A Grading Score for Colon Preservation Injury in the Rat

Abstract

Background: Colon transplantation is rarely performed because of the fear for an advanced ischemic injury that may favor septic complications. Systematic studies on colon preservation are missing. The score used to evaluate the preservation injury of the colon is adapted from that used for the small intestine, despite histological and biological differences between the two organs. We studied sequentially the tissue changes in the rat colon during prolonged cold storage (CS) in histidine-tryptophan-ketoglutarate (HTK) solution and designed a grading score specific for the colon.

Methods: Large bowels of Sprague-Dawley rats (n = 9) were perfused in situ with HTK and stored at 4°C for 6 hours, 12 hours, 18 hours, and 24 hours. Samples from the proximal colon were stained with hematoxylin-eosin and alcian blue. Tight junction protein zonulla occludens (ZO)-1 was also studied.

Results: Minimal subepithelial edema (hallmark of small intestinal preservation injury) was observed throughout the 24 hours of CS. The two major changes observed during the colonic CS were progressive submucosal edema and the depletion of Goblet cells (GC). The submucosal edema was absent at 6 hours, started after 12 hours, and become significant (over 50% of the circumference) after 18 hours of CS. Depletion of GC started in the luminal half of the crypts between 12 and 18 hours of CS, and all samples revealed significant GC depletion only after 24 hours. The overall appearance of the mucosa was little affected under the CS, and ZO-1 expression was frequently maintained throughout the first 18 hours.

Conclusions: The colon is more resilient to cold ischemia than the small bowel and maintains its histological epithelial features longer than the small intestine. On the basis of these serial observations, we suggest the following grading score: grade 0: normal mucosa, repleted GC, mucosa adhering to the muscular layer; grade 1: limited submucosal edema, repleted GC; grade 2: limited submucosal edema, GC depletion in the luminal half of the crypts; grade 3: advanced (>50% of circumference) submucosal edema, GC depletion in the luminal half of the crypts; grade 4: advanced mucosal injury (edema, GC depletion, epithelial breakdown).