title: Pneumatic dissection and storage of donor endothelial tissue for Descemet's membrane endothelial keratoplasty: a novel technique.
abstract: OBJECTIVE
  To investigate the feasibility of pneumatic dissection of donor endothelium and the effect of 7 days of storage in tissue culture medium on the endothelial cell count.
  
  
  METHODS
  Experimental study.
  
  
  METHODS
  We used 20 human donor corneoscleral tissues deemed unsuitable for transplantation.
  
  
  METHODS
  Donor corneas were mounted on an artificial anterior chamber and the anterior stroma removed with a 300-micron microkeratome head. Air was injected into the residual donor tissue with a 30-G needle from the endothelial side to detach Descemet's membrane. The "bubble" was expanded as far as possible into the periphery. A silicone weight was attached to the scleral ring and the prepared tissue was stored in tissue culture medium for 7 days.
  
  
  METHODS
  Complete detachment of Descemet's membrane, size of detachment, pre- and post-storage endothelial cell counts.
  
  
  RESULTS
  Complete detachment of Descemet's membrane was achieved in 19 of 20 (95%) cases. In 12 (60%) cases, this was achieved with a single injection; 7 (35%) cases required repeat injections. In 1 case, Descemet's membrane could not be detached despite repeat air injections. The average size of detachment was 8.11+/-2.0 mm. Endothelial cell loss after 7 days of tissue culture medium storage was 4.44+/-4.3%.
  
  
  CONCLUSIONS
  Descemet's membrane and endothelium can be separated from the overlying stroma with a simple technique using air dissection. An adequate size of graft tissue is obtained without the need to manually handle the tissue. The technique allows storage of the tissue in tissue culture medium with low endothelial cell loss.
authors.affiliation: Villa Serena Hospital, Department of Ophthalmology, Forlì, Italy. mbusin@yahoo.com
authors.forename: Massimo
authors.initials: M
authors.lastname: Busin
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Scorcia
authors.forename: Amit K
authors.initials: AK
authors.lastname: Patel
authors.forename: Gianni
authors.initials: G
authors.lastname: Salvalaio
authors.forename: Diego
authors.initials: D
authors.lastname: Ponzin
mesh_terms: D000328:Adult; D000368:Aged; D000388:Air; D002452:Cell Count; D057111:Descemet Stripping Endothelial Keratoplasty; D004210:Dissection / Q000379:methods*; D004728:Endothelium, Corneal / Q000166:cytology*; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D013997:Time Factors; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods*
chemical_list: D019188:Organ Preservation Solutions