care.

Reverse graft suturing to avoid Descemet's membrane detachment of glycerol-preserved donor cornea used for therapeutic penetrating keratoplasty during COVID-19 to overcome the tissue shortage - A novel surgical technique.

To overcome tissue shortage during pandemic, we switched to 100% glycerol preservation of the donor cornea, which is economical and provides longer duration of storage than the short and intermediate storage mediums we normally use like McCAREY Kaufman (MK) or cornisol. During our initial few cases of therapeutic penetrating keratoplasty using glycerol preserved donor cornea, we faced spontaneous Descemet's detachments resistant to air tamponade. We tried reverse graft suturing and successfully reinforced Descemet's attachment along with air tamponade, in one of the cases after multiple failed air injections. In the subsequent two cases of infective keratitis needing therapeutic penetrating Keratoplasty, we took eight reverse sutures in between the eight cardinals, to anchor the Descemet's membrane of the graft. Both the grafts showed attached Descemet's and maintained good graft clarity. The reverse corneal suturing technique has not been described to the best of our knowledge and hope this helps our corneal fraternity.

Utilization rate of corneal tissue obtained from donors over 75 years of age in Western India for keratoplasty.

PURPOSE: To examine the utilization patterns of cornea procured from diseased individuals ≥75 years of age at an eye bank in western India. METHODS: In this retrospective study, data from 1,217 eyes of 653 donors with age ≥75 years were reviewed from October 2008 to December 2019. Donor age, lens status, endothelial cell count (ECD), utilization of the tissue for transplantation or non-clinical purposes (e.g., research, training/discarded), and causes of non-utilization were noted. RESULTS: The mean age of the donors was 80.9 ± 4.6 years and the tissue utilization rate was 36.5% (445 out of 1,217 eyes). The eyes used for keratoplasty procedures had a lower donor age (79.6 ± 5.7 vs. 81.5 ± 5.1; P < 0.001), a higher endothelial cell count (2493 ± 531 vs. 2034 ± 581; P < 0.001), and were more often phakic (61% vs. 36.6%) compared to the unused group. A multivariable logistic regression analysis showed that the likelihood of tissue utilization for keratoplasty was 13% higher with every 100-cell increment in donor ECD (odds ratio [OR] = 1.13, 95% CI = 1.10-1.16, P < 0.001) and 33% lower with having a pseudophakic status in the donor eye (OR = 0.67, 95% CI = 0.52-0.87, P = 0.03). Age was not a significant determinant of tissue utilization when used in the same multivariable model. CONCLUSION: More than one-third of the eyes (36.5%) can be utilized even when the donors are above 75 years of age. Eyes that were more likely to be utilized for keratoplasty were phakic and had a significantly higher ECD; age was not a determinant in tissue utilization.

Reliability and efficiency of corneal thickness measurements using sterile donor tomography in the eye bank.

To evaluate the reliability and efficiency of sterile pachymetric measurements of donor corneas based on tomographic data using two different methods: a "manual" and a "(semi-)automated" method. Twenty-five (25) donor corneas (50%) stored in MI and 25 (50%) in MII were imaged 5 times consecutively using an anterior segment OCT (AS-OCT). The central corneal thickness (CCT) was measured both with the manual measurement tool of the AS-OCT (= CCTm) and with a MATLAB self-programmed software allowing (semi-)automated analysis (= CCTa). We analyzed the reliability of CCTm and CCTa using Cronbach´s alpha (α) and Wilcoxon signed-Rank Test. Concerning CCTm, 68 measurements (54.4%) in MI and 46 (36.8%) in MII presented distortions in the imaged 3D-volumes and were discarded. Concerning CCTa, 5 (4%) in MI and 1 (0.8%) in MII were not analyzable. The mean (± SD) CCTm was 1129 ± 6.8 in MI and 820 ± 5.1 µm in MII. The mean CCTa was 1149 ± 2.7 and 811 ± 2.4 µm, respectively. Both methods showed a high reliability with a Cronbach´s α for CCTm of 1.0 (MI/MII) and for CCTa of 0.99 (MI) and 1.0 (MII). Nevertheless, the mean SD of the 5 measurements was significantly higher for CCTm compared to CCTa in MI (p = 0.03), but not in MII (p = 0.92). Sterile donor tomography proves to be highly reliable for assessment of CCT with both methods. However, due to frequent distortions regarding the manual method, the (semi-)automated method is more efficient and should be preferred.

Gram stain and addition of amphotericin B to improve the microbial safety of human donor corneas.

To determine the effectiveness of two methods to improve the microbial safety of human corneas preserved in organ culture. We compared the number of positive preservation solutions of corneas in organ culture in which the initial short-term hypothermic corneal maintenance solution was supplemented with amphotericin B 2.5 µg/mL and the historical data of microbial test results (2015-2019). In addition, we appraised the efficacy of Gram stain to detect bacterial or fungal contamination in the organ culture solutions of corneas from at-risk donors compared to the culture tests of corneas from not-at-risk donors. Statistical analysis was performed using STATA and statistical significance set at p < 0.05. The number of positive culture tests after preservation was 15 (0.5%) in 2020 compared to a mean of 37 (1.2%) in the period 2015-2019 (p < 0.01), with 10 (1.0%) positive samples in the cohort of 998 corneas from at-risk donors and 5 (0.2%) in the 2046 corneas from not-at-risk donors (p < 0.01). All corneas from at-risk donors tested positive at Gram stain and the results were available 1-3 days before those of the conventional culture tests. Amphotericin B supplementation in the short-term maintenance solution markedly reduced the number of positive microbial tests after organ culture and the early detection of contaminants, including slow-growing microorganisms, by Gram stain before the standard culture results. This meant fewer corneas being discarded and a greater likelihood of preventing post-graft infections.

Influence of time to procurement, incubation and release of organ cultured donor corneas on graft failure after Descemet Stripping Automated Endothelial Keratoplasty.

PURPOSE: The aim of the present study was to investigate whether the time from death to procurement, to preservation or the storage time of donor corneas preserved in organ culture influenced the clinical outcome of patients undergoing Descemet stripping automated endothelial keratoplasty (DSAEK) for Fuchs endothelial keratoplasty. METHODS: We conducted a registry-based study on 776 patients undergoing DSAEK. Data on time from donor death to cornea retrieval (DRT), time from death to preservation (DPT), the preservation time and donor cornea characteristics: age, sex and endothelial cell density (ECD) at the time of release for surgery, were extracted from The Danish Cornea Bank Registry. Data on recipient follow-up were collected from a corneal graft registry. The primary outcome was presence of graft failure within a period from 2 months to 2 years after surgery. Secondary outcomes were DRT, DPT, ECD ≤2300 and gender mismatch between donor and recipient. RESULTS: Graft failure occurred in 26 patients. The mean preservation time for failed grafts was 34.1 ± 10.0 days (mean ± SD) and 27.3 ± 10.6 days (mean ± SD) for the clear, functional grafts at the 2-year follow-up. A preservation time of >29 days compared with ≤29 days was associated with a lower survival (HR 2.33, 95% CI on 1.06-5.14, p = 0.036) and an increased risk of graft failure (RR 1.53, 95% CI on 1.11-2.10, p = 0.009). For the secondary outcome variables, no difference in the risk of graft failure was observed and did not appear to impact the survival rate of DSAEK patients. CONCLUSION: Preservation time of donor cornea was associated with graft survival and a prolonged preservation time of more than 4 weeks seemed to lower the 2-year survival.

A simple surgical technique for splitting a single donor cornea for performing deep anterior lamellar keratoplasty and Descemet membrane endothelial keratoplasty without using a microkeratome.

PURPOSE: To describe a simple manual surgical technique for splitting a single-donor eye for performing both deep anterior lamellar keratoplasty (DALK) and Descemet membrane endothelial keratoplasty (DMEK) without using a microkeratome. METHODS: Twenty-three eyes with anterior stromal pathology and 23 eyes with irreversible endothelial dysfunction were evaluated for keratoplasty at a tertiary eye care referral center. Twenty-three healthy donor corneas were split into two parts. The Descemet's membrane was stripped and used for DMEK. The stripped stroma was used for DALK. Best-corrected visual acuity (BCVA) of both DALK and DMEK, endothelial cell density, and endothelial cell loss in DMEK were noted at 1-year follow-up, along with any intraoperative or postoperative complications and failures. RESULTS: In the DALK group, mean BCVA improved from 1.264 ± 0.25 log Mar preoperatively to 0.355 ± 0.27 log Mar at 12 months follow-up. There were no complications and failures. In the DMEK group, mean BCVA improved from 1.537 ± 0.61 log Mar preoperatively to 0.592 ± 0.67 log Mar and the mean donor ECD was 3071.66 (range, 2783-3487) cells/mm2 preoperatively, which was reduced to 1989.33 (range, 1546-2543) cells/mm2 at 12 months follow-up indicating a mean endothelial cell loss of 35%. The failure rate was 21.7%. CONCLUSION: This study demonstrates that with a single donor corneal tissue, both DALK and DMEK can be performed successfully without any complications. Our technique will help corneal surgeons in all developing countries to cost effectively perform more lamellar surgeries and help in reducing the magnitude of corneal blindness without the need for expensive microkeratomes.

How to Predict the Suitability for Corneal Donorship?

BACKGROUND: In Germany, more than one-third of donor corneas harvested are not suitable for transplantation. We evaluated the factors associated with the usability of donor corneas. METHOD: Data from 2032 consecutive donor corneas harvested at the Rhineland-Palatinate Eye Bank in Mainz, Germany, were retrospectively analyzed. Factors of interest were age, sex, lens status, cause of death, cardiopulmonary resuscitation (CPR), death-to-explantation-interval (DEI), and the influence of these factors on the proportion of discarded donor corneas. Factors associated with endothelial cell density (ECD) were analyzed in a linear regression mixed model. RESULTS: Higher donor age, male gender, pseudophakic lens status, and longer DEI were associated with significantly reduced ECD. With respect to DEI, the estimated cell loss was 7 ± 2 cells/mm2/hour (p < 0.001). Age was associated with a lower ECD of 6 ± 2 cells/mm2 per year (p = 0.001). Female ECD was 189 ± 44 cells/mm2 higher than male ECD (p < 0.001). Pseudophakic eyes had 378 ± 42 cells/mm2 less compared with phakic eyes (p < 0.001). Cause of death did not affect the ECD. Of note, 55% and 38% of corneas harvested on the second and third postmortem day, respectively, and 45% of corneas from donors older than 80 years were still suitable for transplantation. CONCLUSIONS: In the context of a growing need for donor corneas, we do not recommend limiting donor age and collection time to 24 h or excluding oncology donors, as is the practice in many countries. Therefore, we propose a mathematical model for better donor preselection.

Corneal Donation: Current Guidelines and Future Direction.

PURPOSE: This review aims to outline current practices and guidelines of corneal donation and eye banking, describes the implications of COVID-19 and emerging diseases on the corneal donor pool, and discusses future trends to improve and increase the efficiency of the processes involved in corneal donation and eye banking. SUMMARY: Corneal screening, preservation, corneal storage, and prevention of systemic disease transmission from donor to recipient have been crucial in shaping the policies of the FDA and eye banks across the world. Eye banks globally have developed varying guidelines and criteria for evaluating the viability of donor corneas. Variables such as the age of the donor, medical history, and potential disease transmission are important screening parameters. While known infectious diseases may be transmissible through the cornea, emerging infectious diseases that are not well studied may be more transmissible than other infections. In particular, coronavirus has impacted corneal transplantation as SARS-CoV-2 expression has been detected in corneal tissue and conjunctiva. In recent years, partial-thickness corneal transplantations have been introduced. Lamellar grafts and other corneal layers are now utilized for transplantation of the specific areas that are damaged.

[Comparison of sterile donor tomography in the eye bank and graft tomography after penetrating keratoplasty]. / Vergleich der sterilen Spendertomographie in der Hornhautbank mit der Tomographie des Transplantates nach perforierender Keratoplastik.

BACKGROUND: Since 2018 we routinely use sterile donor tomography in the eye bank to avoid refractive surprises after keratoplasty. The purpose of this study was to compare donor tomography parameters measured with anterior segment optical coherence tomography (AS-OCT) with tomography parameters of the graft after penetrating keratoplasty (PKP). METHODS: This study enrolled 193 eye bank donor corneal tissues used for PKP (graft diameter 8.2 ± 0.7 mm). Preoperative measurements were taken using the AS-OCT Casia 2 (Tomey, Nagoya, Japan). The measurements were repeated postoperatively after 5 ± 4 months with all sutures in place and after 22 ± 4 months after removal of all sutures. Preoperative and postoperative values were compared using a Wilcoxon signed-rank test. RESULTS: Postoperatively, with all sutures in place (or without sutures), the dioptric power (P) at the anterior surface (a) of the cornea remained unchanged (-0.2 D, p = 0.78) (2.7 D bigger, p < 0.01) in the steep meridian (S) (PaS) and 4.5 D (2.8 D, p < 0.01) smaller in the flat meridian (F) (PaF) in comparison with the donor tomography values. The astigmatism (a) was 4.3 D (5.4 D, p < 0.01) bigger. At the posterior surface (p) the PpS was 0.9 D (0.9 D, p < 0.01) and PpF 0.3 D (p < 0.01) (0.1 D, p = 0.42) smaller, while the astigmatism (p) was 0.7 D (0.9 D) bigger (p < 0.01). The central corneal thickness was 55.7 µm (p < 0.01) (27.5 µm, p = 0.01) smaller. The total power of the transplant after removal of all sutures did not change significantly in comparison with the eye bank measurements. CONCLUSION: Central corneal thickness, dioptric power (P) and astigmatism all changed postoperatively compared to donor tomography values, except P at the steep meridian of the anterior corneal surface with all sutures in place and P at the flat meridian of the posterior corneal surface after removal of all sutures; however, the total power of the transplant after removal of all sutures did not change significantly. This information may be of value for IOL power calculation in the classical triple procedure.

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.

Corneal transplantation and eye banking practices during COVID-19-related lockdown period in India from a network of tertiary eye care centers.

PURPOSE: The purpose of this study was to discuss the guidelines and modification of practices with respect to corneal transplantation and eye banking during the COVID-19 pandemic lockdown period and beyond, at a network of tertiary care centers in India. METHODS: Descriptive study of the challenges faced in eye banking during the lockdown, and practices adopted to overcome the critical aspects in the clinical care of patients who presented with emergency corneal diseases requiring keratoplasty. RESULTS: Complete lockdown orders from the Indian government, as a strategy to control the Coronavirus pandemic, resulted in drastic reduction of all types of elective corneal transplants and eye banking activities from March 24, 2020 to May 31, 2020. The sudden cessation of eye banking resulted in an acute demand and supply imbalance of fresh donor corneas for transplants during this time. Our network of eye banks addressed this issue by adopting glycerol preservation of donor corneas, which were subsequently utilized for tectonic penetrating keratoplasty. The donor cornea retrieval was resumed in a strategized manner 3 weeks prior to the date of exit of the lockdown, with modified guidelines on donor suitability, screening, retrieval, processing, and harvesting from various sources. A triage of keratoplasty priority was formulated to tide over the post lockdown shortage of corneas. We performed 31 therapeutic keratoplasties during the nationwide lockdown among our network of tertiary eye care centers. CONCLUSION: The study highlights the approach and strategies to manage and tide over an unprecedented crisis situation faced by corneal surgeons in general and, specifically, the eye banking community.

Corneae from body donors in anatomy department: valuable use for clinical transplantation and experimental research.

BACKGROUND: Explanted corneae are highly needed for the surgical management of patients with severe corneal diseases. The aim of this study was to determine whether the body donors from the Institute of Anatomy are a suitable source of donor corneae. METHODS: At the Institute of Anatomy at Saarland University Medical Center in Homburg, corneae are prelevated from body donors who had consented to the removal of tissues for transplantation purposes during their lifetime. Following the report of death, the LIONS Eye Bank is informed and the contraindications of corneal explantation are clarified. Obtaining a blood sample within 24 h postmortem is mandatory. RESULTS: The Institute of Anatomy had 150 body donors in the time period from January 2018 to June 2019. Out of these, 68 (45.3%) were reported to the Eye Bank. The age of the donors (median 82 years (range: 57-96)) is not critical since the quality of the corneae depends on the number of endothelial cells (mean: 2109 ± 67 cells/mm2 (range: 511-2944 cells/mm2)). Contraindications were present in 19 (12.6%) cases. The corneae were extracted from 49 (32.7%) body donors. Out of these 98 corneae, 46 (46.9%) were successfully transplanted. Of all non-transplanted corneae, 6 (6.1%) were microbiologically contaminated, 10 (10.2%) had a positive serology, 22 (22.5%) had an endothelial cell count < 2000 cells/mm2 and 6 (6.1%) are at time of this analysis still in culture medium. The non-transplanted tissues were used for research. CONCLUSIONS: Explanted corneae from the Institute of Anatomy are a valuable option in obtaining grafts for corneal transplantation, which is why we are working toward on expanding cooperation with this department.

A new storage solution for the hypothermic preservation of corneal grafts: an experimental study.

In this experimental study we used for the first time Tiprotec® as a solution for corneal preservation and cold storage. We compared the resultant endothelial cell morphology and viability with this obtained after preservation of the ex-vivo corneas with both usual standard techniques: conventional cold storage (using Eusol-C) and organ culture. This prospective, in vitro, 3-armed parallel study was performed with the use of 90 porcine corneas (examined for their endothelial quality and transparency) randomly selected for preservation in three storage methods (each 30 corneas): organ culture, standard cold storage (Eusol-C) and experimental cold storage (Tiprotec®) Endothelium cell quantity and quality as well as corneal opacification were assessed. The degree of endothelial transparency was significantly reduced over time with all preservation media, without any significant difference among the three groups at any point of time. A reduction in endothelial cell density was also observed with all three preservation media after 30 days of storage without statistically significant differences between groups. The number of hexagonal and pentagonal endothelium cells was significantly reduced overtime in all media with significantly more hexagonal and pentagonal in the organ culture group compared to the cold storage groups. We could show that the cryopreservation medium Tiprotec®, used until now for the preservation of vascular grafts, was of similar quality compared to the medium Eusol-C for the hypothermic storage of corneal tissue for an extended period of time up to 30 days. In comparison to organic culture with culture medium KII, both Tiprotec® and Eusol-C were found less effective in preserving endothelial cell quality, as assessed by the morphometric analysis, and viability, as assessed by the degree of vacuolization at least up to the 30th day of storage. However, both, Tiprotec®- and Eusol-C-preserved corneas demonstrated a certain capacity to recover after their submission in organ culture.