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The main goals for a successful small bowel transplantation (SBTx) are the control of acute rejection and maintenance of the mucosal barrier, which plays a key role in preventing bacterial translocation and preserving absorptive capacity. According to recent evidence that sustaining enteral nutrition (EN) as rehabilitative therapy improves the integrity of the mucosal barrier after SBTx, we studied the trophic effect of a new elemental enteral solution whose proteinic supply is represented by oligomeric-aminoacidic chains. In a swine SBTx model we studied three groups, divided by the different postoperative feeding: group 1 (n = 5): standard swine chow, group 2 (n = 5): polymeric enteral solution, group 3 (n = 5): elemental enteral solution (Peptamen, Nestlè Corp). All animals were immunosuppressed with a tacrolimus/FK778 combined oral therapy. The nutritional indices evaluated were: body weight, episodes of diarrhea, D-xylose absorption test, and histopatological and villi morphometric analysis. Three pigs died before the end of the study, two in group 1 (pneumonia and sepsis), one in group 2 (pneumonia). Mean days of diarrhea were 15, 10, and 3 in groups 1, 2, and 3, respectively (P < .05). The final/starting weight ratio was 1.08 for group 3 and 0.92 for group 2 (P < .05); the D-xylose curves showed a statistically significant difference for group 3 versus the groups 2 and 1 (P < .05), as well as for the villi height (P < .01) and width (P < .05). In conclusion, elemental enteral solution, with its basic protein supply, does not require a very complex enzymatic system to be metabolized. Thus, it may contribute to a faster recovery of the mucosal barrier and to limit the hypercatabolic state.

Unlabelled: This study was conducted to assess apoptosis and nuclear proliferation in rat small bowel submitted to hypothermic hyperbaric oxygenation for preservation., Methods: Twenty two-month-old, male Wistar rats, weighing 250 g were divided into two groups: group I (n = 10), in which the small bowel was preserved for 12 hours, and group II (n = 10) in which the small bowel was preserved for 24 hours. After vascular and intraluminal perfusion, 3-cm segments were maintained in Ringer's solution at 2 degrees to 4 degrees C under normobaric conditions (groups Ia and IIa) or conditioned in a small hyperbaric metal chamber with 100% oxygen at 5.5 absolute atmospheres (groups Ib and IIb). After 12 or 24 hours, apoptotic and mitotic indices were evaluated by immunohistochemical methods., Results: The apoptotic index was significantly higher in small bowel segments in groups Ia and IIa compared with groups Ib and IIb. The mitotic index was significantly higher among group IIb., Conclusion: Hypothermic hyperbaric oxygenation reduced intestinal epithelial apoptosis and increased nuclear proliferation during rat small bowel preservation.

Introduction: Most centers maintain isolated human islet preparations in tissue culture to improve the safety as well as the practicality of islet transplantation. However, maintaining viability and recovery of islets remains a challenge. Extracellular matrix (ECM) is one of the most important components of the islet microenvironment. Reconstruction of the cell-matrix relationship seems to be necessary to sustain the structure and function of differentiated islets. Small intestinal submucosa (SIS), a natural ECM, is well known to promote wound healing, tissue remodeling, and cell growth. The purpose of this study was to evaluate recovery and function of isolated rat pancreatic islets during in vitro culture with SIS., Methods: Pancreatic islets isolated from Wistar rats following intraductal collagenase distension, mechanical dissociation, and EuroFicoll purification were cultured in plates coated with multilayer SIS (SIS-treated group) or without (standard cultured group) for 7 or 14 days in an islet culture media of RPMI 1640 (Gibco). The islets from both experimental groups were stained and counted with dithizone. Islet recovery following culture was determined by the ratio of counts after culture to the yield of islets immediately following islet isolation. The viability of the islets was assessed by a glucose challenge test with low glucose (2.7 mmol/L), high glucose (16.7 mmol/L), and high glucose solution supplemented with 50 micromol/L 3-isobutyl-1-methylxanthine solution. The apoptosis of islet cells was measured by relative quantification of histone-complexed DNA fragments by using enzyme-linked immunosorbent assay., Results: After 7 or 14 days of in vitro tissue culture, the recovery in SIS-treated islets group was about double of that cultured in the plates without SIS coating. In the SIS-treated group, there was no significant difference between the short- and the long-term periods of culture (95.8%+/-1.0% vs 90.8%+/-1.5%, P>.05). Following incubation with high glucose (16.7 mmol/L) solution, the insulin secretion in the SIS-treated group showed a greater increase than the control group after 14 days of culture (20.7+/-1.1 mU/L vs 11.8+/-1.1 mU/L, P<.05). When islets were placed in the high glucose solution containing IBMX, the stimulated insulin secretion was more increased in the SIS-treated than in the control group despite the duration of the culture. The calculated stimulation index of SIS-treated group was about two to three times greater than the control group. In addition, the stimulation index of the SIS-treated group remained constant regardless of short-term versus long-term culture (9.5+/-0.2 vs 10.2+/-1.2, P>.05). Much less apoptosis of islet cells occurred in the SIS-treated than in the control group., Conclusion: Coculture of isolated rat islets with native sheetlike small intestinal submucosa seemed to build an ECM for islets providing possible biotrophic and growth factors that promote the recovery and subsequent function of islets.

Aim: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) mediates the homing of lymphocytes to gut-associated tissues (GALT). We performed a semiquantitative analysis of MAdCAM-1 expression during small bowel graft rejection in rat treated with FTY720., Methods: Orthotopic small bowel transplantations (SBT) were performed from BN rats to LEW rats. Isografted animals served as controls. Three groups of SBT animals were studied on days 3, 5, 7 after operations (Isograft, untreated allograft, allograft with FTY720). FTY720 was orally administered by gavage (1 mg/kg/d) to allograft models on 7 consecutive days. Cryostat sections were prepared from grafts, including Peyer's patches (PPs). Indirect immunoperoxidase staining was performed using mAbs against MAdCAM-1. The degree of vascular endothelial staining on high endothelial venules (HEV) in the PPs was graded from 1 (low levels) to 5 (high levels), and in the vessels of the lamina propia from 1 (faint), to 2 (low at the base of villi), 3 (low to the middle of villi), 4 (high to the middle of villi), to 5 (high to villi tip)., Results: The graft survival was prolonged in the FTY720-treated group. MAdCAM-1 expression on HEVs in PPs was down-regulated during rejection. In contrast its expression on endothelial cells of vessels in the lamina propria was up-regulated during rejection. In the FTY720-treated groups, MAdCAM-1 expression on HEVs in PPs was up-regulated and its expression on endothelial cells of vessels in the lamina propria was down-regulated compared with untreated allograft group., Conclusions: Alteration in MAdCAM-1 expression may be associated with the development of SB graft rejection. The vessels at the base of villi, which are associated with lymphocyte recruitment, may become sites of intestine immune reactivity during the early phase of small bowel allograft rejection. FTY720 was found to prevent the down-regulation of MAdCAM-1 expression on HEVs in PPs and the up-regulation of its expression on endothelial cells of vessels in the lamina propria while also prolonging small bowel allograft survival.

The implantation of fragmented rat intestinal epithelium into the omentum of syngeneic animals results in the formation of a cyst containing neointestine. The purpose of our project was to study the evolution of this neointestine-containing cyst over time. Harvested jejunum and ileum of neonatal DA rats (6 to 8 days old) was digested with collagenase type XI and dispase at room temperature. The resulting organoid units, containing clusters of intestinal epithelium with stem cells were seeded onto a polyglactin polymer mesh (100000 units per mesh). The absorbable mesh was implanted in the omentum or peritoneal wall of an adult syngeneic animal. Animals were sacrificed at weekly intervals to harvest the neointestinal cysts. The lumen of the neointestine cysts was full of mucous while the wall of the cyst was covered by intestinal mucosa. H&E staining of the cyst demonstrated the morphology of intestinal epithelium; PAS staining identified goblet cells. The size of the cyst was maximal between 4 and 8 weeks postimplantation tending to regress thereafter. Neointestinal cysts are a consistent finding after implantation of intestinal epithelium organoids into the omentum or peritoneal wall in the rat model. The cysts reach a maximal size at 4 to 8 weeks postimplantation, tending to regress thereafter.

The demand for glutamine increased only in the preserved intestine in the early postoperative period (3 days after transplantation). Glutamine demand of the preserved grafts returned to control and immediate levels 7 and 14 days after transplantation. Three days after intestinal transplantation, when the intestinal mucosa was actively regenerating, the demand for glutamine was markedly enhanced. The enhanced demand for glutamine was met by increased output of glutamine by the liver and skeletal muscle. Glutamine uptake by the intestinal graft was enhanced by a brief infusion of glutamine. Thus, we believe exogenous glutamine supplementation may be beneficial for the recovery of intestinal grafts with severe mucosal injury.

Diarrhea is a frequent complication of small bowel transplantation in patients and experimental models. Although there are many potential causes, the underlying basis for most are not fully understood. Improved therapeutic approaches will come about through better understanding of mechanisms of bowel adaptation and regulation.