title: Normothermic preservation of the rat hind limb with artificial oxygen-carrying hemoglobin vesicles.
abstract: BACKGROUND
  For managing major limb amputation, it is important to consider ischemic time and reperfusion injury by free radicals after the blood supply is reestablished. State of preservation during transplant surgery is crucial for the survival and function of the tissue, graft, or organ. In this study, we confirmed the effect of intermittent blood flow in rat ischemic hind limb and developed a new oxygenic preservation method using artificial oxygen carrying hemoglobin vesicles (HbVs).
  
  
  METHODS
  We first compared a continuous ischemic model and an intermittent reflow model on rat hind limb. At postoperative day 7, hind limbs were evaluated. Next, we performed total amputation, normothermic preservation by perfusion with extracellular-trehalose-Kyoto (ETK) solution or HbV, and microsurgical replantation of the left hind limb. Venous efflux was analyzed, the amputated limb evaluated after 6 hr perfusion, and the replantation outcome of each model was compared.
  
  
  RESULTS
  In our early study, 24 hr continuous ischemic model necrotized, but intermittent reflow model almost survived except for partial necrosis at postoperative day 7. Scar tissue on the right limb showed myonecrosis and infiltration of inflammatory cells. Skeletal muscle on the right limb was structurally well maintained. Hemoglobin vesicle-treated limbs appeared to have much better oxygenation than ETK-treated limbs. Aerobic respiration remained in the amputated limb, gastrocnemius muscle was well maintained, and the overall replantation was successful in the limb preserved using HbV.
  
  
  CONCLUSIONS
  These studies demonstrated that oxygenic preservation is effective for rat ischemic limb, suggesting that this method may be useful for other replantation and transplantation surgeries.
authors.affiliation: 1 Department of Plastic Surgery, University of Tokyo, Tokyo, Japan. 2 Artificial Red Cells Group, Waseda Bioscience Research Institute in Singapore (WABIOS), Biopolis, Singapore. 3 Department of Chemistry, School of Medicine, Nara Medical University, Nara, Japan. 4 Department of Plastic Surgery, Saitama Medical Center, Saitama Medical University, Saitama, Japan. 5 Department of Plastic Surgery, University of Yamanashi Hospital, Yamanashi, Japan. 6 Department of Anatomy, Aichi Medical University, Aichi, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Araki
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sakai
authors.forename: Dai
authors.initials: D
authors.lastname: Takeuchi
authors.forename: Yu
authors.initials: Y
authors.lastname: Kagaya
authors.forename: Kensuke
authors.initials: K
authors.lastname: Tashiro
authors.forename: Munekazu
authors.initials: M
authors.lastname: Naito
authors.forename: Makoto
authors.initials: M
authors.lastname: Mihara
authors.forename: Mitsunaga
authors.initials: M
authors.lastname: Narushima
authors.forename: Takuya
authors.initials: T
authors.lastname: Iida
authors.forename: Isao
authors.initials: I
authors.lastname: Koshima
mesh_terms: D000671:Amputation, Surgical; D000818:Animals; D001802:Blood Substitutes / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D004195:Disease Models, Animal; D006085:Graft Survival; D006614:Hindlimb / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D007511:Ischemia / Q000097:blood / Q000503:physiopathology*; D008081:Liposomes; D008297:Male; D009336:Necrosis; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000378:metabolism / Q000494:pharmacology*; D010100:Oxygen / Q000097:blood*; D010108:Oxyhemoglobins / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D017208:Rats, Wistar; D020127:Recovery of Function; D012039:Regional Blood Flow; D015427:Reperfusion Injury / Q000097:blood / Q000503:physiopathology / Q000517:prevention & control; D012092:Replantation; D013997:Time Factors; D016138:Walking
chemical_list: D001802:Blood Substitutes; D008081:Liposomes; D019188:Organ Preservation Solutions; D010108:Oxyhemoglobins; D010100:Oxygen