RESUMOObjetivo: Avaliar a ação do dimetil-sulfóxido (DMSO) na necrose distal de retalhos randômicos isquêmicos em ratos. Métodos: Foram utilizados 30 ratos machos, linhagem Wistar, peso entre 220 e 363g e idade média de 3 meses. O retalho cutâneo dorsal (8x2cm) com pedículo cranial foi descolado, reposto em seu leito e suturado com poliamida 4.0. O grupo controle-CT (n=10) não recebeu nenhuma medicação, o grupo simulado-SM (n=10) recebeu o volume de 1mL de solução salina subcutânea, dividida em dez aplicações ao longo do retalho, o grupo experimento-EX (n=10) recebeu a injeção de 1ml de DMSO 5%. Após sete dias foram avaliadas as áreas de necrose distal e colhido material para o estudo histológico. Resultados: As medidas das áreas de necrose (CT=47,99, SM=58,78, EX=41,57) e as porcentagens das áreas de necrose (CT=29,98, SM=36,73, EX=23,99) mostraramse menores no grupo Ex (p< 0,05). O estudo histológico qualitativo mostrou, no grupo Ex, maior presença de neovascularização, menor desestruturação dos anexos e do estroma conjuntivo e presença de fibroblastos em período mais precoce que nos dois outros grupos. Conclusão: O DMSO teve ação benéfica sobre os retalhos cutâneos randômicos em ratos, expressos pela menor área de necrose distal e pelo aspecto histológico de reparação tecidual mais precoce. Descritores: Retalhos cirúrgicos. Isquemia. Pele. Ratos. ABSTRACT Purpose:To evaluate through the morphologic study (macro and microscopic) the effectiveness of DMSO in the genesis of the distal necrosis of random skin flaps in rats. Methods: 30 male rats were used, lineage Wistar, weight between 220 and 363g and 3 months medium age .The skin flap (8x2cm) with remained cranial vessels it was unstuck, restored at his bed and sutured with polyamide 4.0. The control-group CT (n=10) don't received any medication, the simulated group -SM (n=10) it received the volume of 1mL of subcutaneous saline solution, divided in ten applications along the skin flap, the experiment group (n=10) it received the injection of 1ml of DMSO5%. After seven days they were appraised the areas of distal necrosis and picked material for the histology study. Results: The measures of the necrosis areas (CT=47.99, SM=58.78, EX=41.57) and the percentages of the necrosis areas (CT=29.98, SM=36.73, EX =23.99) they were shown smaller in the EX group (p < 0,05). The qualitative histology study showed, in the group EX, larger angiogenic presence, smaller destruction of the enclosures and of the conjunctive stroma and presence of fibroblasts in more precocious period than in the two other groups. Conclusion: The analysis of the collected data can infer that DMSO had beneficial action on the random skin flaps, expressed by the smallest area of distal necrosis and for the histology aspect of repairing tissue more precocious.
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.
Previous studies described controversial opinions about pediatric tracheotomy concerning type of tracheal incision and long-term results, which remain as important research subjects. Experimental studies on rat tracheas are scarce, probably because of technical difficulties related to the structures' small dimensions. As many rat organ and system operative procedures were studied successfully by using microsurgical techniques, we decided to develop a pediatric tracheotomy model in growing rats which would permit long-term studies. Forty-four Wistar EPM-1 growing rats weighing 86 g and aged 35 days were divided into three groups: submitted to longitudinal, transverse, and segment excision of the trachea. Under sterile technique and intramuscular anesthesia (ketamine/xylazine), the trachea was exposed and incised, according to group, and a hand-made endotracheal cannula was inserted into the organ. This cannula was assembled using a segment of 1.5-cm-long 3 French silicone catheter passed through hexagonal-shaped silicone screen. The tracheal cannula was removed after 7 days, when we evaluated body weight, secretions, and dehiscence. In conclusion, this microsurgical tracheotomy model in growing rats is feasible, allowing studies on long-term repercussions of pediatric tracheotomy.
PURPOSE: To compare morphologically three different types of tracheotomy in growing rats, applying microsurgical technique. METHODS: EPM-1 Wistar growing rats (n=57) weighing 88gm and aged 35 days were randomized in four groups, according tracheotomy incision type (longitudinal, transverse and tracheal segment excision), and sham group. Following intramuscular anesthesia with ketamine and xylazine, the trachea was exposed and incised, according to the group, and a hand-made endotracheal cannula was inserted into the organ, under sterile conditions. This cannula was removed after 7 days, and animals have been sacrificed 30 days later. Tracheas samples were submitted to histological study, stained by hematoxylin-eosin and Masson trichrome, evaluating fibrosis, inflammatory infiltrate and epidermoid metaplasia. RESULTS: There was more frequency of inflammatory infiltrate at the tracheal epithelium in the tracheal segment excision group (87%) compared to the longitudinal (40%) and transverse (36%) incision groups (p=0.009). Evaluating epidermoid metaplasia, tracheal segment excision and the longitudinal groups presented 33% and 40%, respectively, compared to 0% of the transverse group (p=0.03). Concerning to fibrosis, in a global comparison (p=0.1) among the three groups there was no difference, however, compared to the longitudinal group the transverse group showed lower level of fibrosis (p=0.04). Sham group did not present any relevant morphologic alterations and it was used as reference pattern. CONCLUSION: Taken together, our data show that tracheal segment excision promotes more epithelium aggression and transverse tracheal incision shows less morphologic alterations.
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. (
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.
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