Acta Cirúrgica Brasileira, Volume: 28, Issue: 8, Published: 2013
  • Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients Original Articles

    Gragnani, Alfredo; Müller, Bruno Rafael; Silva, Ismael Dale Contrim Guerreiro da; Noronha, Samuel Marcos Ribeiro de; Ferreira, Lydia Masako

    Abstract in English:

    PURPOSE: To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. METHODS: Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. RESULTS: The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). CONCLUSIONS: The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.
  • Ethyl-pyruvate reduces lung injury matrix metalloproteinases and cytokines and improves survival in experimental model of severe acute pancreatitis Original Articles

    Matone, Jacques; Moretti, Ana Iochabel Soares; Apodaca-Torrez, Franz Robert; Goldenberg, Alberto

    Abstract in English:

    PURPOSE: To investigate if the ethyl-pyruvate solution could reduce mortality in AP and/or diminish the acute lung injury. METHODS: Forty male rats, weighing between 270 to 330 grams were operated. An experimental model of severe AP by injection of 0.1ml/100g of 2.5% sodium taurocholate into the bilio-pancreatic duct was utilized. The rats were divided into two groups of ten animals each: CT - control (treatment with 50ml/kg of Ringer's solution, intraperitoneal) and EP (treatment with 50ml/kg of Ringer ethyl- pyruvate solution, intra-peritoneal), three hours following AP induction. After six hours, a new infusion of the treatment solution was performed in each group. Two hours later, the animals were killed and the pulmonary parenchyma was resected for biomolecular analysis, consisting of: interleukin, myeloperoxidase, MDA, nitric oxide, metalloproteinases and heat shock protein. In the second part of the experiment, another, 20 rats were randomly divided into EP and CT groups, in order to evaluate a survival comparison between the two groups. RESULTS: There were no significant differences in IL-1B,IL-10, MMP-9, HSP70, nitric oxide, MPO, MDA (lipidic peroxidation) concerning both groups. The levels of IL-6 were significantly diminished in the EP group. Furthermore, the MMP-2 levels were also reduced in the EP group (p<0.05). The animals from the EP treatment groups had improved survival, when compared to control group (p<0.05). CONCLUSION: The ethyl-pyruvate diminishes acute lung injury inflammatory response in acute pancreatitis and ameliorates survival when compared to control group, in the experimental model of necrotizing acute pancreatitis.
  • Oxidative stress evaluation of ischemia and reperfusion in kidneys under various degrees of hypothermia in rats Original Articles

    Santos, Emanuel Burck dos; Koff, Walter José; Grezzana Filho, Tomaz de Jesus Maria; De Rossi, Samanta Daiana; Treis, Lisiane; Bona, Silvia Regina; Pêgas, Karla Laís; Katz, Betina; Meyer, Fabíola Schons; Marroni, Norma Anair Possa; Corso, Carlos Otávio

    Abstract in English:

    PURPOSE:To design an animal model of ischemia-reperfusion (I/R) in kidneys and evaluate the role that predetermined ranges of local hypothermia plays on markers of stress-oxydative as well as on histologic sections. METHODS: Twenty eight male rats Wistar, under general anesthesia, undergone right nephrectomy (G0, control group) followed by left kidney ischemia during 40 min. Four temperatures groups were designed, with seven animals randomized for each group: normothermic (G1, ±37ºC), mild hypothermia (G2, 26ºC), moderate hypothermia (G3, 15ºC) and deep hypothermia (G4, 4ºC). Left kidney temperature was assessed with an intraparenchymal probe. Left nephrectomy was performed after 240 min of reperfusion. After I/R a blood sample was obtained for f2-IP. Half of each kidney was sent to pathological evaluation and half to analyze CAT, SOD, TBARS, NO3, NO2. RESULTS:Histopathology showed that all kidneys under I/R were significantly more injured than the G0 (p<0.001). TBARS had increased levels in all I/R groups compared with the G0 (p<0.001). CAT had a significant difference (p<0.03) between G1 and G4. Finally, no difference was found on SOD, NO3, NO2 nor on f2-IP. CONCLUSION: This model of I/R was efficient to produce oxidative-stress in the kidney, showing that 4ºC offered significant decrease in free radicals production, although tissue protection was not observed.
  • Effect of autologous stem cells on regenerated bone during distraction osteogenesis by Ilizarov technique in the radius of dogs: histomorphometric analysis Original Articles

    Rolim Filho, Epitácio Leite; Larrazabal, Marcelo Correia de Araújo; Costa Júnior, Lauri Ferreira da; Santos, Saulo Monteiro dos; Santos, Ricardo Monteiro dos; Aguiar, José Lamartine de Andrade

    Abstract in English:

    PURPOSE:To investigate by histomorphometry the distraction osteogenesis by Ilizarov technique in dog radius with the use of autologous stem cells in regenerated bone. METHODS:Ten dogs (20 radiuses) underwent the osteotomy of 20% of extension of their radiuses, and osteogenic distraction using the Ilizarov technique after this procedure at rate of 1mm per day divided into 0.5mm every 12 hours. The elongation was performed until the regeneration reached 20% of the total length of the radius. The stem cells were isolated, concentrated and injected in the regenerated bone, when it reached 10% of the length of the entire radius. The regenerated bone was evaluated using histomorphometric analysis when the elongation was 20% the size of radius. RESULTS: The bone formation was evidenced by histomorphometric indices were significantly greater in the study group. In the histology evaluation the type of healing was mixed in 80% (intra membrane and endocondral) in both groups; the osteoblastic activity from moderate to intense was greater in the study group; and the space occupied by the newly-formed bone tissue was more evident in the study group. CONCLUSIONS:The histomorphometric indices in this study expressing the microarchitecture, trabecular thickness, trabecular spacing, number of trabecula and quantity of bone that was significant in the group study. These data suggest that the use of undifferentiated stem cells autologous bone marrow in the regenerate bone induces osteogenesis and bone quality.
  • Effects of acupuncture and electroacupuncture on estradiol-induced inflammation and oxidative stress in health rodents Original Articles

    Santos, Elba Lucia Wanderley; Dias, Bruno Hállan Meneses; Andrade, Ana Carolina Rodrigues de; Pascoal, Angélica Maria Holanda; Vasconcelos Filho, Francisco Eugênio de; Medeiros, Francisco das Chagas; Guimarães, Sergio Botelho

    Abstract in English:

    PURPOSE:To investigate the effects of classical acupuncture (Ac) and electroacupuncture (EAc) on estradiol-induced inflammation and oxidative stress in health rodents. METHODS: Twenty-four eight-week old female rats were treated with estradiol valerate (EV) 4.0 mg i.m. single dose and randomly assigned to four groups (n=6): G1(control), G2 (Ac), G3 (EAc 2 Hz) and G4 (EAc 100 Hz). After 60 days all rats were anesthetized with chloral hydrate 10% (0.1 ml/30 g weight of the animal) and submitted to Ac/EAc for twenty minutes. The procedures were repeated on days three, five, seven and nine of the study. The equivalent of the human right ST-36 (Zusanli) and SP-6 (Sanyinjiao) acupoints were chosen for needling and electrical stimulation. On the 10th day of the experiment, all rats were anesthetized for collection of blood and tissues (ovaries) samples for biochemical analysis and histological examination. RESULTS:Glutathione (GSH) and malonaldehyde (MDA) concentrations increased significantly in all groups (plasma and ovary) while myeloperoxidase (MPO) activity decreased significantly in all groups compared with control group (G1). CONCLUSIONS:Both classical acupuncture and electroacupuncture decrease systemic and local oxidative stress and ovary inflammation in healthy rats exposed to estrogenic stimulation. EAc enhances lipid peroxidation at systemic and local levels in female rats exposed to estrogenic stimulation.
  • The effect of carbon dioxide therapy on composite graft survival Original Articles

    Durães, Eliana Ferreira Ribeiro; Durães, Leonardo de Castro; Carneiro, Fabiana Pirani; Lino Júnior, Ruy de Souza; Sousa, João Batista de

    Abstract in English:

    PURPOSE: To investigate the effect of carboxytherapy in auricular composite grafts in rabbits. METHODS: An experimental study was conducted using 20 rabbits randomly assigned to a treatment group of carboxytherapy or a control group of saline solution. In each ear, a circular graft with 1.5 cm or 2 cm of diameter was amputated and reattached. Animals underwent carbon dioxide or saline injection four times during the experiment. We analyzed clinical evolution of the animals, grafts survival, histopathology features and histomorphometry of collagen. RESULTS:The treated group had a significantly lower weight gain (p=0.038). Histopathology was not significantly different between groups. There was an increase in amount of collagen in 2 cm grafts submitted to carbon dioxide therapy (p=0.003). Carboxytherapy didn't influence graft survival rate for 1.5 cm grafts or 2 cm grafts (p=0.567 and p=0.777, respectively). CONCLUSIONS:Carbon dioxide therapy increased the amount of collagen in 2 cm grafts. CO2 was not significantly different from saline infusion on composite grafts survival, but this study suggests that there is a mechanical effect caused by distension which favored graft survival.
  • Cenostigma macrophyllum Tul. on the healing of skin wounds in rats with Diabetes mellitus Original Articles

    Coelho, Nayana Pinheiro Machado de Freitas; Nogueira, Veruska Cronemberger; Cardoso, Maria Angélica Gargione; Lopes, Luciano da Silva; Nascimento, Paulo Pedro do; Rocha, Evair dos Santos; Silva, Caio Leôncio Probo da; Arisawa, Emília Ângela Loschiavo

    Abstract in English:

    PURPOSE: To investigate the effect of Cenostigma macrophyllum Tul. in the tissue repair process of skin lesions in rats with induced Diabetes mellitus. METHODS: Sixty-three male Wistar rats were distributed into three groups: control (C), diabetic (D), and diabetic treated with an oil-in-water emulsion of the plant (DPL) subdivided according to time of observation (seven, 14, and 28 days). Diabetes was induced by administration of by injection of streptozotocin (50 mg/kg, after a 12-h fast) into the penile vein and confirmed by glucose levels exceeding 240 mg/dL. Held surgical lesion (2.5 cm in diameter) on the back of the animals. The DPL received topical application of group of the oil-in-water emulsion plant (0.5ml). To evaluate the levels of nitric oxide, was collected 5ml of blood from the abdominal aorta, after his euthanasia. The samples of interest were sent for routine histological processing. RESULTS:A reduction in the percentage of inflammatory cells and increased numbers of fibroblasts in the group DPL, seven days, compared to the other groups. At 14 days, the DPL group also showed a higher concentration of nitric oxide (p<0.01) than in groups C and D. CONCLUSION: The oil-in-water emulsion of C. macrophyllum Tul accelerated wound healing in diabetic rats.
  • Zinc sulphate administered by transdermal iontophoresis improves breaking strength of surgical wounds in skin of alloxan-induced diabetic rats Original Articles

    Cassettari, Lucas Langoni; Dias, Pedro Colli Rocha; Lucchesi, Amanda Natália; Arruda, Maurício Ferraz de; Spadella, César Tadeu

    Abstract in English:

    PURPOSE:To investigate the effect of zinc sulphate administered by transdermal iontophoresis (TDI) on mechanical resistance of surgical wounds performed in the skin of diabetic rats. METHODS:One hundred and sixty male Wistar rats weighing approximately 250g were submitted to an incision surgery at the anterior region of abdomen and randomly distributed into four experimental groups with 40 non-diabetic control animals (G1) and 40 untreated diabetic animals (G2), both without any treatment of incisions; 40 non-diabetic animals (G3) and 40 untreated diabetic animals (G4), both with incisions treated with zinc sulphate, administered for a period of four consecutive days after surgery, in sessions of ten minutes duration, using a continuous-current electrostimulator (Zn + TDI). Each experimental group was further divided into four subgroups with ten rats each to be evaluated on the 4th, 7th, 14th, and 21st day after surgery. In each period were analyzed clinical and laboratory from the animals, and measured the breaking strength and hydroxyproline content (OH-P) of the skin scars. RESULTS: Breaking strength (BS) was significantly reduced (p<0.05) in skin scars of untreated diabetic rats (G2) on the 7th, 14th, and 21st postoperative days when compared to non-diabetic control rats (G1). In contrast, BS in skin scars of non-diabetic and untreated diabetic rats (G3, G4) treated with Zn + TDI showed significant increase (p<0.05) in those periods when compared with their respective controls with untreated incisions. The OH-P content of the scars did not show statistically significant variation in all studied groups at four different times evaluated after surgery. CONCLUSIONS: Zinc sulphate administered by transdermal iontophoresis had beneficial effect on the mechanical resistance of scars produced in the skin of diabetic rats. This therapeutic may have potential to reduce the complications observed in surgical wounds of the skin in diabetic subjects, mainly in most vulnerable stages of incisions to dehiscences, leakages and infections.
  • Use of defatted flaxseed meal reduces precancerous colon lesions in C57BL/6 mice Original Articles

    Gomides, Antônio Frederico de Freitas; Paula, Sérgio Oliveira de; Rosa, Damiana Diniz; Oliveira, Leandro Licusi de; Comastri, Débora Silva; Peluzio, Maria do Carmo Gouveia

    Abstract in English:

    PURPOSE: To investigate the hemopreventive effect of defatted flaxseed meal in C57BL/6 mice after induction of precancerous colon lesions with 1.2-dimethylhydrazine (DMH). METHODS: Thirty-six 12-week-old C57BL/6 mice were divided into three treatment groups(n=12 in each group): (1) diet with 10% defatted flaxseed meal; (2) diet with defatted flaxseed meal and precancerous colon lesions induced by DMH; and (3) precancerous colon lesions induced by DMH, without defatted flaxseed meal. The incidence of aberrant crypt foci (ACF), oxidative processes, expression of tumor suppressor proteins and cyclins, as well as the profile of short-chain fatty acids (SCFA) in animal feces were investigated in the presence and absence of DMH. RESULTS: The rats consuming defatted flaxseed meals showed lesions with lower multiplicity and a reduced incidence of lesions. No changes in the expression of tumor suppressor proteins and those involved in cell cycle control were detected. CONCLUSION: Defatted flaxseed meal protected the distal colon of mice from precancerous lesions.
  • Evaluation of immunohistochemical expression of ghrelin-producing rectal cells in Wistar rats underwent to the cafeteria diet Original Articles

    Lyra Junior, Humberto Fenner; Schiavon, Leonardo de Lucca; Vieira, Daniella Serafin Couto; Teive, André Muller; Costa, Aline; Onzi, Tiago Rafael; Nau, Angélica Luciana; d'Acampora, Armando José

    Abstract in English:

    PURPOSE:To investigate the impact of cafeteria diet on ghrelin expression in rectal tissue and identify the morphologic cell type. METHODS:Twenty-four male Wistar rats were divided into four subgroups of six animals each: RC1 (rat chow 1) and CAF1 (cafeteria diet 1) for a period of 30 days; RC2 (rat chow 2) and CAF2 (cafeteria diet 2) for a period of 60 days. The animal and rectal weight, the number and the type of immunoreactive ghrelin cells were recorded and compared between the subgroups. The statistical study was established by ANOVA and Student's t test. RESULTS:There was no difference in the total of immunoreactive cells (p=0.685) between the subgroups nor between weight and presence or absence of ghrelin expression (p=0.993). All the immunoreactive cells identified were closed-type. CONCLUSION:The cafeteria diet did not have influence on the amount of immunoreactive rectal cells of ghrelin and only one type (closed-type) of immunoreactive cells was expressed in the rectum.
  • Bench surgical training with lyophilized esophageal segments Original Articles

    Sotres-Vega, Avelina; Osorio-Necoechea, María Elba; Salas-Galindo, Gabriela; González-Ramón, Soemi Careli; Guadarrama-Sánchez, Isabel; Villalba-Caloca, Jaime; Santibáñez-Salgado, José Alfredo

    Abstract in English:

    PURPOSE: To present lyophilized esophageal segments that can be used to learn surgical skills. METHODS: Four esophagus were harvested from four non-esophagus related research dogs at the moment of euthanasia. Each esophagus was trimmed in 3 cm long segments. They were lyophilized and stored during 30 days. The day programmed for surgical skills practice, they were rehydrated. RESULTS: Sixteen segments have been used. After rehydrating, all the segments kept their normal anatomic shape and structural integrity. One incision was made on every esophageal segment and sutured with running stitches of 3-0 polyglactin 910. There were no complications, such as tissue tears, nor esophageal hardening. CONCLUSIONS: The lyophilized esophagus is a high fidelity, practical, reproducible, portable, low-cost bench model. It allows general surgery apprentices to learn how to handle an esophagus, as well as to perfect their surgical and suture abilities before applying them on real patient's esophagus.
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