IN-VIVO FLUORESCENCE MICROSCOPY FOR THE ASSESSMENT OF MICROVASCULAR REPERFUSION INJURY IN SMALL-BOWEL TRANSPLANTS IN RATS

With the use of in vivo fluorescence microscopy we have analyzed microvascular reperfusion injury of small bowel isograft transplants in rats. Following 1 hr cold storage in University of Wisconsin solution, the small bowel was transplanted heterotopically, and the intestinal microcirculation was quantitatively analyzed 20-60 min after onset of reperfusion. The intestinal grafts' capillary perfusion of both the mucosa and the circular and longitudinal muscles was not found altered when compared with the intestinal capillary perfusion of sham-operated controls. In contrast, leukocyte-endothelial cell interaction, including leukocyte rolling (40+/-5%) and sticking (280+/-100 mm(-2)) in submucosal postcapillary venules, was significantly increased when compared with nontransplanted controls (12+/-8% and 20+/-10 mm(-2), P<0.01 and P<0.05, respectively). Leukocyte-endothelial cell interaction was associated with a marked alteration of lymphatic capillary drainage, as indicated by the low functional density of lymphatic microvessels of 10.2+/-6.1 cm (P<0.01 vs. sham-operated controls (39.2+/-6.1 cm(-1)). From these results we propose that leukocyte-endothelial cell interaction, not capillary ''no-reflow,'' is the primary step in the manifestation of microvascular reperfusion injury following a short period of cold ischemia in small bowel grafts.