Organ Procurement in Experimental Pancreas Transplantation with Minimal Microcirculatory Impairment

Abstract

Background: Ischemia-reperfusion injury has been shown to deteriorate microcirculation in experimental pancreas transplantation. However, minor concern was taken on the impact of organ procurement in this condition. We examined the impact of a standardized technique of organ procurement on microcirculation and apoptosis in experimental pancreas transplantation. Methods: Male Lewis rats were divided into three groups: sham-operated animals without dissection of the pancreas served as controls (n = 5); animals undergoing nearly total process of organ procurement with the pancreas pedunculated on the aorta and the hepatoduodenal ligament (n = 7), and animals receiving pancreaticoduodenal transplantation. Pancreatic grafts were preserved for 6 h in cold University of Wisconsin solution (n = 7). At 1 and 2 h reperfusion and in time-matched controls, microcirculation was assessed by means of intravital fluorescence microscopy. Tissue samples were obtained after 2 h measurement and DNA breaks of acinar cells were detected by in situ nick end-labeling (TUNEL assay). The apoptotic index (apoptotic cells per high- power fields; hpf) was quantified by microscopic counting of at least 50 hpf. Results: Assessment of functional capillary density (FCD) in animals undergoing subtotal process of organ procurement revealed a slight non-significant decrease at 1 and 2 h compared with controls. In addition, leukocyte sticking to postcapillary venules (LAV) as well as the apoptotic index were found slightly increased after organ procurement compared with controls (p > 0.05). However, after pancreas transplantation the apoptotic index and the LAV were significantly increased and the FCD significantly decreased compared with both groups of non-transplanted animals (p < 0.01). Conclusions: Our validated technique of organ procurement does not negatively impact microcirculation and apoptosis in experimental pancreas transplantation.

In vivoquantification of oxygen-free radical release in experimental pancreas transplantation

“…Major care was taken to minimize the damage to the pancreas during procurement, as the generation of ROS is known to occur even at this early stage of a transplantation procedure [25]. We therefore closely followed the technique described by our group [19] which ensures that only minimal changes in microcirculation are noted after transplantation of a well‐preserved organ with short ischaemic time. To achieve clinically relevant times of ischaemia, the cold storage period for organs in this study was lengthened to 18 h. This time span has been found to have a marked effect on organ viability after reperfusion [26].…”

“…Male inbred Lewis rats (330 g) (Charles River WIGA, Sulzfeld, Germany) were used as donors and recipients. We used a modified technique [19] of syngeneic heterotopical pancreaticoduodenal transplantation firstly described by Lee et al. [20].…”