Influence of tobacco, alcohol and diabetes on the collagen of cremaster muscle in patients with inguinal hernias / Influência do tabaco, álcool e diabete sobre a interação célula-colágeno em músculo cremaster de humanos com hérnias inguinais

ABSTRACT Background: New findings point out that the mechanism of formation of the hernias can be related to the collagenous tissues, under activity of aggressive agents such as the tobacco, alcohol and diabetes. Aim: To analyze the collagen present in the cremaster muscle in patients with inguinal hernias, focusing the effect of tobacco, alcohol, and diabetes. Methods: Fifteen patients with inguinal hernia divided in three groups were studied: group I (n=5) was control; group II (n=5) were smokers and/or drinkers; and group III (n=5) had diabetes mellitus. All subjects were underwent to surgical repair of the inguinal hernias obeying the same pre, intra and postoperative conditions. During surgery, samples of the cremaster muscle were collected for analysis in polarized light microscopy, collagen morphometry and protein. Results: The area occupied by the connective tissue was higher in groups II and III (p<0.05). The collagen tissue occupied the majority of the samples analyzed in comparison to the area occupied by muscle cells. The content of total protein was higher in groups II and III compared to the control group (p<0.05). Conclusion: The tobacco, alcohol and diabetes cause a remodel the cremaster muscle, leading to a loss of support or structural change in this region, which may enhance the occurrences and damage related to inguinal hernias.

RESUMO Racional: Estudos recentes sinalizam que o mecanismo de formação das hérnias pode estar relacionado aos tecidos colagenosos, sob a ação de agentes agressores como o tabaco, o álcool e o diabete. Objetivo: Avaliar o colágeno presente no músculo cremaster em pacientes com hérnias inguinais enfocando o efeito do tabaco, álcool e diabete. Métodos: Foram estudados 15 pacientes com hérnias inguinais divididos em: grupo I (n=5) controles; grupo II (n=5) indivíduos fumantes e/ou etilistas; e grupo III (n=5) indivíduos que apresentavam diabete melito. Todos foram submetidos à correção cirúrgica das hérnias inguinais obedecendo às mesmas condições pré, intra e pós-operatórias. Durante o procedimento cirúrgico, amostras do músculo cremaster foram coletadas para análises em microscopia de luz polarizada, morfometria do colágeno e de proteínas. Resultados: A área ocupada por tecido conjuntivo foi maior nos grupos II e III (p<0,05). O tecido colágeno ocupou a maior parte das amostras analisadas, em comparação à área ocupada pelas células musculares. O conteúdo de proteínas totais foi maior nos grupos II e III, quando comparado com o grupo controle (p<0,05). Conclusão: O tabaco, o álcool e o diabete ocasionam remodelação no músculo cremaster, levando à perda de suporte ou alteração estrutural nesta região, podendo intensificar as ocorrências e os danos relacionados às hérnias inguinais.

Oral or topical administration of L-arginine changes the expression of TGF and iNOS and results in early wounds healing

ABSTRACT PURPOSE: To evaluate the contribution of L-arginine oral or topical rout of administration in the surgical wound healing process. METHODS: L-arginine was orally or topically administrated to mice after a laparotomy model procedure. The wounds were analyzed to evaluate the granulation tissue by HE analysis, collagen deposition, iNOS and cytokines production by immunochemisyry on wound progress. Mice used in this model were healthy, immunosupressed or diabetic and all of them were treated with different concentration of L-arginine and rout of administration. RESULTS: Suggested that groups treated with L-arginine orally or topically improved wound repair when compared with non-treatad mice. L- arginine treatment stimulated TGF-β and restricted NO production leading to a mild Th1 response and collagen deposition in injured area, when it was orally administrated. Topical administration decreased IL-8 and CCR1 expression by wound cells but did not interfere with TNF-α and IL-10 production, ratifying the decrease of inflammatory response, the oral administration however, presented a higher iNOS and TGF-β expression then. L-arginine treatment also improved the improved the wound healing in immunosupressed or diabetic mice. CONCLUSION: L-arginine administrated orally or topically can be considered an important factor in the recuperation of tissues.

Low serum Insulin Like Growth Factor - 1 in patients with Stress Urinary Incontinence

ABSTRACT Objective: SUI, involuntary loss of urine, occurs when intra abdominal pressure exceeds urethral pressure in women. Recent animal study has shown that there are therapeutic effects of Insulin-like growth factors (IGF-1) on stress urinary incontinence in rats with simulated childbirth trauma. IGF-1 is an important mediator of cell growth, differentiation and transformation in various tissues and stimulates fibroblast proliferation and enhances collagen synthesis. The purpose of the current study was to determine the association between IGF-1 levels and SUI. Materials and Methods: All patients were evaluated for SUI and divided into two groups: 116 women with SUI and 76 women without SUI. Diagnosis of SUI was based on the International Consultation on Incontinence Questionnaire-Short Form (ICIQSF). Levels of IGF-1 were measured in serum by enzyme-linked immunosorbent assay. The relationship between IGF-1 levels and SUI in patients was evaluated statisticaly. Results: The mean age of patients wiyh SUI was 49.9±8.6 and 48.7±7.8 in control group. Plasma IGF-1 levels were significantly lower in SUI than in control group (106.5±26.4 and 133.3±37.1ng/mL, respectively, P <0.001). Body mass indexes were higher in women with SUI than women without SUI. Conclusion: In this study lower serum IGF-1 levels were found to be associated with SUI. Serum IGF-1 level appears to be a specific predictor of SUI, and it may be used in early prediction of SUI in female population.

Use of silicon for skin and hair care: an approach of chemical forms available and efficacy

Abstract Silicon is the second most abundant element on Earth, and the third most abundant trace element in human body. It is present in water, plant and animal sources. On the skin, it is suggested that silicon is important for optimal collagen synthesis and activation of hydroxylating enzymes, improving skin strength and elasticity. Regarding hair benefits, it was suggested that a higher silicon content in the hair results in a lower rate of hair loss and increased brightness. For these beneficial effects, there is growing interest in scientific studies evaluating the efficacy and safety of using dietary supplements containing silicon. Its use aims at increasing blood levels of this element and improving the skin and its annexes appearance. There are different forms of silicon supplements available and the most important consideration to be made in order to select the best option is related to safety and bioavailability. Silicon supplements are widely used, though there is wide variation in silicon bioavailability, ranging from values below 1% up to values close to 50%, depending on the chemical form. Therefore, the aim of this study was to evaluate the scientific literature related to the different chemical forms of silicon supplements available and the limitations and recent progress in this field. According to reported studies, among the different chemical forms available, the orthosilicic acid (OSA) presents the higher bioavailability, whereas the others forms have absorption inversely proportional to the degree of polymerization. However, clinical studies evaluating safety and efficacy are still lacking.

Radiofrequency for the treatment of skin laxity: mith or truth

AbstractThe nonablative radiofrequency is a procedure commonly used for the treatment of skin laxity from an increase in tissue temperature. The goal is to induce thermal damage to thus stimulate neocollagenesis in deep layers of the skin and subcutaneous tissue. However, many of these devices haven't been tested and their parameters are still not accepted by the scientific community. Because of this, it is necessary to review the literature regarding the physiological effects and parameters for application of radiofrequency and methodological quality and level of evidence of studies. A literature search was performed in MEDLINE, PEDro, SciELO, PubMed, LILACS and CAPES and experimental studies in humans, which used radiofrequency devices as treatment for facial or body laxity, were selected. The results showed that the main physiological effect is to stimulate collagen synthesis. There was no homogeneity between studies in relation to most of the parameters used and the methodological quality of studies and level of evidence for using radiofrequency are low. This fact complicates the determination of effective parameters for clinical use of this device in the treatment of skin laxity. The analyzed studies suggest that radiofrequency is effective, however the physiological mechanisms and the required parameters are not clear in the literature.

Instrumento para avaliação da qualidade do Time de Resposta Rápida em um hospital universitário público / Instrument for assessing the quality of the Rapid Response Team at a university public hospital / Instrumento para la evaluación de la calidad del Equipo de Respuesta Rápida en un hospital universitario público

O artigo descreve a construção de um questionário para avaliação da qualidade do atendimento de um Time de Resposta Rápida, em Hospital Universitário de Londrina-PR, fundamentado no modelo conceitual de Donabedian (estrutura-processoresultado). A coleta de dados ocorreu no mês de março de 2012 e o processo de adequação do questionário foi desenvolvido com a aplicação da Técnica Delphi com a participação de 15 especialistas. Ao término do estudo obteve-se um questionário com 37 enunciados, sendo alcançado índice de concordância final na pesquisa com valores superiores a 80% para todos os conceitos. Espera-se que as contribuições do grupo de especialistas tornem o instrumento confiável e seja aplicado em outros serviços semelhantes. Aplicações futuras deste instrumento poderão trazer subsídios para melhor avaliação da qualidade dos serviços de equipes de Resposta Rápida.

The paper describes the construction of a questionnaire to assess the quality of care of a Rapid Response Team at the University Hospital of Londrina, based on the conceptual model of Donabedian (structure-process-outcome). Data collection occurred in March 2012 and the process of adjusting the questionnaire was developed with the application of the Delphi technique involving 15 experts. At the end of the study the questionnaire contained 37 statements, achieving final compliance level higher than 80% in all concepts. It is hoped that the contributions of the expert group produce a more reliable questionnaire to be applied in other similar services. Future applications of this instrument may provide information to better assess the quality of teams of Rapid Response services.

El artículo describe la construcción de un cuestionario para la evaluación de la calidad de la atención de un Equipo de Respuesta Rápida en un Hospital Universitario de Londrina, Paraná, basado en el modelo conceptual de Donabedian (estructura-procesoresultado). La recolección de datos ocurrió durante el mes de marzo de 2012 y el proceso de ajuste del cuestionario fue desarrollado por medio de la Técnica Delphi con la participación de 15 especialistas. Al término del estudio se obtuvo un cuestionario con 37 enunciados, alcanzándose un índice de concordancia final en la investigación con valores superiores al 80% para todos los conceptos. Se espera que las contribuciones del grupo de especialistas afiancen la confiabilidad del instrumento y que el cuestionario sea utilizado en otros servicios semejantes. Las aplicaciones futuras podrán traer subsidios para evaluar mejor la calidad de los servicios de los equipos de Respuesta Rápida.

Low transformation growth factor-β1 production and collagen synthesis correlate with the lack of hepatic periportal fibrosis development in undernourished mice infected with Schistosoma mansoni

Undernourished mice infected (UI) submitted to low and long-lasting infections by Schistosoma mansoni are unable to develop the hepatic periportal fibrosis that is equivalent to Symmers’ fibrosis in humans. In this report, the effects of the host’s nutritional status on parasite (worm load, egg viability and maturation) and host (growth curves, biology, collagen synthesis and characteristics of the immunological response) were studied and these are considered as interdependent factors influencing the amount and distribution of fibrous tissue in hepatic periovular granulomas and portal spaces. The nutritional status of the host influenced the low body weight and low parasite burden detected in UI mice as well as the number, viability and maturation of released eggs. The reduced oviposition and increased number of degenerated or dead eggs were associated with low protein synthesis detected in deficient hosts, which likely induced the observed decrease in transformation growth factor (TGF)-β1 and liver collagen. Despite the reduced number of mature eggs in UI mice, the activation of TGF-β1 and hepatic stellate cells occurred regardless of the unviability of most miracidia, due to stimulation by fibrogenic proteins and eggshell glycoproteins. However, changes in the repair mechanisms influenced by the nutritional status in deficient animals may account for the decreased liver collagen detected in the present study.

Role of Triticum aestivum aqueous extract in glucocorticoid induced osteoporosis in rats.

Administration of aqueous extract of T. aestivum (200 and 400 mg/kg/day, po, for 30 days) and risedronate (20 mg/kg, sc, five times a week for 30 days) following methyl prednisolone sodium succinate (10 mg/kg, sc, thrice a week for 4 weeks) induced osteoporosis in Wistar rats showed an increase in the serum levels of bone mineral content markers, decrease in the serum and urinary levels of bone resorption markers. An incline in strength of femur and tibia was seen particularly with 400 mg/kg of T. aestivum. Maintenance of calcium homeostasis, formation of collagen and scavenging of free radicals can plausibly be the mode of action of aqueous extract of T. aestivum thereby combating osteoporosis induced by glucocorticoids.

Análise ultraestrutural e implicação do RNA de interferência na fibrilogênese do colágeno V em pacientes com esclerodermia / Ultrastructural analysis and implication of RNA interference in collagen V fibrillogenesis in patients with scleroderma

Introdução: Recentemente muitas são as funções atribuídas ao colágeno V em condições fisiológicas normais e em algumas doenças, como na esclerodermia. Esta proteína apresenta características estruturais singulares de regulação do diâmetro das fibrilas heterotípicas, e imunogenicidade, sendo capaz de desencadear uma resposta imune independente. Em descobertas recentes, desenvolvidas por nosso grupo, ficou evidenciado que o colágeno V apresenta histoarquitetura anômala, aumentando a sua imunoexpressão na pele e pulmão em estágios iniciais da doença, assim como, aumento do RNAm de suas cadeias, principalmente alfa2(V). Por esta razão e com o intuito de entender sobre os processos moleculares e ultraestruturais que correlacionam o COL V à fibrose na ES, a proposta do presente estudo foi realizar análise morfológica e ultraestrutural das cadeias, alfa1(V) e alfa2(V), da pele de pacientes com ES, assim como avaliar a influência do gene COL5A2 na fibrilogênese. Pacientes e Métodos: As análises foram desenvolvidas utilizando fragmento de pele obtidas de biópsia, tanto de pacientes como controles, sob aprovação do comitê de ética (CAPPesq 0331/10) e de acordo com Declaração de Helsinque. Participaram do estudo 7 pacientes (homens=3, mulheres=4) diagnosticados com ES há dois anos ou menos, e indivíduos voluntários saudáveis (n=7) utilizados como grupo controle. A biópsia das peles foi dividida para dois grupos de análises: estudos histológicos e cultura de fibroblastos; sob solução de formol a 10%, foram realizadas análises histomorfométricas [coloração de Hematoxilina e Eosina (H e E), Tricrômico de Masson, imunofluorescência (IM) para cadeias alfa1(V) e alfa2(V), e reconstrução tridimensional (Zeiss LSM 510 META/UV)], e em solução de glutaraldeído 2% para análise ultraestrutural (microscopia eletrônica de transmissão com imunomarcação com ouro coloidal). A quantificação das cadeias foi realizada por histomorfometria, através de análise de imagem utilizando o software...

Introduction: Recently many functions are attributed to type V collagen in normal physiological conditions and in some diseases, such as scleroderma. This protein presents unique structural features for regulating the diameter of fibrils heterotypic and immunogenicity being capable of eliciting an immune response independent. In recent discoveries, developed by our group, it was evident that collagen V presents anomalous histoarchitecture, increasing the immunoexpression in the skin and lung in the early stages of the disease, as well as increased mRNA of his chains, mainly alpha2(V). For this reason and in order to understand the molecular and ultrastructural processes that correlate COL V fibrosis in SSc, the purpose of this study was to perform morphological and ultrastructural analysis of the chains alpha1(V) and alpha2(V) of the patient´s skin with ES, as well as to assess the influence of the COL5A2 gene in fibrillogenesis. Patients and Methods: The analyses were developed using skin fragment obtained from biopsy, both of patients and controls, under the approval of the ethics committee (CAPPesq 0331/10) and in accordance with the Declaration of Helsinki. The study included 7 patients (male = 3, female = 4) diagnosed with ES for two years or less, and healthy volunteers (n = 7) used as a control group. The biopsy of the skin was divided in two groups of analyzes: histological studies and cultured fibroblasts; Under formaldehyde solution 10%, histomorphometric analysis [staining with hematoxylin and eosin (H e E), Masson's trichrome, immunofluorescence (IM) to alpha1(V) and alpha2(V) chains, and three-dimensional reconstruction (Zeiss LSM 510 META were performed/UV), and in a solution of 2% glutaraldehyde for ultrastructural analysis (transmission electron microscopy with immunostaining with colloidal gold). Quantitation was performed by the chain histomorphometry by image analysis using Image Pro-Plus 6.0 software. Already a fibroblast culture...

Effect of copaiba oil on correction of abdominal wall defect treated with the use of polypropylene/polyglecaprone mesh

PURPOSE: To evaluate the effects of copaiba oil on the correction of abdominal defect treated with the use of polypropylene/polyglecaprone mesh in rats. METHODS: A defect in the abdominal wall was created and corrected with polypropylene/polyglecaprone mesh in 36 rats. They were randomly distributed into three groups: control, copaiba by oral administration (gavage) and copaiba oil dip in the mesh. Euthanasia was performed after seven, 14 and 21 post-operative days. The healing process was analyzed regarding the meshes and macroscopic and microscopic aspects. RESULTS: All animals had abdominal adhesions, which were smaller in the copaiba (gavage) group (p<0.05). In microscopy, all animals had an acute inflammation stage and the inflammatory response was best characterized by foreign body-type granulomas around the mesh fragments, which was not found in the mesh fragments within the copaiba dip group. There was a greater area of necrosis and fibrosis in the copaiba dip group compared to the control group (p<0.05). The copaiba (gavage) group had a greater quantity of collagen fibers compared to the control group. CONCLUSION: Copaiba oil administered by gavage decreased the amount of abdominal adhesions, besides accelerating the process of collagen fibers formation, without damages within the early stages of healing. However, when used by dip directly on the mesh, it had corrosive effects compromising the healing process of the abdominal wall.

The effects of ghrelin on colonic anastomosis healing in rats

OBJECTIVES: In addition to its roles in the stimulation of growth hormone secretion and the regulation of appetite and metabolism, ghrelin exerts immunomodulatory, anti-inflammatory and antioxidant actions in several organ systems. In this study, we investigated the effects of ghrelin on the healing of experimental colonic anastomoses. METHODS: Wistar rats were randomly divided into two groups (n = 10 in each). A segment of colon was excised, and an end-to-end anastomosis was performed in the distal colon. The Ghrelin Group received 10 ng/kg/day IP ghrelin for seven days postoperatively, whereas the Control Group received an identical volume of saline. On the seventh postoperative day, the anastomotic bursting pressures and hydroxyproline levels were measured, and adhesion formation around the anastomoses was examined. Histopathological analyses were performed to evaluate inflammatory cell infiltration, fibroblast infiltration, collagen density and neovascularization. RESULTS: In the Ghrelin Group, the bursting pressure and hydroxyproline levels were significantly higher than in the Control Group. The adhesion formation scores were lower in the Ghrelin Group than in the Control Group. Although the inflammatory cell infiltration was diminished in the Ghrelin Group, the degrees of fibroblast infiltration, collagen density and neovascularization were not significantly different between the groups. CONCLUSION: Our results indicate that ghrelin improves the healing of colonic anastomoses in rats.

Luz intensa pulsada no fotoenvelhecimento: avaliação clínica, histopatológica e imuno-histoquímica / Intense pulsed light in photoaging: a clinical, histopathological and immunohistochemical evaluation

FUNDAMENTOS: A luz intensa pulsada tem sido muito utilizada no tratamento do fotoenvelhecimento sem completo conhecimento de seu mecanismo de ação. OBJETIVO: Estudar a ação da luz intensa pulsada no fotoenvelhecimento e na resposta imunológica cutânea por meio de estudo clínico, histopatológico, avaliando células de Langerhans (CD1), expressão da molécula de adesão intercelular, de linfócitos CD4 e CD8 e quantificação de colágeno e fibras elásticas. MÉTODOS: Um total de 26 pacientes, com idades entre 40 e 65 anos, com fototipos II a III de Fitzpatrick, foram tratadas do fotoenvelhecimento usando LIP, em 5 sessões, com intervalo mensal, durante o ano de 2006. Todas as pacientes foram submetidas à avaliação histológica e imuno-histoquímica 6 meses após o tratamento. RESULTADOS: Ao término do tratamento, houve melhora clínica em 76,92 por cento dos casos, estando relacionada ao aumento significante de fibras colágenas (51,33 por cento) e elásticas (44,13 por cento). O tratamento com luz intensa pulsada promoveu redução de linfócitos CD4 e não alterou a intensidade de linfócitos CD8. Além disso, promoveu aumento significante de pequenos vasos sanguíneos, não ectásicos, molécula de adesão intercelular positivos. CONCLUSÃO: O tratamento facial com luz intensa pulsada promoveu intensa melhora clínica que foi comprovada pelo estudo histopatológico da pele, constituindo boa opção de tratamento para o fotoenvelhecimento cutâneo, por ser técnica não- ablativa, segura e eficaz.

BACKGROUND: Intense pulsed light has been used in the treatment of photoaging without a full understanding of its mechanism of action. OBJECTIVE: To study the effect of intense pulsed light on photoaging and on the skin immune response by means of a clinical and histopathological study, evaluating Langerhans cells (CD1), expression of intercellular adhesion molecule, of CD4 and CD8 lymphocytes and quantification of collagen and elastic fibers. METHODS: In 2006 a total of 26 patients, aged 40 to 65 years, with phototypes II to III (Fitzpatrick scale), were treated for photoaging using intense pulsed light in five sessions with a monthly interval. All the patients were subjected to histological and immunohistochemical evaluation 6 months after treatment. RESULTS: At the end of the treatment clinical improvement was observed in 76.92 percent of cases. This improvement was associated to a significant increase of collagen (51.33 percent) and elastic (44.13 percent) fibers. Intense pulsed light treatment led to a reduction of CD4 lymphocytes and did not alter the amount of CD8 lymphocytes. It also led to a significant increase of small, nonectatic blood vessels, positive intercellular adhesion molecule. CONCLUSION: Facial treatment with intense pulsed light promoted major clinical improvement that was confirmed by histological examination of the skin. This technique is a good treatment option for skin photoaging because it is non-ablative, safe and effective.

Influence of ingaalp laser (660nm) on the healing of skin wounds in diabetic rats / Influência do laser ingaalp (660nm) na cicatrização de feridas cutâneas em ratos diabéticos

PURPOSE: To determine the influence of low-power laser (660 nm) on the collagen percentage and macrophages in skin wounds in diabetic rats. METHODS: 30 male Wistar rats were used, distributed in two groups: laser treated diabetic (n= 15); untreated diabetic (n = 15). The diabetes was induced by intravenous injection of alloxan into the dorsal vein of the penis, at a rate of 0.1 ml of solution per 100 g of body weight. A wound was made on the back of all the animals. Groups 2 were treated with Aluminium Gallium Indium Phosphide - InGaAlP type diode laser (Photon Laser III DMC®) with a continuous output power of 100 mW and wavelength (?) of 660 nm (4 J/cm²) for 24 s. five animal from each group was sacrificed on the 3rd, 7th and 14th days after wounding. Samples were taken, embedded in paraffin, stained with hematoxylin-eosin, Masson's trichrome, and immunohistochemical macrophage. morphometrically analyzed using the Image Pro Plus 4.5 software. The percentages of collagen fibers and macrophages were determined from the samples from the euthanasia animals. RESULTS: The data were treated statistically using analysis of variance (ANOVA) and the Post-hocTukey test. The significance level was set at 0.05 or 5 percent. CONCLUSION: The low-power laser (660 nm) was shown to be capable of influencing the collagen percentage in skin wounds by increasing the mean quantity of collagen fibers and macrophages.

OBJETIVO: Determinar a influência do diodo laser InGaAlP (660 nm) sobre o percentual de colágeno e macrófagos em feridas cutâneas de ratos diabéticos. MÉTODOS: Para tanto 30 ratos machos Wistar foram distribuídos em dois grupos: diabético tratado com laser (n=15), diabético não tratado (n = 15). O diabetes foi induzido por injeção intravenosa de aloxana na veia dorsal do pênis, a uma taxa de 0,1 ml de solução por 100 g de peso corporal. A lesão foi confeccionada no dorso de todos os animais utilizando um punch de 8mm. 2 grupos foram tratados com o diodo laser arsenieto de gálio e alumínio dopado com, fosforeto de índio - InGaAlP (tipo Photon Laser III DMC®) com uma potência contínua de 100 mW e comprimento de onda (?) de 660 nm (4 J/cm²) por 24 s. cinco animais de cada grupo foi sacrificado no 3, 7 e 14 dias após a lesão. As amostras obtidas foram emblocadas em parafina, coradas com hematoxilina-eosina, tricrômico de Masson, e para a analise dos macrófagos foi realizado um imunohistoquímico. Os resultados foram avaliados morfometricamente pelo programa Image Pro Plus 4.5. RESULTADOS: Os dados foram tratados estatisticamente pela análise de variância (ANOVA) e pós-teste de Tukey. O nível de significância foi fixado em 0,05 ou 5 por cento. CONCLUSÃO: O laser de baixa potência (660 nm) mostrou-se capaz de influenciar o percentual de colágeno em feridas cutâneas, aumentando a quantidade média de fibras colágenas e macrófagos.

Angiogenesis, fibrinogenesis and presence of synechiae after exeresis of a swine vocal fold mucosal microflap and use of topical mitomycin-C / Estudo da angiogênese, fibrogênese e presença de sinéquias após exérese de fragmento de mucosa de pregas vocais de suínos utilizando instrumental frio e mitomicina-C tópica

PURPOSE: To evaluate swine vocal fold healing in a period of 30 days after topical mitomycin-C application. METHODS: Twelve swine underwent exeresis of mucosal flaps from the free edge of the anterior third of both vocal folds with a cold instrument (laryngeal scissors). The animals were divided into two groups: EG (Experimental Group), consisting of 6 animals undergoing topical MMC application (4 mg/dL) on the operated area for 4 minutes; CG (Control Group), 6 animals undergoing topical saline solution application on the operated area for 4 minutes. After 30 days, the animals were sacrificed and the larynges were collected and examined for the presence of synechiae as well as a histological immunohistochemical assessment of immature and mature collagen deposition, number of blood vessels and myofibroblasts. RESULTS: Mature collagen deposition in the EG was 452.12 μm² and 1332.31μm² in the CG; immature collagen deposition was 1511.73μm² in the EG and 1020.61μm² in the CG. The number of myofibroblasts was 1.556 in the EG and 3.583 in the CG. The number of blood vessels was 2.565 in EG and 6.917 in the CG. There were no synechiae in the two studied groups. CONCLUSIONS: There was an increase in immature collagen deposition in the experimental group when compared with the control group. There was a decrease in mature collagen deposition in the experimental group when compared with the control group. There was a decrease in the number of myofibroblasts in the experimental group when compared with the control group. A decrease in blood vessels was observed in the experimental group when compared with the control group. There was no synechia formation in either studied group.

OBJETIVO: Avaliar a cicatrização de pregas vocais de suínos utilizando mitomicina-C tópica, em 30 dias. MÉTODOS: Doze suínos foram submetidos à exérese de mucosa do bordo livre do terço anterior de ambas as pregas vocais com instrumental frio (tesoura curva) e divididos em dois grupos: Grupo E (GE), composto por seis animais com utilização tópica de mitomicina-C(4 mg/dL) na área cruenta por quatro minutos e Grupo Controle (GC), composto por seis animais com utilização tópica de soro fisiológico na área cruenta por quatro minutos. Após 30 dias os animais foram sacrificados e coletadas amostras das pregas vocais para avaliação de sinéquias e análise histológica com quantificação da deposição de colágeno maduro e imaturo, quantificação do número de vasos sanguíneos e miofibroblastos, por método imunoistoquímico. RESULTADOS: A deposição do colágeno maduro no GE foi de 452,12 μm² e 1332,31μm² no GC. A do colágeno imaturo foi de 1511,73μm² no GE e de 1020,61μm² no GC. O número de miofibroblastos foi de 1,556 no GE e de 3,583 no GC. O número de vasos sanguíneos foi de 2,565 no GE e de 6,917 no GC. Não houve formação de sinéquias nos grupo s estudados. CONCLUSÕES: No grupo experimento, quando comparado ao controle, houve significativamente aumento da deposição do colágeno imaturo e diminuição da deposição do colágeno maduro, do número de miofibroblastos e de vasos sanguíneos. Não houve formação de sinéquias em ambos os grupos.

Tracheal suture in rats with hypothyroidism: wound healing study / Traqueorrafia em ratos com hipotireoidismo: estudo da cicatrização

PURPOSE: To evaluate the influence of hypothyroidism in tracheal wound healing process. METHODS: A controlled study was designed with 48 male Wistar rats, divided into two groups: study and control groups. In the first one hypothyroidism was surgically induced and 10 weeks after, a tracheal opening followed by suture was performed in both groups, subdivided into 7, 14, and 21 days in accordance with the date of animals death. A laboratorial evaluation was performed to prove the decreased in thyroid function in the study group. Also a macroscopic evaluation through a stablished protocol and a microscopic analysis with Hematoxylin-eosin and Sirius-Red staining methods were done. RESULTS: The laboratorial evaluation certified suppressed thyroid function in the study group. The macroscopic evaluation showed the presence of suture blockade in the study group in all the evaluated days. Microscopic analysis showed a prolongated inflammatory process and less collagen with delay in organization in the study group comparing to control group. All these data were statistic significant. CONCLUSION: Hypothyroidism had an influence in tracheal wound healing process, promoting delay in the inflammatory and organization processes and diminished tissue collagen quantity.

OBJETIVO: Investigar a influência que o hipotireoidismo exerce no processo cicatricial da traquéia. MÉTODOS: Estudo controlado, com 48 ratos Wistar machos, divididos em dois grupos, denominados: grupo estudo e grupo controle. No primeiro induziu-se hipotireoidismo cirurgicamente e após 10 semanas, realizou-se nos dois grupos o procedimento de abertura traqueal seguida de sutura. Houve subdivisão destes grupos em 7, 14 e 21 dias, de acordo com a data de morte dos animais. Foi realizada avaliação laboratorial para comprovar o hipotireoidismo no grupo estudo, avaliação macroscópica e microscópica através das colorações de hematoxilina-eosina e Sirius Red. RESULTADOS: A avaliação laboratorial atestou que o grupo estudo encontrava-se em estado de hipotireoidismo. A avaliação macroscópica demonstrou predomínio de bloqueio da sutura no grupo estudo em todos os momentos avaliados. A microscopia comprovou prolongamento da fase inflamatória e diminuição da quantidade total de colágeno bem como atraso na sua organização no grupo estudo em relação ao grupo controle. Todos estes dados apresentaram significância estatística. CONCLUSÃO: O hipotireoidismo influencia no processo de cicatrização traqueal prolongando a fase inflamatória, diminuindo a quantidade de colágeno e atrasando sua maturação e organização.

Anastomotic healing in ileum and colon of alloxan-induced diabetic rats / Cicatrização de anastomoses realizadas no íleo e cólon de ratos diabéticos induzidos pela aloxana

PURPOSE: To investigate if diabetes mellitus may alter breaking strength (BS) and collagen content in ileum and colon anastomoses in rats. METHODS: Three-hundred Wistar rats were randomly assigned to 5 experimental groups, 60 per group: normal controls surgically manipulated (G1); normal controls submitted to ileum (G2) and colon (G3) anastomotic construction; diabetic rats submitted to ileum (G4) and colon (G5) anastomotic construction. Each group was further divided into 6 subgroups with 10 rats each for sacrifice at 0, 4, 7, 14, 21, and 30 days after surgery. All surgical procedures were performed 3 months after alloxan diabetes induction. BS was measured in all intestinal anastomoses. Fragments of ileum and colon anastomoses were taken for hydroxyproline concentration (HP) and total tissue protein (TP) dosages. RESULTS: Anastomotic BS was significantly decreased (P<0.05) in ileum and colon of G4 and G5 diabetic groups up to 7 and 14 days after surgery, respectively, compared with G2 and G3 normal control groups. Anastomotic HP and TP content did not significantly differ between diabetic and normal control operated groups in ileum or colon at all evaluation times. CONCLUSION: Experimental diabetes leads to impaired intestinal anastomotic strength during early surgical wound repair, but does not appear to be implicated with collagen synthesis capacity.

OBJETIVO: Investigar se o diabetes mellitus pode alterar a força de ruptura (FR) e o conteúdo de colágeno em anastomoses realizadas no íleo e cólon de ratos. MÉTODOS: 300 ratos Wistar foram distribuídos por sorteio em 5 grupos experimentais com 60 animais cada: controle normal manipulado cirurgicamente (G1); normais controles submetidos a anastomoses no íleo (G2) e cólon (G3); ratos diabéticos submetidos a anastomoses no íleo (G4) e cólon (G5). Cada grupo foi dividido em 6 subgrupos com 10 ratos cada para sacrifícios com 0, 4, 7, 14, 21 e 30 dias após as operações. Os procedimentos cirúrgicos foram realizados 3 meses após a indução do diabetes com aloxana. A FR foi medida em todas anastomoses intestinais. Fragmentos de anastomoses do íleo e cólon foram retirados para dosagens de hidroxiprolina (HP) e proteína tecidual total (PT). RESULTADOS: A FR teve significante redução (P<0,05) nos grupos diabéticos G4 e G5, até 7 e 14 dias após a operação, respectivamente, quando comparada à observada nos grupos controles G2 e G3. Não foram observadas diferenças significantes nas dosagens de HP e PT em ratos diabéticos e controles, tanto operados no íleo como no cólon, em todos os momentos de avaliação. CONCLUSÃO: O diabetes conduz a alterações da força de ruptura de anastomoses intestinais durante a fase inicial da reparação da ferida cirúrgica, porém, este fato parece não estar relacionado à capacidade de sintetizar colágeno.

Effect of thalidomide on the healing of colonic anastomosis, in rats / Efeito da talidomida na cicatrização de anastomoses colônicas em ratos

PURPOSE: Thalidomide, because of its anti-inflammatory properties, as re-emerged as an option for the treatment of Crohn's disease refractory to standard therapy. We studied the effect of thalidomide on the healing of colonic anastomosis. METHODS: Sixty male rats (Rattus norvegicus), were divided into 3 groups of 20 animals each, respectively receiving 0.5 or 1.0 mg/kg thalidomide by the oral route for 7 days, or saline solution (control). All animals were submitted to continuous end-to-end anastomosis with 6-0 Prolene sutures. After sacrifice the anastomoses were analyzed macroscopically and submitted to determination of hydroxyproline, to histology and to immunohistochemistry for metalloproteinase 1, metalloproteinase 1 inhibitor and vascular endothelial growth factor (VEGF). RESULTS: Statistical analysis of the data showed no significant difference in macroscopic aspect or hydroxyproline determination (p= 0.5403). In the immunohistochemical analysis, the following p values were obtained: p = 0.5817 for VEGF, p = 0.1854 for metalloproteinase 1, and p = 0.0023 for metalloproteinase 1 inhibitor, with this last value being considered statistically significant. CONCLUSION: We conclude that thalidomide influenced collagen maturation. There was a stronger action of metalloproteinases, possibly indicating a negative tendency for the healing process.

OBJETIVO: Sabe-se que agentes farmacológicos podem influenciar no processo de cicatrização. A talidomida, devido às suas propriedades antiinflamatórias, tem ressurgido como uma opção no tratamento da doença de Crõhn refratária à terapêutica convencional. Neste trabalho, estudamos o efeito da talidomida na cicatrização de anastomoses colônicas no rato. MÉTODOS: Foram utilizados 60 animais Rattus norvegius, com peso médio de 300g. Organizou-se 3 grupos de 20 animais, sendo um grupo controle (AC), um grupo (BD), com administração de talidomida 0,5 mg/kg por 7 dias e um grupo (AD) com administração de talidomida 1,0 mg/kg por 7 dias. Foi realizada anastomose término-terminal contínua de prolene 6-0. O sacrifício foi no 7º. dia pós operatório e as anastomoses foram analisadas quanto a aspecto macroscópico, dosagem de hidroxiprolina, histologia em hematoxilina-eosina e imuno-histoquímica para metaloproteinase 1, inibidor de metaloproteinase 1 e VEGF. RESULTADOS: Não houve diferença estatisticamente significante para a observação macroscópica e para dosagem de hidroxiprolina (p=0,5403). Na análise imunohistoquímica, para VEGF houve p=0,5817, para metaloproteinase 1, p=0,1854 e para inibidor de metaloproteinase, p=0,0023, considerado estatisticamente significante. CONCLUSÃO: Concluímos que a talidomida influenciou na maturação do colágeno. Notou-se maior ação das metaloproteinases, que pode significar uma tendência negativa para o processo cicatricial.

An experimental model for the study of drug effects on cutaneous healing

PURPOSE: Immunosuppressive agents are known to interfere with the healing of surgical wounds. The increasing use of these drugs warrants a better understanding of their effects on wound healing. The aim of this study was to develop an experimental model that would allow for a reliable and rapid assessment of drug effects on cutaneous wound healing. METHODS: Thirty syngeneic Lewis rats underwent surgical incision on their dorsal region, in the presence or absence of a three-week regimen of immunosuppressant drug therapy (i.e., cyclosporin, azathioprine, and prednisone). Surgical site tissue was collected at intervals over 21 days after surgery and analyzed for cell number and collagen fiber content. Both of these quantitative assessments were performed using digital image capture with the Image Pro Plus 4.5 software. RESULTS: Computerized histomorphometric analyses revealed an apparent inhibition of cellular responses and collagen fiber production in drug-treated animals compared to control animals. CONCLUSION: The experimental model was reproducible, easy to perform, and allowed for quantitative histological evaluations. It may be useful for the study of surgical healing in the presence of other drug classes.

OBJETIVO: Várias drogas podem interferir no processo de cicatrização de feridas cirúrgicas, incluindo-se aqui agentes imunossupressores. Devido ao crescimento do número de pessoas tratadas com estas drogas, um modelo experimental, baseado em um sistema computadorizado de avaliação histológica, para verificar os efeitos destas drogas sobre a cicatrização cirúrgica pode se mostrar útil e permitir, também, o estudo das ações de outras drogas sobre este processo. MÉTODOS: Trinta ratos Lewis isogênicos foram submetidos a incisões cirúrgicas na região dorsal e metade deles foi imunossuprimida utilizando-se ciclosporina, azatioprina e prednisona por 3 semanas. Em cinco intervalos de tempo pré-fixados, avaliações histológicas quantitativas de células e fibras colágenas dos sítios operatórios de animais tratados e controles foram realizadas, utilizando-se captura digital das imagens por meio do programa Image Pro Plus 4.5. RESULTADOS: A avaliação dos sítios cirúrgicos mostrou, nos animais sob uso de drogas imuossupressoras, aparente diminuição no número de células e na produção de fibras colágenas. CONCLUSÕES: O modelo experimental proposto mostrou-se viável por exigir recursos pouco sofisticados, pela fácil realização e por fornecer avaliações histológicas quantitativas. Este modelo poderá ser utilizado para o estudo da ação de outras classes de drogas sobre a cicatrização cirúrgica.

Estudo da pele humana fotoenvelhecida após tratamento com terapia fotodinâmica associada ao ácido 5 delta-aminolevulínico tópico: avaliação imunoistoquímica, do colágeno e do tecido elástico / Study of photo aged human skin after photodynamics therapy treatment with topic 5 delta aminolevulinic acid: immunohistochemical, colagenous and elastic fibre analysis

A TFD atualmente é considerada uma modalidade terapêutica no fotoenvelhecimento.A destruição seletiva do tecido alvo ocorre através de uma reação química, onde se utiliza um agente fotossensibilizante,determinado comprimento de onda de luz e oxigênio. Foram estudadas 13 pacientes com fotoenvelhecimanto clinico que foram submetidas a três sessões de TFD-ALA tópico associado com luz vermelha (630nm) com intervalo quinzenal. Após 21 dias da ultima aplicação 12 pacientes apresentaram melhora clinica do fotonvelhecimento e aumento do tecido colágeno e fibra elástica.Em relação a modulação do sistema imune houve diminuição da população de linfócitos CD4 e CD8, aumento da IL-4 na epiderme e derme, diminuição da TNF-α na epiderme e aumento na derme.A população de células de Langerhans e o INF-γ não apresentaram alterações.A TFD atualmente é considerada uma modalidade terapêutica no fotoenvelhecimento.A destruição seletiva do tecido alvo ocorre através de uma reação química, onde se utiliza um agente fotossensibilizante,determinado comprimento de onda de luz e oxigênio. Foram estudadas 13 pacientes com fotoenvelhecimanto clinico que foram submetidas a três sessões de TFD-ALA tópico associado com luz vermelha (630nm) com intervalo quinzenal. Após 21 dias da ultima aplicação 12 pacientes apresentaram melhora clinica do fotonvelhecimento e aumento do tecido colágeno e fibra elástica.Em relação a modulação do sistema imune houve diminuição da população de linfócitos CD4 e CD8, aumento da IL-4 na epiderme e derme, diminuição da TNF-α na epiderme e aumento na derme.A população de células de Langerhans e o INF-γ não apresentaram alterações.

PDT is nowadays considered one of the therapies for photo-aging.The selective destruction of target tissue occurs through a chemical reaction, by using a photosensitizing, a certain light with a wave length and oxygen. Thirteen pacients with clinical photo-aging have been studied, submitted to three sections of topic ALA-PDT associated with red light (630nm) at fortnight’s intervals.After 21 days of the last treatment, 12 patients showed improved clinic photo-aging and an increase of colagenous and elastic fibre tissue. Regarding the modulation of the immune system, there was a decrease of the lymphocytes CD4 and CD8 population, an increase of interleucin-4 in the epidermis and in the dermis, decrease of TNF-α in the epidermis and increase in the dermis. The Langerhans cells poulation and the INF-y did not present alterations.