title: Reproducibility of graft preparations in Descemet's membrane endothelial keratoplasty.
abstract: OBJECTIVE
  To assess the reproducibility of manual graft preparation and evaluate the incidence rate and nature of structural anomalies of Descemet's membrane (DM) preventing successful graft preparation in DM endothelial keratoplasty (DMEK).
  
  
  METHODS
  Prospective, single-center, nonrandomized, consecutive case series.
  
  
  METHODS
  We analyzed 350 corneoscleral buttons from donors aged 18-95 years stored in Optisol-GS or Dulbecco's modified Eagle's medium and used for DMEK surgery in 343 consecutive patients with Fuchs' endothelial dystrophy or pseudophakic bullous keratopathy.
  
  
  METHODS
  Residual endothelial cell-DM complexes obtained after successful DM stripping for DMEK and whole donor corneas obtained after unsuccessful DM stripping were examined by transmission electron microscopy and immunohistochemistry.
  
  
  METHODS
  Accuracy of the cleavage plane between DM and corneal stroma and structural abnormalities of the DM-stroma interface.
  
  
  RESULTS
  Uneventful manual separation without any disruption of DM was achieved in 335 of 350 donor corneas (95.7%) by use of a previously established bimanual submerged preparation technique. Correspondingly, the peeled DM specimens revealed a regular and smooth cleavage plane exposing the amorphous interfacial matrix on their anterior surface. Although 8 of 350 donor corneas (2.3%) showed focal adhesions of DM to the corneal stroma and developed isolated tears during stripping, preparation of the graft could be successfully completed. However, 7 of the 350 donor corneas (2.0%) showed extremely strong adhesion and multiple tears of DM, preventing successful preparation of the graft. These specimens revealed either ultrastructural (peg-like interlockings) or biochemical abnormalities (increased staining intensities for adhesive glycoproteins) along the DM-stroma interface.
  
  
  CONCLUSIONS
  Using an appropriate technique, manual preparation of grafts for DMEK with reproducible tissue qualities is possible in the vast majority (98%) of donor corneas. Although a relatively rare phenomenon, interindividual variations in DM structure and composition may be responsible for failure of graft preparation in about 2% of donor corneas.
  
  
  BACKGROUND
  The authors have no proprietary or commercial interest in any of the materials discussed in this article.
authors.affiliation: Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany. ursula.schloetzer-schrehardt@uk-erlangen.de
authors.forename: Ursula
authors.initials: U
authors.lastname: Schlötzer-Schrehardt
authors.forename: Bjoern O
authors.initials: BO
authors.lastname: Bachmann
authors.forename: Theofilos
authors.initials: T
authors.lastname: Tourtas
authors.forename: Claus
authors.initials: C
authors.lastname: Cursiefen
authors.forename: Matthias
authors.initials: M
authors.lastname: Zenkel
authors.forename: Kathrin
authors.initials: K
authors.lastname: Rössler
authors.forename: Friedrich E
authors.initials: FE
authors.lastname: Kruse
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D003886:Descemet Membrane / Q000378:metabolism / Q000648:ultrastructure*; D057111:Descemet Stripping Endothelial Keratoplasty / Q000379:methods*; D004728:Endothelium, Corneal / Q000378:metabolism / Q000637:transplantation / Q000648:ultrastructure*; D016326:Extracellular Matrix Proteins; D005125:Eye Banks; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D008875:Middle Aged; D009924:Organ Culture Techniques; D015676:Osteonectin / Q000378:metabolism; D011446:Prospective Studies; D012029:Refraction, Ocular / Q000502:physiology; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods / Q000592:standards; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods / Q000592:standards*; D016212:Transforming Growth Factor beta; D014792:Visual Acuity / Q000502:physiology; D019096:Vitronectin / Q000378:metabolism; D055815:Young Adult; D000099222:betaIG-H3 Protein
chemical_list: D015415:Biomarkers; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D015676:Osteonectin; D016212:Transforming Growth Factor beta; D019096:Vitronectin; D000099222:betaIG-H3 Protein