raffinose and Delayed-Graft-Function

Deceased donor kidneys are exposed to cold ischemic insult which makes them particularly susceptible to the effects of cold ischemic injury during hypothermic preservation resulting in high rates of delayed graft function. Bone morphogenetic protein-7 (BMP-7) is a valuable reagent in the field of tissue regeneration and preservation under ischemic conditions. Following these insights, we investigated the effect of recombinant human BMP-7 (rhBMP-7) on graft preservation during cold ischemia. The study was conducted on an experimental model of kidney cold ischemia in rats. Kidneys were perfused with University of Wisconsin (UW) saline solution, rhBMP-7, or rhBMP-7 + UW, and exposed to cold ischemia for 6, 12, and 24 hours. In tubular epithelial cells of kidneys perfused with rhBMP-7 and rhBMP-7+UW solution, the expression of BMP-7 and E-cadherin was observed after 24 hours of cold ischemia. In kidneys not perfused with rhBMP-7, high expression of transforming growth factor-β and α-smooth muscle actin was found. Also, in kidneys perfused with rhBMP-7 solution, statistically higher levels of Smad1, Smad5, and Smad8 messenger RNA expressions were proven. BMP-7 maintains the morphology of kidney tissue better than UW solution during 24 hours of cold ischemia. BMP-7 prevents epithelial to mesenchymal transformation and consequently maintains epithelial phenotype of tubular cells.

Topics: Adenosine; Allopurinol; Animals; Bone Morphogenetic Protein 7; Cold Ischemia; Delayed Graft Function; Glutathione; Insulin; Kidney Transplantation; Male; Organ Preservation Solutions; Raffinose; Rats; Reperfusion Injury

Kidneys from extended criteria donors are associated with higher rates of delayed graft function (DGF). Hypothermic machine perfusion (MP) for storage is associated with more favourable outcomes.. A retrospective analysis was performed in 93 patients where the kidney was stored using hypothermic MP (LifePort(®)) and compared to an age-matched control group where the kidney was stored in cold static storage (CSS) using University of Wisconsin solution.. Median age was similar in both groups (59.2 years in MP vs 59.9 years in CSS, p = 0.5598). Mean cold storage time was 15.6 h in MP vs 17.9 h in CSS. Post transplant mean serum creatinine was as follows; MP group-144.7 μmol/L at 1 month; 138.3 μmol/L at 3 months and 129.5 μmol/L at 12 months. In the CSS group-163 μmol/L at 1 month; 154.9 μmol/L at 3 months and 140.2 μmol/L at 12 months. There was a statistically significant difference at 1 month (p = 0.0096) and 3 months (p = 0.0236). DGF was defined as the need for haemodialysis within 7 days post transplant. In the MP group, DGF occurred in 17.2 % patients with mean of 6 days (range 1-18). In the CSS group, 25.8 % patients with mean of 8.1 days (range 3-25). One-year graft survival rate was better in the MP group (97.85 vs 96.77 %).. Our experience to date recommends the use of hypothermic MP for storage of kidneys from extended criteria deceased heart-beating donors.

Topics: Adenosine; Aged; Allopurinol; Cryopreservation; Delayed Graft Function; Female; Glutathione; Graft Survival; Humans; Insulin; Kidney Transplantation; Male; Middle Aged; Organ Preservation; Organ Preservation Solutions; Perfusion; Raffinose; Retrospective Studies; Tissue and Organ Procurement

Appropriate hypothermic packaging techniques are an essential part of organ procurement. We present a case in which deviation from standard packaging practice may have caused sub-zero storage temperatures during transport, resulting in a clinical picture resembling PNF. An 18-month-old male with alpha-1-antitrypsin deficiency underwent liver transplant from a size-matched pediatric donor. Upon arrival at the recipient hospital, ice crystals were noted in the UW solution. The transplant proceeded uneventfully with short ischemia times. Surprisingly, transaminases, INR, and total bilirubin were markedly elevated in the postoperative period but returned to near normal by discharge. Follow-up of over five yr has demonstrated normal liver function. Upon review, it was discovered that organ packaging during recovery included storage in the first bag with only 400 mL of UW solution, and pure ice in the second bag instead of slush. This suggests that the postoperative delayed graft function was related to sub-zero storage of the graft during transport. This is the first report of sub-zero cold injury, or frostbite, following inappropriate packaging of an otherwise healthy donor liver. The clinical picture closely resembled PNF, perhaps implicating this mechanism in other unexpected cases of graft non-function.

Topics: Adenosine; Allopurinol; alpha 1-Antitrypsin Deficiency; Bilirubin; Cold Temperature; Delayed Graft Function; Frostbite; Glutathione; Humans; Ice; Infant; Insulin; International Normalized Ratio; Ischemia; Liver Transplantation; Male; Organ Preservation Solutions; Postoperative Period; Raffinose; Tissue and Organ Procurement; Treatment Outcome

The aim of this study was to compare 2 preservation solutions in kidney transplant recipients in the same center during the same period since initiation of the use of High Na+; low K+ solution (Celsior).. From January 1999 to April 2011, 610 consecutive renal transplantations were done in our department with deceased donor kidneys. Data were collected prospectively. Organ procurement was performed in our center for 305 kidneys. We washed and preserved 409 kidneys in UW, and 201 in Celsior solution.. Donors criteria were worse in the Celsior group for age, male sex, creatinemia, and cold ischemia. Populations of recipients were comparable. There were no differences at 1 and 12 months in creatinine levels (p=0.9 and 0.8, respectively) and in number of delayed graft functions (DGF) (p=0.8 and relative risk =0.9) between groups. There were no differences in post-transplantation outcomes for all variables. At 5 years, graft survival was 90.4% for UW and 93.5% for Celsior (p=0.44).. Our retrospective study did not succeed in demonstrating superiority of a High Na+; low K+ solution compared to a UW type reference solution. Celsior has the same effectiveness as UW during kidney cold storage.

Topics: Adenosine; Adolescent; Adult; Aged; Allopurinol; Delayed Graft Function; Disaccharides; Electrolytes; Female; France; Glutamates; Glutathione; Graft Survival; Histidine; Humans; Insulin; Kaplan-Meier Estimate; Kidney Transplantation; Male; Mannitol; Middle Aged; Organ Preservation Solutions; Prospective Studies; Raffinose; Time Factors; Treatment Outcome; Young Adult

Prolonged cold preservation frequently causes delayed renal graft function resulting from tubular epithelial injury. Inhibition of signal transduction downstream from protein kinase C (PKC) may reduce renal ischemia-reperfusion injury and confer renal graft protection. We therefore evaluated the effect of sotrastaurin, a small-molecule inhibitor of Ca²⁺-dependent and Ca²⁺-independent PKC isoforms, in comparison with mycophenolic acid (MPA) on rat renal transplants with prolonged cold preservation.. Donor kidneys from male Lewis rats were cold stored in University of Wisconsin solution for 24 hr before syngeneic grafting. Recipients received sotrastaurin (30 mg/kg twice daily), MPA (20 mg/kg/day), or vehicle through gavage starting 1 hr after surgery. Renal function was evaluated by serum creatinine and histology on day 2 (acute injury) and day 7 (repair phase) after transplantation. Postreperfusion inflammation was determined by real-time polymerase chain reaction of proinflammatory genes and histology. Signaling mechanisms were studied by Western blotting and immunohistochemistry.. Sotrastaurin enhanced immediate transplant function, attenuated epithelial injury, and accelerated renal function recovery compared with MPA. Despite the stronger anti-inflammatory capacity of MPA, only sotrastaurin treatment achieved significant cellular protection with persisting reduced apoptosis of tubular epithelial cells. Decreased phosphorylation of extracellular signal-regulated protein kinase and p66Shc adaptor protein, both involved in cellular stress and apoptosis, were likely the responsible mechanism of action.. The PKC inhibitor sotrastaurin effectively ameliorated ischemia-reperfusion organ damage and promoted cytoprotection in a clinically relevant model of extended renal cold preservation followed by transplantation. Pharmacologic targeting of PKC may be beneficial for recipients receiving renal transplants at risk for delayed graft function.

Topics: Adenosine; Allopurinol; Animals; Apoptosis; Biomarkers; Blotting, Western; Cell Proliferation; Cold Temperature; Creatinine; Cytokines; Cytoprotection; Delayed Graft Function; Glutathione; Immunohistochemistry; Inflammation Mediators; Insulin; Kidney; Kidney Transplantation; Male; Mycophenolic Acid; Organ Preservation; Organ Preservation Solutions; Protein Kinase C; Protein Kinase Inhibitors; Pyrroles; Quinazolines; Raffinose; Rats; Rats, Inbred Lew; Real-Time Polymerase Chain Reaction; Reperfusion Injury; Signal Transduction; Time Factors

Histidine-Tryptophan-Ketoglutarate (HTK) solution is increasingly used to flush and preserve organ donor kidneys, with efficacy claimed equivalent to University of Wisconsin (UW) solution. We observed and reported increased graft pancreatitis in pancreata flushed with HTK solution, which prompted this review of transplanting HTK-flushed kidneys. We analyzed outcomes of deceased-donor kidneys flushed with HTK and UW solutions with a minimum of 12 months follow-up, excluding pediatric and multi-organ recipients. We evaluated patient and graft survival and rejection rates, variables that might constitute hazards to graft survival and renal function. Two-year patient survival, rejection, renal function and graft survival were not different, but early graft loss (<6 months) was worse in HTK-flushed kidneys (p < 0.03). A Cox analysis of donor grade, cold ischemic time, panel reactive antibodies (PRA), donor race, first vs. repeat transplant, rejection and flush solution showed that only HTK use predicted early graft loss (p < 0.04; relative risk = 3.24), almost exclusively attributable to primary non-function (HTK, n = 5 (6.30%); UW, n = 1 (0.65%); p = 0.02). Delayed graft function and early graft loss with HTK occurred only in lesser grade kidneys, suggesting it should be used with caution in marginal donors.

Topics: Adenosine; Adult; Aged; Allopurinol; Cadaver; Delayed Graft Function; Female; Glucose; Glutathione; Humans; Insulin; Kidney Function Tests; Kidney Transplantation; Male; Mannitol; Middle Aged; Organ Preservation Solutions; Potassium Chloride; Procaine; Raffinose; Renal Dialysis; Tissue Donors

Histidine-tryptophan-ketoglutarate (HTK) is replacing University of Wisconsin (UW) solution as the preservation fluid for renal allografts in many centers, but recent large-scale data to support this transition are lacking. We conducted a retrospective analysis of patient and graft outcomes after renal transplantation at our center, comparing 475 consecutive living donor and 317 deceased donor transplants since the adoption of HTK with equal numbers of grafts preserved using UW solution. Data collected included donor and recipient age, race, sex, comorbidities and graft ischemia time. Graft and patient survival, as well as the incidence of delayed graft function (DGF), were studied by Kaplan-Meier and Cox regression analysis. No significant difference was seen in either patient or graft survival. Deceased donor kidneys in the HTK group had a higher incidence of DGF than the UW cohort, whereas this trend was reversed in the case of living donor organs. In multivariate analysis, HTK was associated with a significant risk reduction on the incidence of DGF. Prolonged preservation with HTK compared to UW was not associated with excess risk to the graft or patient. In summary, HTK demonstrated efficacy similar to UW in terms of patient and graft survival.

Topics: Adenosine; Adult; Allopurinol; Delayed Graft Function; Female; Glucose; Glutathione; Graft Survival; Humans; Incidence; Insulin; Kidney Transplantation; Male; Mannitol; Middle Aged; Organ Preservation; Organ Preservation Solutions; Potassium Chloride; Procaine; Raffinose; Retrospective Studies