Este trabalho foi suportado por uma concessão do &quot;Deutsche Forschungsgemeinschaft&quot; para SU Eisenhardt (EI 866/1-1).

<ol>
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Não há conflitos de interesse declarados.

Leucócito-endotelial interacção, a produção de espécies reactivas de oxigénio e de activação do sistema do complemento são as principais características de IRI induzida por disfunção do tecido. 26 microcirculação A do tecido afectado é considerado como o local integral para o início inflamatória. Para além de experimentações ex vivo, tais como ensaios de câmara de fluxo 27, 28, é obrigatória a existência de modelos bem-estabelecidos de imagem intravital para avaliar melhor a relevância in vivo. Embora IRI tem sido implicado em vários sistemas de órgãos, nós aqui descrever um método para analisar sistematicamente leucócito-endotelial de interacção em lesão de reperfusão do tecido do músculo estriado que foi primeiro descrito por Baez et al 13, portanto, particularmente relevante em cirurgia de retalho (Figura 1). 29 Como é assumido que o pós-isquémica lesão tecidual de outros órgãos envolve os mesmos mecanismos inflamatórios, o modelo descrito pode ser usado para gain percepções princípio em mecanismos fisiopatológicos da IRI, relevantes em doenças como enfarte do miocárdio, acidente vascular cerebral ou isquemia intestinal. Os animais utilizados O modelo aqui apresentado é adequado para a maioria dos roedores, incluindo ratos e camundongos. Para o longo período de imagem que é necessário neste modelo que geralmente preferem ratos como traqueotomia e volume de respiração controlada pode ser facilmente realizados. Em combinação com a imagem cardiovascular através canulação arterial, os ratos podem ser mantidos circulatório e homeostaticamente estável para horas (Figura 2). Além disso tecido cremastérico é mais extensa do que em ratos é em ratinhos que saem mais opções para imagiologia postcapillary. Embora a preparação do tecido pode ser mais fácil executada em ratos lager, deve-se considerar que o tecido cremastérico e especialmente a fáscia cremastérico é mais fino em ratos jovens (100 -150 g), reduzindo assim a fluorescência de fundo e sobreposição de addititecido fascial onal. No entanto uma vantagem da utilização de ratinhos para microscopia intravital é a disponibilidade de animais transgénicos, valiosa para elucidar o papel de genes individuais em eventos microcirculatórias modulação. Tipos de tecidos Uma vez que descrito pela primeira vez, a microscopia intravital tem sido utilizado em uma variedade de preparações microcirculatórias incluindo a bolsa da bochecha mestre, orelha de coelho, mesentério do roedor, e cremaster. Uma desvantagem principal associado com a maior parte destas preparações é a activação potencial das células por manipulação cirúrgica acompanhada com um aumento transiente em leucócitos de rolamento e aderente que tem sido bem descrito para o mesentério. Preparação cremastérico 30 tem, portanto, ser efectuada com extremo cuidado. Para nossa experiência, humidificante frequente do tecido exposto e cauterização restritiva, bem como um período de estabilização suficiente do tecido de pelo menos 30 minutos ajuda a evitar ou reduzir de leucócitosestimulação. Uma grande vantagem da imagem cremastérico sobre imagiologia mesentérica é a fácil acessibilidade que poupa abertura da cavidade abdominal. Trauma cirúrgico geral é reduzido e montagem em qualquer um Plexiglas viram palco para iluminação transmitida ou, se utilizados em nosso protocolo, uma plataforma de alumínio térmico controlado para epi-illumintaion é facilmente realizada. Além disso, a gordura que recolhe cerca de vénulas pós-capilares em animais mais velhos limitantes de imagem mesentérica está ausente em volta do tecido cremastérico. Rotulagem de leucócitos Reagentes tais como rodamina 6G 31, laranja de acridina ou vermelho de acridina qual os núcleos de manchas e / ou organelos intracelulares que não são encontrados em células vermelhas do sangue acumular selectivamente em leucócitos (e até certo ponto em plaquetas e células endoteliais). 32 Nós preferimos usar 6G rodamina para a detecção de leucócitos, em concentrações de 0,4 mg de peso corporal / kg, que tem previously sido demonstrado que não têm nenhum efeito significativo sobre a activação de neutrófilos. 33 laranja de acridina mostrou fundo muito mais mancha e fototoxicidade por meio de vasoconstrição arteriolar (observações não publicadas). A utilização de LEDs para imagiologia de fluorescência fornece espectros de excitação precisa e permite que as aplicações mais complexas, tais como a combinação de diferentes comprimentos de onda, por exemplo para a detecção de activação de leucócitos e vazamento capilar, ao mesmo tempo e devem, portanto, ser o método de escolha no futuro do em in vivo de imagem. IRI no tecido cremastérico Aplicando isquemia no músculo cremaster pode ser facilmente realizado e reflete perfeitamente as condições clinicamente relevantes, tais como a cirurgia de retalho livre. Além disso, as modificações tais como a isquemia fria, quente ou isquemia de superfusão cremastérico com substâncias que influenciam a inflamação pode ser facilmente aplicado. Injecção Intrascrotal pode ser o método de Choice quando o tratamento do tecido cremastérico é necessária horas ou dias antes da gravação. Uma variedade de cursos de tempo para IRI pode ser utilizado, dependendo do nível de prejuízo requeridos. Nós aqui descrever um modelo de isquemia 30 minutos para produzir níveis significativos de rolamento após duas horas de reperfusão (Figura 3) através da contagem de glóbulos brancos que passam de um ponto fixo através do vaso. Com o aumento no rolamento é moderado deixa bastante potencial para estimular leucócitos no IRI através de outras drogas de interesse. No entanto, uma variedade de cursos de tempo podem ser utilizados dependendo do nível de activação de leucócitos necessário 21, 25 a adesão de leucócitos é mais frequentemente medido por contagem de células claramente visíveis na parede do vaso em um 100 -. 200 uM de estiramento que permanecem estacionários durante 30 s , embora este tempo pode variar entre laboratórios. Observou-se um aumento marcado no número de leucócitos aderentes em IRI que obtiveram significativa após 60 minutos. Em conclusão microscopia de epifluorescência intravital no IRI de tecido muscular estriado pode ser bem aplicado em tecidos de ratos cremastérico uma vez que fornece dados reprodutíveis de ativação em leucócitos in vivo para pesquisa inflamação.

<table border="1"><tbody><tr><td> Nome do equipamento: </td><td> Empresa: </td><td> N º de Catálogo: </td><td> Comentários: </td></tr><tr><td> Forene 100% (V / V) </td><td> Abade </td><td> B506 </td><td> API isoflurano </td></tr><tr><td> Terylene Sutura </td><td> Serag Weissner </td><td> OC108000 </td><td></td></tr><tr><td> Portex fina Diâmetro Tubos de polietileno </td><td> Smiths Medical </td><td> 800/100/100 </td><td> 0,28 mm de diâmetro interno </td></tr><tr><td> 0,9% de solução salina </td><td> Fresinus Kabi </td><td> 808771 </td><td></td></tr><tr><td> Mudanças-A ponta de luxo kit cauterização </td><td> Bovie Medical </td><td> DEL1 </td><td></td></tr><tr><td> Abbocath-T 14G </td><td> Venisystems </td><td> G713- A01 </td><td> utilizado como tubo de lente </td></tr><tr><td> Ventilador Servo 900C </td><td> Maquet </td><td></td><td> utilizado como ventialtor animais </td></tr><tr><td> Transdutor de pressão lógico </td><td> Smiths Medical </td><td> MX1960 </td><td></td></tr><tr><td> Sirecust 404 monitor </td><td> Siemens </td><td></td><td></td></tr><tr><td> ABL 700 bancada Analyzer </td><td> Radiômetro </td><td></td><td> para medição de gás de sangue </td></tr><tr><td> Almofada de aquecimento </td><td> Effenberger </td><td> 8319 </td><td></td></tr><tr><td> Estágio de alumínio </td><td> Alfun </td><td> AW7022 </td><td></td></tr><tr><td> Microscópio cirúrgico OPMI 6-SDFC </td><td> Carl Zeiss </td><td></td><td></td></tr><tr><td> Microcirúrgica instrumentos de laboratório set </td><td> S &amp; T </td><td> 767 </td><td></td></tr><tr><td> Biemer clipe de navio </td><td> Diener </td><td> 64,562 </td><td></td></tr><tr><td> Aplicando fórceps </td><td> Diener </td><td> 64,568 </td><td> para Biemer clipe de navio </td></tr><tr><td> Rodamina 6G </td><td> Sigma-Aldrich </td><td> R4127 </td><td></td></tr><tr><td> Vaseline branco DAB </td><td> Winthrop </td><td> 2726853 </td><td></td></tr><tr><td> Cubra óculos </td><td></td><td></td><td> 32x32 milímetros </td></tr><tr><td></td><td></td><td></td><td> Configuração intravital </td></tr><tr><td> Zeis Axio Scope A-1 MAT </td><td> Carl Zeis </td><td> 490036 </td><td> microscópio de epifluorescência </td></tr><tr><td> 470 nm LED </td><td> Carl Zeis </td><td> 423052 </td><td> fonte de luz de fluorescência </td></tr><tr><td> Colibri Sistema 2 </td><td> Carl Zeis </td><td> 423052 </td><td></td></tr><tr><td> W Plano Apochromat-20x / 1,0 DIC </td><td> Carl Zeis </td><td> 421452 </td><td> objetiva de imersão de água </td></tr><tr><td> AxioCam MRM Rev. 3 FireWire </td><td> Carl Zeis </td><td> 426509 </td><td> de alta resolução da câmera digital </td></tr><tr><td> Axio LE visão software </td><td> Carl Zeis </td><td> 410130 </td><td> utilizar para análise offline </td></tr></tbody></table>

real-time digital imaging of leukocyte-endothelial interaction in ischemia-reperfusion injury (iri) of the rat cremaster muscle

Isquemia-reperfusão (IRI) tem sido implicado em uma grande variedade de condições patológicas tais como acidente vascular cerebral, enfarte do miocárdio, isquemia intestinal, assim como após o transplante, e cirurgia cardiovascular. 1 reperfusão do tecido isquémico anteriormente, enquanto essencial para a prevenção de irreversível lesão tecidual, elicia inflamação excessiva do tecido afectado. Adjacente à produção de espécies reactivas de oxigénio, activação do sistema do complemento e aumento da permeabilidade microvascular, a activação de leucócitos é um dos agentes princípio na cascata patológico do dano tecidual inflamatória durante a reperfusão. 2, 3 activação dos leucócitos é um processo em várias etapas consistindo de laminagem, adesão firme e transmigração e é mediada por uma interacção complexa entre moléculas de adesão, em resposta a chemoattractants tais como factores do complemento, quimiocinas, ou factor de activação de plaquetas. 4 &lt;p = classe &quot;jove_content&quot;&gt; Enquanto circulante de leucócitos em vénulas pós-capilares é predominantemente mediado pela interacção das selectinas 5 com os seus ligandos de balcão, adesão firme dos leucócitos ao endotélio é-selectina controlada através da ligação a moléculas de adesão intercelular (ICAM) e celular vascular As moléculas de adesão (VCAM). 6, 7 Padrão-ouro para a observação em vivo de leucócito-endotelial interação é a técnica de microscopia intravital, descrita pela primeira vez em 1968 8. Embora vários modelos de IRI (isquemia-reperfusão) têm sido descritos para vários órgãos, 9-12 apenas poucos são adequados para a visualização directa do recrutamento de leucócitos no leito microvascular em um elevado nível de qualidade de imagem. 8 Nós aqui promover a microscopia de epifluorescência intravital digitais da vênula postcapillary na microcirculação cremastéricodo rato 13 como um método conveniente para qualitativamente e quantitativamente analisar o recrutamento de leucócitos para a pesquisa IRI-no tecido do músculo estriado e fornecer um manual detalhado para a realização da técnica. Nós ilustrar ainda mais as armadilhas comuns e fornecer dicas úteis que deverão permitir ao leitor a apreciar verdadeiramente, e executar com segurança o método. Em um passo a passo protocolo que descrevem como começar com anestesia respiração controlada sob monitorização suficiente para manter o animal anestesiado com firmeza por longos períodos de tempo. Em seguida, descrevem a preparação cremastérico como uma folha fina e plana para a resolução óptica surpreendente e fornecer um protocolo para imagiologia de leucócitos no IRI que tem sido bem estabelecidas nos nossos laboratórios.

Watch this Scientific Journal Video about Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle at JoVE.com

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Microscopia de epifluorescência intravital Digital de vênulas na microcirculação cremastérico é um método conveniente para obter insights sobre leucócito-endotelial interação<em&gt; In vivo</em&gt; Na isquemia-reperfusão (IRI) do tecido muscular estriado. Nós aqui fornecer um protocolo detalhado para executar com segurança a técnica e discutir suas aplicações e limitações.

Real-time Digital Imaging de leucócito-endotélio Interação em isquemia-reperfusão (IRI) do músculo cremaster de rato

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

real-time-digital-imaging-leukocyte-endothelial-interaction-ischemia

Nanyang Technological University

Research

JoVE Journal

Medicine

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

15.1K visualizações.  University of Freiburg Medical Centre. Microscopia de epifluorescência intravital Digital de vênulas na microcirculação cremastérico é um método conveniente para obter insights sobre leucócito-endotelial interação In vivo Na isquemia-reperfusão (IRI) do tecido muscular estriado. Nós aqui fornecer um protocolo detalhado para executar com segurança a técnica e discutir suas aplicações e limitações.

Vídeo: Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite tissue. Ischemia reperfusion injury (IRI) is a known cause of partial free flap failure and has no effective treatment. Establishing a laboratory model of this injury can prove costly both financially as larger mammals are conventionally used and in the expertise required by the technical difficulty of these procedures typically requires employing an experienced microsurgeon. This publication and video demonstrate the effective use of a model of IRI in rats which does not require microsurgical expertise. This procedure is an in situ model of a transverse abdominis myocutaneous (TRAM) flap where atraumatic clamps are utilized to reproduce the ischemia-reperfusion injury associated with this surgery. A laser Doppler Imaging (LDI) scanner is employed to assess flap perfusion and the image processing software, Image J to assess percentage area skin survival as a primary outcome measure of injury.

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

Free tissue transfer is widely employed in reconstructive surgery to restore form and function following oncological resection and trauma. Preconditioning this tissue prior to surgery may improve outcome. This article describes an in situ transverse rectus abdominis myocutaneous flap (TRAM) in rats as a means for testing preconditioning strategies.

In situ transversal reto abdominal miocutâneo: um modelo de rato de miocutâneo Isquemia Traumatismo por Reperfusão

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite ...

Medicina

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

Isquemia-reperfusão modelo de lesão renal aguda e Pós Lesão fibrose em camundongos

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

Indução e Avaliação de isquemia-reperfusão em Langendorff-perfundidos Rato Corações

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Modelo murino de isquemia-reperfusão

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent re-oxygenation of the tissues (reperfusion). In the skin, ischemia-reperfusion (IR) is the main contributing factor to the pathophysiology of pressure ulcers. While the cascade of events leading up to the inflammatory response has been well studied, the spatial and temporal responses of the different subsets of immune cells to an IR injury are not well understood. Existing models of IR using the clamping technique on the skin flank are highly invasive and unsuitable for studying immune responses to injury, while similar non-invasive magnet clamping studies in the skin flank are less-than-ideal for intravital imaging studies. In this protocol, we describe a robust model of non-invasive IR developed on mouse ear skin, where we aim to visualize in real-time the cellular response of immune cells after reperfusion via multiphoton intravital imaging (MP-IVM).

This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury.

Induzindo a lesão de isquemia-reperfusão na pele da orelha de rato por Intravital Multiphoton imagem de respostas imunes

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

Modelo de Mouse de dois vasos oclusão de isquémia-reperfusão Cerebral

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

Avaliação da reserva de fluxo coronariano após reperfusão de isquemia miocárdica em ratos

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Um modelo pré-clínico do rato para o estudo da lesão da Isquemia-reperfusion na microcirurgia reconstrutiva

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

Entrega intramiocárdica atrasada de células-tronco após lesão de reperfusão de Isquemia em um modelo murino

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

Lesão por Isquemia-Reperfusão de Órgãos simulando alterações hemodinâmicas no modelo de transplante de fígado de rato

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...