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... Show moreAim: 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. Show less
Vasiliauskaite, I.; Jong, M. de; Quilendrino, R.; Wees, J. van der; Oellerich, S.; Melles, G.R.J. 2021
Purpose: To evaluate the suitability of corneas from septic donors for transplantation by analyzing the discard rate in the eye bank and the clinical outcome of Descemet membrane endothelial... Show morePurpose: To evaluate the suitability of corneas from septic donors for transplantation by analyzing the discard rate in the eye bank and the clinical outcome of Descemet membrane endothelial keratoplasty (DMEK) using organ-cultured corneal grafts from septic versus nonseptic donors. Methods: This retrospective study included 1554 corneas of which 456 corneas (29%) were from septic and 1072 corneas (69%) from nonseptic donors [for 26 corneas (2%) sepsis status was unknown]. The clinical outcome at 6 months after DMEK was evaluated for 82 grafts (26 from septic and 56 from nonseptic donors). Outcome measures were endothelial cell density, central corneal thickness, and postoperative complications. Results: Primary discard rates were higher for corneas from septic than from nonseptic donors (32.9% vs. 24.5%, P = 0.001). The main discard reason was poor endothelial cell quality for both septic (13.8%) and nonseptic (11.8%) donor corneas. Eye bank contamination rates for septic and nonseptic donor corneas were 1.1% and 1.7%, respectively (P = 0.102). After DMEK, donor endothelial cell density at 6m postoperatively was comparable between grafts from septic and nonseptic donors (1410 +/- 422 cells/mm(2) vs. 1590 +/- 519 cells/mm(2), P = 0.140). No differences in 6m central corneal thickness and in the rebubbling rate were observed between the 2 groups (P = 0.780 and P = 0.396, respectively). None of the cases had graft rejection nor endophthalmitis in both groups. Conclusions: Provided strict adherence to donor screening and evaluation protocols, the use of organ-cultured corneas from septic donors for DMEK does not seem to increase the risk for recipients and allows for expansion of the donor pool for corneal tissue. Show less
Groeneveld-van Beek, E.A.; Parker, J.; Lie, J.T.; Bourgonje, V.; Ham, L.; Dijk, K. van; ... ; Melles, G.R.J. 2016
