Modified procedure for Loading "Flat" DMEK Grafts Into an Injector.

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.

Ink Retention and Endothelial Cell Viability After the Application of an Orientation Stamp Over an Air Bubble During Descemet Membrane Endothelial Keratoplasty Graft Preparation.

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.

Minimizing Endothelial Cell Loss Caused by Orientation Stamps on Preloaded Descemet Membrane Endothelial Keratoplasty Grafts.

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.

Measuring Endothelial Cell Loss on DMEK Grafts After Transplantation in Human Cadaveric Whole Eyes: Description of the Technique and Pilot Study.

PURPOSE: To describe a novel method for analyzing Descemet membrane endothelial keratoplasty (DMEK) graft damage after implantation into human cadaveric donor eyes and to compare results achieved by performing DMEK with a surgeon's long-established technique compared with those of an unfamiliar technique. METHODS: Eight DMEK grafts were implanted into previously frozen human cadaveric eyes. Four grafts were implanted using a Straiko injector and tap technique familiar to the surgeon (C.S.S., 3-yr experience), and 4 grafts were implanted using the Tan EndoGlide and "donor mat device" pull-through technique new to the surgeon. After implanting a DMEK graft and attaching it to the recipient stroma with an air bubble tamponade, the corneoscleral cap was "recovered" from the cadaveric globe using standard techniques. The DMEK graft was stained with Calcein-AM. After staining, a 9.5-mm stromal "carrier button" was punched, and the carrier and graft were transferred to a microscope slide. Grafts were imaged and analyzed using FIJI trainable segmentation. RESULTS: Donor graft characteristics were similar between both groups. Grafts implanted using the surgeon's routine technique showed an average endothelial cell loss (ECL) of 31% ± 4% (n = 3). Grafts implanted using the technique unfamiliar to the surgeon showed an average ECL of 47% ± 24%, but with a trend toward improvement (1 = 76%, 2 = 65%, 3 = 32%, 4 = 17% ECL). CONCLUSIONS: Our proof-of-principle experiment shows that this imaging approach enables quantification of ECL caused by different instruments and surgical techniques after graft implantation. We have used this method to visualize the learning curve of 1 surgeon when learning a new surgical technique.

Partitioning an Artificial Anterior Chamber With a Latex Diaphragm to Simulate Anterior and Posterior Segment Pressure Dynamics: The "DMEK Practice Stage," Where Surgeons Can Rehearse the "DMEK Dance".

PURPOSE: To present a novel apparatus for simulating the anterior and posterior segment pressure dynamics involved in executing Descemet membrane endothelial keratoplasty (DMEK) surgery when using a chamber-shallowing technique. METHODS: An artificial anterior chamber (AAC), 18-mm trephine, latex glove, two 3-mL syringes, and one donor cornea comprising an intact corneoscleral cap from which a DMEK tissue was peeled and punched are required for the model. After making the corneal incisions with the corneoscleral cap mounted on the AAC in the usual fashion, the corneoscleral cap is remounted onto the dried AAC over an 18-mm latex diaphragm. The space between the latex diaphragm and the cornea is filled with saline to pressurize the anterior chamber, and the posterior segment is pressurized with air from a syringe. The resulting apparatus comprises a posterior segment and anterior chamber that exert pressure on each other by way of a distensible latex diaphragm. RESULTS: A novice and experienced DMEK surgeon and 2 eye bank technicians were able to assemble the apparatus and perform the routine steps of a DMEK procedure, including maneuvers that require shallowing the anterior chamber and lowering its pressure. Only one cornea was required per apparatus. CONCLUSIONS: We present a novel in vitro model of the human eye that more closely mimics the anterior and posterior segment pressure dynamics of in vivo DMEK surgery than average human and animal cadaveric globes. The model is easy to assemble, inexpensive, and applicable to a range of teaching environments.

Evaluation and Quality Assessment of Prestripped, Preloaded Descemet Membrane Endothelial Keratoplasty Grafts.

PURPOSE: To determine graft quality and feasibility of Descemet membrane endothelial keratoplasty (DMEK) grafts that are prestripped and preloaded into injectors by eye bank technicians before shipping to surgeons. METHODS: DMEK grafts (n = 31) were prepared from donor corneas and preloaded into Straiko Modified Jones tubes and set inside viewing chambers filled with 20 mL of Optisol-GS. Preloaded grafts were evaluated using specular microscopy and slit-lamp biomicroscopy. Endothelial cell loss (ECL) was captured by vital dye staining and quantified using FIJI. A subset of preloaded tissues was subjected to a shipping validation and 5-day storage assay. Fourteen additional DMEK grafts (not preloaded) were examined to quantify damage resulting from prestripping alone. RESULTS: Specular microscopy was able to be performed for all preloaded tissues. Average ECL for preloaded tissues quantified by vital dye staining and FIJI after overnight storage was 16.8% ± 5.9%, and differed from slit-lamp ECL estimation by an average of 5.3% ± 3.6%. The average damage caused by prestripping alone was 9.3% ± 5.9%, and it was significantly less than that of preloaded tissues (P < 0.01). Average ECL for preloaded tissues subjected to round-trip shipping events was 18.5% ± 12.4%, and ECL for tissues stored at 4°C for 5 days after preloading was 13.1% ± 9.5%. CONCLUSIONS: It is possible to prepare, evaluate, and ship DMEK grafts loaded inside a glass carrier and viewing chamber. The ability to evaluate tissues after processing allows for adherence to the Eye Bank Association of America Medical Standards, and for surgeons to receive the most accurate tissue information.

The S-stamp in Descemet Membrane Endothelial Keratoplasty Safely Eliminates Upside-down Graft Implantation.

PURPOSE: To present 6-month clinical outcomes from a series of 165 consecutive Descemet membrane endothelial keratoplasty (DMEK) procedures before and after the introduction of a novel stromal-sided S-stamp preparation technique that has decreased the incidence of iatrogenic primary graft failure by eliminating upside-down grafts. DESIGN: Retrospective nonrandomized comparative case series. PARTICIPANTS: We included 165 consecutive eyes that had undergone DMEK surgery for Fuchs' or pseudophakic bullous keratopathy. These cases were divided into 2 cohorts: the first cohort comprised 31 cases that used unstamped tissue before the S-stamp was introduced, and the second cohort comprised 133 cases after the S-stamp was incorporated into the standardized technique. A single unstamped DMEK case was performed after the introduction of the S-stamp for a total of 32 unstamped cases. METHODS: Donor materials were prepared at a single eye bank using a standardized technique, which subsequently incorporated the addition of a dry ink gentian violet S-stamp to the stromal side of Descemet membrane. All surgeries were performed at a single clinical site by 5 surgeons (2 attending surgeons and 3 fellows). Two of the 165 DMEK cases were performed for pseudophakic bullous keratopathy (2 cases, 1 in each cohort), and the remaining cases were for Fuchs' endothelial dystrophy. Primary outcome measures were assessed at 6 months and maintained in a prospective institutional review board-approved study. MAIN OUTCOME MEASURES: We analyzed the 6-month endothelial cell density, incidence of iatrogenic primary graft failure, upside-down graft implantation, and rebubble events. RESULTS: The S-stamp eliminated upside-down graft implantations (0/133 S-stamped vs 3/32 unstamped) and did not significantly alter 6-month endothelial cell loss (31±17% S-stamped vs 29±14% unstamped; P = 0.62) or frequency of rebubble (17/133 S-stamped vs 1/32 unstamped; P = 0.20). CONCLUSION: The incorporation of a stromal-sided S-stamp eliminates iatrogenic primary graft failure owing to upside-down implantation of DMEK grafts, without adversely affecting early postoperative complications or 6-month endothelial cell loss.

Stamping an S on DMEK Donor Tissue to Prevent Upside-Down Grafts: Laboratory Validation and Detailed Preparation Technique Description.

PURPOSE: To report endothelial cell loss (ECL) caused by a novel S-stamp preparation technique for Descemet membrane endothelial keratoplasty (DMEK). METHODS: Six cadaveric human corneas were prepared for DMEK transplantation using a single standardized technique, including the application of a dry ink gentian violet S-stamp to the stromal side of Descemet membrane. Endothelial cell death was evaluated and quantified using computerized analysis of vital dye staining. RESULTS: ECL caused by the S-stamp was 0.6% (range 0.1%-1.0%), which comprised less than one-tenth of the total ECL caused by our preparation of the DMEK graft from the start to finish, including recovery, prestripping, S-stamping, and trephination (13.7% total ECL, range 9.9%-17.6%). CONCLUSIONS: Our novel S-stamp donor tissue preparation technique is intuitive to learn and holds the promise of preventing iatrogenic primary graft failure due to upside-down grafts without causing unacceptable increases in ECL.