Search

Purpose:: To evaluate the effect of remote ischemic preconditioning (IPC-R) in the fetal small bowel transplantation model., Methods:: Two groups were constituted: The Isogenic transplant (ISO, C57BL/6 mice, n=24) and the allogenic transplant (ALO, BALB/c mice, n=24). In each group, the animals were distributed with and without IPC-R. It was obtained the following subgroups: Tx, IPC-R, Fk, IPC-Fk, in both strains. Intestinal grafts were stained with hematoxylin and eosin and immunohistochemically., Results:: The graft development evaluation in ISO group showed that IPC-R reduced the development compared with ISO-Tx (5.2±0.4 vs 9.0±0.8) and IPC-R-Fk increased the graft development compared with IPC-R (11.2±0.7 and 10.2±0.8). In ALO group, IPC-Fk increased the development compared with ALO-Tx and ALO with IPC-R (6.0±0.8, 9.0±1.2, 0.0±0.0, 0.5±0.3). The PCNA expression was increased in ISO group treated with Fk and IPC-R compared to other groups (12.2±0.8 vs Tx: 8.8±0.9, IPC-R: 8.0±0.4 and Fk: 9.0±0.6). The graft rejection was lower in groups treated with IPC-R (-18%), Fk (-68%) or both (-61%) compared with ALO-Tx., Conclusion:: Remote ischemic preconditioning showed benefic effect even associate with Tacrolimus on the development and acute rejection of the fetal small bowel graft in the Isogenic and Allogenic transplants.

Background: Intestinal obstruction is an abdominal disease associated to mortality, especially if complicated with sepsis. Resuscitation increases survival, although controversies remain concerning to therapeutic strategy., Methods: To assess the effects of hypertonic saline and pentoxifylline on the inflammatory response and oxidative stress, Wistar rats underwent a laparotomy loop intestinal obstruction and ischemia. After 24 h, the intestinal segment was resected (IO) without any other treatment and resuscitation/pentoxifylline were administered according to the group: Ringer's lactate (RL); hypertonic saline (HS); pentoxifylline (PTX); Ringer's lactate with pentoxifylline (RL + PTX); hypertonic saline with pentoxifylline (HS + PTX) and the control group (CG) that was not submitted to ischemia and obstruction. Mean arterial pressure (MAP) was recorded 4 times, and euthanasia was done 3 h after the resuscitation to obtain lung tissue, for malondialdehyde (MDA) by thiobarbituric acid reactive substances (TBARS) method, inflammatory cytokines were assessed using ELISA and NF-κΒ by Western blotting., Results: The initial MAP levels were higher in the RL and HS groups than in the others; however, the last measurement was similar among all the groups. IL-1β, IL-6 and CINC-1 (Cytokine-Induced Neutrophil Chemoattractant-1) were lower in the HS, PTX and HS + PTX groups compared with the IO and RL groups. IL-10 was lower in the HS + PTX group than in the IO group. NF-κB in the HS, PTX and HS + PTX groups were lower than in the IO group; NF-κB in the HS + PTX group was lower than in the RL group. MDA in the lung was lower in the HS + PTX group compared with other groups., Conclusion: Hypertonic saline and pentoxifylline, both alone and in combination, attenuated oxidative stress and the activation of NF-κB, leading to a decrease in the inflammatory response., (Copyright © 2014 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Purpose: To investigate the clinical evolution of orthotopic small bowel transplantation in outbred rats., Methods: Seventy-two outbred Wistar rats weighting from 250 to 300g were used as donor and recipient in 36 consecutives ortothopic small intestine transplantation without immunosuppression. The graft was transplanted into the recipient using end-to-side aortic and portacaval microvascular anastomosis. Procedure duration, animal clinical course and survival were evaluated. Survival shorter than four days was considered technical failure. Recipients were sacrificed with signs of severe graft rejection or survival longer than 120 days. Necropsies were performed in all recipients to access histopathological changes in the graft., Results: Median time for the procedure was 107 minutes. Six recipients (16.7%) presented technical failure. Twenty-seven recipients were sacrificed due to rejection, being nineteen (52.7%) between 7(th) and 15(th) postoperative day and eight (22.2%) between 34(th) and 47(th) postoperative day. Graft histology confirmed severe acute cellular rejection in those recipients. Uneventful evolution and survival longer than 120 days without rejection were observed in three recipients (8.3%)., Conclusion: Intestinal transplantation in outbred rats without immunosuppressant regiment accomplishes variable clinical evolution.

Purpose: To evaluate the effects of ischemic preconditioning (IPC) associate with different preservation solutions, in the protecting of gut., Methods: Four groups of 14 rats underwent laparotomy and collecting 20 cm of ileum, for preservation, at 4ºC, in Belzer (Belz), Ringer (RL), Celsior (Cs) and Custodiol (Cust) solutions, for 24 hours. Prior to collection, half of the animals in each group were subjected to IPC. During preservation, in the periods of zero, 12, 18 and 24 hours, were conducted evaluating the degree of mucosal injury and dosage of malondialdehyde acid (MDA)., Results: In all periods the RL group, with and without IPC, presented MDA values higher than the Belz and Cs. The degree of mucosal injury in the non-ipc RLgroup with 12h preservation was higher than the others; with 18 and 24h, the RL and Cust had higher degrees of damage than Cs and Belz. With IPC, in all periods, the group Cs and Belz had lower degrees of injury., Conclusion: The Celsior and Belzer solutions had better protective effects on the gut and these effects were enhanced by IPC.

Purpose: The purpose of this study was to establish morphologically the best time of vascular occlusion to induce ischemic preconditioning (IPC) for rat small bowel undergoing ischemia and reperfusion injury., Methods: After approval by the Ethics Committee, 36 EPM-1 young adult Wistar rats from 300-350 g were distributed into 6 groups: sham (S); ischemia and reperfusion (IR), with 50 minutes of cranial mesenteric artery occlusion and 30 minutes of reperfusion; IPC with 1 cycle of 2 minutes (IPC-2), 5 minutes (IPC-5), 10 minutes (IPC-10), or 15 minutes (IPC-15), followed by sustained IR. The animals anesthetized with ketamine (60 mg/kg) and xylazine (10 mg/kg) intramuscular (IM), were maintained on mattress heat, hydrated with saline (80 mL/kg), and injected with 100 IU heparin. Samples of jejunum were stained with hematoxylin and eosin (HE) and classified according to Park et al. Statistical analysis of results was performed using Kruskal-Wallis tests (P < .05)., Results: The histological evaluation showed no difference between IR and IPC15 rats (5.2 and 5, respectively; P = .84). Greater jejunal injury was observed with IPC15 (5) compared with other groups (IPC2 = 3, P = .03; IPC5 = 3.2, P = .05; IPC10 = 2.8, P = .02, respectively). There was no difference between groups IPC2 x IPC5 x IPC10., Conclusion: Morphologically, IPC with short times promoted greater intestinal protection against the IR lesion in rats.

Purpose: To evaluate tissue lesions, especially those of the intestinal innervation, in an excluded jejunal loop subjected to ischemia and reperfusion in rats., Methods: To evaluate the role of ischemia and reperfusion lesions in an excluded intestinal loop, four groups of 20 rats were set up: control group (GCEI7) and three experimental groups (GIREI7, GIREI14 and GIREI28). They were all subjected to exclusion of an intestinal segment of six centimeters in length, at a distance of 10 centimeters from the Treitz angle. The 60 animals in the three experimental groups were additionally subjected to ischemia of the vascular pedicle for 30 minutes. The control group and the experimental group GIREI7 were evaluated on the 7th day after the operation. The groups GIREI14 and GIREI28 (which also underwent ischemia) were utilized to evaluate the evolution of the lesion over time, on the 14th and 28th days after the operation, respectively. From the intestinal excluded loop, we take one ring of 0,5 cm distal and proximal, that were fixed in formaline 10% solution in order to do histological (HE) and immuno-hystochemial (PS-100) evaluation (enteric nervous system.) The distal loop was exteriorized in stoma and the proximal part closed with polipropilene 6-0., Results: It was observed a decrease in the number of ganglionic cells in the myenteric plexus in the group subjected to ischemia and reperfusion (GIREI7), in relation to the control group (GCEI7) at the 7th post-operative day (Mann-Whitney test: p = 0.0173 *. Comparing the numbers of ganglionic cells in the myenteric plexus before and after jejunal loop exclusion GCEI7 - (Wilcoxon test: p = 0.0577). GIREI7 - Comparing the numbers of ganglionic cells in the myenteric plexus before and after ischemia (*p = 0.0399). Comparing the percentage variations in ganglionic cells in the myenteric plexus on the 7th, 14th and 28th days after the procedure, in the groups GIREI7, GIREI14 and GIREI28, it was observed that there were no significant alterations. Kruskal-Wallis test: p = 0.6501., Conclusion: There was a decrease in the number of ganglionic cells in the myenteric plexus due to ischemia and reperfusion that did not recover in the late post-operative period.

Morphometric evaluation of the fetal intestinal graft in the late post-transplant period was done by using C57BL/6 mice. Under anesthesia, after hysterotomy, a transversal laparotomy was done in the fetus to harvest all intestines. Grafts, cleaned from its mesentery, and divided were transplanted into the preperitoneum space, from male mice, C57BL/6 strain. Thirty days after, mice were killed, and the graft was analyzed with HE stain and S-100 protein. All grafts as well as fetal and adult intestines were evaluated as controls. All developed grafts showed muscular layer and positivity for S-100 protein as the adult intestines. The goblet cells, absent in fetal intestines, were in larger number in isogeneic grafts than in adult intestines, even with lower villus. These findings showed sharp progress of the isogeneic graft from fetal to adult intestine, suggesting that they can be viable and might be functionally appropriate in the late period after transplantation., (Copyright (c) 2007 Wiley-Liss, Inc. Microsurgery 2007.)

To examine the apoptosis expression in the intestinal mucosa in accordance of different periods of hyperbaric oxygen (HBO) treatment, rats were submitted to 60 min of mesenteric artery and vein ischemia and 60 min of reperfusion occlusion. A group (G-IR) was the control and HBO was applied in the ischemia (GHBO-I), reperfusion (GHBO-R), and ischemia and reperfusion time (GHBO-IR). After 60 min of reperfusion, samples of small bowel were prepared for immunohistochemical expression of caspase-3. The expression of caspase-3 was significantly inferior when HBO was administered in the ischemia (0.16 +/- 0.01) in comparison with the control (0.70 +/- 0.08), but HBO in the further reperfusion (0.84 +/- 0.03) or both ischemia and reperfusion time (0.42 +/- 0.05) was significantly worse. There was a connection between HBO, small bowel I/R injury, and mucosa apoptosis. The favorable effect was obtained when HBO was administered early in the ischemia time., (Copyright (c) 2007 Wiley-Liss, Inc. Microsurgery 2007.)

In previous work, it was shown that gangliosides (Gang) have an inhibitory effect on lymphocyte proliferation as well as on delayed-type hypersensitivity response and mixed lymphocyte reaction. Therefore, we decided to examine the effect of gangliosides in acute allorejection after fetal intestinal transplantation. We used two female C57BL/6 mice on pregnancy day 19 as a source of fetal intestine. All animals were anesthetized with ketamine (70 mg/kg) and xylazine (10 mg/kg), intramuscularly. We harvested intestinal segments of 1 cm to transplant into BALB/c and C57BL/6 mice (male, weighing around 20 g) used as recipients. They were divided into groups of six animals each: isogeneic and allogeneic without treatment, or treated with tacrolimus 1 mg/kg/day, or gangliosides 3 and 9 mg/kg/day, during 7 days posttransplantation, intramuscularly. On postoperative day 7, intestinal grafts were collected and fixed in 10% formalin solution. Using an anesthetic overdose as euthanasia, we removed the intestinal grafts. Tissue samples were stained with hematoxylin-eosin for histological analysis regarding grafts development (D) and rejection (R) aspects. Data were analyzed by the Kruskal-Wallis test, considering P < or = 0.05 as significant. In the isogeneic and tacrolimus groups, we observed a very good degree of development (D = 9 +/- 0.5; D = 9 +/- 0.4, respectively), but a severe degree of rejection (R = 15 +/- 1.3) and a low degree of development (D = 1 +/- 0.8) in animals without treatment. The ganglioside groups showed D = 5 +/- 1.6 and R = 13 +/- 3.3, and D = 7 +/- 2.9 and R = 9 +/- 1.9, for the 3-mg and 9-mg groups, respectively. There was a statistically significant difference between the ganglioside groups and allogeneic groups without treatment. Based on the above data, we conclude that avascular fetal intestine transplantation is a good experimental model for studying immunological events, and that gangliosides only partially modulate the allorejection response, allowing intestinal development, mainly at the highest ganglioside dose. Maybe immunomodulation would be better observed by using isolated types of gangliosides or association with other immunosuppressive drugs., (Copyright 2006 Wiley-Liss, Inc.)

We investigated histopathological changes following murine fetal intestinal transplantation. Fetal intestine, obtained from a pregnant C57BL/6 mouse, was transplanted into BALB/c and C57Bl/6 mice. Recipients were divided into three groups: isogeneic, and allogeneic treated with 3 mg/kg/day gangliosides (Allo-a) or 9 mg/kg/day (Allo-b). One week after transplant, all grafts showed good viability, confirmed by cellular mitosis in the mucosa and a well-defined propria muscular layer. Isogeneic grafts showed a thicker muscular layer than in the Allo-a (P = 0.02) and Allo-b (P = 0.004) groups. There was no difference in number of mitotic cells among groups. Goblet cells were significantly reduced in allografts treated with 3 mg gangliosides (P = 0.013) or 9 mg gangliosides (P = 0.002) compared to isografts. Villi height was similar in all studied groups. There was no difference in positivity of the enteric nervous system among groups. Atrophy was more common in the allogeneic groups, suggesting that isografts had better development than allografts treated with gangliosides. (, (Copyright 2006 Wiley-Liss, Inc.)

Objective: To study the effects of extensive intestinal resection on growing rats, with regard to animal weight and histomorphometry of the remaining small intestine., Methods: Forty growing rats were allocated according to the extent of small intestine resection: 60%, 70%, 80%, or 90%. The animals were weighed every week and observed for 30 days. Following sacrifice the remaining small intestine was resected, fixed in 10% formol for 24 hours, embedded in paraffin, and stained using hematoxylin and eosin. The histological changes in the remaining small intestine were assessed for the length and thickness of villi, the thickness of the muscle layer, and the number of intestinal glands., Results: All growing rats showed a fall in body weight, although it was more significant with the largest intestinal resection (80% and 90%). Villus length and muscle thickness increased after 30 postoperative days in all rats, but the number of intestinal glands remained unaltered., Conclusion: Growing rats with greatest resection of small intestine (80% and 90%) had better intestinal adaptation and slower recovery of body weight.

The objective of this study was to evaluate the effect of ischemic preconditioning upon lesions produced by ischemia-reperfusion of the small intestine. Thirty EPM-1 Wistar rats were randomly distributed into three groups: ischemic preconditioning (IPC; n = 12), ischemia-reperfusion (I/R; n = 12), and control (C; n = 6). Laparotomy permitted isolation of the mesenteric artery for clamping. The animals were heparinized and hydrated. IPC was induced by: 10 minutes of ischemia followed by 10 minutes of reperfusion and then 50 minutes ischemia followed by another 30 minutes reperfusion. Group I/R was submitted to the same protocol except for the 20 minutes of preconditioning. Group C animals underwent only laparotomy for 100 minutes. After reperfusion small intestine fragments were examined histologically. Blood samples were obtained to measure LDH and lactate prior to euthanasia. Lactate values were significantly lower in the IPC as compared to I/R group, 39 versus 67 mg/dL, respectively (P < or =.05). However, neither IPC (grade 3) lesions of the mucosa versus I/R (grade 4) nor LDH values (PCI = 680, I/R = 873 U/L) were statistically different. Thus No morphological evidence of protection was observed following ischemic preconditioning.

Our objective was to evaluate intestinal ischemia-reperfusion injury in growing rats, modulated by hypothermia (I/RH) and N-acetylcysteine (NAC). We used 30 EPM-1 Wistar male rats, aged around 35 days, weighing 90 g. Rats were randomized into 5 groups with 6 animals in each: I/RH group, intestinal ischemia under hypothermia for 40 min and reperfusion for 30 min; I/RH-NAC group, same procedure but adding NAC (150 mg x kg(-1)), previously with ischemia; S-H group, topic hypothermia for 40 min, and observation for 30 min; I/R H-Ve group; and S-NAC group, NAC administration and observation for 70 min. All animals were heparinized and anesthetized with ketamine (60 mg kg(-1)) and xylazine (10 mg kg(-1)) intramuscularly. Surgical procedures were done under microsurgical technique (augmentation, 10x). After laparotomy, the superior mesenteric artery was dissected and clamped to promote ischemia. Topic hypothermia was obtained by using plastic bags at 4 degrees C, changed every 10 min. Rats were sacrificed by exsanguination, and blood samples were utilized to measure D(-)lactate. Intestinal fragments were removed for morphological study. Statistical analysis was done with nonparametric tests (P

The aim of this study was to evaluate the early morphological development and acute rejection process in fetal intestine allografts. Grafts from C57BL/6 fetal intestines were implanted in an avascular form in BALB/C recipients. A syngeneic group of animals was used to compare the evolution. The allogeneic recipients were distributed in 6 groups, according to the day of sacrifice (3rd, 4th, 5th, 6th, 7th, and 10th postoperational day (POD)) and the control group on the 2nd, 5th, and 7th POD. These grafts were stained with hematoxylin and eosin for histological evaluation, in agreement with the classification of Auber et al. (Chirurgie 123:122-130, 1998). Data showed a progressive development of the graft until POD 5. On POD 3 and 4, a top grade of development and an initial rejection were observed. From POD 5-7 and on POD 10, the acute rejection reaction was more important than the development process. The higher level of rejection was observed on POD 10, and it was similar to the 7th POD. Our results showed good graft development until POD 5. After that, the acute rejection response impeded analysis of the development process., (Copyright 2003 Wiley-Liss, Inc.)