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.

Cumulative Endothelial Cell Loss in Descemet Membrane Endothelial Keratoplasty Grafts From Preparation Through Insertion With Glass Injectors.

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.

Good syndrome and other causes of cytomegalovirus retinitis in HIV-negative patients-case report and comprehensive review of the literature.

We describe a 65-year-old Thai woman who developed cytomegalovirus retinitis (CMVR) in the setting of Good syndrome-a rare, acquired partial immune deficiency caused by thymoma. The patient subsequently developed vitritis with cystoid macular edema (CME) similar to immune recovery uveitis (IRU) despite control of the retinitis with antiviral agents. A comprehensive review of the literature through December, 2014, identified an additional 279 eyes of 208 patients with CMVR in the absence of human immunodeficiency virus (HIV) infection. Including our newly reported case, 9 of the 208 patients (4.3 %) had Good syndrome. Twenty-one of the 208 patients (10.1 %) had CMVR related to intraocular or periocular corticosteroid administration. The remaining 178 patients (85.6 %) acquired CMVR from other causes. Within the subset of patients who did not have Good syndrome or did not acquire CMVR followed by intraocular or periocular corticosteroid administration, there were many other factors contributing to a decline in immune function. The most common included age over 60 years (33.1 %), an underlying malignancy (28.7 %), a systemic autoimmune disorder requiring systemic immunosuppression (19.1 %), organ (15.2 %) or bone marrow (16.3 %) transplantation requiring systemic immunosuppression, and diabetes mellitus (6.1 %). Only 4.5 % of the patients had no identifiable contributor to a decline in immune function. While the clinical features of CMVR are generally similar in HIV-negative and HIV-positive patients, the rates of moderate to severe intraocular inflammation and of occlusive retinal vasculitis appear to be higher in HIV-negative patients.