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RESUMEN: Los concentrados plaquetarios han emergido como un potencial material regenerativo, utilizado de forma aislada o como andamiaje para otros materiales de injerto. Son extractos de sangre, obtenidos después de procesar una muestra de sangre completa, mediante centrifugación. El primer reporte data de 1970, con un CP utilizado como pegamento para mejorar cicatrización de heridas de piel. En 1998, se usaron en cirugía oral y maxilofacial. Desde entonces, se han desarrollado diferentes técnicas y una variedad de preparaciones. Entre ellas, cabe destacar el plasma rico en plaquetas, fibrina rica en plaquetas y leucocitos (L-PRF) y plasma rico en factores de crecimiento (PRGF). El desarrollo de estos biomateriales, se debe en parte, a la posibilidad de alterar la concentración de mediadores químicos liberados en una lesión que provoque la formación de un coágulo, que pueda madurar conforme transcurran las fases del proceso inflamatorio y concluya con la regeneración íntegra del tejido dañado. El objetivo de este manuscrito fue describir las principales vías de señalización intracelular que se activan en presencia del L-PRF en cirugía oral, y sus efectos en la regulación del ciclo celular.

ABSTRACT: Platelet concentrates (PC) have emerged as a potential regenerative material, used in isolation or as scaffolding for other graft materials. They are blood extracts, obtained after processing a sample of whole blood, by centrifugation. The first report dates from 1970, with a PC used glue to improve the healing of skin wounds. In 1998, they were used in oral and maxillofacial surgery. Since then, different techniques and a variety of preparations have been developed. These include platelet-rich plasma, fibrin rich in platelets and leukocytes (L-PRF) and plasma rich in growth factors (PRGF). The development of these biomaterials, is due in part to the possibility of altering the concentration of chemical mediators released in a lesion that causes the formation of a clot, which can mature as the phases of the inflammatory process pass and conclude with the complete regeneration of the damaged tissue. The aim of this manuscript was to describe the main intracellular signaling pathways that are activated in the presence of LPRF in oral surgery, and its effects on the regulation of the cell cycle.

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This study aimed to investigate the effects of gallium-aluminum-arsenium (GaAlAs) (670 nm) laser therapy on neoangiogenesis and fibroplasia during tissue remodelling. Forty male Wistar rats underwent cutaneous surgery and were divided into 2 experimental groups: the Control and Laser group (9 mW, 670 nm, 0.031 W/cm2 , 4 J/cm2 ). After 14, 21, 28, and 35 days, the animals were euthanised. Descriptive and quantitative analyses were performed in sections stained with haematoxylin-eosin and Sirius Red, respectively. The amounts of VEGF+ and CD31+ cells were evaluated by immunohistochemistry and histomorphometric analysis, respectively. Statistical analysis was performed using the Mann-Whitney, Friedman, and Spearman correlation test, P < 0.05. The collagen expression was significantly higher in the laser group compared with the control group on days 14 and 21 after the creation of the skin wound (P = 0.008; P = 0.016) and in the control group between 14 and 28 and 14 and 35 days (P = 0.001; P = 0.007). There were more blood vessels in three periods of the study only in the (Laser) treated group, with statistical significance at day 14 (P = 0.016). There was no statistically significant difference in VEGF+ cell count in the different experimental groups throughout the study, although a positive correlation was shown with the area of collagen on days 14 and 28 (P = 0.037). Laser treatment had a positive effect in the late course of healing, particularly with regards to collagen expression and the number of newly formed vessels. VEGF+ cells were present in both experimental groups, and VEGF appeared to influence fibroplasia in the treated group.

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Purpose: To investigated the inflammatory, angiogenic and fibrogenic activities of the Schinus terebinthifolius Raddi leaves oil (STRO) on wound healing. Methods: The excisional wound healing model was used to evaluate the effects of STRO. The mice were divided into two groups: Control, subjected to vehicle solution (ointment lanolin/vaseline base), or STRO- treated group, administered topically once a day for 3, 7 and 14 days post-excision. We evaluated the macroscopic wound closure rate; the inflammation was evaluated by leukocytes accumulation and cytokine levels in the wounds. The accumulation of neutrophil and macrophages in the wounds were determined by assaying myeloperoxidase and N-acetyl--D-glucosaminidase activities. The levels of TNF-, CXCL-1 and CCL-2 in wound were evaluated by ELISA assay. Angiogenesis and collagen fibers deposition were evaluated histologically. Results: We observed that macroscopic wound closure rate was improved in wounds from STRO-group than Control-group. The wounds treated with STRO promoted a reduction in leucocyte accumulation and in pro-inflammatory cytokine. Moreover, STRO treatment increased significantly the number of blood vessels and collagen fibers deposition, as compared to control group. Conclusion: Topical application of STRO display anti-inflammatory and angiogenic effects, as well as improvement in collagen replacement, suggesting a putative use of this herb for the development of phytomedicines to treat inflammatory diseases, including wound healing.(AU)

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Abstract Purpose: To investigated the inflammatory, angiogenic and fibrogenic activities of the Schinus terebinthifolius Raddi leaves oil (STRO) on wound healing. Methods: The excisional wound healing model was used to evaluate the effects of STRO. The mice were divided into two groups: Control, subjected to vehicle solution (ointment lanolin/vaseline base), or STRO- treated group, administered topically once a day for 3, 7 and 14 days post-excision. We evaluated the macroscopic wound closure rate; the inflammation was evaluated by leukocytes accumulation and cytokine levels in the wounds. The accumulation of neutrophil and macrophages in the wounds were determined by assaying myeloperoxidase and N-acetyl-β-D-glucosaminidase activities. The levels of TNF-α, CXCL-1 and CCL-2 in wound were evaluated by ELISA assay. Angiogenesis and collagen fibers deposition were evaluated histologically. Results: We observed that macroscopic wound closure rate was improved in wounds from STRO-group than Control-group. The wounds treated with STRO promoted a reduction in leucocyte accumulation and in pro-inflammatory cytokine. Moreover, STRO treatment increased significantly the number of blood vessels and collagen fibers deposition, as compared to control group. Conclusion: Topical application of STRO display anti-inflammatory and angiogenic effects, as well as improvement in collagen replacement, suggesting a putative use of this herb for the development of phytomedicines to treat inflammatory diseases, including wound healing.

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Purpose: To investigated the inflammatory, angiogenic and fibrogenic activities of the Schinus terebinthifolius Raddi leaves oil (STRO) on wound healing. Methods: The excisional wound healing model was used to evaluate the effects of STRO. The mice were divided into two groups: Control, subjected to vehicle solution (ointment lanolin/vaseline base), or STRO- treated group, administered topically once a day for 3, 7 and 14 days post-excision. We evaluated the macroscopic wound closure rate; the inflammation was evaluated by leukocytes accumulation and cytokine levels in the wounds. The accumulation of neutrophil and macrophages in the wounds were determined by assaying myeloperoxidase and N-acetyl--D-glucosaminidase activities. The levels of TNF-, CXCL-1 and CCL-2 in wound were evaluated by ELISA assay. Angiogenesis and collagen fibers deposition were evaluated histologically. Results: We observed that macroscopic wound closure rate was improved in wounds from STRO-group than Control-group. The wounds treated with STRO promoted a reduction in leucocyte accumulation and in pro-inflammatory cytokine. Moreover, STRO treatment increased significantly the number of blood vessels and collagen fibers deposition, as compared to control group. Conclusion: Topical application of STRO display anti-inflammatory and angiogenic effects, as well as improvement in collagen replacement, suggesting a putative use of this herb for the development of phytomedicines to treat inflammatory diseases, including wound healing.

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Abstract Background: Crocin is reported to have a wide range of biological activities such as cardiovascular protection. Recent epidemiologic studies have shown that exercise reduces cardiovascular morbidity and mortality in the general population. Objective: The aim of this study was to evaluate the effect of crocin and voluntary exercise on miR-126 and miR-210 expression levels and angiogenesis in the heart tissue. Methods: Animals were divided into 4 groups: control, exercise, crocin, and exercise-crocin. Animals received oral administration of crocin (50 mg/kg) or performed voluntary exercise alone or together for 8 weeks. Akt, ERK1/2 protein levels, miR-126 and miR-210 expression were measured in the heart tissue. Immunohistochemical method was used to detect CD31 in the heart tissue. Results: Akt and ERK1/2 levels of the heart tissue were higher in crocin treated group and voluntary exercise trained group after 8 weeks. Combination of crocin and exercise also significantly enhanced Akt and ERK1/2 levels in the heart tissue. MiR-126, miR-210 expression and CD31 in the heart increased in both crocin and voluntary exercise groups compared with control group. In addition, combination of exercise and crocin amplified their effect on miR-126 and miR-210 expression, and angiogenesis. Conclusion: Crocin and voluntary exercise improve heart angiogenesis possibly through enhancement of miR-126 and miR-210 expression. Voluntary exercise and diet supplementation with crocin could have beneficial effects in prevention of cardiovascular disease.

Resumo Fundamentos: A crocina tem uma vasta gama de atividades biológicas, tais como a proteção cardiovascular. Estudos epidemiológicos recentes demonstraram que o exercício reduz a morbidade e a mortalidade cardiovasculares na população em geral. Objetivo: O objetivo deste estudo foi avaliar o efeito da crocina e do exercício voluntário nos níveis de expressão miR-126 e miR-210 e na angiogênese no tecido cardíaco. Métodos: Os animais foram divididos em 4 grupos: controle, exercício, crocina e exercício-crocina. Os animais receberam a administração oral de crocina (50 mg/kg) ou realizaram exercício voluntário sozinhos ou em conjunto durante 8 semanas. Os níveis de proteína Akt, ERK1/2, e a expressão de miR-126 e miR-210 foram medidos no tecido cardíaco. O método imunohistoquímico foi utilizado para detectar CD31 no tecido cardíaco. Resultados: Os níveis de Akt e ERK1/2 do tecido cardíaco foram maiores no grupo tratado com crocina e no grupo de exercício voluntário após 8 semanas. A combinação de crocina e exercício também aumentou significativamente os níveis de Akt e ERK1/2 no tecido cardíaco. A expressão de MiR-126, miR-210 e CD31 no coração aumentou tanto em no grupo de crocina como no grupo de exercício voluntário em comparação com o grupo de controle. Além disso, a combinação de exercício e crocina amplificou seu efeito na expressão de miR-126 e miR-210 e angiogênese. Conclusão: A Crocina e o exercício voluntário melhoram a angiogênese cardíaca possivelmente através do aumento da expressão de miR-126 e miR-210. O exercício voluntário e a suplementação dietética com crocina podem ter efeitos benéficos na prevenção de doenças cardiovasculares.

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La regulación fisiológica de la función vascular es esencial para la salud cardiovascular y depende de un adecuado control de mecanismos moleculares desencadenados por células endoteliales en respuesta a estímulos mecánicos y químicos inducidos por flujo sanguíneo. La disfunción endotelial es uno de los principales factores de riesgo de enfermedad cardiovascular, donde un desequilibrio entre la síntesis de moléculas vasodilatadoras y vasoconstrictoras constituye uno de sus principales mecanismos. En este contexto, el estrés de flujo es uno de los estímulos más importantes para mejorar la función vascular, gracias a que la mecanotransducción endotelial generada por la estimulación de diversos mecanosensores endoteliales induce la generación de estímulos intracelulares que culmina con un incremento en la biodisponibilidad de moléculas vasodilatadoras como el óxido nítrico y, a largo plazo, con la inducción de mecanismos angiogénicos. Estos mecanismos permiten proporcionar el sustento fisiológico a los efectos del ejercicio físico sobre la salud vascular. En la presente revisión se discuten los mecanismos moleculares implicados en la respuesta vascular modulada por estrés de flujo inducido por ejercicio y su impacto en la reversión del daño vascular asociado a las enfermedades cardiovasculares más prevalentes en nuestra población.

The physiological regulation of vascular function is essential for cardiovascular health and depends on adequate control of molecular mechanisms triggered by endothelial cells in response to mechanical and chemical stimuli induced by blood flow. Endothelial dysfunction is one of the major risk factors for cardiovascular disease, where an imbalance between synthesis of vasodilator and vasoconstrictor molecules is one of its main mechanisms. In this context, the shear stress is one of the most important mechanical stimuli to improve vascular function, due to endothelial mechanotransduction, triggered by stimulation of various endothelial mechanosensors, induce signaling pathways culminating in increased bioavailability of vasodilators molecules such as nitric oxide, that finally trigger the angiogenic mechanisms. These mechanisms allow providing the physiological basis for the effects of exercise on vascular health. In this review it is discussed the molecular mechanisms involved in the vascular response induced by shear stress and its impact in reversing vascular injury associated with the most prevalent cardiovascular disease in our population.

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The physiological regulation of vascular function is essential for cardiovascular health and depends on adequate control of molecular mechanisms triggered by endothelial cells in response to mechanical and chemical stimuli induced by blood flow. Endothelial dysfunction is one of the major risk factors for cardiovascular disease, where an imbalance between synthesis of vasodilator and vasoconstrictor molecules is one of its main mechanisms. In this context, the shear stress is one of the most important mechanical stimuli to improve vascular function, due to endothelial mechanotransduction, triggered by stimulation of various endothelial mechanosensors, induce signaling pathways culminating in increased bioavailability of vasodilators molecules such as nitric oxide, that finally trigger the angiogenic mechanisms. These mechanisms allow providing the physiological basis for the effects of exercise on vascular health. In this review it is discussed the molecular mechanisms involved in the vascular response induced by shear stress and its impact in reversing vascular injury associated with the most prevalent cardiovascular disease in our population.

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A Doença arterial periférica (DAP) é uma manifestação clínica da aterosclerose, quando esta afeta principalmente as artérias que irrigam os membros inferiores. O exercício aeróbico provoca nos membros afetados pela doença um ciclo de reperfusão-isquemia que desencadeia uma resposta sistêmica aguda caracterizada por aumento do estresse oxidativo, inflamação e disfunção endotelial. Assim, uma terapia antioxidante pode ser uma terapia alternativa para esses pacientes. Os microRNAs (miRNAs) foram recentemente reconhecidos como reguladores pós-transcricionais, e a identificação desses pode elucidar mecanismos gênicos adicionais pelos quais o exercício é atuante, levando a identificação de genes que são modulados, abrindo perspectivas de abordagens de terapia gênica que podem levar à reversão do quadro da doença arterial periférica. Objetivo: Verificar o efeito de uma sessão aguda de exercício aeróbico máxima com e sem uso do antioxidante Nacetilcisteína (NAC) sobre a expressão de microRNAs e marcadores inflamatórios e de estresse oxidativo circulantes em pacientes com DAP. Métodos: Foram recrutados pacientes com DAP estágio II do Ambulatório da Disciplina de Cirurgia Vascular do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. Os pacientes foram submetidos a duas sessões experimentais após a suplementação de NAC ou Placebo. Foi analisado o perfil de expressão de microRNAs circulantes dos indivíduos em repouso e após o exercício máximo, e confirmado a expressão gênica dos miRNAs alterados após o exercício e de seus alvos; e dos níveis plasmáticos de endotelina-1 (ET-1), proteína quimiotática de monócitos-1 (MCP-1), molécula de adesão intercelular-1 (ICAM-1), molécula de adesão celular vascular-1 (VCAM-1), substâncias reativas ao ácido tiobarbitúrico, 8- isoprostano e glutationa. Resultados: O tratamento com NAC não alterou o xvii tempo de caminhada, as respostas hemodinâmicas e cardiopulmonar dos...

Peripheral arterial disease (PAD) is a clinical manifestation of atherosclerosis, when it mainly affects the arteries supplying the lower limbs. Aerobic exercise causes the member affected by the disease cycle of reperfusion-ischemia triggers an acute systemic response characterized by increased oxidative stress, inflammation and endothelial dysfunction. Thus, an antioxidant therapy is an alternative therapy for these patients. MicroRNAs (miRNAs) have been recognized as posttranscriptional regulators, and identifying these may elucidate additional gene mechanisms by which the exercise is active, leading to identification of genes that are modulated, opening prospects of gene therapy approaches that can lead to picture reversal of peripheral arterial disease. Aim: To determine the effect of an acute bout of maximal aerobic exercise with and without the antioxidant N-acetylcysteine (NAC) on the expression of microRNAs and inflammatory markers and circulating oxidative stress in patients with PAD. Methods: We recruited patients with PAD stage II Clinic of Vascular Surgery of Hospital das Clinicas, Faculty of Medicine, University of São Paulo. The patients underwent two experimental sessions after supplementation of NAC or placebo. the expression profile of circulating microRNAs of individuals at rest and after maximal exercise was analyzed and confirmed the gene expression of miRNAs changed after exercise and its targets; and plasma levels of endothelin-1 (ET-1), monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1) cell vascular cell adhesion molecule-1 (VCAM-1) , reactive substances to thiobarbituric acid, 8-isoprostane and glutathione. Results: Treatment with NAC did not change the walking time, and cardiopulmonary hemodynamic responses of patients with PAD. The rest blood flow in the leg of these patients was higher after the completion of the exercise and treatment with NAC did not change this...

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La vasculogénesis es controlada por una serie de mecanismos que se activan en función del tiempo y del espacio durante el desarrollo embrionario. Múltiples son las vías de señalización implicadas en las etapas del proceso vasculogénico, las que se inician con estímulos angiogénicos desde el mesodermo o desde el endodermo para dar origen a los angioblastos (células progenitoras endoteliales). Proteínas como el factor de crecimiento vascular endotelial (VEGF), factor de crecimiento fibroblastico 2 (FGF2), entre otras, constituyen factores claves en la inducción de este proceso. Posteriormente, los angioblastos deben migrar para dar origen a los vasos primitivos, proceso en el que participan factores atrayentes y repulsivos que orientarán la dirección de su migración. Adicionalmente, los mecanismos de diferenciación arterio-venosa, regulados por la vía de señalización Hedgegog, VEGF y Notch, son determinados antes del inicio de la circulación, lo que sugiere que el destino de la célula endotelial se encuentra genéticamente determinado. Por su parte, los procesos de remodelación y proliferación vascular post natal, son generados a través de la formación de nuevos vasos a partir de vasos pre existentes (angiogénesis). El factor angiogénico que induce los cambios morfológicos y funcionales en las células endoteliales es el VEGFA, las cuales, adquieren la capacidad de direccionar al nuevo vaso en desarrollo. Uno de los principales estímulos físicos que modifica el patrón de crecimiento de los lechos vasculares es el estrés de flujo, el cual, es susceptible de ser modificado por situaciones de estrés como el ejercicio físico. En la presente revisión, se abordan los principales mecanismos implicados en la regulación fisiológica de la vasculogénesis y angiogénesis. Adicionalmente, se discutirán los mecanismos que sustentan la respuesta vascular inducida por estrés de flujo, considerando su rol en el establecimiento de los patrones de crecimiento vascular.

Vasculogenesis is controlled by a number of mechanisms that are activated as a function of time and space during embryonic development. Multiple signaling pathways are involved in the stages of vasculogenic process, which start with angiogenic stimuli from the mesoderm or the endoderm to give rise to angioblasts (endothelial progenitor cells). Proteins such as vascular endothelial growth factor (VEGF), fibroblast growth factor 2 (FGF2), among others, are key factors in the induction of this process. Subsequently, the angioblasts must migrate to give birth to primitive vessels, a process that involves attractive and repulsive factors that guide the direction of their migration. Additionally, arterial and venous differentiation regulated hedgegog signaling pathway, VEGF and Notch are determined before the start of circulation, suggesting that the endothelial cell fate is determined genetically. On the other hand, the processes of remodeling and postnatal vascular proliferation are generated through the formation of new vessels from pre-existing vessels (angiogenesis). The angiogenic factor that induces morphological and functional changes in the endothelial cells is the VEGFA, these vessels acquire the ability to address the new developing vessel. One of the main physical stimuli that modify the growth pattern of the vascular beds is the shear stress, which is modified by exercise. In this review, the main mechanisms involved in the physiological regulation of vasculogenesis and angiogenesis are addressed. Additionally, the mechanisms underlying the vascular response induced by shear stress will be discussed, considering its role in establishing patterns of vascular growth.

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Sepsis is a systemic uncontrolled inflammatory response in the presence of an infection. It remains a major cause of morbidity and mortality in hospitalized patients. According to its severity, sepsis can progress to three different states: severe sepsis, septic shock, and multiple organ dysfunction syndrome, related to organ dysfunction and/or tissue hypoperfusion. Different processes underlie its pathophysiology; among them are oxidative stress, endothelial and mitochondrial dysfunction, and angiogenesis-related factors. However, no studies have integrated these elements in sepsis. The main difficulty in sepsis is its diagnosis. Currently, the potential of inflammatory biomarkers in septic patients remains weak. In this context, the research into new biomarkers is essential to aid with sepsis diagnosis and prognostication. Furthermore, even though the current management of severe forms of sepsis has been effective, morbimortality remains elevated. Therefore, it is essential to explore alternative approaches to therapy development. The aim of this review is to present an update of evidence supporting the role of oxidative stress and angiogenesis-related factors in the pathophysiology of the different forms of sepsis. It proposes a novel convergence between both elements in their role in the disease, and it will cover their utility as new diagnostic tools, predictors of outcome, and as novel therapeutic targets.

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A neutropenia febril (NF) em pacientes com neoplasias hematológicas é caracterizada pelo alto risco de sepse e choque séptico. Embora a utilização de escores clínicos como o MASCC permita a identificação de pacientes de baixo risco, este escore é menos informativo em pacientes de alto risco, onde se encaixam a maioria dos pacientes com neoplasias hematológicas, além daqueles submetidos a esquemas intensivos de quimioterapia. Ao mesmo tempo, a aplicação de biomarcadores de gravidade como a procalcitonina, validados em pacientes não-neutropênicos, é controversa em pacientes com NF. A quebra da barreira endotelial é um elemento chave no choque séptico, de modo que proteínas envolvidas neste processo são candidatos atrativos como biomarcadores de gravidade na sepse. Neste estudo, avaliamos prospectivamente o valor da dosagem de VEGF-A, sFlt-1, Ang-1 e Ang-2 como biomarcadores da evolução para choque séptico em 120 pacientes com NF. Pacientes internados nas enfermarias de Hematologia e Transplante de Medula Óssea do HC da UNICAMP para tratamento de NF entre março de 2011 e 2012 foram convidados a participar. As amostras foram coletadas na manhã seguinte à entrada no estudo, junto com a coleta de exames de rotina. O estudo foi desenhado com o objetivo de mimetizar as condições de coleta e processamento das amostras, que seriam encontradas na prática clínica real. Foi avaliada a evolução para choque séptico e mortalidade em 28 dias. Os resultados foram comparados com marcadores de prognóstico clássicos como proteína C reativa, e escores MASCC e SOFA. No total, 99 pacientes preencheram os critérios de inclusão, dos quais 19,8% evoluíram com choque séptico. Não foram observadas diferenças clínicas e demográficas entre os pacientes com NF não-complicada e choque séptico, exceto pelo escore SOFA, significativamente mais elevado no segundo grupo.

Febrile neutropenia (FN) in patients with hematologic malignancies is characterized by a high risk of sepsis complications and septic shock. Although the use of clinical scores such as the MASCC allows the identification of low-risk patients, this score is much less informative in high-risk patients, a category in which most patients with hematologic malignancies, and those undergoing intensive chemotherapy regimens, fit in. At the same time, the use of classical biomarkers such as procalcitonin in non-neutropenic patients is controversial in patients with FN. Endothelial barrier breakdown is a key element in septic shock, so that proteins involved in this process are attractive candidates as biomarkers of sepsis severity. In this study, we prospectively evaluated the value of VEGF-A, sFlt-1, Ang-1 and Ang-2 serum levels as biomarkers of progression to septic shock in 120 patients with FN. Patients hospitalized in the Hematology and Bone Marrow Transplantation in-patient units of a university hospital (HC-UNICAMP) for the treatment of FN between March 2011 and March 2012 were invited to participate. Samples were collected in the following morning after study entry, along with the collection of routine labs. The study was designed to mimic the conditions of blood sample collection and processing that would be encountered in "real-world" clinical practice. Clinical outcomes were (1) progression to septic shock and (2) death within 28 days from fever onset. Results were compared with classical prognostic markers such as C-reactive protein, and MASCC and SOFA scores. In total, 99 patients met the inclusion criteria, of which 19.8% progressed to septic shock. No differences clinical and demographic differences were observed between patients with uncomplicated-FN or septic shock, except for a higher SOFA scores in the latter group.

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A sinalização celular por PAR¹ tem mostrado influenciar uma ampla gama de respostas patofisiológicas, incluindo ativação plaquetária, crescimento tumoral, inflamação e metástases. Baseando-se nisto, o objetivo do presente estudo foi avaliar o efeito do Parstatin, droga que tem ação biológica oposta àquela desencadeada pela ativação do PAR¹, durante o processo de indução e reparo da inflamação. Foi utilizado um modelo de periodontite experimental em ratos através da instalação de ligaduras de fio de algodão nos segundos molares superiores. Para isto, 72 ratos foram separados aleatoriamente em 9 grupos com 8 animais cada e receberam as ligaduras e injeção de veículo ou Parstatin nos períodos de 7 e 14 dias para observar a ação da inibição deste receptor nos períodos de indução de inflamação e reparo. Após estes períodos, os animais foram sacrificados e tiveram as maxilas removidas, dissecadas e divididas ao meio para avaliação histológica e radiográfica a fim de caracterizar infiltrado de células inflamatórias e perda óssea ao redor dos dentes.

PAR¹ cell signaling has been shown to be involved in several pathophysiological responses including platelet activation, tumor growth, inflammation and metastasis. Based on this, the aim of the present study was to evaluate the influence of Parstatin, a drug that presents a biological effect opposed to that of PAR¹ receptor activation, on inflammation induction and repair processes. Rats were subjected to cotton ligature-induced periodontitis bilaterally on the second upper molar teeth. Seventy-two rats were randomly assigned to 9 groups (n=8/group) and received ligatures and injection of vehicle or Parstatin for 7 or 14 days for both inflammation and repair induction. After that, the animals were sacrificed and their maxilla removed, dissected and divided in two for histologic and radiographic evaluation to characterize inflammatory cell infiltrate and bone loss around teeth.

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BACKGROUND: Osteopontin (OPN) has been verified to be closely associated with oncogenesis and remodeling processes. But this cytokine was rarely assessed in the presence of aortopathies, especially acute aortic dissection. The aim of the present study was to evaluate the expressions of OPN by way of molecular biological approaches so as to offer a better understanding of the possible mechanisms of the aortopathies. METHODS: Consecutive patients with type A acute aortic dissection (20 patients), aortic aneurysm (nine patients) or coronary artery disease (21 patients) referred to this hospital for surgical operations were enrolled into this study. Blood samples of the surgical patients after systematic heparinization, and control fast morning blood samples drawn from 21 young healthy volunteers who had no evidence of any healthy problems were investigated for enzyme linked immunosorbent assay (ELISA). The surgical specimens of the aortic tissues collected from the surgical patients during the operations were obtained for quantitative realtime reverse transcription polymerase chain reaction (RT-PCR) for OPN mRNA, western blot assay for OPN protein, and for immunohistochemical staining of OPN. Ascending aortic tissues from the autopsies of the healthy individuals dying of accident were obtained as controls of immunohistochemistry. RESULTS: By quantitative RT-PCR, the expressions of OPN mRNA were all upregulated in all three surgical groups. The quantitative results did not reveal any intergroup differences. Western blot assay revealed that OPN was positive with similar intensities of expressions in all three surgical groups. Quantitative western blot analyses of OPN expressions did not show any significance between groups. The OPN expressions by ELISA in the aortic tissue were 3.09311 ± 1.65737, 3.40414 ± 1.15095, and 1.68243 ± 0.31119 pg/mg protein in the aortic dissection, aortic aneurysm, and coronary artery disease groups, respectively. The OPN level of the patients with coronary artery disease was much lower than those with aortic dissection (P = 0.033) or with aortic aneurysm (P = 0.019). By unparametric tests, there were significant differences in the aortic OPN contents among aortic dissection, aortic aneurysm and coronary artery disease groups (P < 0.01). A significant direct correlation was present between plasma OPN concentration and the time interval from the onset to surgery of aortic dissection (Y = 0.1420X + 2.4838, r² = 0.5623, r = 0.750, P = 0.032). By immunohistochemistry, OPN was expressed in the aortic cells: in the intima, it was weaker in all three surgical groups in comparison with the healthy control; in the media, it was weak in the aortic dissection, intense positive in aortic aneurysm, focal positive in the coronary artery disease, but evenly positive in the healthy control groups; and in the adventitia, it was positive in the aortic dissection, coronary artery disease and healthy control groups, but weak positive in the aortic aneurysm group. CONCLUSION: These data may provide evidences that OPN may play a role in the pathogenesis of aortopathies including aortic dissection, aortic aneurysm, and coronary artery disease. OPN might be of potential perspective as a clinically diagnostic tool in the evaluations of the complex remodeling process incorporating vascular injury and repair.

OBJETIVOS: A osteopontina (OPN) está estreitamente associada com os processos de oncogênese e remodelação. Entretanto, essa citocina era raramente avaliada na presença de aortopatias, especialmente na dissecção aórtica aguda. O objetivo do presente estudo foi avaliar a expressão de OPN por meio de abordagens moleculares biológicas, de modo a oferecer uma melhor compreensão dos possíveis mecanismos das aortopatias. MÉTODOS: Pacientes consecutivos com um tipo de dissecção aguda da aorta (20 pacientes), aneurisma da aorta (nove pacientes) ou doença arterial coronária (21 pacientes) foram incluídos neste estudo. As amostras de sangue depois da heparinização sistemática e de 21 voluntários jovens e saudáveis não apontaram nenhuma evidência de qualquer problema ao serem investigados por ensaio imunoenzimático (ELISA). Os espécimes cirúrgicos dos tecidos aórtica coletados dos pacientes durante as operações foram obtidos para a reação de transcrição reversa quantitativa em tempo real em cadeia da polimerase (RT-PCR) para OPN mRNA, técnica de Western blot para a proteína OPN, e imunohistoquímica de OPN. Amostras da aorta de indivíduos saudáveis que morreram de acidente foram obtidos para controle imunohistoquímico. RESULTADOS: Com uso do RT-PCR quantitativo, as expressões de OPN mRNA foram suprarreguladas em todos os três grupos cirúrgicos. Os resultados quantitativos não revelaram quaisquer diferenças intergrupais. Western blot revelou que OPN foi positiva com intensidade semelhante de expressões em todos os três grupos. As análises quantitativas Western blot de expressões OPN não apresentaram significâncias entre os grupos. As expressões OPN medidas pelo teste ELISA no tecido aórtico foram 3,09311 ± 1,65737, 3,40414 ± 1,15095 e 1,68243 ± 0,31119 pg/mg de proteína na dissecção de aorta, aneurisma da aorta, e grupos de doença arterial coronariana, respectivamente. O nível de OPN dos pacientes com doença arterial coronariana foi muito menor do que aqueles com dissecção aórtica (P = 0,033) ou com aneurisma da aorta (P = 0,019). Testes não-paramétricos apontaram diferenças significativas nos teores de aorta OPN entre dissecção aórtica, aneurisma da aorta e grupos com doença arterial coronariana (P <0,01). Uma correlação direta significativa estava presente entre a concentração plasmática OPN e o intervalo de tempo entre o início da cirurgia de dissecção de aorta (Y = 2,4838 + 0,1420X, r² = 0,5623, r = 0,750, P = 0,032). Pela imunohistoquímica, a OPN foi expressa em células aórticas: na íntima, foi fraca em todos os três grupos cirúrgicos em comparação ao grupo saudável; na média, era fraca na dissecção aórtica, positiva intensa no aneurisma de aorta, focal positivo na doença arterial coronariana, mas igualmente positiva no grupo controle; e na adventícia, positiva para a dissecção da aorta, doença arterial coronariana e grupos de controle saudáveis, mas fraca positiva no grupo de aneurisma da aorta. CONCLUSÃO: Estes dados fornecem evidências de que a OPN pode desempenhar um papel na patogênese da aortopatias, incluindo dissecção aórtica, aneurisma da aorta R e doença arterial coronariana. OPN tem perspectiva potencial como ferramenta de diagnóstico clínico nas avaliações do processo de remodelação complexa, incluindo lesão vascular e de reparação.

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A pré-eclâmpsia (PE) é a principal causa de morbimortalidade materno-fetal, acometendo de 5 a 7% das gestantes no mundo. Apesar de exaustivamente estudada, ainda não existe tratamento satisfatório, além da interrupção da gestação nos casos mais graves. Além da divisão clássica da doença entre leve e grave, os estudos mais recentes sugerem uma nova classificação, subdividindo a PE em precoce (com aparecimento dos sintomas antes de 34 semanas de gestação) e tardia (sintomas após 34 semanas de gestação). As etapas fisiopatológicas diferem de tal forma entre esses dois grupos que alguns autores chegam a propor que se trata de entidades patológicas distintas. O objetivo deste estudo foi realizar uma revisão bibliográfica dos métodos de predição de PE atualmente pesquisados, dando ênfase a exames que podem ser realizados ainda no primeiro trimestre de gestação e àqueles que melhor identificam a paciente com risco aumentado de desenvolver PE precoce, devido ao pior prognóstico materno-fetal dessa condição.

Pre-eclampsia (PE) is the main cause of maternal and fetal morbidity and mortality, reaching 5 to 7% of pregnant women in the world. Although it has been broadly studied, there is still no satisfactory treatment other than the interruption of pregnancy in the most serious cases. In addition to the classical division of the disease in mild and severe, more recent studies suggest a new classification, subdividing the PE in early (with the appearance of symptoms before 34 weeks of gestation) and late (symptoms showing after 34 weeks of gestation). Physiopathology events differ so between these groups that some authors propose that those pathological entities are distinct. The goal of this study was to perform a bibliographic review of methods for predicting PE currently in research, emphasizing the exams that can be performed even in the first trimester of pregnancy and those that best recognize the patient with increased risk of developing PE early due to the worse prognosis of this condition.

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INTRODUÇÃO: O crescimento e a progressão de diversos tipos de tumores dependem da angiogênese, isto é, a formação de novos vasos sanguíneos. Fatores da família do VEGF (VEGF-A, VEGF-C, Flk-1/KDR) são potentes mediadores da angiogênese e níveis elevados destes fatores podem ser detectados em vários tumores humanos. Outras proteínas como NDRG1, osteonectina, HIF-1 e galectina-3 encontram-se associadas, de modo indireto, à angiogênese. MÉTODOS: O objetivo deste estudo foi analisar a expressão imunoistoquímica de NDRG1, osteonectina, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1 e galectina-3 em 265 lesões benignas e malignas da tireóide por tissue microarray (TMA), incluindo tireóide normal, bócio, adenoma folicular, carcinoma papilífero, carcinoma folicular e metástases de carcinoma papilífero e folicular da tireóide. Lâminas em diferentes níveis do bloco de TMA foram submetidas à reação imunoistoquímica utilizando-se anticorpos específicos contra NDRG1, osteonectina, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1, galectina-3. Foi realizada uma avaliação semiquantitativa da expressão destas proteínas, analisando-se a intensidade de coloração e a porcentagem de células positivas para cada marcador. RESULTADOS: De um modo geral, a expressão de NDRG1, osteonectina, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1 e galectina-3 foi maior nos carcinomas de tireóide em relação ao bócio e adenoma folicular. Dentre as proteínas estudadas, a galectina-3 foi aquela que apresentou melhor acurácia diagnóstica na distinção entre lesões benignas e malignas da tireóide, com uma sensibilidade 64,1% e especificidade de 94,4%. Nos carcinomas de tireóide, a expressão de NDRG1, VEGF-A, VEGF-C e HIF-1 apresentou correlação estatística significativa com estádio TNM mais avançado e alto risco pelo método AMES. A análise multivariada revelou que a idade 45 anos, o tamanho tumoral maior do que 4,0 cm, o estádio IV do TNM e a perda de expressão de galectina-3 foram fatores que contribuíram para a recorrência...

INTRODUCTION: The growth and progression of different types of tumors depend on angiogenesis, i.e., the formation of new blood vessels. Members of the VEGF family (VEGF-A, VEGF-C, Flk-1/KDR) are potent mediators of angiogenesis and elevated levels of these factors can be detected in various human tumors. Other proteins such as NDRG1, osteonectin, HIF-1 and galectin-3 are indirectly associated with angiogenesis. METHODS: The objective of this study was to investigate the immunohistochemical expression of NDRG1, osteonectin, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1 and galectin-3 in 265 benign and malignant thyroid lesions by tissue microarray (TMA), including normal thyroid, goiter, follicular adenoma, papillary carcinoma, follicular carcinoma, and papillary and follicular thyroid carcinoma metastases. Slides obtained from different levels of the TMA block were submitted to immunohistochemistry using specific antibodies against NDRG1, osteonectin, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1, and galectin-3. The expression of these proteins was analyzed semiquantitatively, evaluating the intensity of staining and the percentage of positive cells for each marker. RESULTS: In general, the expression of NDRG1, osteonectin, VEGF-A, VEGF-C, Flk-1/KDR, HIF-1 and galectin-3 was higher in thyroid carcinomas when compared to goiter and follicular adenoma. Among the proteins studied, galectin-3 presented the best diagnostic accuracy in the distinction between benign and malignant thyroid lesions, with a sensitivity of 64.1% and specificity of 94.4%. In thyroid carcinomas, the expression of NDRG1, VEGF-A, VEGF-C and HIF-1 was significantly correlated with a more advanced TNM stage and AMES high risk. Multivariate analysis revealed that age 45 years, tumor size > 4.0 cm, TNM stage IV and loss of expression of galectin-3 were associated with recurrence or death in patients with thyroid carcinoma. CONCLUSIONS: In thyroid carcinomas, the expression of NDRG1, VEGF-A, VEGF-C and HIF-1...

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RACIONAL: O câncer gástrico continua sendo objeto de grande número de estudos que tentam entender melhor sua gênese, e conseqüentemente propor novos tratamentos. Na atualidade destaque especial está sendo dado na marcação imunoistoquímica onde estão sendo utilizados marcadores diversos sem se saber ainda qual ou quais são os mais efetivos no diagnóstico. OBJETIVO: Identificar e quantificar por citometria de imagem a expressão dos marcadores de angiogênese Fator VIII e CD-34 em tumores gástricos. MÉTODOS:Foram utilizados 29 casos de adenocarcinomas gástricos, todos oriundos de material arquivado e conservado em blocos de parafina. Após a desparafinização, realizou-se coloração com marcadores imunoistoquímicos de angiogênese Fator VIII e CD-34, e as lâminas coradas foram submetidas a estudo citofotométrico de imagem em sistema informatizado SAMBA 4000. RESULTADOS: Foram comparados os dois parâmetros oferecidos pelo método, índice de marcagem e densidade óptica, apenas nos 17 casos em que ocorreu identificação de ambos marcadores. Observou-se expressão numericamente significativa de ambos, porém o Fator VIII apresentou melhor média de densidade óptica, enquanto o CD-34 melhor resultado quanto ao índice de marcagem. CONCLUSÃO:A citometria de imagem foi capaz de identificar e quantificar a expressão do Fator VIII e CD-34 de maneira confiável e satisfatória demonstrando presença de angiogênese, sendo que o Fator VIII marcou menores áreas com melhor qualidade e o CD-34 marcou maiores áreas com menor qualidade.

BACKGROUND: Gastric cancer remains a subject of great interest in several studies searching for understanding its genesis and, therefore, proposing new treatments. Currently special credits have been given to immunohistochemistry as various markers are being tested with uncertain efficacy on diagnosis. AIM: To identify and quantify the expression of Factor VIII and CD-34, angiogenesis markers, in gastric tumors by imaging cytometry. METHODS: Twenty-nine 29 gastric adenocarcinomas sampled-tissue in paraffin blocks underwent immunohistochemical angiogenesis markers evaluation for factor VIII and CD-34 and the marked slides were submitted to SAMBA 4000 reading. RESULTS:The investigation method offered two comparison parameters, marking index and optical density, showing up both in 17 cases. Significant numerical expression had been observed for both markers, but Factor VIII showed a better optical density average while CD-34 presented a better result for marking index. CONCLUSIONS: Image cytometry identified and quantified Factor VIII and CD-34 expressions in a reliable and satisfactory manner, revealing the presence of angiogenesis. Factor VIII enhanced in smaller areas with better quality while CD-34 enhanced greater areas with lower quality.

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Luxações dentárias resultam em injúrias ao feixe vásculo-nervoso, o que pode causar redução significante no nível de oxigênio das polpas dentárias, ou mesmo morte deste tecido conjuntivo especializado. O conhecimento dos mecanismos envolvidos na resposta das células pulpares à hipóxia pode melhorar o prognóstico dos tratamentos das luxações dentárias. O objetivo deste trabalho foi avaliar o comportamento de células-tronco (DPSC) e de fibroblastos (HDPF) de polpas dentárias humanas em condições de hipóxia e a capacidade destas em estimular a proliferação e formação de tubos capilares por células endoteliais microvasculares dérmicas humanas (HDMEC). As análises do western blot e imunocitoquímica foram realizadas para avaliar o efeito da hipóxia na ativação do fator de transcrição induzido pela hipóxia 1 (HIF-1alfa). A expressão dos fatores de crescimento vascular endotelial (VEGF) e fibroblástico básico (bFGF) foi avaliada pelos testes de imunoensaio ELISA. O efeito do HIF-1alfa na expressão do VEGF foi avaliado pelo tratamento das células DPSC e HDPF com 40 μM YC-1, inibidor do HIF-1alfa. Para a avaliação da angiogênese in vitro, os testes de proliferação celular (WST-1) e de formação de tubos capilares em gel de colágeno foram realizados após tratamento das células HDMEC com os meios de acondicionamento das células DPSC ou HDPF, em condições de normóxia ou hipóxia.. A análise dos dados obtidos foi realizada por testes estatísticos não-paramétricos com nível de significância de 5%. Aumento da atividade do HIF-1alfa e da expressão de VEGF foram observados nas células DPSC e HDPF em condições de hipóxia. Entretanto, não foram observadas alterações nos níveis de expressão de bFGF. A expressão do HIF-1alfa foi inibida...

Dental luxations can result in the severing of the dental pulp and the creation of a hypoxic environment that might lead to the death of the pulp tissue. Improved understanding of the mechanisms underlying the response of dental pulp cells to hypoxic conditions might lead to better treatment of dental luxations. The purpose of this work was to evaluate the behavior of Dental Pulp Stem Cells (DPSC) and Human Dental Pulp Fibroblasts (HDPF) under hypoxia and their ability to stimulate proliferation and sprouting of endothelial cells. Western blots and immunohistochemistry were used to evaluate the effect of hypoxia on the activation of the transcriptional factor HIF-1α. Vascular Endothelial Growth Factor (VEGF) and Basic Fibroblastic Growth Factor (bFGF) expression was evaluated by ELISA. The effect of HIF-1αon VEGF expression was evaluated by treating DPSC or HDPF with 40 μM YC-1, an inhibitor of HIF-1α. To evaluate the angiogenic potential of dental pulp cells in vitro, human dermal microvascular endothelial cells (HDMEC) were treated with conditioned medium from DPSC or HDPF in normoxia or hypoxia. Sprouting and WST-1 assays were used to evaluate capillaries tubes-like formation and cell proliferation rate,respectively. The statistic analysis was performed by non-parametric tests. HIF-1αactivity and VEGF expression were up-regulated in DPSC and HDPF under hypoxic conditions. However, no changes in the expression levels of bFGF were observed. YC-1 40μM was able to inhibit HIF-1alpha in both cell lines and it also reduced VEGF expression in DPSCs in hypoxia. HDPF under hypoxia increased HDMEC sprouting by 50% when compared to HDPF in normoxia, and by 30% when compared to DPSC in hypoxia. Notably, HDPF under hypoxia stimulated HDMEC proliferation by 30% when compared to DPSC in hypoxia. It was concluded...

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