title: Single-Pass Dissection of Ultrathin Organ-Cultured Endothelial Lamellae Using an Innovative Microkeratome System.
abstract: OBJECTIVE
  To determine the feasibility, dissection accuracy, and endothelial viability of ultrathin endothelial lamellae harvested from organ-cultured corneas using a single-pass with an innovative motor-driven linear microkeratome system.
  
  
  METHODS
  Forty-eight (n = 48) paired organ-cultured human corneas were randomly assigned to dissection (study eyes, n = 24) with fellow eyes serving as the control (fellow eyes, n = 24). After organ culture and deswelling in a medium containing 6% dextran, endothelial lamellae with a target thickness ≤100 μm were dissected using a motor-driven linear microkeratome system (SLc, Gebauer, Neuhausen, Germany) equipped with 400-μm (n = 4), 450-μm (n = 10), 500-μm (n = 5), or 550-μm (n = 5) heads. Central corneal thickness (CCT) and posterior and anterior lamellar thicknesses were measured using ultrasound pachymetry (Pachette 3; DGH Technology Inc, PA) and anterior segment optical coherence tomography (Casia SS-1000; Tomey, Nagoya, Japan). Endothelial viability [endothelial cell density (ECD)] was measured using trypan vital staining.
  
  
  RESULTS
  CCT measured 595 ± 66 μm (n = 48) on arrival, 846 ± 131 μm (n = 48) after organ culture, and 565 ± 58 μm (n = 48) after deturgescence. CCT did not differ between study and control eyes. Posterior lamellar thickness measured 88 ± 18 μm (n = 24) immediately after dissection, 126 ± 30 μm (n = 24) 1 hour after dissection, and 131 ± 41 μm (n = 24) 2.3 ± 0.6 days after dissection. ECD measured 2637 ± 264 cells per square millimeter (n = 48) on arrival, 2524 ± 232 cells per square millimeter (n = 48) after organ culture, 2493 ± 253 cells per square millimeter (n = 48) after dissection, and 2311 ± 218 cells per square millimeter (n = 48) 2.3 ± 0.6 days after dissection. ECD did not differ between study and control eyes at all time points.
  
  
  CONCLUSIONS
  Single-pass motor-driven linear microkeratome dissection provides an accurate and safe alternative for harvesting ultrathin endothelial lamellae from organ-cultured donor corneas.
authors.affiliation: *Maastricht University Medical Center, University Eye Clinic Maastricht, Maastricht, the Netherlands; and †Euro Cornea Bank, Beverwijk, the Netherlands.
authors.forename: Mor M
authors.initials: MM
authors.lastname: Dickman
authors.forename: Pieter Jan
authors.initials: PJ
authors.lastname: Kruit
authors.forename: Frank J H M
authors.initials: FJ
authors.lastname: van den Biggelaar
authors.forename: Tos T J M
authors.initials: TT
authors.lastname: Berendschot
authors.forename: Rudy M M A
authors.initials: RM
authors.lastname: Nuijts
mesh_terms: D002452:Cell Count; D003316:Corneal Diseases / Q000175:diagnosis / Q000601:surgery; D063171:Corneal Pachymetry; D057111:Descemet Stripping Endothelial Keratoplasty / Q000379:methods*; D004728:Endothelium, Corneal / Q000166:cytology* / Q000637:transplantation; D004867:Equipment Design; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009924:Organ Culture Techniques; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000295:instrumentation*; D041623:Tomography, Optical Coherence