title: Effect of Cold Preservation on Chronic Rejection in a Rat Hindlimb Transplantation Model.
abstract: BACKGROUND
  Previous studies on solid organ transplantation have shown that cold ischemia contributes to the development of chronic allograft vasculopathy. The authors evaluated the effect of cold ischemia on the development of chronic rejection in vascularized composite allotransplantation.
  
  
  METHODS
  Thirty rat hindlimbs were transplanted and divided into two experimental groups: immediate transplantation and transplantation after 7 hours of cold ischemia. The animals received daily low-dose immunosuppression with cyclosporine A for 2 months. Intimal proliferation, arterial permeability rate, leukocyte infiltration, and tissue fibrosis were assessed. The CD3, CD4, CD8, CD20, and CD68 cells per microscopic field (200×) were counted, and C4d deposition was investigated. Cytokine RNA analysis was performed to measure tumor necrosis factor-α, interleukin-6, and interleukin-10 levels.
  
  
  RESULTS
  Significant differences were found in the intimal proliferation and arterial permeability rate between the two groups (p = 0.004). The arterial permeability rate worsened in the most distal and small vessels (p = 0.047). The numbers of CD3, CD8, CD20, and CD68 were also statistically higher in the cold ischemia group (p < 0.05, all levels). A trend toward significance was observed with C4d deposition (p = 0.059). No differences were found in the RNA of cytokines.
  
  
  CONCLUSIONS
  An association between cold ischemia and chronic rejection was observed in experimental vascularized composite allotransplantation. Chronic rejection intensity and distal progression were significantly related with cold ischemia. The leukocyte infiltrates in vascularized composite allotransplantation components were a rejection marker; however, their exact implication in monitoring and their relation with cold ischemia are yet to be clarified.
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: Jorge
authors.initials: J
authors.lastname: Bonastre
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: Luis
authors.initials: L
authors.lastname: Landín
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: Pedro
authors.initials: P
authors.lastname: Bolado
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: César
authors.initials: C
authors.lastname: Casado-Sánchez
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: Eduardo
authors.initials: E
authors.lastname: López-Collazo
authors.affiliation: Madrid, Spain.|From the Division of Plastic and Reconstructive Surgery, Tumor Immunology Lab and Innate Immunity Group, IdiPAZ, and the Biostatistics Section, University Hospital "La Paz."
authors.forename: Jesús
authors.initials: J
authors.lastname: Díez
mesh_terms: D000818:Animals; D002199:Capillary Permeability / Q000502:physiology; D050377:Cold Ischemia / Q000009:adverse effects*; D006084:Graft Rejection / Q000209:etiology*; D006614:Hindlimb / Q000637:transplantation; D007165:Immunosuppression Therapy; D012313:RNA / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D014021:Tissue Preservation / Q000379:methods*; D063986:Vascularized Composite Allotransplantation
chemical_list: D012313:RNA