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The scope of eye banking activities has been expanding with the advances and techniques of keratoplasty. With the popularity of descemet membrane endothelial keratoplasty (DMEK) in the recent decade, there is a need to adopt the preparation of DMEK tissues in the eye banks. This necessitated surgical training of the eye bank technicians, development of infrastructure in the eye bank, innovative methods of graft preparation, and delivery for the surgery at distant surgery centers. Eye banks across various countries have devised and adopted different methods of providing DMEK grafts based on the demands and preferences of corneal surgeons. This review focuses on the strategies of DMEK graft preparation and its distribution at the eye banks., (Copyright © 2024 Indian Journal of Ophthalmology.)

Objective: To investigate endothelial cell loss (ECL) associated with Descemet membrane endothelial keratoplasty (DMEK) donor tissues preloaded in the DMEK RAPID transport system after 1 and 5 days and to compare prestamping with 2 different F-mark inks., Methods: DMEK donor tissues were stripped, marked with gentian violet dye applied as an F-mark, trephined, stained with trypan blue, and then preloaded into the DMEK RAPID transport system by an eye bank technician. Preloaded DMEK tissues were then unfolded and stained with calcein AM after 1 or 5 days of storage. Tissues were imaged, analyzed for total tissue ECL, and immunostained for corneal endothelium markers zonular occludens-1 and xCD166. Additionally, ECL and the intensity of an F-mark caused by 2 different inks were quantified., Results: Preloaded DMEK tissues displayed an average ECL of 11.9% ± 4.5% (n = 8) at 1 day and 9.9% ± 4.2% (n = 9) at 5 days. No difference was found between the 2 groups. Zonular occludens-1 and activated leukocyte cell adhesion molecule (ALCAM; also know as CD166) staining showed that the corneal endothelial monolayer remained intact on preloaded tissues. On 5-day preloaded DMEK tissues, the average ECL and mean grayscale caused by the Keir Surgical ink F-mark and the Cardinal Health ink F-mark were 4.3% ± 0.8% and 158.5 ± 13.9% and 5.0% ± 1.1% and 142.9% ± 20.0%, respectively. No difference was found between the F-mark inks., Conclusion: Preloaded DMEK donor tissues resulted in an acceptable ECL range after 1 and 5 days of storage and were deemed suitable for transplantation. Both F-mark inks are acceptable for prestamping preloaded DMEK tissues prior to surgical transplantation with comparable ECL and intensities., Competing Interests: Footnotes and Disclosure The authors have no proprietary or commercial interest in any materials discussed in this article. Supported by: This work was funded by research grants from Labtician Ophthalmics Inc, the Independent Order of Odd Fellows, and the Rebekah Assembly of Ontario., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Purpose: ReLEx (Refractive Lenticule Extraction) Small Incision Lenticule Extraction (SMILE), the second generation of ReLEx Femtosecond Lenticule Extraction (FLEx), is a minimally invasive, flapless procedure designed to treat refractive errors such as myopia, hyperopia, presbyopia, and astigmatism. This review aims to provide a comprehensive overview of the methods for preserving SMILE-derived lenticules and discusses their potential future applications., Methods: A narrative literature review was conducted using PubMed, Scopus, and Web of Science databases, focusing on articles published up to January 2024 and available in English. The authors also evaluated the reference lists of the collected papers to identify any additional relevant research., Results: No standardized protocols currently exist for the storage or clinical application of SMILE-derived lenticules. However, these lenticules present a promising resource for therapeutic uses, particularly in addressing the shortage of donor corneal tissues. Their potential applications include inlay and overlay additive keratoplasty, as well as other ocular surface applications. Further research is needed to establish reliable protocols for their preservation and clinical use., Conclusion: SMILE-derived lenticules offer significant potential as an alternative to donor corneal tissues. Standardizing their storage and application methods could enhance their use in clinical settings., (© 2024. The Author(s).)

Endothelial cell density (ECD) is a crucial parameter for the release of corneal grafts for transplantation. The Lions Eye Bank of Baden-Württemberg uses the "Rhine-Tec Endothelial Analysis System" for ECD quantification, which is based on a fixed counting frame method considering only a small sample of 15 to 40 endothelial cells. The measurement result therefore depends on the frame placement and manual correction of the cells counted within the frame. To increase the sample size and create higher objectivity, we developed a new method based on "deep learning" that automatically detects all visible endothelial cells in the image. This study aims to compare this new method with the conventional Rhine-Tec system. 9375 archived phase-contrast microscopic images of consecutive grafts from the Lions Eye Bank were evaluated with the deep learning method and compared with the corresponding archived analyses of the Rhine-Tec system. Means, Bland-Altman and correlation analyses were compared. Comparable results were obtained for both methods. The mean difference between the Rhine-Tec system and the deep learning method was only - 23 cells/mm 2 (95% confidence interval - 29 to - 17). There was a statistically significant positive correlation between the two methods, with a correlation coefficient of 0.748. What was striking in the Bland-Altman analysis were clustered deviations in the cell density range between 2000 and 2500 cells/mm 2 - with higher values in the Rhine-Tec system. The comparable results for cell density measurement values underline the validity of the deep learning-based method. The deviations around the formal threshold for graft release of 2000 cells/mm 2 are most likely explained by the higher objectivity of the deep learning method and the fact that measurement frames and manual corrections were specifically selected to reach the formal threshold of 2000 cells/mm 2 when the full area endothelial quality was good. This full area assessment of the graft endothelium cannot currently be replaced by deep learning methods and remains the most important basis for graft release for keratoplasty., Competing Interests: Die Autorinnen/Autoren geben an, dass kein Interessenkonflikt besteht./The authors declare that there is no conflict of interest., (Thieme. All rights reserved.)

Purpose: The aim of this study was to examine endothelial cell loss (ECL) associated with preloaded Descemet stripping automated endothelial keratoplasty (DSAEK) grafts loaded into 3 carriers of different size and shape., Methods: Thirty-six donor corneas were prepared for DSAEK and loaded into an EndoGlide Ultrathin (control) (2.0 mm × 3.5 mm lumen, 4.5 mm/4.9 mm incision for scleral tunnel/clear corneal insertion), Descemet membrane endothelial keratoplasty EndoGlide (experimental) (1.1 mm × 1.7 mm lumen, 2.65 mm incision), or round glass Jones tube (experimental) (1.8 mm lumen, 3.0 mm incision). Control grafts were stored for 6 days in Optisol-GS and experimental grafts stored for 24 hours in Life4C before analysis using Calcein AM staining. Grafts were imaged and ECL was analyzed by FIJI segmentation. The statistical significance of ECL was determined using 1-way ANOVA and Tukey post hoc analysis., Results: There were no significant differences in donor characteristics for grafts in each cohort. ECL for grafts loaded into the EndoGlide Ultrathin was 10.3% ± 2.3% (graft thickness: 60-189 μm, n = 9). ECL for grafts loaded into the Descemet membrane endothelial keratoplasty EndoGlide was 22.2% ± 7.1% (graft thickness: 38-63 μm, n = 9). ECL for thin grafts (34-60 μm, n = 9) loaded into the Jones tube was 24.0% ± 5.0%. ECL for thick grafts (92-119 μm, n = 9) loaded into the Jones tube was 34.2% ± 6.1% ECL ( P < 0.001). Combined regression analysis revealed that graft thickness is directly correlated to ECL ( P < 0.01)., Conclusions: The size and shape of the carrier can influence the cell viability of preloaded DSAEK grafts. Compared with a larger carrier, smaller lumen carriers are associated with greater ECL. In smaller lumen carriers, ECL increases with increasing DSAEK graft thickness., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Objective: To explore the factors influencing the quality of donor corneal endothelium. Methods: A retrospective case series study was conducted. Data from 568 donor corneas obtained from the Shandong Eye Bank between July 1, 2020, and June 30, 2021, were collected for analysis. The corneal endothelium of the donor corneas was observed using corneal endothelial microscopy to assess corneal endothelial cell density (ECD), coefficient of variation, and hexagonal cell ratio (HEX). Relevant factors of corneal donors were collected, including gender, age, cause of death, season of death, time from death to corneal retrieval, and methods of corpse preservation, to investigate their impact on the quality of donor corneal endothelium. The age factor was divided into five age groups: 0-20 years, 21-40 years, 41-60 years, 61-80 years, and >80 years. The time of corneal retrieval was divided into three periods based on the time elapsed since the donor's death: <6 hours, 6-12 hours, and >12 hours. The relationship between these factors and corneal endothelial conditions was analyzed. Results: The 568 donor corneas were obtained from 288 donors, including 225 males (78.13%) and 63 females (21.87%). The mean age was 51.77±18.48 years. The causes of death among donors were as follows: cardiovascular diseases 54.58% (275 individuals), cancer 17.96% (74 individuals), organ failure 14.26% (49 individuals), and accidents 13.20% (64 individuals). The mean time of corneal retrieval after donor death was 140 (76, 400) minutes (ranging from 30 minutes to 45 hours). Among the 145 corneas (25.53%) that had their initial corneal endothelial microscopy examination, the images were not clear, and after thorough rewarming, 106 corneas (18.7%) still had unclear images and could not be analyzed. Among the 462 corneas (81.3%) with clear images, the ECD was (2 602.23±318.40) cells/mm², the coefficient of variation was 36.61%±4.81%, and the HEX was 52.73%±7.15%. The ECD of corneas from older donors was lower compared to younger donors, and the differences between age groups were statistically significant ( P <0.001). Corneas from donors who died due to accidents had a higher ECD [(2 829.88±313.90) cells/mm²] compared to those who died from cancer, cardiovascular diseases, and organ failure, and the differences were statistically significant ( P <0.001). The ECD was highest when corneas were retrieved within 6 hours after death, and the difference was statistically significant ( P <0.001). Older donors had higher coefficients of variation but lower HEX values (both P <0.05). Corneas retrieved after a longer time from death had higher coefficients of variation, and the difference was statistically significant ( P <0.05), but there was no statistically significant difference in HEX ( P >0.05). Organ failure, cryopreservation, and corneal retrieval time >12 hours were risk factors for unclear corneal endothelial imaging (all P <0.001). Among the 136 corneal endothelial images (23.94%), circular, oval, or band-shaped dark areas were observed, and corneas with dark areas had lower ECD ( P <0.05). The longer the time elapsed from death to corneal retrieval, the more dark areas were observed ( P <0.001). The presence of dark areas did not affect the coefficient of variation and HEX ( P >0.05). Conclusion: Advanced donor age, death due to chronic diseases, longer time elapsed from death to corneal retrieval, and cryopreservation of the body lead to a decrease in the quality of donor corneal endothelium.

Objective: To compare subjective and objective outcomes of 4 different Descemet membrane endothelial keratoplasty (DMEK) peeling techniques performed by novice surgeons at different stages in their surgical career., Design: An ex vivo prospective study., Methods: In the first round, 2 DMEK peeling techniques were pitched against each other: the peripheral scoring and Sinskey dissection technique with the peripheral scoring and microhoe dissection and the peripheral blunt microhoe dissection against the scleral spurectomy and microhoe dissection. Three surgeons with different operative experience performed the peeling. Outcome measures included graft peeling time, surgeon's peeling difficulty grading (on a scale of 1-10, 1 being the easiest and 10 the hardest), number of radial and circumferential tears before and after trephination, and tissue loss. The 2 techniques that performed the best from the first round proceeded to the final round to identify the best overall technique., Results: In total, 90 tissues (45 pairs) were peeled by 3 surgeons. Following the first-round results, the peripheral scoring and Sinskey dissection and peripheral blunt microhoe dissection proceeded to the final round. There were no significant differences between the groups in terms of peeling times, subjective feeling of difficulty, post-trephination tears, and peeling success rates (P > 0.05 for all). However, the peripheral scoring and Sinskey dissection technique had significantly fewer pretrephination radial tears (1.3 ± 1.3 vs 6.1 ± 5.2, P = 0.007) and circumferential tears (0.6 ± 0.9 vs 1.8 ± 2.1, P = 0.02)., Conclusions: This study demonstrates that the learning curve can be overcome quickly with appropriate DMEK peeling techniques. The peripheral scoring and Sinskey dissection peeling technique allows efficient peeling with fewer related tears., (Copyright © 2022 Canadian Ophthalmological Society. Published by Elsevier Inc. All rights reserved.)

Purpose: To analyze the impact on eye donation and corneal transplantation during the COVID-19 pandemic in a tertiary eye hospital in south India., Methods: A retrospective analysis of the donor and recipient records during the study period from January 2020 to May 2021 was conducted and tabulated in Microsoft Excel 2013. Demographic details of the donor, utility rate, cause of death, culture characteristics, storage methods, wet lab usage, and the surgical donor outcomes were evaluated. Additionally, the postoperative workup of the recipients, diagnosis, graft infection and rejection episodes, development of COVID-19 postoperatively, and outcome in terms of visual acuity at one, three, and six months were also noted., Results: A total of 466 eyes from 249 donors were received during the study period. The mean age of the donor population was 62.43 years (20.9). The corneal transplantation utility rate was 36.4% (n = 170). Fifty-one percent of the total transplant surgeries were for therapeutic purposes. This was followed by penetrating optical keratoplasty (34%), Descemet's stripping endothelial keratoplasty (9%), and patch grafts (3%). Seventeen (10%) graft rejection episodes were noted and nine (53%) had complete resolution after medical treatment., Conclusion: Proper preventive measures are key to carrying out safe and efficient eye banking activities even during a deadly pandemic, as COVID-19 transmission via transplantation is rare., Competing Interests: None

Purpose: The purpose of this study was to assess the impact of ongoing waves of the COVID-19 pandemic and resulting guidelines on the corneal donor pool with resumption of clinical operations., Methods: A retrospective analysis of donors deemed eligible for corneal transplantation at an eye bank from July 1, 2020, through December 31, 2021. Donors ineligible due to meeting Eye Bank Association of America (EBAA) COVID-19 guidelines or a positive postmortem COVID-19 testing were examined. The correlation between COVID-19 rule outs and state COVID positivity was calculated. The number of scheduled surgeries, suitable corneas, imports, and international exports was compared with a pre-COVID period. Postmortem testing was reduced for the final 5 months of the study, and numbers were compared before and after the policy change., Results: 2.85% of referrals to the eye bank were ruled out because of EBAA guidelines. 3.2% of postmortem tests were positive or indeterminate resulting in an ineligible tissue donor (0.42% of referrals). Over the 18-month period, there was a 4.30% shortage of suitable corneas compared with transplantation procedures. There was a significant correlation between postmortem testing and state COVID-19 positivity (r = 0.37, P <0.01), but not with EBAA guidelines (r = 0.19, P = 0.07). When postmortem testing was reduced, significantly more corneas were exported internationally., Conclusions: Although corneal transplant procedures were back to normal levels, there was a shortage of suitable corneal tissue. The discontinuation of postmortem testing was associated with a significant increase in international exports of corneal donor tissue., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Corneal transplantation is one of the most common forms of tissue transplantation worldwide. Donor corneal tissue used in transplantation is provided by eye banks, which store the tissue in culture medium after procurement. To date, the effects of cell culture on human corneal tissue have not been fully elucidated. Using the 3' RNA sequencing method for massive analysis of cDNA ends (MACE), we show that cultivation of corneal tissue leads to significant changes in a variety of molecular processes in human corneal tissue that go well beyond aspects of previously known culture effects. Functionally grouped network analysis revealed nine major groups of biological processes that were affected by corneal organ culture, among them keratinization, hypoxia, and angiogenesis, with genes from each group being affected by culture time. A cell type deconvolution analysis revealed significant modulations of the corneal immune cell profile in a time dependent manner. The results suggest that current culture conditions should be further refined and that prolonged cultivation may be detrimental. Recently, we showed that MACE enables transcriptional profiling of formalin-fixed and paraffin-embedded (FFPE) conjunctival tissue with high accuracy even after more than 10 years of storage. Here we demonstrate that MACE provides comparable results for native and FFPE corneal tissue, confirming that the technology is suitable for transcriptome analysis of a wide range of archived diseased corneal samples stored in histological archives. Finally, our data underscore the feasibility of bioinformatics cell-type enrichment analysis in bulk RNA-seq data to profile immune cell composition in fixed and archived corneal tissue samples, for which RNA-seq analysis of individual cells is often not possible.

Purpose: The aim of this study was to evaluate the feasibility of Descemet membrane endothelial keratoplasty (DMEK) tissue preparation using a glasses-assisted 3-dimensional (3D) display system and to compare it with a conventional surgical microscope., Methods: Healthy pairs of human corneas suitable for penetrating keratoplasty surgery were selected for this study. The tissues were randomly divided into 2 groups. Each pair of corneas had 1 cornea (group 1) prepared with NGENUITY (Alcon) with a 5-second staining time with vision blue, and the fellow cornea (group 2) was prepared using a OPMI Lumera 700 surgical microscope (Carl Zeiss Meditec, Jena, Germany) with a 30-second staining time. DMEK graft preparation time, speed of stripping, graft width, and endothelial cell loss were evaluated., Results: Twenty-eight pairs of corneas were included in this study. The graft preparation time was significantly higher in the 3D group than in the conventional group (498 ± 147 vs. 418 ± 85 seconds, P value = 0.031). The mean speed of stripping was 0.59 ± 0.081 mm/s in group 1 and 0.089 ± 0.005 mm/s in group 2 ( P value = 0.024). The mean endothelial cell density in group 1 and group 2 before tissue preparation was 2162 ± 115.21 and 2153 ± 122.45, respectively ( P value > 0.1). After tissue preparation, the endothelial cell density reduced to 1911 ± 150.72 in group 1 and 1998 ± 90.72 in group 2 ( P value = P value > 0.05). The graft width was 5.05 ± 0.71 mm in group 1 and 4.92 ± 0.23 mm in group 2 ( P value > 0.05)., Conclusions: DMEK tissue preparation with 3D display system NGENUITY is feasible with a slightly increased preparation time. The improved visualization allows a reduced staining time that could be beneficial for eye banks because it may reduce the toxic effect of staining colorants., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: The purpose of this study was to evaluate the safety, efficacy, and efficiency of a Descemet membrane endothelial keratoplasty (DMEK) graft preparation device, DescePrep, through measurement of graft viability, yield, and preparation time in both healthy and diabetic (high-risk) donor eyes., Methods: Twenty nondiabetic and 10 diabetic donor corneas were processed using DescePrep, which standardizes the liquid bubble technique. Corneas were stained with trypan blue and then processed. Cell counts through specular microscopy, optical coherence tomography imaging, and slit-lamp analysis were used for the evaluation of graft separation and viability in 5 nondiabetic corneas. The remaining 25 corneas (15 nondiabetic and 10 diabetic) were evaluated for preparation success rate and processing time. Ten corneas (5 nondiabetic and 5 diabetic) were randomly selected for further evaluation of global cell loss through staining., Results: Ninety-seven percent of corneas (29 of 30) were prepared successfully with DescePrep. The average preparation time was 2.83 ± 1.8 minutes. There was no significant difference in the time of preparation between the nondiabetic and diabetic groups (P = 0.077). The overall average cell death after processing was 7.9% ± 3.7% for all corneas. There was no significant difference in cell viability between diabetic and nondiabetic tissues after DescePrep processing (P = 0.769)., Conclusions: DescePrep is a new DMEK preparation technique that can process both nondiabetic and diabetic donor corneas at high yields in minutes. High-yield preparation of diabetic corneas may offer eye banks access to a larger donor pool, which is important because the demand for DMEK grafts continues to rise worldwide., Competing Interests: K. D. Solley, C. Diaz, B. T. Ostrander, E. Ruppel, A. Haileyesus, Y. Dou, S. Pramanik, and K. S. Parikh have ownership interest in Eyedea Medical, Inc. K. S. Parikh serves on the Board of Directors and owns company stock, which is subject to certain restrictions under Johns Hopkins University policy. The terms of this arrangement are being managed by Johns Hopkins University in accordance with its conflict-of-interest policies. The remaining authors have no conflicts of interest to disclose., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: The aim of this study was to determine whether loading a Descemet membrane endothelial keratoplasty (DMEK) graft using a drop-in procedure results in more endothelial cell loss (ECL) than the standard suction procedure., Methods: Pairs of donor corneas with equivalent preprocessing endothelium were prepared using the standard protocol of our eye bank. One member of each pair was loaded into an injector using the standard suction protocol. The mate graft was loaded using a drop-in protocol, in which the edge of the graft was gently grasped with a forceps, lifted to the edge of the injector, and dropped inside. Grafts were evaluated for ECL and examined for grab marks or other loading-associated damage., Results: There was no difference in mean ECL of grafts prepared for DMEK using the standard protocol (20.6% ± 4.5%) compared with that of mate grafts prepared using the drop-in loading protocol (19.5% ± 4.8%, P = 0.59). There was no consistent pattern of damage in the drop-in-loaded grafts, as grab marks or other tissue damage associated with the drop-in loading protocol were not consistently identified by a trained corneal surgeon., Conclusions: ECL was not significantly different in grafts prepared using a drop-in loading procedure compared with grafts prepared using the standard suction protocol. The drop-in loading protocol may be particularly useful to surgeons who load their own grafts and eye bank processing technicians who encounter a "flat" DMEK graft that does not scroll or a loosely scrolled DMEK graft., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Aim : To evaluate the effect of graft preparation and organ-culture storage on endothelial cell density (ECD) and viability of Descemet membrane endothelial keratoplasty (DMEK) grafts. Materials and methods : DMEK grafts (n = 27) were prepared at Amnitrans EyeBank Rotterdam from 27 corneas (15 donors) that were eligible for transplantation but could not be allocated due to the Covid-19-related cancellation of elective surgeries. Cell viability (by Calcein-AM staining) and ECD of five grafts originally scheduled for transplantation were evaluated on the originally planned surgery day, whereas 22 grafts from paired donor corneas were evaluated either directly post-preparation or after 3-7 days of storage. ECD was analyzed by light microscopy (LM ECD) and Calcein-AM staining (Calcein-ECD). Results : Light microscopy (LM) evaluation of all grafts showed an unremarkable endothelial cell monolayer directly after preparation. However, median Calcein-ECD for the five grafts initially allocated for transplantation was 18% (range 92-73%) lower than median LM ECD. For the paired DMEK grafts, Calcein-ECD determined by Calcein-AM staining on the day of graft preparation and after 3-7 days of graft storage showed a median decrease of 1% and 2%, respectively. Median percentage of central graft area populated by viable cells after preparation and after 3-7 days of graft storage was 88% and 92%, respectively. Conclusion : Cell viability of most of the grafts will not be affected by preparation and storage. Endothelial cell damage may be observed for some grafts within hours after preparation, with insignificant additional ECD changes during 3-7 days of graft storage. Implementing an additional post-preparation step in the eye bank to evaluate cell density before graft release for transplantation may help to reduce postoperative DMEK complications.

Competing Interests: The author has no conflicts of interest to disclose.

Competing Interests: The author has no funding or conflicts of interest to disclose.

Purpose: To investigate stamp visibility and endothelial cell loss (ECL) after the application of an orientation mark to Descemet membrane endothelial keratoplasty (DMEK) grafts supported by an air bubble., Methods: Eighteen DMEK grafts were prepared at an eye bank using a technique where an orientation mark was applied to the stromal surface of a DMEK graft that was supported by a small air bubble placed at the edge of the 2 endothelial surfaces of the graft. Grafts were evaluated at 2 and 5 days for stamp visibility and at 5 days with calcein-AM staining for ECL. Nine grafts underwent cross-country shipping, and the ECL of shipped and nonshipped grafts was compared using unpaired t test., Results: All 18 DMEK grafts exhibited a single, solid, readily visible orientation mark 2 and 5 days after preparation with a mean ECL of 13.5% ± 4.9%. Shipping conditions had no effect on stain retention or ECL., Conclusions: The application of an orientation stamp to a DMEK graft over an air bubble in an eye bank setting results in a single, solid orientation mark that is readily visible within the period in which most eye bank-prepared tissue is used. This technique produces no further ECL compared with the methods where the orientation stamp is applied through a stromal window. Eye bank technicians and surgeons can be confident that this modified preparation technique results in transplant-quality DMEK grafts with the additional benefit of conserving the stromal cap for use in other anterior lamellar procedures, thereby making efficient use of donor tissue., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: To examine tissue loss rates, processing time, and primary graft failure (PGF) of "prestripped-only" Descemet membrane endothelial keratoplasty (DMEK) grafts at a single eye bank and how these parameters changed after the introduction of steps to preload tissue among experienced processors., Methods: Tissue loss and processing time during DMEK graft preparation as well as PGF were analyzed retrospectively at a single eye bank between 2012 and 2018. Outcomes were assessed in consecutive grafts before and after the introduction of preloading to the eye bank's standard operating procedure., Results: A total of 1326 grafts were analyzed, composed of the first 663 preloaded DMEK grafts and, for comparison, the 663 DMEK grafts processed immediately before starting the preloaded service. Mean processing time increased from 17.0 ± 3.9 minutes to 26.0 ± 5.4 minutes with the advent of preloading (P < 0.01). Initially, average processing time increased dramatically, with a maximum processing time of 51 minutes, before regressing to the average. No significant difference in the rate of tissue wastage was observed before versus after the implementation of preloaded DMEK (1.2% vs. 1.7%, P = 0.48). PGF occurred in 7 grafts before the preloaded service and 10 grafts after starting the service (1.6% vs. 2.3%, P = 0.47)., Conclusions: Preloading does not affect tissue wastage for experienced technicians or the PGF rate but increases processing time. Eye banks that are considering adding preloading to their standard operating procedure may need to account for longer processing times in their daily operations., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: To report the management of precut Descemet stripping automated endothelial keratoplasty (DSAEK) lenticules unsuitable for transplantation because of irregular anterior profile after microkeratome cutting., Methods: After preparation for DSAEK, 20 tissues were considered unsuitable for transplantation because of nonhomogeneous posterior stromal thickness. To convert them into suitable tissues for surgery, manual stromal delamination was performed by removing the excess stromal layers after the indications obtained through optical coherence tomography. These tissues were further transplanted as ultrathin DSAEK., Results: Nineteen tissues were delaminated successfully. The average reduction in thickness in the center (63 ± 69 μm; P = 0.0101) and periphery (129 ± 39 μm; P < 0.0001) before and after delamination was significantly different. One tissue showed signs of perforation during manual dissection and therefore considered unsuitable for transplantation. Primary graft failure was reported in one case, but it was not correlated with the tissue preparation. No other clinical complications were observed after surgery., Conclusions: Manual delamination of the stroma because of irregular microkeratome cutting is a viable option to obtain a uniform graft thickness required for DSAEK surgeries. This technique can further reduce tissue wastage that is observed after microkeratome cutting errors., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Introduction: Since the beginning of the COVID-19 pandemic there has been some debate regarding the risk of transmission through tissue transplantation and tissue banking processes., Aim of the Study: To analyze the changes that SARS-CoV-2 has caused regarding the harvesting of corneal donor tissue and eye bank activities in Germany., Methods: A questionnaire was provided to 26 eye banks in Germany, consisting of questions about adaptations made in the screening of potential donors and the harvesting of corneal tissue following the pandemic spread of SARS-CoV-2., Results: Eighteen eye banks actively reduced recruitment of donors and two banks ceased all activity. Additional diagnostic screening was performed in eight banks, using conjunctival swabs and/or nasopharyngeal swabs. In six eye banks, additional protective measures, such as FFP2 masks and/or facial shields, were implemented. Overall, a mean reduction in the number of obtained donor tissues of 17% was observed., Discussion: Conjunctival and/or nasopharyngeal swabs of donors have been implemented by a minority. Reasons for not performing additional tests may be moderate sensitivity and lack of validation for postmortem use of RT-PCR testing. Also, the hazard of SARS-CoV-2 entering the corneal donor pool with subsequent transmission might be perceived as theoretical. Face shields provide a sufficient barrier against splash and splatter contamination but may be insufficient against aerosols. Additional face masks would provide support against aerosols, but it remains debatable if corneal harvesting can be considered an aerosol-producing procedure. In the future we expect to see changes in current guidelines because of a surge in scientific activities to improve our understanding of the risks involved with cornea donation in the COVID-19 pandemic, and because current practice may reduce the availability of donor corneas due to new exclusion criteria while the demand remains unchanged.

Purpose: To describe 2 insertion techniques, outcomes, and complications of preloaded, trifolded Descemet membrane endothelial keratoplasty (DMEK) cases using the DMEK EndoGlide inserter., Methods: This retrospective, consecutive case series analyzed the first 35 cases using the DMEK EndoGlide performed between October 2018 and October 2019 at a single center. Preloaded, trifolded DMEK tissues were delivered through a fluid-injected or pull-through technique. To inject the tissue, a burst of fluid was delivered into the lumen of the injector with a second instrument. Postoperatively, best-spectacle corrected visual acuity (BSCVA), pachymetry, graft survival, and complications were assessed., Results: Thirty-five eyes of 29 patients underwent DMEK alone (n = 11), with cataract surgery (n = 21), or with additional surgeries (n = 3). Of these, 19 (54.3%) grafts were injected. Video analysis revealed a median time of 3.5 minutes from graft insertion to opening for gas insertion. Median preoperative BSCVA of 0.398 logMAR improved to 0.097 logMAR (P = 0.02) at 9 months. Median pachymetry decreased from 619 μm to 551 μm (P = 0.03) at 9 months. Median donor endothelial cell count of 2890.5 cells/mm2 reduced to 2123 cells/mm2 (26.6% endothelial cell loss; P = 0.008) 6 months postoperatively. One (2.9%) graft failed due to inverted marking at the eye bank and subsequent reverse implantation., Conclusions: Pre-loaded, tri-folded tissues can be implanted with acceptable levels of endothelial cell loss. We describe a no-touch method of injecting pre-loaded, tri-folded tissue and highlight incorrect marking as a potential complication. This may not be identifiable intraoperatively due to lack of scroll formation., Competing Interests: A. O. Eghrari has ownership interest in Treyetech and LuckyVision, LLC. The remaining authors have conflicts of interest to disclose., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: Heterogeneity in scrolling behavior of Descemet membrane endothelial keratoplasty (DMEK) grafts complicates DMEK surgery. This prospective observational study assessed scrolling axes of DMEK grafts relative to the donor's eye., Methods: The eye bank randomly marked the rim of corneoscleral donor buttons during trephination and recorded the position relative to the donor's axis. Surgeons were masked to the absolute position of the eye bank marking and recorded the scrolling axis relative to the eye bank marking and DMEK upside-down orientation. The scrolling axis was categorized as vertical (0 to 30 degrees and 150 to 180 degrees), oblique (>30 to 60 degrees and 120 to <150 degrees), and horizontal (>60 to <120 degrees). Scrolling patterns of corneas from the same donor were assessed., Results: Scrolling patterns of 202 donor corneas from 149 donors were determined. The donor graft scrolled predominantly vertically to the donor's cornea [75%; 95% confidence interval (CI), 68%-80%]. Horizontal axes (11%) and oblique axes (14%) were less common. The median deviation in scrolling axes after unfolding the grafts was 0 degrees from the original scrolling axis (interquartile range, 0-15), indicating that scrolling axes were stable. Fellow eyes of 46 donors had 3.55 times higher odds of a nonvertical scrolling pattern if the first eye had a nonvertical scrolling pattern (95% CI, 1.37-9.20), suggesting that donor factors influencing both eyes could contribute to scrolling patterns., Conclusions: DMEK grafts have a natural and stable scrolling tendency at vertical axis of donor's cornea. Anticipating scrolling axes might help improve preparation techniques for DMEK grafts and outcomes.

Purpose: To compare the one-year outcomes of preloaded Descemet membrane endothelial keratoplasty (pDMEK) and non-preloaded DMEK (n-pDMEK) in patients with Fuchs endothelial corneal dystrophy (FECD)., Methods: This retrospective comparative cohort study consecutively included 68 eyes with Fuchs endothelial corneal dystrophy who underwent either pDMEK (n = 38) or n-pDMEK (n = 30) performed by cornea fellows with an experienced surgeon between 2016 and 2018 at the Massachusetts Eye and Ear Infirmary. Exclusion criteria were previous surgery (other than uncomplicated cataract surgery) and any documented evidence of macular or other corneal diseases. Corrected distance visual acuity (CDVA), central corneal thickness, intraocular pressure, patient characteristics, postprocessing endothelial cell count, donor graft data, and complications were compared., Results: CDVA showed similar results for pDMEK (0.12 ± 0.11 logarithm of the minimal angle of resolution [LogMAR]) and n-pDMEK (0.13 ± 0.13 LogMAR) (P = 0.827). Sixty-six percent of the pDMEK eyes and 57% of the n-pDMEK eyes achieved a VA of ≥0.1 LogMAR, and 95% and 97%, respectively, achieved a CDVA ≥0.3 LogMAR. The preoperative central corneal thickness of pDMEK and n-pDMEK (644 ± 62.2 μm, 660.5 ± 56.2 μm) decreased significantly after surgery (525.1 ± 43.6 μm, 526.5 ± 45.2 μm, P < 0.001), with no difference between groups (P = 0.840). The postprocessing endothelial cell count did not differ between pDMEK (2959.2 ± 182.9 cells/mm2) and n-pDMEK (2939.3 ± 278.7 cells/mm2) (P = 0.484). Complication rates were comparable with just the rebubbling performed in a minor procedure room showing a lower rate for pDMEK (13.16%) compared with n-pDMEK (33.33%) (P < 0.045)., Conclusions: One-year clinical outcomes were similar between pDMEK and n-pDMEK procedures, rendering eye bank-prepared pDMEK tissues a useful tool in the treatment of endothelial dysfunction., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: To investigate a cluster of corneoscleral rim cultures positive for Achromobacter species over a 6-month period at Massachusetts Eye and Ear., Methods: An increased rate of positive corneal donor rim cultures was noted at Massachusetts Eye and Ear between July and December 2017. Positive cultures were subjected to identification and antimicrobial susceptibility testing by phenotypic (MicroScan WalkAway) and genotypic (16S rDNA sequencing) methods. Samples of the eye wash solution (GeriCare) used in the eye bank were also evaluated. Antimicrobial activity of Optical-GS against Achromobacter spp. at 4°C and 37°C was assessed by time-kill kinetics assay., Results: Of 99 donor rims cultured, 14 (14.1%) grew bacteria with 11 (78.6%) due to uncommon nonfermenting Gram-negative bacilli. These had been identified by standard automated methods as Achromobacter (n = 3), Alcaligenes (n = 3), Ralstonia (n = 2), Pseudomonas (n = 2), and Stenotrophomonas (n = 1). Eight of these 11 isolates were subsequently available for molecular identification, and all were identified as Achromobacter spp. Six bottles of eyewash solution were evaluated and were positive for abundant Achromobacter spp. (3.4 × 105 ± 1.1 CFU/mL). Optisol-GS had no bactericidal activity against Achromobacter spp. at 4°C after 24-hour incubation but was bactericidal at 37°C. None of the patients who had received the contaminated corneas developed postoperative infection., Conclusions: An eyewash solution arising from a single lot was implicated in the contamination of donor rims by Achromobacter spp. The isolates were able to survive in the Optisol-GS medium at the recommended storage temperature. This highlights the need to continue improving protocols for tissue preparation and storage., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: Corneal transplantation is the most frequently performed transplant procedure. In much of the world, the demand for donor tissue heavily outstrips supply. With developments within lamellar corneal graft surgery, the use of split corneal donor tissue to increase donor tissue supply seems a pragmatic solution to reduce the supply and demand mismatch. This is especially important with tissue supply expected to be affected by the COVID-19 pandemic., Methods: A literature review of techniques was performed, enabling multiple transplants to be derived from a single donor and simulation of a model to quantify the number of corneas potentially saved., Results: Studies on splitting corneal donor tissue have demonstrated that up to 5 recipients may benefit from 1 donor scleral button. The impact of splitting donor tissue may provide a saving of up to 25.3% of donor graft tissue., Conclusions: Splitting and preparing the donor tissue within an eye bank will improve tissue validation and donor tissue availability and may increase surgeon efficiency.

Prestripped Descemet membrane endothelial keratoplasty is a relatively new technique that offers advantages in reduced surgical time and reduced tissue wastage compared with surgeon-stripped graft. Here, we report the following 2 types of prestripped Descemet membrane endothelial keratoplasty graft complications and their management: 1 free floating in the transport media and the other graft mispositioned.

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., Competing Interests: None

Purpose: This study compares the effect of the transport of conventionally prestripped Descemet membrane endothelial keratoplasty (DMEK) tissue with the DMEK revolutionary advanced Preloadable Injection Device (RAPID) preloaded transport system from Geuder AG (Heidelberg, Germany). Endothelial cell loss, tissue integrity, endothelial cell phenotype, and viability were assessed and compared., Methods: Twelve DMEK grafts were prestripped by the cornea bank and transported using the following 2 conditions: conventional flask (n = 6) or a preloaded transport cartridge (DMEK RAPID, n = 6). After transport, tissues were analyzed for cell density; denuded areas; immunolocalization of corneal endothelial markers, such as ZO-1, CD166, and Na/K ATPase; histology analysis; and cell viability staining with Hoechst, calcein AM, and ethidium homodimer., Results: Endothelial cell loss (10.35% vs. 9.15%) did not differ between transport conditions. Histological analysis confirmed the integrity of the Descemet membrane and endothelial cell layer with both transport conditions. Similarly, the corneal endothelial cell mosaic was conserved in both conditions. The ZO-1 tight junctions confirmed the integrity of the confluent corneal endothelial cell monolayer. CD166 and Na/K ATPase detection with immunofluorescence was also comparable. A similar percentage of dead cells was reported in both conditions (18.1% vs. 16.73%). Moreover, the surface covered with calcein-positive cells (59.02% vs. 61.95%) did not differ between transport conditions., Conclusions: Our results suggest that DMEK grafts can be prestripped or preloaded into a novel transport cartridge and shipped to the clinic with comparable endothelial cell loss, phenotypical marker expression, and viability to the conventional prestripped donor tissue.

Purpose: The purpose of this study was to compare the clinical outcomes from using eye bank-prepared, endothelium-out preloaded Descemet membrane endothelial keratoplasty (DMEK) tissue with those obtained with endothelium-out surgeon-loaded DMEK tissue using the same surgical technique at 1 site., Methods: This study retrospectively reviewed 400 consecutive cases of DMEK from March 2016 to April 2018. The last 200 cases using surgeon-loaded tissue were compared with the first 200 cases using preloaded tissue. Statistical analysis was performed using the Wilcoxon signed-rank test, binomial logistic regression, Kruskal-Wallis 1-way analysis of variance, Student t test, or Pearson χ tests., Results: Comparing surgeon-loaded versus preloaded DMEK tissue, respectively, no statistical difference was found in the mean 6-month postoperative values for endothelial cell loss (32.9% ± 18.5% vs. 29.9% ± 16.4%, P = 0.31), best corrected visual acuity (20/26 vs. 20/25, P = 0.54), or change in central corneal thickness (-14.4% ± 8.9% vs. -15.6% ± 11.7%, P = 0.43). The mean 1-year endothelial cell loss was also not significantly different (37.6% ± 17.2% vs. 33.2% ± 14.8%, P = 0.07). Overall, the rebubble rate for surgeon-loaded tissue was 17.5% and 12.5% for preloaded tissue, a statistically nonsignificant difference. Operative outcomes for mean tissue scroll tightness (1-4) and tissue unscroll time (minutes) for surgeon-loaded and preloaded tissue were similar between groups (2.4 vs. 2.2, P = 0.12 and 3.5 vs. 3.3 minutes, P = 0.50)., Conclusions: Tissue that is trephinated, stained, and loaded into an injector by the eye bank and then shipped to the surgeon had no difference in clinical outcomes compared with tissue where the surgeon performs these steps. The safety and speed of using preloaded tissue should be considered by DMEK surgeons.

Purpose: To ascertain whether offsetting the Descemet membrane endothelial keratoplasty (DMEK) punch can expand the donor pool in conjunction with prepunched and preloaded services by recapturing the corneas otherwise excluded by the conventional central clear zone criteria., Methods: In this retrospective review of corneas recovered and processed for DMEK by a single eye bank between March 2017 and October 2018, corneas failing to meet the conventional central clear zone requirement during initial evaluation (defined as an area in the central cornea where an 7.5- to 8.0-mm diameter graft can be obtained free of previous surgical scars, Descemet tears, or confined areas of endothelial defects) were further evaluated for offset punching. Corneas with a central endothelial cell density of at least 2000 cells/mm at the initial screening (average of 3 specular images assessed with the center dot method) that had a clear zone of 7.5- to 8.0-mm diameter where a graft could be obtained were designated as eligible for offset punching for either prepunched or preloaded DMEK., Results: A total of 2607 corneas were found to be suitable for DMEK using the conventional central clear zone criteria. An additional 62 corneas were deemed DMEK suitable by offsetting the punch, yielding a 2.4% increase in the availability of DMEK suitable corneas., Conclusions: Offsetting the DMEK punch can recapture corneas otherwise excluded from the DMEK donor pool because of a failure to meet the conventional central clear zone criteria, and by our estimation may help eye banks meet the growing demand for DMEK tissue while maximizing the transplant potential of every cornea.

Purpose: To describe and evaluate the efficacy and safety of a novel technique to prepare Descemet membrane endothelial keratoplasty (DMEK) donor grafts using a newly designed partial-thickness hinge punch., Methods: The novel punch has a circular guarded blade missing 1 clock hour, creating an uncut hinge on the donor cornea. In addition, 2 straight cuts are made by the punch perpendicular to the edge of trephination toward the trabecular meshwork in the hinge area. After the donor corneoscleral rim is positioned endothelial side up, a partial-thickness trephination is performed avoiding any rotational movements. Descemet membrane is lifted from Schwalbe line in the hinge area, and DMEK graft is peeled after desired marking without further preparation., Results: Three surgeons of different experience levels on DMEK (senior/independent/fellow) initially applied the new technique in 18 research corneas, divided into equal groups. Two failures in graft preparation were noted, defined as radial tears extending ≥0.5 mm. The mean preparation time was 6.21 ± 1.45 minutes. No statistically significant differences were noted in success rate, duration, and endothelial cell loss (ECL) between surgeons (P > 0.05). ECL was evaluated as an average of 5 readings on randomly selected graft areas, not including graft periphery. Fifteen additional research corneas were stripped by 1 single user in an eye bank setting. No tissue loss was recorded, whereas ECL and mortality rate remained unaffected after preparation (P = 0.64 and P = 0.72, respectively)., Conclusions: This new DMEK graft preparation technique, simulating the opening of a yogurt cup, seems to be a safe and an efficient method, providing shorter preparation time and low failure rates independent of surgeon's experience level.

Background: Corneal storage for the very long term, without degradation, would make it possible to optimize a very limited resource worldwide. We previously demonstrated the superiority, compared to conventional 4-week passive organ culture (OC), of an active storage machine (ASM) that restores intraocular pressure and medium renewal. Here, we investigate eye banking for up to 3 months., Methods: In a randomized preclinical trial with 24 paired corneas, 1 was stored in OC and the other in ASM, using the same medium. Assessments were done on the second day and at 3 months: endothelial cell density (ECD in cells/mm), corneal transparency and thickness. At day 86, OC corneas were deswelled in a common hyperosmotic medium, but not the ASM corneas, which had remained thin. In addition, at day 88, viable ECD was measured using a live/dead assay, and endothelial expression of Na/K ATPase, Cox IV, ZO-1, N-CAM, and CD166 was observed., Results: The ASM extended storage to 3 months with unprecedented endothelial cell quality: no OC corneas remained suitable for transplantation, but one-third of ASM corneas were compliant (ECD > 2000/mm). Given that corneas with ECD > 1600/mm were also usable for emergency, 58% of ASM corneas were usable versus 33% in OC. EC survival was 53% higher in ASM (P < 0.001), structural and functional proteins of ECs were much better preserved in ASM, and it prevented the constant major edema of OC., Conclusions: By extending graft survival to 3 months, the ASM will optimize eye banking and open up new perspectives in experimental research.

Pre-Descemet endothelial keratoplasty (PDEK) is an alternative technique to Descemet membrane endothelial keratoplasty (DMEK). The preparation of PDEK tissue by pneumatic dissection is simple and reproducible. The PDEK clamp helps to consistently obtain a type 1 big bubble. The mean size of type 1 big bubble is 7.255 ± 0.535 × 6.745 ± 0.668 mm. The volume of air required to obtain type 1 big bubble is 0.14 to 0.37 mL. Dissection of PDEK tissue can be achieved by trephination or manual excision. Insertion of tissue into the recipient eye can be by injection or pulling. Unfolding techniques used for PDEK are similar to those used in DMEK. Unlike DMEK, PDEK tissue is easier to handle and unscroll and allows use of younger donors. It could help surgeons converting to endothelial keratoplasty, with significant advantages in preparation, handling, and unscrolling in the eye.

This study aimed to evaluate the possibility to extend the storage of unused organ-cultured donor corneas. After 28 days of corneal culture in TISSUE-C (AL.CHI.MI.A. S.R.L., Italy) and 5-day storage in transport/deswelling medium CARRY-C (AL.CHI.MI.A. S.R.L., Italy), 25 corneas that were deemed suitable for transplantation were transferred in fresh TISSUE-C at 31 °C for additional 7 days and then in fresh CARRY-C at room temperature for 24 h. Tissues were assessed for endothelial cell density (ECD), endothelial mortality and morphology after the standard and the extended corneal storage. In addition, the effect of donor age < 85 years and ≥ 85 years on corneal characteristics was assessed. After the extended storage, 6 out of 25 tested corneas (24%) showed ECD values below the acceptance limit (< 2000 cells/mm 2 ). 19 corneas (76%) were still suitable for transplantation and showed a 5.9% loss in ECD, which was not statistically significant (P = 0.0949) compared to standard storage period. The two donor age groups did not show statistically significant differences in any tested parameter, although a trend for lower ECD and higher mortality in Descemet's folds after standard storage was observed in the ≥ 85 age donor group. Thus, the attempt of the current study to provide new sight-restorative options for unused tissues and increasing the availability of corneas in case of shortage gave encouraging results. Although a higher vulnerability of corneas from very old donors could not be statistically demonstrated in the present study, higher sample size could be required for prolonging the shelf life of these tissues.

Purpose: This study compares endothelial cell loss (ECL) between donor grafts loaded in the Geuder Glass Cannula and the Coronet EndoGlide made specifically for Descemet membrane endothelial keratoplasty (DMEK)., Methods: Prestripped tissues using the submerged cornea using backgrounds away (SCUBA) technique were obtained from an eye bank. Donor grafts were loaded with the Geuder Glass Cannula or Coronet DMEK EndoGlide. Grafts were then ejected directly onto a glass slide, stained, and imaged with confocal fluorescent microscopy. Trainable Fiji (ImageJ) segmentation software was used to quantify ECL., Results: Twenty-three total tissue samples were tested. Eleven grafts were loaded using the Geuder, and 12 were loaded with the Coronet. Preloading graft endothelial cell density was comparable between Geuder (2436 ± 581 cells/mm) and Coronet (2577 ± 483 cells/mm) groups with P = 0.56. Other baseline characteristics including donor age and time from death to loading were comparable (P = 0.73 and P = 0.66, respectively). Average ECL was 11.3% ± 3.0% in the Geuder group versus 7.2% ± 7.0% in the Coronet group and not significantly different (P = 0.07). When observing patterns of cell loss, grafts loaded in the Coronet DMEK EndoGlide resulted in more focal forceps grabs and trifold patterns. Grafts loaded with the Geuder Glass Cannula yielded more diffuse stippled stress lines., Conclusions: Our results suggest comparable ECL between Geuder (endothelium-out) and Coronet (endothelium-in) DMEK EndoGlide tissue injection techniques. Patterns of ECL in the Coronet group correlated with forceps grabs, but this study suggests that additional manipulation of tissue does not result in higher ECL.

We describe a rapid and easy technique for the preparation of pre-stripped Descemet membrane (DM) endothelial keratoplasty (DMEK) tissue in the eye bank, without the use of dye or trephining by the eye bank technician. After drawing 2 radial marks 90° apart on the scleral rim of the cornea, a circumferential incision was made internal to the trabecular meshwork and the edge of the DM opposite to the marked area was scored and gently peeled towards the marked area. Finally, the peeled DM was gently unrolled and attached to the posterior stromal bed via absorbing excess liquid from the periphery. The prepared DMEK tissue while reattached on the posterior aspect of the cornea was subjected to slit-lamp biomicroscopic and specular microscopic examinations. In case of proper preparation of DMEK tissue and suitability of endothelial cells for DMEK surgery, the tissue was distributed for transplantation. At the time of surgery, trypan blue staining was performed by the cornea surgeon before DM trephination. Duration of DMEK tissue preparation, uneventful preparation, post-preparation endothelial cell loss, surgeon's satisfaction, and intra- and post-operative attachment of the tissue were investigated variables in this study. Over a 2-year period, 28 DMEK tissues were successfully prepared from 30 corneas using our new technique. The median duration of DMEK preparation was 100 s. The area of endothelial cell loss was 1.8% ± 1.2% centrally and 6.2% ± 1.8% in areas close to the forceps contacts. Seven out of the 28 (25%) DMEK tissues were ruined by surgeons during surgery and the overall rate of re-bubbling in 19 transplanted DMEK tissues was 16%. The presented method, which uses no trephining or vital dye, has simplified and expedited the preparation process of pre-stripped DMEK tissue in the eye banks.

Purpose: To determine the feasibility of using telemedicine consultations in the evaluation of recovered donor corneas for transplant suitability., Methods: This study aims to establish and test the minimum imaging requirements for telemedical consultations of corneal tissue by remote eye bank medical directors. Digital images from the slit lamp, optical coherence tomography, and/or specular microscope were assembled into telemedical consults and emailed to 4 eye bank medical directors (M.A.T., J.W., C.S.S., N.K.R.). Feedback on the minimum image requirements for each corneal finding was collected. After establishing a standardized imaging and presentation protocol, test cases were presented to the medical directors to examine the validity of these remote consults. To establish a benchmark for the study's parameters, one medical director (J.W.) examined each case in person after his initial remote review. Examiners were masked to each other's responses., Results: Minimum image requirements for determination of corneal findings were defined and were specific to each anatomic layer of the cornea (epithelial, stromal, or endothelial). Using a defined set of digital images for a set of common corneal findings, the rate of agreement between remote evaluators, eye bank staff, and the in-person evaluator was 100% (11 of 11 examples). For ambiguous test cases, remote evaluators agreed on 80% of the cases (4 of 5)., Conclusions: Results from this pilot study suggest that telemedical review of corneal tissue using high-quality digital images may be adequate for accurate identification of specific corneal findings commonly encountered by eye banks.

Purpose: To examine postoperative reports of infection rates for eye bank-prepared and non-eye bank-prepared corneas from January 1, 2006 to December 31, 2017, from a single eye bank., Method: A retrospective review of reported fungal and bacterial infections with corneal transplant surgeries using corneas distributed by our eye bank was conducted. The reported number of infections for corneas that underwent eye bank preparation (pre-cut and pre-stripped corneas) and for those distributed without eye bank preparation was quantified. The potential association between infection rates in tissue prepared by the eye bank and those in corneas that had no additional eye bank processing was also examined., Results: Four of 17,035 corneas distributed during the study period were associated with fungal infections (1 eye bank-prepared and 3 non-eye bank-prepared corneas) and were attributed to the tissue after investigation by eye bank medical directors. There was no ascending trend of infections reported with eye bank-prepared corneas in the first 3 years (2 of 1054 corneas, 0.19%) compared with that in the last 3 years of the study period (6 of 3500 corneas, 0.17%; P = 0.901) when the eye bank distributed 3 times more prepared corneas than non-eye bank-prepared corneas. A significant increase in the numbers of reported infections for non-eye bank-prepared corneas was observed between these 3-year intervals (0.1% in the first 3 years to 1.58% in the last 3 years; P = 0.001)., Conclusions: Reports of infections remained low despite increased use of eye bank-prepared tissue. These results suggest that factors other than eye bank tissue preparation should be considered when investigating potential sources of pathogen contamination in donor corneas.

Purpose: To compare corneal endothelial damage associated with 2 techniques for preloaded Descemet membrane endothelial keratoplasty (DMEK): a tri-folded graft stored in a plastic cartridge designed for DMEK and a scrolled graft stored in a modified Jones Tube, at the time of preparation and after shipping., Methods: DMEK grafts were prepared at the Rocky Mountain Lions Eye Bank. The grafts were either tri-folded and loaded in a plastic cartridge or scrolled and loaded into a modified Jones Tube. In each group, the grafts were then either immediately removed from the cartridges or shipped for 48 hours. The grafts were then stained with Calcein AM and imaged using a fluorescent microscope. Endothelial cell loss (ECL) was determined using trainable segmentation in Fiji by 2 graders. At each time point, rates of ECL loss were compared across the 2 groups. To explore the role of donor characteristics, a multivariable regression model was produced to account for method (tri-folding vs. scroll), donor age, donor gender, death-to-preservation time, death-to-preparation time, and shipping., Results: A total of 40 grafts were prepared, processed, imaged, and analyzed. No significant difference in cell loss was seen between groups at either time point alone. In the multivariate model, no significant increase in cell loss was associated with either tri-folding (3.7% less ECL; P = 0.051) or shipping (4.3% less ECL; P = 0.049)., Conclusions: All techniques used resulted in clinically acceptable levels of ECL. Tri-folded tissue in a plastic cartridge did not result in ECL inferior to a scroll when prepared either immediately or preloaded for 48 hours.

Purpose: To quantify endothelial cell loss (ECL) caused by orientation stamps on prestripped and preloaded Descemet membrane endothelial keratoplasty (DMEK) grafts, and to examine a method for reducing ECL using a smaller stamp., Methods: Ten prestripped and 10 preloaded DMEK grafts were prepared with S-stamps. Ten additional preloaded DMEK grafts were prepared with both an S-stamp and a smaller F-stamp in different paracentral areas of the graft. The footprint of each stamp was measured using ink on cardstock. DMEK grafts were stored in viewing chambers filled with 20 mL of Optisol-GS for 3 days at 4°C. ECL was quantified using Calcein-AM staining and FIJI Weka Segmentation., Results: S-stamps on prestripped DMEK grafts contributed an average ECL of 1.1% ± 0.5% (range: 0.6%-2.2%) toward total graft damage, whereas S-stamps on preloaded DMEK grafts contributed approximately twice that amount (average ECL: 2.0% ± 0.7%, range: 1.3%-3.1%, P = 0.004). Overall ECL for prestripped grafts (average: 7.1% ± 3.3%, range: 3.3%-13.7%) and preloaded grafts (average: 11.3% ± 4.2%, range: 6.9%-19.4%) was similar to previous reports. The footprint of the S-stamp was approximately 45% larger than that of the F-stamp. In 10 preloaded grafts marked with both stamps, the S-stamp caused an average ECL of 1.9% ± 0.6% (range: 1.2%-3.2%), whereas the smaller F-stamp caused an average ECL of 1.0% ± 0.2% (range: 0.8%-1.4%, P = 0.0002)., Conclusions: Loss of endothelial cells associated with graft-stamping was greater in preloaded tissue than in prestripped tissue and was less with a smaller F-stamp than with a larger S-stamp. Using a smaller stamp could help minimize ECL in prestripped and preloaded DMEK grafts.

Purpose: Preloaded, trifolded grafts in Descemet membrane endothelial keratoplasty require transfer of the trifolding process from the corneal transplant surgeon to the eye bank technician. We sought to assess whether trifolding may be safely conducted by an eye bank technician with cell loss comparable to standard peeling and lifting., Methods: A total of 10 grafts were stained, peeled, and transferred directly onto a bed of Calcein-AM and Amvisc Plus by an eye bank technician. Five grafts were removed and stained as a scroll, and 5 grafts were trifolded with the endothelium in before transfer. Photographs were acquired with an inverted fluorescence microscope, and image segmentation was performed. A t test was conducted to compare differences in endothelial cell loss across groups., Results: Mean cell loss in the scroll group was 18.5% [95% confidence interval (CI): 15.2%-21.9%] compared with 7.6% of the trifolded group (95% CI: 1.7%-13.5%). A 2-tailed t test indicated decreased cell loss in the trifolded group (P = 0.013)., Conclusions: Despite additional manipulation of the graft, trifolding of Descemet membrane and endothelium may be performed by an eye bank technician without significantly increased cell loss relative to graft preparation as a scroll.

Purpose: To investigate the learning curve of Descemet membrane endothelial keratoplasty (DMEK) graft preparation in an eye bank., Methods: Four operators prepared 645 DMEK grafts using the stripping technique between 2014 and 2017 at the Veneto Eye Bank Foundation, Italy. Endothelial cell loss (ECL) and tissue wastage were recorded retrospectively after DMEK preparation and correlated with the number of tissues prepared each year by each operator. On average, our operators performed 1 donor preparation a week over the course of this study. Only donors older than 60 years were used in this study, and approximately 10% of donors had diabetes. The Wilcoxon test for paired data and 1-way ANOVA were used for checking statistical significance with the Tukey test as post hoc analysis. P < 0.05 was considered statistically significant., Results: ECL did not change significantly over time from operator 1. Significant ECL drop was noted from operator 2 between years 2014-2016 (P = 0.0049) and 2017 (P = 0.0094); from operator 3 between years 2015-2016 (P = 0.0288) and 2017 (P = 0.0097); and from operator 4 between 2015-2016 (P = 0.0469) and 2017 (P = 0.0331). Operators 1 and 3 did not show a significant difference, considering every 50 grafts prepared by each operator. Operator 2 showed significant ECL drop between 1 to 50 and 51 to 100 (P = 0.0002) and 1 to 50 and 101 to 150 (P = 0.0001) grafts. Operator 4 showed significant ECL drop between 1 to 50 and 101 to 150 (P = 0.002) and 51 to 100 and 101 to 141 (P = 0.0207) grafts. No intraoperator difference was observed per 50 grafts (P > 0.05)., Conclusions: There is a learning curve for DMEK graft preparation. ECL and tissue wastage can be reduced with practice and skills. However, each operator may be limited to his or her own learning capability.

Purpose: Evaluation of cumulative Descemet membrane endothelial keratoplasty endothelial cell loss (ECL) from preparation through injection using 2 different glass injectors., Methods: Eighteen Descemet membrane endothelial keratoplasty grafts with "S" stamps were prepared by eye bank technicians. Nine grafts were assigned to injection with a modified glass Jones tube injector with a 2.4-mm opening and 9 were assigned to injection with the DORC glass pipette injector (<1.5-mm opening). The grafts were prepared and loaded into the injectors using the standard surgical technique, ejected onto a bed of viscoelastic on a glass slide, and unscrolled using viscoelastic. The grafts were stained with the vital dye Calcein-AM, then digitally imaged and analyzed using FIJI. The percentage of ECL was calculated by measuring the area of nonfluorescent pixels and dividing it by the total graft area pixels. A statistical comparison was performed using a 2-tailed unpaired t test., Results: Grafts injected using the DORC injector versus Jones tube injector had ECL of 29.2% ± 8.5% [95% confidence interval (CI)] versus 23.0% ± 5.1% (95% CI), respectively. This difference was not statistically significant (P = 0.17); however, the patterns of ECL on the grafts was different between injectors. Peripheral ECL caused by trephination and surgeon grasp sites accounted for 7.5% ± 1.2% (95% CI)., Conclusions: There was no statistical difference in ECL between the 2 injectors. There were characteristic differences in patterns of ECL seen between injectors, which may be clinically relevant and indicate the types of stress that grafts are exposed to during passage through various injectors.

Purpose: The purpose of this study was to develop a nomogram to predict postcut thickness of corneal grafts prepared at an eye bank for Descemet stripping automated endothelial keratoplasty (DSAEK)., Methods: Retrospective chart review was performed of DSAEK graft preparations by 3 experienced technicians from April 2012 to May 2017 at the Eye Bank of Canada-Ontario Division. Variables collected included the following: donor demographics, death-to-preservation time, death-to-processing time, precut tissue thickness, postcut tissue thickness, microkeratome head size, endothelial cell count, cut technician, and rate of perforation. Linear regression models were generated for each microkeratome head size (300 and 350 μm)., Results: A total of 780 grafts were processed during the study period. Twelve preparation attempts resulted in perforation (1.5%) and were excluded. Mean precut tissue thickness was 510 ± 49 μm (range: 363-670 μm). Mean postcut tissue thickness was 114 ± 22 μm (range: 57-193 μm). Seventy-nine percent (608/768) of grafts were ≤130 μm. The linear regression models included precut thickness and donor age, which were able to predict the thickness to within 25 μm 80% of the time., Conclusions: We report a nomogram to predict thickness of DSAEK corneal grafts prepared in an eye bank setting, which was accurate to within 25 μm 80% of the time. Other eye banks could consider performing similar analyses.

Surgical use of donor corneal tissue from victims of water submersion (drowning or submersion secondary to death) remains controversial due to limited evidence about the quality of these tissues. To assess the safety of donor corneal tissue from victims of water submersion, an investigation of eye banks' practice patterns and tissue outcomes was conducted. All 79 Eye Bank Association of America accredited eye banks were contacted for a phone interview of practices regarding tissue from victims of water submersion. A retrospective review of corneal tissues from 2014 to 2016 from a large eye bank network was performed to identify all donors submerged in water. Corneal epithelial integrity, endothelial cell density (ECD), rim cultures, and adverse events were analyzed for associations with water submersion characteristics. 49 eye banks (62% response) participated in the survey. 55% of these eye banks had specific, written protocol for tissue eligibility from donors submerged in water. With or without specific protocol, eye banks reported considering water type (84%) and length of time submerged (92%) to determine eligibility. 22% of eye banks reported medical director involvement when eligibility determination was unclear. 79 tissues from 40 donors who were submerged were identified in 2014-2016 eye bank data. No donor tissues had pre-processing corneal infiltrates, positive rim cultures, or adverse events post-keratoplasty. Corneal epithelial integrity and ECD were not associated with water type or length of time submerged. In conclusion, data from a large eye bank network showed no adverse events or outcomes, indicating these tissues may be safe.

Purpose: The purpose of this study is to analyze the impact of death causes and documented donor diseases on initial endothelial cell counts (after retrieval) and the development of corneal graft endothelia during organ culture., Methods: The retrospective statistic analyses was conducted on a data set of 10,185 human corneas prepared at the Hamburg Eye Bank., Results: Although we observed that death by gunshot trauma or alcoholism seems to be associated with marginally higher endothelium cell counts (independently from donor age), we could prove that only donor age is a relevant predictive parameter for the initial cell-density of the endothelium and its development in vitro., Conclusion: We conclude that an extension of prospective quality parameters for donor selection additional to donor age (such as individual causes of death) is not necessary.

Purpose: To provide an overview of the current literature on donor tissue preparation for Descemet membrane endothelial keratoplasty (DMEK)., Methods: A comprehensive database search without date restrictions was performed in PubMed and in The Cochrane Library in May, 2017. Keywords included Descemet membrane endothelial keratoplasty, corneal transplantation, graft, harvest, dissection, preparation, endothelial cell, and endothelial cell density. Articles aiming to describe or evaluate a technique for DMEK graft preparation were considered eligible and were included in this review., Results: A graft dissection technique that provides consistent tissue qualities and a low risk of preparation failure is essential for surgeons and eye banks preparing DMEK tissue. Various techniques have been described aiming to facilitate DMEK graft dissection, including manual dissection, pneumatic dissection, and hydrodissection. All show a trend toward a no-touch technique, for example, without direct physical tissue manipulation during tissue harvesting, as a potential ideal approach to minimize graft damage., Conclusions: An overview of the current harvesting techniques available for DMEK may benefit corneal surgeons and eye banks in choosing the best approach for each specific user.