We are grateful to Dr. Clowes for first developing and describing this method. We are also thankful to Dr. Tulis for his detailed protocol which has been fundamentally helpful to our previous, current and future work. This work was supported by grants R01HL097111 and R01HL123364 from the NIH to M.T., and by American Heart Association grant 14GRNT18880008 to M.T.
We would like to thank Rachel Newton for her expert technical support and for her valuable help during the filming process.

NOTA: O uso de animais para os seguintes experimentos tem sido revisado e aprovado pelo cuidado e uso Comitê Institucional Animal (IACUC). 1. Procedimentos pré-operatórios <ol style="margin-left: 40px;"><li> Esterilizar instrumentos cirúrgicos antes de usar. <ol><li> Autoclave todos os instrumentos cirúrgicos 24 horas ou menos antes da cirurgia. Se várias cirurgias são realizadas no mesmo dia, esterilizar os instrumentos por um esterilizador talão seca entre cirurgias. </li></ol></li><li> Filtro-esterilizar a solução salina antes da utilização. </li><li> Pesa-se o rato e o cálculo da dose de anestésicos (cetamina 80 mg / kg de xilazina e 7 mg / kg). </li><li> Administrar as drogas anestésicas intra-peritoneal (ip). <ol><li> Verificar a adequação da sedação por toe pitada de 5-10 min. Administrar uma pequena dose adicional de drogas (cetamina 7 mg / kg e xilazina 0,6 mg / kg), se a sedação não está completa. </li><li> Certifique-se de que a agulha é profundo o suficiente para entrega de drogas intra-peritoneally porque falha na entrega de toda a solução de um fármaco para a cavidade peritoneal irá causar sedação inadequada. NOTA: As úlceras e perda de cabelo na pele no local da injecção será visíveis vários dias após a cirurgia, devido a inadvertida por via subcutânea (sc) de injecção de solução de fármaco. </li></ol></li><li> Injectar 3 ml de solução salina estéril solução por via subcutânea (sc). Usando uma mecha de algodão estéril, aplicar uma pequena quantidade de pomada oftálmica para ambos os olhos para evitar as córneas de secar. </li><li> Prepare o equipamento de aquecimento. Almofadas de calor Pré-quentes por microondas ou banho-maria. </li><li> Coloque o animal indolentemente na plataforma cirúrgica. <ol><li> Retire o cabelo na região do pescoço ventral. Aplique o removedor de pêlos com um cotonete, espere 30 segundos e limpe completamente com gaze. </li><li> Esfregue o pescoço com esfoliação iodopovidona e 70% de etanol. </li></ol></li><li> Utilizar equipamento de proteção pessoal, incluindo vestido, capa de cabelo, máscara cirúrgica e óculos. Coloque em sterile luvas cirúrgicas no final antes de manusear os instrumentos cirúrgicos estéreis e suprimentos. </li><li> Armar o rato com um lençol cirúrgico estéril com apenas a região do pescoço exposto. </li></ol> 2. Procedimentos Cirúrgicos <ol style="margin-left: 40px;"><li> Durante o procedimento cirúrgico, verifique a profundidade da sedação de subprodutos animais pitada toe em cada 15 min. Se um animal responde a toe pitada, adicionar pequena dose adicional (10% da dose inicial) da cetamina e xilazina. </li><li> Dissecar artéria carótida comum esquerda (CCA) <ol><li> Usar um bisturi para fazer uma incisão longitudinal recta no meio do pescoço. O comprimento aproximado de incisão é de 1,5-2 cm, com a finalidade de isolar uma porção de 1,5-2 cm da artéria. O comprimento pode variar depende da finalidade do estudo. </li><li> Bluntly dissecar o tecido conjuntivo da pele. Mantenha as dicas fórceps-se e certifique-se para não perfurar a pele ou o tecido subjacente. </li><li> Dissecar camadas musculares longitudinalmente Along do lado esquerdo da traquéia. </li><li> Ao abrir-se a camada muscular, visualize a CCA esquerda com o nervo vago intimamente ligados. Sem rodeios dissecar ao lado da artéria carótida esquerda com extrema cautela para separar o nervo vago com o mínimo de alongamento. </li><li> Dissecar CCA distal até a bifurcação. Dissecar cuidadosamente a bifurcação e dois ramos - artéria carótida interna (ACI) e artéria carótida externa (ECA). </li><li> Manter dissecando em torno do CCA, até aproximadamente uma porção de 1,5-2 cm da artéria é isolado a partir dos tecidos circundantes. </li></ol></li><li> Lesão por balão <ol><li> Permanentemente fazer uma ligadura sobre o ECA em aproximadamente 5 mm de distância da bifurcação. Permanentemente ligadura do ramo occipital da ECA, que é próximo à bifurcação da ECA e ICA. Também ligar permanentemente outros ramos, se houver, localizando entre a bifurcação e ECA ligadura - por exemplo, o ramo da tireóide superior. Sutura que é utilizado para todas as ligaduras é 4-0 pretoseda. Grampo na extremidade proximal do CCA e a extremidade distal da ICA. NOTA: Agora, o fluxo sanguíneo foi interrompido permanentemente através da ligadura (on ECA) ou temporariamente por meio de recorte (no CCA e ICA). Conteúdo luminal na área de bifurcação ter sido isolado a partir da circulação sistémica. </li><li> Faça uma incisão arteriotomy no ECA por pequenos produtores, micro tesoura. Certifique-se de que a incisão fica perto do nó de sutura distal. Sangue limpo com soro fisiológico e cotonetes. </li><li> Insira o cateter balão insuflado 2F para o lúmen do TCE. Avançar o cateter balão proximal ao lúmen CCA. Mantenha o avanço do cateter proximal até sua ponta chega onde o clipe fica. </li><li> Conectar o dispositivo de insuflação do balão para um luer lock fêmea num torneira de 3 vias e ligar o bloqueio Luer macho da torneira de 3 vias para o cateter de balão. </li><li> Lentamente inflar o balão com aproximadamente 1,5 atm de pressão, de modo a distender a artéria carótida para 1,5 vezes o diâmetro. </li&gt; <li> Puxe cuidadosamente o balão rotativo de volta para a bifurcação. </li><li> Desinflar o balão e avançá-lo de volta para a extremidade proximal. Encha-o novamente e repita o procedimento puxando-back mais duas vezes. </li><li> Retirar o cateter balão do lúmen da artéria. </li></ol></li><li> Administração Intraluminal de reagentes (por exemplo, siRNA, drogas) NOTA: Aqui, o reagente utilizado foi uma solução contendo partículas lentivirais que codificam quer estromais da molécula alvo shRNA Interacção 1 (STIM1) ou não-alvo de controlo de shRNA. STIM1 é um retículo endoplasmático (ER) única proteína transmembranar que é um sensor de ER Ca 2+ controlando a activação da membrana plasmática canais de Ca2 + e é regulado para cima durante a desdiferenciação do músculo liso vascular em um fenótipo proliferativo migratório 12,30-33. <ol><li> Anexar um cateter intravascular sobre-a-agulha de uma seringa (24 L, 1,6 cm). Aspirar solução de 30 ul do ensaio reagentos. Inserir o cateter na mesma incisão no ECA. </li><li> Avance a ponta do cateter no CCA e amarrar um pedaço de fio de sutura com um único nó no ECA para fixar o cateter e fechar a incisão temporariamente. </li><li> Injectar a solução reagente de teste para o lúmen do CCA. Conservar a solução do lúmen do vaso para 30 min. Mantenha o tecido exposto úmido com solução salina e cobri-lo com um pedaço de gaze molhada. </li><li> Após a incubação, aspirar a solução restante. Solte que nó simples e retirar o cateter. </li><li> Amarre o ECA com um pedaço de fio de sutura proximal à incisão arteriotomy. Adicione o nó tão próximo quanto possível da bifurcação. </li></ol></li><li> Feche acima da ferida. <ol><li> Uma vez que a lesão por balão e introdução do cateter pode causar um vazamento ou punção do vaso sanguíneo, remover o clipe na ICA e verifique se há algum vazamento. Se o sangramento for observado, aplicar um pedaço de gaze e aplique uma leve pressão para parar o sangramento. </li><li> Removero outro clipe no CCA. </li><li> Certifique-se de que não há sinais de hemorragia, e depois remover todas as braçadeiras e outros instrumentos cirúrgicos. Corte o excesso de suturas. </li><li> Fechar ferida com suturas de pele (4-0 seda preta). Cotonete em todos os lados da ferida fechada com o Povidon-iodo ou outros agentes anti-séptico / agente bactericida / virucida. Injectar 3 ml de solução salina estéril sc </li></ol></li></ol> 3. Os procedimentos pós-operatórios <ol style="margin-left: 40px;"><li> Mantenha rato na almofada de calor após a cirurgia. Administrar uma dose de 0,05 mg / kg de buprenorfina por via de injecção intramuscular (im) para o rato. Durante o processo de recuperação, mantenha os olhos ea boca dos animais úmido. Monitorar o animal até que ele está acordado e ambulante. </li><li> Colocar o animal em uma gaiola limpa, sem qualquer roupa de cama até que esteja totalmente recuperado. Após a recuperação, o animal será devolvido para a sala de animais e alojados individualmente. </li></ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+carotid+artery+balloon+injury+model.">Rat carotid artery balloon injury model.</a> Methods In Molecular Medicine. 139, 1-30 (2007).</li><li>Zhang, W., Trebak, M., Szallasi, A., B&#237;r&#243;, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon+Injury+in+Rats+as+a+Model+for+Studying+TRP+Channel+Contribution+to+Vascular+Smooth+Muscle+Remodeling.">Balloon Injury in Rats as a Model for Studying TRP Channel Contribution to Vascular Smooth Muscle Remodeling.</a> T TRP Channels in Drug DiscoveryMethods in Pharmacology and Toxicology. , 101-111 (2012).</li><li>Tulis, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+and+morphometric+analyses+for+rat+carotid+balloon+injury+model).">Histological and morphometric analyses for rat carotid balloon injury model).</a> Methods In Molecular Medicine. 139, 31-66 (2007).</li></ol>

The authors declare that they have no competing financial interests.

O rato da lesão do balão carótida foi bem descrito por Tulis em 2007 34. Tem sido exaustivamente discutido todos os detalhes deste procedimento pelo Dr. Tulis. Os leitores que estão interessados ​​em realizar este procedimento são altamente recomendados para ler protocolo Tulis &#39;. No entanto, há uma coisa que não está concordando com o Dr. Tulis: Em vez de inflar o balão com soro fisiológico ou qualquer tipo de líquido, sugerimos para inflá-lo com o ar. De acordo com nossa experiência pessoal, inflando com l?...

Os cateteres de balão são dispositivos médicos utilizados no procedimento da angioplastia, com a finalidade de alargamento local obstruído (s) de ateroma ou trombo num vaso sanguíneo. Os lúmen do vaso estreitadas é forçado a abrir pelo balão inflado e fornecimento de sangue seria restaurada sequencialmente para aliviar os sintomas de isquemia a jusante, tais como angina, infarto do miocárdio e dor na perna. No entanto, o grande sucesso da angioplastia foi diminuída por complicações pós-operatórias, tais como resultados de força causando barotrauma vascular (lesão por balão), nomeadamente a remodelação da parede do vaso e, em muitos casos re-estreitamento da luz do vaso (reestenose) 1. Uma série de modelos animais têm sido desenvolvidas imitando o procedimento de angioplastia para ajudar os investigadores a compreender os mecanismos subjacentes a embarcação remodelação parede relacionados com o balão-lesão 2. Entre todas as espécies de animais utilizados para a modelagem, o rato é o mais utilizado. Compared a coelhos, cães e porcos, as vantagens de os ratos são seu baixo custo, sua relativa facilidade de uso e o conhecimento atual da fisiologia rato. Embora os ratos têm uma vantagem adicional em uma ampla gama de estirpes geneticamente manipuladas, o navio ratinhos é muito pequeno para inserir um cateter balão. Ao longo das últimas três décadas, os ratos experimentais têm permitido aos pesquisadores obter uma melhor compreensão dos mecanismos moleculares e celulares que sustentam a formação da neoíntima e remodelação vascular 3-6. Além da lesão do balão, o remodelamento vascular também estão envolvidos na maioria das doenças vasculares importantes, como a aterosclerose 7,8, hipertensão 9 e 10 de aneurisma. Assim, o conhecimento adquirido através do modelo de lesão balão é, em geral, benéfica para estudos globais da doença parede vascular. O objectivo geral do modelo de lesão por balão de rato não é apenas de compreender ainda as doenças vasculares, mas também para testar a potência de novos agentes parao controle da doença 11,12. Tratamento medicamentoso clínico atual para reestenose é aplicado por stents farmacológicos efectuadas através do lúmen do vaso logo após a angioplastia. Em modelos animais, uma forma eficiente ainda mais econômica para novos testes agente é um método de perfusão intraluminal locais bem desenvolvida. Os agentes candidatos que foram testados através deste método incluem drogas de moléculas pequenas 13,14, citocinas ou factores de crescimento 15,16, agentes de manipulação de genes (clones de cDNA, siRNA, etc.), 17-20, e novas formulações farmacêuticas 21,22. Até agora, o modelo de lesão balão rato continua sendo um dos modelos mais úteis para o estudo de doenças / distúrbios vasculares. É o passo fundamental da bancada de cabeceira, geralmente como o primeiro passo, no sentido de in vitro para in vivo, mas não deve ser o último. O resultado de experiências com ratos precisa ser deliberado e ainda caracterizado antes tradução humanautilização clínica, devido à diferença de leitos vasculares e a anatomia do vaso, bem como as diferenças intrínsecas entre as espécies humana e de rato 23-26. No entanto, ainda é uma ferramenta essencial na investigação médica translacional. Embora essa pesquisa utilizou a ser limitada pela falta de ratos geneticamente modificados, que deixou de ser um problema desde abordagens genômicas inéditas, como zinc-finger nucleases 27, 28 e TALENS CRISPR-Cas 29 fizeram ratos knockout facilmente acessível.

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vascular balloon injury and intraluminal administration in rat carotid artery

O modelo de lesão por balão da artéria carótida em ratos tem sido bem estabelecida há mais de duas décadas. Mantém-se um método importante para estudar os mecanismos moleculares e celulares envolvidos na desdiferenciação do músculo liso vascular, a formação da neoíntima e remodelação vascular. Ratos machos Sprague-Dawley são os animais mais utilizados para este modelo. Ratas não são preferidos como hormônios femininos protegem contra doenças vasculares e, assim, apresentar uma variação para este procedimento. A carótida esquerda é tipicamente feridos com o carótida direita que serve como um controlo negativo. Lesão carótida esquerda é causada pelo balão insuflado que desnuda o endotélio e distender s parede do vaso. Após lesão, potenciais estratégias terapêuticas tais como a utilização de compostos farmacológicos e qualquer gene ou shRNA transferência pode ser avaliada. Tipicamente para gene ou transferência shRNA, a seção de feridos do lúmen do vaso é transduced localmente por 30 min com vIRAL partículas que codificam uma proteína ou shRNA ou para a entrega e expressão na parede do vaso lesionado. Espessamento neointimal que representa as células musculares lisas vasculares proliferativas normalmente picos a 2 semanas após a lesão. Os vasos são principalmente colhidas neste ponto de tempo para análise celular e molecular de vias de sinalização celular, bem como a expressão do gene e proteína. As embarcações também podem ser colhidas a pontos de tempo anteriores para determinar o início da expressão e / ou activação de uma proteína ou via específica, de acordo com os objectivos pretendidos experimentais. Os navios podem ser caracterizadas e avaliadas por meio da coloração histológica, imunohistoquímica, ensaios de proteína / mRNA, e ensaios de atividade. A artéria carótida direita intacto do mesmo animal é um controle interno ideal. Mudanças induzidas pelo prejuízo, em parâmetros moleculares e celulares pode ser avaliada comparando a artéria ferido à artéria controle direito interno. Da mesma forma, as modalidades terapêuticas pode ser avaliada comparando a firad e tratado artéria para o controle lesionado só artéria.

Duas semanas após a lesão, artérias carótidas são colhidas, seccionados e sujeitos a análise morfológica. As artérias são cross-seccionados e corados com H &amp; E (Figuras 1, 2B, C e 3). Da parede da artéria carótida de rato contém quatro camadas de lâmina elástica, que aparecem como linhas de rosa. A área entre a lâmina externa, lâmina elástica externa (EEL) e a lâmina mais interior, da lâmina elástica interna (IEL) é a camada de músculo liso meios (Figura 1). A...

Watch this Scientific Journal Video about Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery at JoVE.com

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Este protocolo utiliza um cateter balão para causar uma lesão intraluminal na artéria carótida de rato e, doravante, provocar hiperplasia neointimal. Este é um modelo bem estabelecido para o estudo dos mecanismos de remodelação vascular em resposta a lesão. É também amplamente utilizada para determinar a validade de potenciais abordagens terapêuticas.

Lesão Balloon Vascular e Administração Intraluminal em Rat Carótida

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

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Research

JoVE Journal

Medicine

16.2K visualizações.  State University of New York College of Nanoscale Science and Engineering (SUNY CNSE).  Este protocolo utiliza um cateter balão para causar uma lesão intraluminal na artéria carótida de rato e, doravante, provocar hiperplasia neointimal. Este é um modelo bem estabelecido para o estudo dos mecanismos de remodelação vascular em resposta a lesão. É também amplamente utilizada para determinar a validade de potenciais abordagens terapêuticas. 

Vídeo: Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

Treinar um técnica microcirúrgica sofisticado: Interposição de enxerto de veia jugular externa na artéria carótida comum em ratos

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

Medicina

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema otimizado para Monitorização de perfusão cerebral no modelo do rato do curso de oclusão da artéria cerebral média intraluminal

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

Cateterismo da artéria carótida e jugular para executar as medidas hemodinâmicas, infusões e Sangue de amostragem em um Rat Consciente Modelo

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

Infusões artéria carótida para farmacocinéticas e farmacodinâmicas Análise de taxanos em Ratos

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Férrico Thrombosis Modelo rato na artéria carótida e Mesentery Vessel induzida por Chloride

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

A Rat carótida Balão Injury modelo para testar Therapeutics remodela Anti-vasculares

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Drug Delivery intraluminal para o Rato Fístula Arteriovenosa endotélio

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the industrialized world. New Zealand White (NZW) rabbits are widely used as an animal model for the study of atherosclerosis. They develop spontaneous lesions when fed with atherogenic diet; however, this requires long time of 4 - 8 months. To further enhance and accelerate atherogenesis, a combination of atherogenic diet and mechanical endothelial injury is often employed. The presented procedure for inducing atherosclerotic plaques in rabbits uses a balloon catheter to disrupt the endothelium in the left iliac artery of NZW rabbits fed with atherogenic diet. Such mechanical damage caused by the balloon catheter induces a chain of inflammatory reactions initiating neointimal lipid accumulation in a time dependent fashion. Atherosclerotic plaque following balloon injury show neointimal thickening with extensive lipid infiltration, high smooth muscle cell content and presence of macrophage derived foam cells. This technique is simple, reproducible and produces plaque of controlled length within the iliac artery. The whole procedure is completed within 20 - 30 min. The procedure is safe with low mortality and also offers high success in obtaining substantial intimal lesions. The procedure of balloon catheter induced arterial injury results in atherosclerosis within two weeks. This model can be used for investigating the disease pathology, diagnostic imaging and to evaluate new therapeutic strategies.

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

O modelo de coelho da aterosclerose acelerada: uma perspectiva metodológica da artéria ilíaca balão lesão

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Imagens de ressonância magnética fase contraste na artéria carótida comum de rato

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

Trombose da artéria carótida canino induzida por cloreto férrico: Um modelo Animal grande de lesão Vascular

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...