title: Descemet membrane endothelial keratoplasty tissue preparation from donor corneas using a standardized submerged hydro-separation method.
abstract: OBJECTIVE
  To standardize a novel submerged hydro-separation technique for Descemet membrane endothelial keratoplasty (DMEK) graft preparation from donor corneal tissues.
  
  
  METHODS
  Experimental study, laboratory investigation.
  
  
  METHODS
  
  
  
  METHODS
  The Veneto Eye Bank Foundation, Venice, Italy.
  
  
  METHODS
  Fifty-four random human donor corneal tissues unsuitable for transplantation.
  
  
  METHODS
  Donor corneas were laid in a sterile basin partially filled with tissue culture medium. A 25 gauge needle with 1 mL mounted syringe was filled with the tissue culture medium. The needle (with bevel up) was bent to 90 degrees and was inserted in the posterior cornea initiating beneath the trabecular meshwork. It was further advanced toward the midperiphery, ensuring that only the bevel was inserted, considering it as a threshold of insertion. The liquid was injected with a medium to high pressure into the posterior stroma or in the Descemet membrane-stroma interface to create the bubble. The tissues were preserved for 7 days in tissue culture medium at 31°C. Parametrical, physiological and histological analyses were carried out.
  
  
  METHODS
  Larger-diameter tissue, no tissue wastage, reproducibility, and preshipment evaluation.
  
  
  RESULTS
  Complete detachment was achieved in all the cases without any tissue wastage. Average diameter of the excised graft was 10.80 (±0.28) mm and endothelial cell loss post preservation was 11.48%. Expression of tight junction protein and regular morphology was observed post preservation. No signs of cell apoptosis were seen. Histological analysis showed elimination of residual stroma in most of the cases.
  
  
  CONCLUSIONS
  The submerged hydro-separation method reduces tissue wastage. It allows preshipment evaluation, thus allowing a validated tissue to be transported from the eye banks to the surgeon. Because of the liquid interface, the peeling of the DMEK graft becomes easy for transplantation.
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy. Electronic address: mohit.parekh@fbov.it.
authors.forename: Mohit
authors.initials: M
authors.lastname: Parekh
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ruzza
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Salvalaio
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferrari
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Camposampiero
authors.affiliation: Department of Ophthalmology, Villa Serena Hospital, Forlì, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Busin
authors.affiliation: International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Diego
authors.initials: D
authors.lastname: Ponzin
mesh_terms: D003316:Corneal Diseases / Q000601:surgery*; D003886:Descemet Membrane / Q000601:surgery* / Q000648:ultrastructure; D057111:Descemet Stripping Endothelial Keratoplasty / Q000379:methods*; D004728:Endothelium, Corneal / Q000637:transplantation* / Q000648:ultrastructure; D005125:Eye Banks; D005260:Female; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D009924:Organ Culture Techniques; D015203:Reproducibility of Results; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods*