RESUMEN
BACKGROUND: Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is needed. HTK is a widely used preservation solution for static cold storage in liver transplantation. The present study was to investigate the beneficial effect of warm HTK donor pretreatment on liver preservation. METHODS: Male inbred Wistar rats (weighing 230-260 g) served as donors and recipients (n=6/group). Donors of treatment groups received i.v. 0.01 mL/g body weight (BW) warm (21 degree centigrade) HTK systemically 15 minutes prior to cold perfusion. Control groups received 0.01 mL/g BW warm (21 degree centigrade) NaCl 0.9%. Following pretreatment, donors were flushed with 4 degree centigrade cold HTK, livers were explanted and stored in 4 degree centigrade HTK for six hours. Thereafter orthotopic liver transplantation was performed. Recipients were harvested four hours, two and five days after reperfusion and blood and liver tissue samples were obtained. Blood samples were analyzed for AST, ALT, lactate dehydrogenase and bilirubin. Liver histological analysis as well as tissue analysis for pro-MMP2, MMP2 and pro-MMP9 using zymography was conducted. RESULTS: Treatment groups showed significantly lower ALT and lactate dehydrogenase levels as well as significantly lower activities of pro-MMP2, MMP2 and pro-MMP9. Histological analysis revealed only minor damage in all groups. CONCLUSIONS: The new concept of warm HTK pretreatment significantly reduced ischemia-reperfusion injury. The reduced ischemia-reperfusion injury was due to MMP inhibition. Warm HTK donor pretreatment is easy to handle and could further improve HTK's potency in liver preservation.
Asunto(s)
Isquemia Fría/efectos adversos , Hepatectomía , Trasplante de Hígado/métodos , Hígado/efectos de los fármacos , Hígado/cirugía , Soluciones Preservantes de Órganos/administración & dosificación , Preservación de Órganos/métodos , Daño por Reperfusión/prevención & control , Temperatura , Administración Intravenosa , Animales , Biomarcadores/sangre , Esquema de Medicación , Precursores Enzimáticos/metabolismo , Gelatinasas/metabolismo , Glucosa/administración & dosificación , Hígado/metabolismo , Hígado/patología , Trasplante de Hígado/efectos adversos , Masculino , Manitol/administración & dosificación , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Preservación de Órganos/efectos adversos , Cloruro de Potasio/administración & dosificación , Procaína/administración & dosificación , Ratas Wistar , Daño por Reperfusión/sangre , Daño por Reperfusión/etiología , Daño por Reperfusión/patología , Factores de TiempoRESUMEN
BACKGROUND: Although non-heart-beating donors have the potential to increase the number of available organs, the livers are used very seldom because of the risk of primary non-function. There is evidence that machine perfusion is able to improve the preservation of marginal organs, and therefore we evaluated in our study the influence of the perfusate temperature during oxygenated machine perfusion on the graft quality. METHODS: Livers from male Wistar rats were harvested after 60-min warm ischemia induced by cardiac arrest. The portal vein was cannulated and the liver flushed with Lifor (Lifeblood Medical, Inc.) organ preservation solution for oxygenated machine perfusion (MP) at 4, 12 or 21 degrees C. Other livers were flushed with HTK and stored at 4 degrees C by conventional cold storage (4 degrees C-CS). Furthermore two groups with either warm ischemic damage only or without any ischemic damage serve as control groups. After 6h of either machine perfusion or cold storage all livers were normothermic reperfused with Krebs-Henseleit buffer, and functional as well as structural data were analyzed. RESULTS: Contrary to livers stored by static cold storage, machine perfused livers showed independently of the perfusate temperature a significantly decreased enzyme release of hepatic transaminases (ALT) during isolated reperfusion. Increasing the machine perfusion temperature to 21 degrees C resulted in a marked reduction of portal venous resistance and an increased bile production. CONCLUSIONS: Oxygenated machine perfusion improves viability of livers after prolonged warm ischemic damage. Elevated perfusion temperature of 21 degrees C reconstitutes the hepatic functional capacity better than perfusion at 4 or 12 degrees C.
Asunto(s)
Frío , Preservación de Órganos/métodos , Perfusión/métodos , Donantes de Tejidos , Animales , Trasplante de Hígado , Masculino , Soluciones Preservantes de Órganos/farmacología , Ratas , Ratas Wistar , Recolección de Tejidos y Órganos/métodosRESUMEN
BACKGROUND: Because of the limited tolerance to portal venous clamping, the model of liver transplantation in rats represents a difficult task, which requires a great proportion of experience. Since techniques that include the introduction of an artificial stent increase the risk of thrombosis, it was our goal to modify the classical vascular hand-sewn venous anastomosis technique by using a modified end-to-end knotless procedure. MATERIALS AND METHODS: Seventy-two animals were randomly assigned to 1 of 2 experimental groups, which differed by the technique for the 3 venous anastomoses (supra- and infrahepatic vena cava, portal vein). Group 1 comprised the established suturing technique for rat liver transplantation, whereas all venous anastomosis of the second group were performed using our modified technique. RESULTS: With our method, average anhepatic time could be significantly reduced from 14 min 10 s (+/-100 s) to 11 min 40 s (+/-60 s) (P < 0.001). Kaplan-Meier survival rates demonstrated a better 7-d survival for the knotless (94%) compared to the classic technique (83%) (not significant, P = 0.137). Biliary complications were low in both groups but tended to be higher in the classical group. CONCLUSIONS: Our modified knotless anastomosis proves to be equally safe in regard to complications, improves timing, and provides excellent results in the model of orthotopic rat liver transplantation.