USE OF DECELLULARIZED HUMAN AMNIOTIC MEMBRANE IN INTESTINAL ANASTOMOSES: A STUDY IN RATS TREATED WITH 5-FLUOROURACIL / USO DE MEMBRANA AMNIÓTICA HUMANA DESCELULARIZADA EM ANASTOMOSES INTESTINAIS: ESTUDO EM RATOS TRATADOS COM 5-FLUOROURACIL

Introduction: currently, the most relevant applications of perinatal derivatives are in wound healing, for example, the use of amniotic membrane (AM) as a biomaterials source for use in different healing processes. The anastomotic healing compromised by antimetabolic drugs, such as 5-fluorouracil (5-FU), is a potential target of AM. Objective: To evaluate the healing effects of the AM use in rats treated with 5-FU at a dose of 20 mg/kg on the 7th postoperative day, regarding the parameters percentage of type I collagen (mature), cell viability, microvascular density and granulation tissue formation. Method: Thirty-two Wistar rats were submitted to colotomy and colorraphy, separated in 4 groups with 8t rats each and received, by 7 days, different treatments intraperitoneally, once daily from the operation day until sacrifice: 1- saline solution (C), 2- 20 mg/kg 5-FU, 3- 20 mg/kg 5-FU and 4- MA, and MA. Results: Tthe treatment with 5-FU (20 mg/kg ­ 7 days) induced adverse effects in the anastomotic healing process evidenced by the decrease of type I collagen (mature), cell viability, percentage microvascular density, granulation tissue formation (leukocyte-fibrin crust and angio-fibroblastic proliferation). The use of AM, under these conditions, induced an improvement in type I collagen (mature) percentage. Conclusion: The treatment with 5-FU induced adverse effects on the healing anastomotic process, and the use of AM, under these conditions, induced improvement in type I (mature) collagen percentage.

Introdução: Atualmente emerge em nosso meio aplicações mais relevantes dos derivados perinatais, por exemplo, a membrana amniótica (MA), como fonte de biomateriais para emprego em diferentes processos cicatriciais. O comprometimento anastomótico por drogas antimetabólicas como o 5-fluorouracil (5-FU) é um potencial alvo da MA. Objetivo: Avaliar os efeitos cicatriciais do uso da MA em ratos tratados com 5-FU na dose de 20 mg/kg ao 7º. dia de evolução pós-operatória quanto aos parâmetros porcentagem de colágeno tipo I (maduro), viabilidade celular, densidade microvascular e formação do tecido de granulação. Método: Utilizaram-se 32 ratos da linhagem Wistar submetidos à colotomia e colorrafia separados em 4 grupos com 8 e receberam diferentes tratamentos diariamente por via intraperitoneal até o dia do sacrifício: Solução fisiológica (C), 20  mg/kg 5-FU, 20 mg/kg 5-FU e MA, MA. Resultados: O tratamento com 20 mg/kg de 5-FU, ao 7º. dia pós-operatório induziu efeitos adversos no processo cicatricial anastomótico evidenciados pela diminuição da porcentagem de colágeno tipo I (maduro), da viabilidade celular, da densidade microvascular, da formação de crosta fibrinoleucocitária e da proliferação angiofibroblástica; o uso da MA nestas condições induziu melhora da porcentagem de colágeno tipo I (maduro). Conclusão: O tratamento com 20 mg/kg de 5-FU, ao 7º. dia pós-operatório induziu efeitos adversos no processo cicatricial anastomótico e o uso da MA nestas condições induziu melhora da porcentagem de colágeno tipo I (maduro).

Effects of arginine on abdominal wall wound healing in Wistar rats. / Efeitos da arginina na reparação cicatricial da parede abdominal em ratos Wistar.

OBJECTIVE: to evaluate the effects of arginine on abdominal wall healing in rats. METHODS: we submitted 20 Wistar rats to laparotomy and divided them into two groups, arginine and control, which then received, respectively, daily intraperitoneal treatment with arginine (300mg/kg/day) and weight-equivalent phosphate buffered solution, during five days. On the seventh postoperative day, we collected blood and scar wall samples from both groups. We evaluated serum nitrate and nitrite levels, wound evolution by tissue hydroxyproline dosages, granulation tissue formation, percentage of mature and immature collagen, myofibroblast density and angiogenesis. We used the ANOVA and the Student's t tests with p=0.05 for comparisons between groups. RESULTS: there were no significant differences between the groups studied for nitrate and nitrite (p=0.9903), tissue hydroxyproline (p=0.1315) and myofibroblast density (p=0.0511). The arginine group presented higher microvascular density (p=0.0008), higher percentage of type I collagen (p=0.0064) and improved granulation tissue formation, with better angiofibroblastic proliferation rates (p=0.0007) and wound edge reepithelization (p=0.0074). CONCLUSION: in the abdominal wall healing evaluation of Wistar rats under arginine treatment, there was no change in serum nitrate and nitrite levels, total collagen deposition and myofibroblast density. There was an increase in type I collagen maturation, microvascular density and improvement in scar granulation tissue formation by better edge reepithelization and angiofibroblastic proliferation.

OBJETIVO: avaliar os efeitos da arginina na cicatrização da parede abdominal de ratos Wistar. MÉTODOS: vinte ratos Wistar foram submetidos à laparotomia e separados em dois grupos (arginina e controle), que receberam tratamento diário por via intraperitoneal com arginina (300mg/kg/dia) e solução tampão fosfato em dose equivalente ao peso, respectivamente, durante cinco dias. No sétimo dia pós-operatório, coletaram-se amostras de sangue e da cicatriz da parede abdominal de ambos os grupos. Avaliaram-se o nível sérico de nitratos e nitritos, a evolução cicatricial pelas dosagens de hidroxiprolina tecidual, formação de tecido de granulação, determinação da porcentagem de colágeno maduro e imaturo, densidade de miofibroblastos e angiogênese. Empregaram-se os testes de ANOVA e t de Student com p=0,05 para as comparações entre os grupos. RESULTADOS: não ocorreram diferenças significantes entre os grupos estudados para dosagens de nitratos e nitritos (p=0,9903), hidroxiprolina tecidual (p=0,1315) e densidade de miofibroblastos (p=0,0511). O grupo arginina apresentou maior densidade microvascular (p=0,0008), maior porcentagem de colágeno tipo I (p=0,0064) e melhora na formação do tecido de granulação, com melhores índices de proliferação angiofibroblástica (p=0,0007) e re-epitelização das bordas (p=0,0074). CONCLUSÃO: na avaliação cicatricial da parede abdominal de ratos Wistar sob tratamento com arginina, não houve alteração do nível sérico de nitratos e nitritos, da deposição de colágeno total e da densidade de miofibroblastos. Verificaram-se aumento da maturação de colágeno do tipo I, da densidade microvascular e melhora na formação do tecido de granulação cicatricial pelas melhores re-epitelização de bordas e proliferação angiofibroblástica.

Efeitos da arginina na reparação cicatricial da parede abdominal em ratos Wistar / Effects of arginine on abdominal wall wound healing in Wistar rats

RESUMO Objetivo: avaliar os efeitos da arginina na cicatrização da parede abdominal de ratos Wistar. Métodos: vinte ratos Wistar foram submetidos à laparotomia e separados em dois grupos (arginina e controle), que receberam tratamento diário por via intraperitoneal com arginina (300mg/kg/dia) e solução tampão fosfato em dose equivalente ao peso, respectivamente, durante cinco dias. No sétimo dia pós-operatório, coletaram-se amostras de sangue e da cicatriz da parede abdominal de ambos os grupos. Avaliaram-se o nível sérico de nitratos e nitritos, a evolução cicatricial pelas dosagens de hidroxiprolina tecidual, formação de tecido de granulação, determinação da porcentagem de colágeno maduro e imaturo, densidade de miofibroblastos e angiogênese. Empregaram-se os testes de ANOVA e t de Student com p=0,05 para as comparações entre os grupos. Resultados: não ocorreram diferenças significantes entre os grupos estudados para dosagens de nitratos e nitritos (p=0,9903), hidroxiprolina tecidual (p=0,1315) e densidade de miofibroblastos (p=0,0511). O grupo arginina apresentou maior densidade microvascular (p=0,0008), maior porcentagem de colágeno tipo I (p=0,0064) e melhora na formação do tecido de granulação, com melhores índices de proliferação angiofibroblástica (p=0,0007) e re-epitelização das bordas (p=0,0074). Conclusão: na avaliação cicatricial da parede abdominal de ratos Wistar sob tratamento com arginina, não houve alteração do nível sérico de nitratos e nitritos, da deposição de colágeno total e da densidade de miofibroblastos. Verificaram-se aumento da maturação de colágeno do tipo I, da densidade microvascular e melhora na formação do tecido de granulação cicatricial pelas melhores re-epitelização de bordas e proliferação angiofibroblástica.

ABSTRACT Objective: to evaluate the effects of arginine on abdominal wall healing in rats. Methods: we submitted 20 Wistar rats to laparotomy and divided them into two groups, arginine and control, which then received, respectively, daily intraperitoneal treatment with arginine (300mg/kg/day) and weight-equivalent phosphate buffered solution, during five days. On the seventh postoperative day, we collected blood and scar wall samples from both groups. We evaluated serum nitrate and nitrite levels, wound evolution by tissue hydroxyproline dosages, granulation tissue formation, percentage of mature and immature collagen, myofibroblast density and angiogenesis. We used the ANOVA and the Student's t tests with p=0.05 for comparisons between groups. Results: there were no significant differences between the groups studied for nitrate and nitrite (p=0.9903), tissue hydroxyproline (p=0.1315) and myofibroblast density (p=0.0511). The arginine group presented higher microvascular density (p=0.0008), higher percentage of type I collagen (p=0.0064) and improved granulation tissue formation, with better angiofibroblastic proliferation rates (p=0.0007) and wound edge reepithelization (p=0.0074). Conclusion: in the abdominal wall healing evaluation of Wistar rats under arginine treatment, there was no change in serum nitrate and nitrite levels, total collagen deposition and myofibroblast density. There was an increase in type I collagen maturation, microvascular density and improvement in scar granulation tissue formation by better edge reepithelization and angiofibroblastic proliferation.

EFFECTS OF RENAL DYSFUNCTION ON HEALING OF COLONIC ANASTOMOSIS: EXPERIMENTAL STUDY IN WISTAR RATS.

BACKGROUND: Chronic kidney disease affects more than 500 million people worldwide. In this context, the uremic toxins present are related to worsening in tissue healing. AIM: Evaluate on healing of colonic anastomosis in uremic rats, serum and anatomopathological indicators, which may be related to the change tissue repair process. METHODS: Twenty Wistar rats, were randomly separated into two groups. In the sham group they were submitted to 5/6 nephrectomy simulation in left kidney, simulation right nephrectomy, median laparotomy, colotomy and colorraphy. In the uremia group, they were submitted to 5/6 nephrectomy of the left kidney, total nephrectomy of the right kidney and median laparotomy, colotomy and colorraphy. Were collected for serum urea, creatinine and CRP dosages and the colonic segments were studied for evaluation of granulation tissue, collagen maturation, microvascular and myofibroblasts density, and cell viability. Through histochemical processing, microvascular density was evaluated by anti-CD34 monoclonal antibody marking, cell viability by cell proliferation nuclear antigen screening and myofibroblasts density with monoclonal anti-α-actin antibody. Computerized histometry was used for evaluations of collagens type I and III by the coloration of picrosirius. RESULTS: The group submitted to nephrectomy 5/6, compared to the sham group, show urea increase (p<0.0000) and higher C reactive protein (p=0.0142). Decrease of granulation tissue formation (border reepithelialization p=0,0196, angiofibroblast proliferation p=0.0379), mean collagen I (p=0,0009) and collagen III (p=0,016), microvascular density (p=0,0074), cell proliferation nuclear antigen (p<0,0000) and myofibroblasts (p<0,0001). CONCLUSION: The uremia induced by nephrectomy 5/6 model establishes negative impact in the colonic wound healing.

Effects of renal dysfunction on healing of colonic anastomosis: experimental study in wistar rats / Efeitos da disfunção renal na cicatrização de anastomoses colônicas: estudo experimental em ratos wistar

ABSTRACT Background: Chronic kidney disease affects more than 500 million people worldwide. In this context, the uremic toxins present are related to worsening in tissue healing. Aim: Evaluate on healing of colonic anastomosis in uremic rats, serum and anatomopathological indicators, which may be related to the change tissue repair process. Methods: Twenty Wistar rats, were randomly separated into two groups. In the sham group they were submitted to 5/6 nephrectomy simulation in left kidney, simulation right nephrectomy, median laparotomy, colotomy and colorraphy. In the uremia group, they were submitted to 5/6 nephrectomy of the left kidney, total nephrectomy of the right kidney and median laparotomy, colotomy and colorraphy. Were collected for serum urea, creatinine and CRP dosages and the colonic segments were studied for evaluation of granulation tissue, collagen maturation, microvascular and myofibroblasts density, and cell viability. Through histochemical processing, microvascular density was evaluated by anti-CD34 monoclonal antibody marking, cell viability by cell proliferation nuclear antigen screening and myofibroblasts density with monoclonal anti-α-actin antibody. Computerized histometry was used for evaluations of collagens type I and III by the coloration of picrosirius. Results: The group submitted to nephrectomy 5/6, compared to the sham group, show urea increase (p<0.0000) and higher C reactive protein (p=0.0142). Decrease of granulation tissue formation (border reepithelialization p=0,0196, angiofibroblast proliferation p=0.0379), mean collagen I (p=0,0009) and collagen III (p=0,016), microvascular density (p=0,0074), cell proliferation nuclear antigen (p<0,0000) and myofibroblasts (p<0,0001). Conclusion: The uremia induced by nephrectomy 5/6 model establishes negative impact in the colonic wound healing.

RESUMO Racional: A doença renal crônica atinge mais de 500 milhões de pessoas em todo o mundo. Neste contexto, as toxinas urêmicas estão relacionadas ao comprometimento da cicatrização tecidual. Objetivo: Avaliar, na cicatrização de anastomoses colônicas de ratos urêmicos indicadores séricos e anatomopatológicos que possam estar relacionados com alteração do processo de reparação tissular. Métodos: Utilizaram-se 20 ratos Wistar divididos aleatoriamente em dois grupos. No grupo simulação eles foram submetidos à simulação da nefrectomia 5/6 do rim esquerdo, simulação de nefrectomia total do rim direito, laparotomia mediana, colotomia e colorrafia. No grupo uremia, eles foram submetidos à nefrectomia 5/6 do rim esquerdo, nefrectomia total do rim direito, laparotomia mediana, colotomia e colorrafia. Coletaram-se amostras de sangue para dosagens séricas da ureia, creatinina e proteína C reativa, e do cólon para processamentos histológicos e histoquímicos na avaliação do tecido de granulação, maturação de colágeno, densidade microvascular e de miofibroblastos, viabilidade celular cicatricial. Empregou-se a histometria computadorizada para as avaliações de colágenos tipos I e III, densidade microvascular pela marcação com anticorpo monoclonal anti-CD34, viabilidade celular pela pesquisa do antígeno nuclear de proliferação celular e a densidade de miofibroblastos com anticorpo monoclonal anti-α-actina. Resultados: O grupo submetido à nefrectomia 5/6, em comparação ao grupo simulação, demonstraram aumentos da ureia sérica (p<0,0000) e proteína C reativa (p=0,0142), redução da formação de tecido de granulação (reepitelização de bordas p=0,0196, proliferação angiofibroblástica p=0,0379), porcentagens de colágeno I (p=0,0009) e colágeno III (p=0,016), densidade microvascular (p=0,0074) e miofibroblastos (p<0,0001) e antígeno nuclear de proliferação celular (p<0,0000). Conclusão: A uremia induzida pelo modelo de nefrectomia 5/6 determina impacto negativo no processo de cicatrização colônico.

Perfil Epidemiológico do Trauma Facial em um Hospital Terciário / Epidemiological Study of Facial Trauma in a Terciary Hospital

O trauma facial é prevalente na emergência e pode trazer múltiplas repercussões para o indivíduo. O objetivo desse trabalho é avaliar o perfil epidemiológico de trauma facial em um hospital terciário. Trata-se de um estudo retrospectivo analítico, com dados de 58 prontuários de vítimas de fraturas faciais traumáticas entre janeiro de 2014 a janeiro de 2016, diagnosticadas através de exame físico e exames de imagem. As variáveis analisadas: sexo, idade, etiologia, topografia das fraturas, lesões associadas, período de internamento e estado geral. Homens foram mais acometidos, o mecanismo de trauma mais frequente foi lesão direta e a maioria das fraturas foi individualizada. O número de fraturas faciais múltiplas variou conforme a idade e com o mecanismo etiológico. O período de internamento foi curto. A pesquisa nessa área tem grande importância no sentido de estruturar os serviços de saúde, de modo que atuem reduzindo danos

Facial trauma is prevalent in the emergency room and can have multiple repercussions for the individual. The objective of this study is to evaluate the epidemiological profile of facial trauma in a tertiary hospital. This is a retrospective analytical study, with data from 58 medical records of victims of traumatic facial fractures between January 2014 and January 2016, diagnosed through physical examination and imaging. The variables: sex, age, etiology, fracture topography, associated lesions, period of hospitalization and general condition. The men were more affected, the most frequent trauma mechanism was direct injury and most of the fractures were individualized. The number of multiple facial fractures varied according to age and etiological mechanism. The period of hospitalization was short. Research in this area has great importance in order to prioritize and structure health services so that they act to reduce damages

Vertical gastrectomy in overweight and non-overweight rats.

PURPOSE:: To compare the two lines suture (total and seromuscular) after partial gastrectomy in normal and overweight rats. METHODS:: Forty Wistar rats were distributed in two groups. Group A received normal diet; group B, normal diet and supplementation with saccharose in the water. When group B progressed to a statistically greater weight than the animals of group A, the experiment (sleeve-like gastrectomy) was conducted with gastrorraphy in two sutures lines (total and seromuscular).The animals were distributed into two subgroups of 10. A1 and A2 subgroups were sacrificed at 7 and 14 days postoperatively as well as B1 and B2. Mortality, morbidity, complications attributed to the gastric suture, biochemical dosages, Lee index, macroscopy, weight of retroperitoneal and gonadal fat, optical microscopy with hematoxylin-eosin and picrosirius-red, were the evaluation parameters. RESULTS:: The overweight group achieved statistically greater weight after 16 weeks in induced obesity; there was no mortality or complications with clinical consequences attributable to morbidity. The overweight group had statistically greater weight of gonadal and retroperitoneal fat. The difference was observed in urea, albumin, total cholesterol and indirect bilirubin. CONCLUSION:: There was no outcome difference between the overweight and non-overweight group in two suture lines in gastrorrhaphy after sleeve-like gastrectomy.

Vertical gastrectomy in overweight and non-overweight rats

ABSTRACT PURPOSE: To compare the two lines suture (total and seromuscular) after partial gastrectomy in normal and overweight rats. METHODS: Forty Wistar rats were distributed in two groups. Group A received normal diet; group B, normal diet and supplementation with saccharose in the water. When group B progressed to a statistically greater weight than the animals of group A, the experiment (sleeve-like gastrectomy) was conducted with gastrorraphy in two sutures lines (total and seromuscular).The animals were distributed into two subgroups of 10. A1 and A2 subgroups were sacrificed at 7 and 14 days postoperatively as well as B1 and B2. Mortality, morbidity, complications attributed to the gastric suture, biochemical dosages, Lee index, macroscopy, weight of retroperitoneal and gonadal fat, optical microscopy with hematoxylin-eosin and picrosirius-red, were the evaluation parameters. RESULTS: The overweight group achieved statistically greater weight after 16 weeks in induced obesity; there was no mortality or complications with clinical consequences attributable to morbidity. The overweight group had statistically greater weight of gonadal and retroperitoneal fat. The difference was observed in urea, albumin, total cholesterol and indirect bilirubin. CONCLUSION: There was no outcome difference between the overweight and non-overweight group in two suture lines in gastrorrhaphy after sleeve-like gastrectomy.

Effects of nutritional supplementation with l-arginine on repair of injuries due to muscle strain: experimental study on rats.

OBJECTIVE: To evaluate the influence of oral supplementation with arginine on regeneration of injuries due to straining of the anterior tibial muscle of rats. METHODS: Twenty-four Wistar rats of weight 492.5 ± 50.45 g were used. Injuries were induced through straining the anterior tibial muscles. The rats were separated into three groups of eight rats each. In the untreated group (UTG), after induction of injuries, the rats were observed for 24 h. In the simulation group (SG) and the arginine group (AG) respectively, the rats received isotonic saline solution and arginine solution via direct gavage, over a seven-day period. At the end of the period, blood samples were collected for serum evaluations of creatine kinase (CK), lactic dehydrogenase (LDH), aspartate aminotransferase (AST) and C-reactive protein (CRP). The right and left anterior tibial muscles were resected for histopathological evaluations on the muscle injuries, investigating edema, hemorrhage and disorganization or morphometric alteration of the muscle fibers. The tissue repair was investigated in terms of proliferation of adipose tissue, angiogenesis and collagen fibers. The ANOVA and Student's t methods were used and p ≤ 0.05 was taken to be statistically significant. RESULTS: In the serum evaluations, the AG showed lower CK assay values and higher AST values. In the histopathological evaluation, the UTG presented edema and hemorrhage compatible with injuries due to strain; the SG presented edema and hemorrhage with proliferation of adipose tissue and collagen fibers; and the AG presented not only the findings of the SG but also, especially, intense angiogenesis. CONCLUSION: Oral supplementation with arginine did not cause any significant metabolic alterations that would contraindicate its use and it induced angiogenesis during the repair of muscles injured due to strain.

OBJETIVO: Avaliar a influência da suplementação oral com arginina na regeneração de lesão por estiramento do músculo tibial anterior de ratos. MÉTODO: Usaram-se 24 ratos Wistar (492,5 ± 50,45 gramas), induzidos com lesão por estiramento dos músculos tibiais anteriores e separados em três grupos com oito ratos cada. No grupo não tratado (GNT), após a indução das lesões, os ratos foram observados por 24 horas, nos grupos simulação (GS) e arginina (GA) receberam, por gavagem diariamente, respectivamente solução salina isotônica e solução de arginina, durante sete dias. Ao término dos períodos foram coletadas amostras de sangue para as avaliações séricas de creatina-quinase (CK), desidrogenase lática (LDH), aspartato-aminotransferase (AST) e proteína C reativa (PCR). Foram ressecados os músculos tibiais anteriores (direitos e esquerdos) para avaliações histopatológicas das lesões musculares e pesquisa de edema, hemorragia, desorganização ou alteração morfométrica das fibras musculares. E foi feita a reparação tecidual, para pesquisa da proliferação de tecido adiposo, angiogênese e fibras colágenas. Empregaram-se os testes ANOVA e t de Student com p ≤ 0,05 para significação estatística. RESULTADOS: Nas avaliações séricas o GA mostrou valores menores nas dosagens de CPK e maiores nas dosagens de AST. Nas avaliações histopatológicas, no GNT foram evidenciados edema e hemorragia compatíveis com lesões por estiramento, no GS edema, hemorragia com proliferação de tecido adiposo e fibras colágenas e no GA. Além dos achados do GS destacou-se intensa angiogênese. CONCLUSÃO: A suplementação oral com arginina não causou alterações metabólicas importantes que contraindiquem seu uso e induziu angiogênese durante a reparação de lesões musculares por estiramento.

Effects of nutritional supplementation with l-arginine on repair of injuries due to muscle strain: experimental study on rats / Efeitos da suplementação nutricional com L-arginina no reparo de lesões por estiramento muscular. Estudo experimental em ratos

Avaliar a influência da suplementação oral com arginina na regeneração de lesão por estiramento do músculo tibial anterior de ratos. MÉTODO: Usaram-se 24 ratos Wistar (492,5 ± 50,45 gramas), induzidos com lesão por estiramento dos músculos tibiais anteriores e separados em três grupos com oito ratos cada. No grupo não tratado (GNT), após a indução das lesões, os ratos foram observados por 24 horas, nos grupos simulação (GS) e arginina (GA) receberam, por gavagem diariamente, respectivamente solução salina isotônica e solução de arginina, durante sete dias. Ao término dos períodos foram coletadas amostras de sangue para as avaliações séricas de creatina-quinase (CK), desidrogenase lática (LDH), aspartato-aminotransferase (AST) e proteína C reativa (PCR). Foram ressecados os músculos tibiais anteriores (direitos e esquerdos) para avaliações histopatológicas das lesões musculares e pesquisa de edema, hemorragia, desorganização ou alteração morfométrica das fibras musculares. E foi feita a reparação tecidual, para pesquisa da proliferação de tecido adiposo, angiogênese e fibras colágenas. Empregaram-se os testes ANOVA e tde Student com p ≤ 0,05 para significação estatística. RESULTADOS: Nas avaliações séricas o GA mostrou valores menores nas dosagens de CPK e maiores nas dosagens de AST. Nas avaliações histopatológicas, no GNT foram evidenciados edema e hemorragia compatíveis com lesões por estiramento, no GS edema, hemorragia com proliferação de tecido adiposo e fibras colágenas e no GA. Além dos achados do GS destacou-se intensa angiogênese. CONCLUSÃO: A suplementação oral com arginina não causou alterações metabólicas importantes que contraindiquem seu uso e induziu angiogênese durante a reparação de lesões musculares por estiramento.

To evaluate the influence of oral supplementation with arginine on regeneration of injuries due to straining of the anterior tibial muscle of rats. METHODS: Twenty-four Wistar rats of weight 492.5 ± 50.45 g were used. Injuries were induced through straining the anterior tibial muscles. The rats were separated into three groups of eight rats each. In the untreated group (UTG), after induction of injuries, the rats were observed for 24 h. In the simulation group (SG) and the arginine group (AG) respectively, the rats received isotonic saline solution and arginine solution via direct gavage, over a seven-day period. At the end of the period, blood samples were collected for serum evaluations of creatine kinase (CK), lactic dehydrogenase (LDH), aspartate aminotransferase (AST) and C-reactive protein (CRP). The right and left anterior tibial muscles were resected for histopathological evaluations on the muscle injuries, investigating edema, hemorrhage and disorganization or morphometric alteration of the muscle fibers. The tissue repair was investigated in terms of proliferation of adipose tissue, angiogenesis and collagen fibers. The ANOVA and Student's tmethods were used and p≤ 0.05 was taken to be statistically significant. RESULTS: In the serum evaluations, the AG showed lower CK assay values and higher AST values. In the histopathological evaluation, the UTG presented edema and hemorrhage compatible with injuries due to strain; the SG presented edema and hemorrhage with proliferation of adipose tissue and collagen fibers; and the AG presented not only the findings of the SG but also, especially, intense angiogenesis. CONCLUSION: Oral supplementation with arginine did not cause any significant metabolic alterations that would contraindicate its use and it induced angiogenesis during the repair of muscles injured due to strain.

Qualitative analysis of the deposit of collagen in bladder suture of rats treated with tacrolimus combined with mycophenolate-mofetil.

PURPOSE: To evaluate the synthesis of type I (mature) and type III (immature) collagen in bladder suture of rats treated with a combination of tacrolimus and mycophenolate mofetil for 15 days. MATERIALS AND METHODS: Thirty rats were divided into 3 groups: the sham, control and experimental groups. All the animals underwent laparotomy, cystotomy and bladder suture in two planes with surgical PDS 5-0 thread. The sham group did not receive treatment. The control group received saline solution, and the experimental group received 0.1mg/kg/day of tacrolimus with 20mg/kg/day of mycophenolate mofetil, for 15 days. From then on, the tacrolimus was dosed. The surgical specimens of the bladder suture area were processed so that the total type I and type III collagen could be measured by the picrosirius red technique. RESULTS: There was a predominance of type I collagen production in the sham and control groups compared to the experimental group, in which type III collagen was predominant. The production of total collagen did not change. CONCLUSION: The association of tacrolimus and mycophenolate mofetil in animals qualitatively changes the production of collagen after 15 days with a predominance of type III collagen.

Qualitative analysis of the deposit of collagen in bladder suture of rats treated with tacrolimus combined with mycophenolate-mofetil

PurposeTo evaluate the synthesis of type I (mature) and type III (immature) collagen in bladder suture of rats treated with a combination of tacrolimus and mycophenolate mofetil for 15 days.Materials and MethodsThirty rats were divided into 3 groups: the sham, control and experimental groups. All the animals underwent laparotomy, cystotomy and bladder suture in two planes with surgical PDS 5-0 thread. The sham group did not receive treatment. The control group received saline solution, and the experimental group received 0.1mg/kg/day of tacrolimus with 20mg/kg/day of mycophenolate mofetil, for 15 days. From then on, the tacrolimus was dosed. The surgical specimens of the bladder suture area were processed so that the total type I and type III collagen could be measured by the picrosirius red technique.ResultsThere was a predominance of type I collagen production in the sham and control groups compared to the experimental group, in which type III collagen was predominant. The production of total collagen did not change.ConclusionThe association of tacrolimus and mycophenolate mofetil in animals qualitatively changes the production of collagen after 15 days with a predominance of type III collagen.

Efeitos do aditivo do leite materno com ferro sobre as propriedades bacteriostáticas do leite materno / Effects of human milk fortifier with iron on the bacteriostatic properties of breast milk

OBJETIVO: Comparar o crescimento bacteriano em colostro puro e colostro com aditivo do leite materno contendo ferro. MÉTODOS: Foram comparadas 78 amostras de colostro puro ou colostro com adição de aditivo do leite materno contendo ferro para avaliar o crescimento de Escherichia coli, Staphylococcus aureus e Pseudomonas aeruginosa. Para a análise qualitativa, discos de papel-filtro foram imersos em amostras de cada grupo e incubados por 48 horas com 10¹ Unidades Formadoras de Colônias/mL de cada cepa. Para a avaliação quantitativa, 1 mL de cada cepa contendo 10(7) Unidades Formadoras de Colônias/mL foi homogeneizado com 1 mL, tanto de colostro puro quanto de colostro com aditivo do leite materno, espalhado em placa de Petri e incubado a 37ºC. O número de Unidades Formadoras de Colônias foi contado 24 horas depois. RESULTADOS: A análise qualitativa não mostrou nenhuma diferença no crescimento bacteriano. Na avaliação quantitativa, o crescimento de Escherichia coli (EC) no grupo C foi de 29,4±9,7 x 10(6) CFU/mL, enquanto no grupo FM85 foi de 31,2±10,8 x 10(6) CFU/mL. A diferença entre o crescimento médio foi de 1,9±4,9 x 10(6) CFU/mL (p = 0,001). Não houve diferenças no crescimento de Staphylococcus aureus e Pseudomonas aeruginosa. CONCLUSÃO: A adição de ferro a essa concentração reduz a ação bacteriostática do leite materno contra Escherichia coli.

OBJECTIVE: To compare bacterial growth in pure colostrum versus colostrum with human milk fortifier (HMF) containing iron. METHODS: The growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in 78 samples of pure colostrum or colostrum with added iron-containing HMF was compared. For qualitative analysis, filter paper discs were immersed in samples from each group and incubated for 48 hours with 10¹ colony forming units (CFUs)/mL of each strain. For quantitative assessment, 1 mL of each strain containing 10(7) CFUs/mL was homogenized with 1 mL of either colostrum or colostrum with human milk fortifier, seeded into a Petri dish, and incubated at 37ºC. Twenty-four hours later, the number of CFUs was counted. RESULTS: The qualitative analysis showed no difference in bacterial growth. In the quantitative evaluation, E. coli growth in the control group was 29.4±9.7 x 10(6) CFU/ mL, while in the HMF group it was 31.2±10.8 x 10(6) CFU/mL. The difference between the average growth was 1.9±4.9 x 10(6) CFU/mL (p = 0.001). There were no differences in S. aureus and P. aeruginosa growth. CONCLUSION: Addition of iron at this concentration reduces breast milk bacteriostatic action against E. coli.

Effects of human milk fortifier with iron on the bacteriostatic properties of breast milk.

OBJECTIVE: To compare bacterial growth in pure colostrum versus colostrum with human milk fortifier (HMF) containing iron. METHODS: The growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in 78 samples of pure colostrum or colostrum with added iron-containing HMF was compared. For qualitative analysis, filter paper discs were immersed in samples from each group and incubated for 48 hours with 10(1) colony forming units (CFUs)/mL of each strain. For quantitative assessment, 1 mL of each strain containing 10(7) CFUs/mL was homogenized with 1 mL of either colostrum or colostrum with human milk fortifier, seeded into a Petri dish, and incubated at 37°C. Twenty-four hours later, the number of CFUs was counted. RESULTS: The qualitative analysis showed no difference in bacterial growth. In the quantitative evaluation, E. coli growth in the control group was 29.4±9.7×10(6)CFU/mL, while in the HMF group it was 31.2±10.8×10(6)CFU/mL. The difference between the average growth was 1.9±4.9×10(6)CFU/mL (p=0.001). There were no differences in S. aureus and P. aeruginosa growth. CONCLUSION: Addition of iron at this concentration reduces breast milk bacteriostatic action against E. coli.

L-arginine action in cutaneous flap evolution under nicotine exposure in rats.

OBJECTIVE: To evaluate whether treatment with L-arginine influences the healing of skin flaps in rats exposed to nicotine. METHODS: 40 male Wistar rats weighing 142.4 ± 10.1 g were separated into four groups: GC: treatment with 7.4 pH phosphate buffer, submitted to skin flap and observation for ten days; GN: exposure to nicotine for four weeks, submitted to skin flap and observation for ten days; GA: treatment with 7.4 pH phosphate buffer for four weeks, submitted to skin flap and arginine treatment for ten days; GAN: exposure to nicotine for four weeks, submitted to skin flap and treatment with arginine for ten days. We evaluated: areas of necrosis, re-epithelialization, inflammatory reaction and formation of granulation tissue by HE stain; the total area of deposition and differentiation of collagens I and III by histometry with picrosirius staining; and the scar vascular density by immunohistochemical staining with monoclonal anti-CD34 antibodies. RESULTS: The percentages of necrotic areas in GN and GNA were higher (p <0.001) than in GC and GA. In histological scores, collagen deposition, and the percentage of type I collagen, GA and GC were similar to each other (p> 0.05), but higher (p <0.001) than GA and GNA; as for vascular densities, they were lower in GN and GAN (p <0.001) than in GC and GA. CONCLUSION: Exposure to nicotine inhibited the effects of arginine and in unexposed mice there was induction of angiogenesis and improvement in the total collagen deposition in the skin flaps.

Ação da l-arginina na evolução de retalhos cutâneos de ratos sob exposição à nicotina / L-arginine action in cutaneous flap evolution under nicotine exposure in rats

OBJETIVO: Avaliar se o tratamento com L-arginina influencia a cicatrização de retalhos cutâneos em ratos expostos à nicotina. MÉTODOS: Foram utilizados 40 ratos Wistar pesando 142,4±10,1g separados em quatro grupos: GC- tratamento com solução tampão fosfatos pH 7,4, confecção de retalho cutâneo, observação por 10 dias; GN- exposição à nicotina por quatro semanas, confecção de retalho cutâneo, observação por dez dias; GA- tratamento com solução tampão fosfatos pH 7,4 por quatro semanas, confecção de retalho cutâneo, tratamento com arginina por dez dias; GAN- exposição à nicotina por quatro semanas, confecção de retalho cutâneo, tratamento com arginina por dez dias. Foram avaliadas as áreas de necrose, re-epitelização, reação inflamatória e formação de tecido de granulação, pela coloração HE, a área de deposição total e a diferenciação de colágenos I e III por histometria com a coloração de picrosirius, e, através da marcação imunoistoquímica com anticorpo monoclonal anti-CD34, a densidade vascular cicatricial. RESULTADOS: As porcentagens de áreas de necrose de GN e GNA foram maiores (p<0,001) do que GC e GA. Nos escores histológicos, a deposição de colágeno e a porcentagem de colágeno tipo I, no GC e GA foram similares (p>0,05) e maiores (p<0,001) do que em GA e em GNA e, nas densidades vasculares, GN e GAN foram menores (p<0,001) do que em GC e em GA. CONCLUSÃO: A exposição à nicotina inibiu os efeitos da arginina, e nos ratos não expostos, induziu melhora na angiogênese e na deposição de colágeno total nos retalhos cutâneos.

OBJECTIVE: To evaluate whether treatment with L-arginine influences the healing of skin flaps in rats exposed to nicotine. METHODS: 40 male Wistar rats weighing 142.4 ± 10.1 g were separated into four groups: GC: treatment with 7.4 pH phosphate buffer, submitted to skin flap and observation for ten days; GN: exposure to nicotine for four weeks, submitted to skin flap and observation for ten days; GA: treatment with 7.4 pH phosphate buffer for four weeks, submitted to skin flap and arginine treatment for ten days; GAN: exposure to nicotine for four weeks, submitted to skin flap and treatment with arginine for ten days. We evaluated: areas of necrosis, re-epithelialization, inflammatory reaction and formation of granulation tissue by HE stain; the total area of deposition and differentiation of collagens I and III by histometry with picrosirius staining; and the scar vascular density by immunohistochemical staining with monoclonal anti-CD34 antibodies. RESULTS: The percentages of necrotic areas in GN and GNA were higher (p <0.001) than in GC and GA. In histological scores, collagen deposition, and the percentage of type I collagen, GA and GC were similar to each other (p> 0.05), but higher (p <0.001) than GA and GNA; as for vascular densities, they were lower in GN and GAN (p <0.001) than in GC and GA. CONCLUSION: Exposure to nicotine inhibited the effects of arginine and in unexposed mice there was induction of angiogenesis and improvement in the total collagen deposition in the skin flaps.

Oxidative stress and inflammatory response increase during coronary artery bypass grafting with extracorporeal circulation.

INTRODUCTION: Thiobarbituric acid-reactive substance is a marker of oxidative stress and has cytotoxic and genotoxic actions. C- reactive protein is used to evaluate the acute phase of inflammatory response. OBJECTIVES: To assess the thiobarbituric acid-reactive substance and C-reactive protein levels during extracorporeal circulation in patients submitted to cardiopulmonary bypass. METHODS: Twenty-five consecutive surgical patients (16 men and nine women; mean age 61.2 ± 9.7 years) with severe coronary artery disease diagnosed by angiography scheduled for myocardial revascularization surgery with extracorporeal circulation were selected. Blood samples were collected immediately before initializing extracorporeal circulation, T0; in 10 minutes, T10; and in 30 minutes, T30. RESULTS: The thiobarbituric acid-reactive substance levels increased after extracorporeal circulation (P=0.001), with average values in T0=1.5 ± 0.07; in T10=5.54 ± 0.35; and in T30=3.36 ± 0.29 mmoles/mg of serum protein. The C-reactive protein levels in T0 were negative in all samples; in T10 average was 0.96 ± 0.7 mg/dl; and in T30 average was 0.99 ± 0.76 mg/dl. There were no significant differences between the dosages in T10 and T30 (P=0.83). CONCLUSIONS: C-reactive protein and thiobarbituric acid-reactive substance plasma levels progressively increased during extracorporeal circulation, with maximum values of thiobarbituric acid-reactive substance at 10 min and of C-reactive protein at 30 min. It suggests that there are an inflammatory response and oxidative stress during extracorporeal circulation.

Cecorraphy in single layer using polypropylene and poliglecaprone 25 threads: comparative study in rats.

PURPOSE: To compare sutures with polypropylene and poliglecaprone 25 after partial cecotomy in rats. METHODS: Thirty six rats divided into two groups, A and B, of 18 animals; each group was also divided into three subgroups of six animals sacrificed at 4(th), 7(th) and 14(th) days after surgery. Were studied the mortality, morbidity, complications attributable to sutures, macroscopy, optical microscopy and measurement of hydroxyproline at the level of the suture. RESULTS: There were no deaths or wound complications such as hematoma, seroma, abscess, evisceration or eventration. On microscopic evaluation reepithelization, coaptation and inflammation in both groups did not differ significantly. The average rate of tissue hydroxyproline found in the samples on the 4(th) day after surgery, was 21.38 mg/g tissue for group A and 16.68 mg/g for group B; on day 7 after surgery, the average was 15.64 mg/g tissue for group A and 26.53 mg/g for group B; on day 14, the average was 8.09 mg/g tissue for group A and 25.07 mg/g for group B. CONCLUSION: There were no differences on clinical evolution, macroscopic aspect, microscopic data and hydroxyproline concentration on both sutures.

Oxidative stress and inflammatory response increase during coronary artery bypass grafting with extracorporeal circulation / Estresse oxidativo e resposta inflamatória aumentam durante cirurgia de revascularização miocárdica com circulação extracorpórea

INTRODUCTION: Thiobarbituric acid-reactive substance is a marker of oxidative stress and has cytotoxic and genotoxic actions. C- reactive protein is used to evaluate the acute phase of inflammatory response. OBJECTIVES: To assess the thiobarbituric acid-reactive substance and C-reactive protein levels during extracorporeal circulation in patients submitted to cardiopulmonary bypass. METHODS: Twenty-five consecutive surgical patients (16 men and nine women; mean age 61.2 ± 9.7 years) with severe coronary artery disease diagnosed by angiography scheduled for myocardial revascularization surgery with extracorporeal circulation were selected. Blood samples were collected immediately before initializing extracorporeal circulation, T0; in 10 minutes, T10; and in 30 minutes, T30. RESULTS: The thiobarbituric acid-reactive substance levels increased after extracorporeal circulation (P=0.001), with average values in T0=1.5 ± 0.07; in T10=5.54 ± 0.35; and in T30=3.36 ± 0.29 mmoles/mg of serum protein. The C-reactive protein levels in T0 were negative in all samples; in T10 average was 0.96 ± 0.7 mg/dl; and in T30 average was 0.99 ± 0.76 mg/dl. There were no significant differences between the dosages in T10 and T30 (P=0.83). CONCLUSIONS: C-reactive protein and thiobarbituric acid-reactive substance plasma levels progressively increased during extracorporeal circulation, with maximum values of thiobarbituric acid-reactive substance at 10 min and of Creactive protein at 30 min. It suggests that there are an inflammatory response and oxidative stress during extracorporeal circulation.

INTRODUÇÃO: Substâncias reativas do ácido tiobarbitúrico são um marcador de estresse oxidativo. A proteína C reativa é usada para avaliar a fase aguda da resposta inflamatória. OBJETIVOS: Avaliar os níveis de substâncias reativas do ácido tiobarbitúrico e da proteína C reativa durante a circulação extracorpórea em pacientes submetidos à cirurgia de revascularização miocárdica. MÉTODOS: Vinte e cinco pacientes consecutivos (16 homens e nove mulheres com idade média de 61,2 ± 9,7 anos) com doença arterial coronária severa diagnosticada por angiografia, escalados para cirurgia de revascularização miocárdica com circulação extracorpórea, foram selecionados. Amostras sanguíneas foram coletadas imediatamente antes de iniciar a circulação extracorpórea (T0), 10 minutos após (T10) e 30 minutos após (T30). RESULTADOS: Os níveis de substâncias reativas do ácido tiobarbitúrico aumentaram após a extracorpórea (P=0,001) com valores médios de 1,5 ± 0,07 em T0; 5,54 ± 0,35 em T10 e 3,36 ± 0,29 mmoles/mg de proteína sérica em T30. Os níveis de proteína C reativa foram negativos em T0 em todas as amostras. Em T10, os valores médios foram de 0,96 ± 0,7 mg/dl e em T30 os valores médios foram de 0,99 ± 0,76 mg/ dl. Não houve diferença significativa entre os valores de proteína C reativa nos tempos T10 e T30 (P= 0,83). CONCLUSÕES: Os níveis de substâncias reativas do ácido tiobarbitúrico e da proteína C reativa aumentam durante a circulação extracorpórea, com máximos valores de substâncias reativas do ácido tiobarbitúrico em 10 minutos e de proteína C reativa em 30 minutos. Estes achados sugerem resposta inflamatória e estresse oxidativo durante a circulação extracorpórea.

Cecorraphy in single layer using polypropylene and poliglecaprone 25 threads: comparative study in rats / Cecorrafia em plano único com fios de polipropileno e poliglecaprone 25: estudo comparativo em ratos

PURPOSE: To compare sutures with polypropylene and poliglecaprone 25 after partial cecotomy in rats. METHODS: Thirty six rats divided into two groups, A and B, of 18 animals; each group was also divided into three subgroups of six animals sacrificed at 4th, 7th and 14th days after surgery. Were studied the mortality, morbidity, complications attributable to sutures, macroscopy, optical microscopy and measurement of hydroxyproline at the level of the suture. RESULTS: There were no deaths or wound complications such as hematoma, seroma, abscess, evisceration or eventration. On microscopic evaluation reepithelization, coaptation and inflammation in both groups did not differ significantly. The average rate of tissue hydroxyproline found in the samples on the 4th day after surgery, was 21.38 mg/g tissue for group A and 16.68 mg/g for group B; on day 7 after surgery, the average was 15.64 mg/g tissue for group A and 26.53 mg/g for group B; on day 14, the average was 8.09 mg/g tissue for group A and 25.07 mg/g for group B. CONCLUSION: There were no differences on clinical evolution, macroscopic aspect, microscopic data and hydroxyproline concentration on both sutures.

OBJETIVO: Comparar a sutura com fio de polipropileno e poliglecaprone 25 após cecotomia parcial em ratos. MÉTODOS: Trinta e seis ratos foram distribuídos em dois grupos A e B de 18 animais, e cada grupo foi dividido em três subgrupos de seis, sacrificados no 4º, 7º e 14º dias do pós-operatório. Estudou-se a mortalidade, morbidade, complicações atribuíveis às suturas, macroscopia, microscopia ótica e dosagem de hidroxiprolina no nível da sutura. RESULTADOS: Não houve mortalidade ou complicações da ferida operatória como hematoma, seroma, abscesso, evisceração ou eventração. Na avaliação microscópica os critérios de re-epitelização, coaptação e processo inflamatório ambos os grupos não apresentaram diferença significativa. A taxa tecidual média da hidroxiprolina encontrada nas amostras no 4º dia de pós-operatório foi de 21,38 mg/g de tecido para o grupo A e 16,68 mg/g para o grupo B; no 7º dia a média foi de 15,64 mg/g de tecido para o grupo A e 26,53 mg/g para o grupo B; no 14º dia ela foi de 8,09 mg/g de tecido para o grupo A e 25,07 mg/g para o grupo B. CONCLUSÃO: Não houve diferença estatística entre a evolução clínica, avaliação macroscópica, microscopia e dosagem de hidroxiprolina entre as suturas realizadas com os fios estudados.