Monitoring of microhemodynamic changes during ex vivo xenogeneic liver perfusion using intravital microscopy

Abstract

The main targets of xenogeneic rejection mechanisms are the endothelial cells of the graft. Their activation and the consequent alteration of the organ's microcirculation lead to the destruction of the xenograft. Microhemodynamic changes occurring during this process are still poorly characterized. The aim of this study was to analyze the microcirculation during xenogeneic ex vivo hemoperfusion of rat livers and to monitor the impact of treatment strategies using intravital fluorescence microscopy. In contrast to the isogeneic control group, blood flow almost completely stopped within the first minutes of xenoperfusion. Simultaneously, perfusion pressure increased and bile production was reduced. Acetylsalicylate (Aspisol) and the platelet-activating factor antagonist WEB 2170 improved the microcirculation and function of the xenoperfused liver. The combination showed a synergistic effect. After apheresis of preformed xenogeneic antibodies, the parameters measured were comparable with those seen in isogeneic experiments. Complement degradation with cobra venom factor revealed a minor improvement in perfusion. A rapid, extensive, and irreversible leukocyte accumulation in terminal portal vessels was observed in all xenogeneic experiments. Blood counts of the perfusate confirmed the early trapping of leukocytes and platelets in the xenoperfused liver, indicating nonimmunological, cellular involvement in this rejection process.