title: Histomorphometric evaluation of ischemia-reperfusion injury and the effect of preservation solutions histidine-tryptophan-ketoglutarate and University of Wisconsin in limb transplantation.
abstract: BACKGROUND
  The effect of cold ischemia (CI) in vascularized composite allotransplantation is unknown. We herein assess tissue-specific damage, acceptable CI time, and the effect of preservation solutions in a syngenic rat hindlimb transplant model.
  
  
  METHODS
  Lewis rat limbs were flushed and stored for 2, 10, or 30 hr CI in saline, histidine-tryptophan-ketoglutarate or University of Wisconsin preservation solution before transplantation. Morphologic alterations, inflammation, and damage of the individual tissues were analyzed on day 10 using histomorphology, confocal, light, and transmission-electron microscopy.
  
  
  RESULTS
  Two-hour CI led to mild inflammation of tissues on day 10, whereas 10-hr and 30-hr CI resulted in massive inflammation and tissue damage. Although muscle was mainly affected after prolonged CI (≥10 hr), nerve was affected in all CI groups. A perineural cell infiltrate, hypercellular appearance, pronounced vacuolization, and mucoid degeneration, appearing as Wallerian degeneration, were observed. Staining with propidium iodide and Syto 16 revealed a decrease in viable muscle cell nuclei in the anterior tibial muscle on day 10 in all groups, which was most pronounced in 10-hr and 30-hr CI animals. Transmission-electron microscopy indicated that a large number of mitochondria were degenerated in the 10-hr and 30-hr CI groups. Histidine-tryptophan-ketoglutarate preservation solution slightly decreased inflammation and tissue damage compared to University of Wisconsin-treated and saline-treated animals, especially in skin and muscle when CI times did not exceed 10 hr.
  
  
  CONCLUSIONS
  Severe inflammation and tissue damage are observed after prolonged CI in muscle and nerve. Ischemia times in vascularized composite allotransplantation should be kept as short as possible and certainly below 10 hr.
authors: 1 Center for Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, Innsbruck, Austria. 2 Department of Anatomy, Histology and Embryology, Division of Clinical and Functional Anatomy, Innsbruck Medical University, Innsbruck, Austria. 3 Department of Anaesthesiology and Critical Care Medicine, Innsbruck Medical University, Innsbruck, Austria. 4 Department of Pathology, Innsbruck Medical University, Innsbruck, Austria. 5 Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine, Baltimore, MD. 6 Address correspondence to: Stefan Schneeberger, M.D., Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, Anichstrasse 35, A-6020 Innsbruck, Austria.
authors: Theresa
authors: T
authors: Hautz
authors: Tilman
authors: T
authors: Hickethier
authors: Michael J F
authors: MJ
authors: Blumer
authors: Mario
authors: M
authors: Bitsche
authors: Johanna
authors: J
authors: Grahammer
authors: Martin
authors: M
authors: Hermann
authors: Bettina
authors: B
authors: Zelger
authors: Franka
authors: F
authors: Messner
authors: Elisabeth J
authors: EJ
authors: Pechriggl
authors: Christoph
authors: C
authors: Krapf
authors: Michael
authors: M
authors: Kimelman
authors: Gerald
authors: G
authors: Brandacher
authors: W P Andrew
authors: WP
authors: Lee
authors: Raimund
authors: R
authors: Margreiter
authors: Johann
authors: J
authors: Pratschke
authors: Stefan
authors: S
authors: Schneeberger
mesh_terms: D000241:Adenosine / Q000737:chemistry; D000493:Allopurinol / Q000737:chemistry; D000818:Animals; D050377:Cold Ischemia; D004305:Dose-Response Relationship, Drug; D005121:Extremities / Q000098:blood supply / Q000637:transplantation*; D005947:Glucose / Q000737:chemistry; D005978:Glutathione / Q000737:chemistry; D007249:Inflammation; D007328:Insulin / Q000737:chemistry; D008297:Male; D008353:Mannitol / Q000737:chemistry; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D018482:Muscle, Skeletal / Q000473:pathology; D009926:Organ Preservation / Q000295:instrumentation* / Q000379:methods; D019188:Organ Preservation Solutions / Q000737:chemistry*; D011189:Potassium Chloride / Q000737:chemistry; D011343:Procaine / Q000737:chemistry; D011887:Raffinose / Q000737:chemistry; D051381:Rats; D011917:Rats, Inbred Lew; D015427:Reperfusion Injury / Q000175:diagnosis*; D012584:Sciatic Nerve / Q000473:pathology; D013997:Time Factors
chemical_list: C039984:Bretschneider cardioplegic solution; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D008353:Mannitol; D011343:Procaine; D000493:Allopurinol; D011189:Potassium Chloride; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose