title: The role of corneal endothelial morphology in graft assessment and prediction of endothelial cell loss during organ culture of human donor corneas.
abstract: OBJECTIVE
  Endothelial assessment is crucial in the release of corneas for grafting. We retrospectively analysed the role of endothelial morphology parameters in predicting endothelial cell loss during organ culture.
  
  
  METHODS
  Human donor corneas were cultured in minimal essential medium with 2% fetal calf serum and antibiotics. Initial endothelial morphology was assessed microscopically using score parameters polymegethism (POL), pleomorphism (PLE), granulation (GRA), vacuolization (VAC), segmentation of cell membranes (SEG), Descemet's folds (DF), trypan blue-positive cells (TBPC) and endothelial cell-free areas (ECFA). Some corneas were primarily rejected based on endothelial assessment. Endothelial cell density (ECD) was assessed at the beginning (I-ECD) and end of culture. Corneas were then placed in dehydration medium (as above + 5% dextran 500). In a subgroup, ECD was reassessed after dehydration. Endothelial cell loss during culture (ECL@Culture) and culture+dehydration (ECL-Culture&Dehydration) were calculated. Data were given as mean ± SD and analysed using multiple linear and logistic regression. Odds ratios (OR) and 95% confidence intervals (CI) were calculated.
  
  
  RESULTS
  I-ECD was 2812 ± 360/mm2(n = 2356). The decision to reject a cornea due to endothelial assessment was associated negatively with I-ECD (OR = 0.77/100 cells, CI 0.7-0.82) and positively with ECFA (OR = 2.7, CI 1.69-4.35), SEG (OR =1.3, CI 1.01-1.68) and donor age (OR = 1.26/decade, CI 1.33-1.41). ECL@Culture was 153 ± 201/mm2(n = 1277), ECL@Culture&Dehydration was 169 ± 183/mm2(n = 918). ECL@Culture was associated positively with donor age, I-ECD, GRA and TBPC, and negatively with PLE, and DF. ECL@Culture&Dehydration was associated positively with age, sex, initial ECD, POL, PLE, VAC and TBPC.
  
  
  CONCLUSIONS
  Morphological parameters displayed associations with the exclusion of corneas from culture and with endothelial cell loss. Appropriate parameter selection for screening purposes may help improve graft quality.
authors.affiliation: Department of Ophthalmology and Aachen Cornea Bank, RWTH Aachen University, Aachen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Hermel
authors.affiliation: Department of Ophthalmology and Aachen Cornea Bank, RWTH Aachen University, Aachen, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Salla
authors.affiliation: Department of Ophthalmology and Aachen Cornea Bank, RWTH Aachen University, Aachen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Fuest
authors.affiliation: Department of Ophthalmology and Aachen Cornea Bank, RWTH Aachen University, Aachen, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Walter
mesh_terms: D002452:Cell Count; D002470:Cell Survival; D055954:Corneal Endothelial Cell Loss / Q000175:diagnosis* / Q000209:etiology; D004728:Endothelium, Corneal / Q000166:cytology*; D005125:Eye Banks; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D008297:Male; D008875:Middle Aged; D009924:Organ Culture Techniques / Q000379:methods*; D009926:Organ Preservation / Q000009:adverse effects / Q000379:methods*; D012189:Retrospective Studies; D013997:Time Factors
keywords: cornea donation; cornea organ culture; cornea preservation; corneal endothelial assessment; ex vivo study; eye banking; retrospective study