Name: a rat carotid balloon injury model to test anti-vascular remodeling therapeutics
Uploaded: 2016-09-19T12:30:00
Description: Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

The work was supported by the intramural research program of the NIH, National Institute on Aging, and by a Priority Research Centers Program grant from the National Research Foundation (NRF-2009-0093812) funded by the Ministry of Science, Information and Communication Technology, &amp; Future Planning, the Republic of Korea (H.T.). We thank Dr. Han-sol Park for putting "material" part together for the manuscript.

NOTA: o uso do modelo de lesão por balão rato e procedimentos associados, incluindo injeção de estudos ultra-sonográficos sRAGE e ultra-som recombinantes foram aprovados pela Comissão de animal Cuidado e Uso (ACUC) do Instituto Nacional sobre Envelhecimento, NIH. 1. Preparações Pré-cirurgia <ol><li> Instrumento, Plataforma cirúrgica e Equipamentos de Proteção Individual. <ol><li> Ver Materiais e Equipamento da Tabela para todos os instrumentos ...

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Houve dois métodos utilizados para insuflar o balão, de modo a gerar a lesão que remove o endotélio mural no lúmen da carótida arteriais. Um é para encher a seringa ligada com o líquido 20, e a outra é a utilização de pressão de ar 21. Nós preferimos usar uma seringa cheia de líquido, porque o volume de líquido exacto (0,02 ml) será usado para cada procedimento. Isto rende uma inflação preciso e reprodutível do balão, o que leva a um nível semelhante de lesões em cada animal s...

A angioplastia é um procedimento endovascular utilizado para alargar as artérias estreitadas ou obstruídas resultantes de condições patológicas tais como a aterosclerose. Uma complicação comum da angioplastia é a hiperplasia neointimal pós-operacional, ou a restenose, que ocorre devido a lesões cirúrgicas e subsequente remodelação vascular induzida por inflamação. Estas condições conduzem à proliferação de células lisas vasculares, e várias consequências fisiopatológicas 1-3. Hiperplasia neointimal re-engrossa o navio, e ocorre em até 60% dos pacientes pós-angioplastia no primeiro ano. Portanto, a restenose é um grande revés do procedimento de angioplastia amplamente utilizado 4. Embora a implantação do stent droga-eluição pode ajudar a prevenir reestenose, os candidatos selecionados apenas pode submeter a esse procedimento caro 5. Ambos os estudos animais e clínicos têm demonstrado que a inflamação crónica gerado por vasculesão lar e / ou ferida cirúrgica serve como o principal estímulo para 2,4 crescimento neointimal pós-angioplastia. As carótidas de ratos lesão por balão modelo mimetiza a situação clínica e, portanto, serve como um sistema modelo útil para identificar factores celulares que envolvem na remodelação vascular e proliferação de células vasculares 6-9. Este sistema modelo também é uma ferramenta muito útil para avaliar e / ou tela de medicamentos e reagentes terapêuticos que suprimem o crescimento neointimal em estudos translacionais pré-clínicos 10-14. Em comparação com o modelo de murino de arame carótida ferimento 15 e o fio artéria femoral modelo de lesão de murino 16, o modelo de lesão carótida em ratos de balão tem as vantagens de ser suficientemente grande em tamanho para a facilidade de procedimento cirúrgico que facilita a reprodutibilidade da lesão infligida. Ele pode fornecer um maior número de células primárias (por exemplo, células do músculo liso vascular, células endoteliais) para additional estudos in vitro para delinear o mecanismo molecular que regula a remodelação vascular. Importante, em comparação com os ratinhos, os ratos são também conhecidos como sendo um melhor modelo para estudos fisiológicos e toxicológicos 17. Embora uma desvantagem ou limitação do modelo de rato, é a falta de modelos genéticos modificados e knockout de genes, esta desvantagem pode ser superada pela disponibilidade da sequência genómica de rato e o desenvolvimento recente de poderosas ferramentas de edição genómicas, tais como tecnologia CRISPR-CAS que torna possível manipulação de vastas faixas de seqüências genômicas em diferentes sistemas modelo 18,19. Embora o modelo de lesão por balão de rato foi utilizado por vários laboratórios e vários protocolos completos foram publicados 20,21, este protocolo visa fornecer mais detalhes em preparações pré-cirurgia e pode orientar investigadores novos para este procedimento para configurar esta prática cirúrgica. Ressaltamos, ainda, o cuidado pós-operatório de tele animais que não permite apenas análises post-mortem patológicos e histomorfológicas dos efeitos terapêuticos sobre o remodelamento arterial, mas também estudos de ultra-som ultra-sonográficos de animais vivos 13,22.

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			<td height="21" style="height:21px;width:287px;">2 F Fogarty balloon embolectomy catheter&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="40" style="height:40px;width:287px;">Small&#160; curved forceps (Large radius Dumont#7shanks curved)&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Atraumatic straight small arterial clamps&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="41" style="height:41px;width:287px;">Retractor&#160; with maximum spread 5.5 cm long blunt teeth</td>
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			<td height="21" style="height:21px;width:287px;">Silk suture (4.0 and&#160; 6.0 )</td>
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			<td height="21" style="height:21px;width:287px;">Syringe (1.0 ml)&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Curity gauze sponges</td>
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			<td height="21" style="height:21px;width:287px;">Cotton tip applicators sterile and non-sterile</td>
			<td style="width:163px;">Puritan Medical Products</td>
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			<td height="21" style="height:21px;width:287px;">Compact hot bead sterilizer</td>
			<td style="width:163px;">Fine Science Tools</td>
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			<td height="21" style="height:21px;width:287px;">Self-regulating heating pad</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">ADS200 anesthesia system/ventilator</td>
			<td style="width:163px;">Paragon Medical</td>
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			<td height="21" style="height:21px;width:287px;">Isoflurane (forane), liquid form</td>
			<td style="width:163px;">Baxter</td>
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			<td height="21" style="height:21px;width:287px;">Sodium chloride 0.9% (Saline)&#160;</td>
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			<td height="41" style="height:41px;width:287px;">Buprenex (buprenorphine)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Reckitt Benckiser Healthcare (UK) Ltd.&#160;</td>
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			<td height="21" style="height:21px;width:287px;">70% alcohol</td>
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			<td height="21" style="height:21px;width:287px;">1: 10 Betadine</td>
			<td style="width:163px;">Fisher</td>
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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.

Duas semanas após a lesão por balão, o rato é sacrificado e as artérias carótidas são isoladas para análise histo-morfológica. Ambos operados artéria carótida esquerda e artéria direita un-operado são cortados transversalmente, processados ​​e embebidos em parafina. As amostras são, em seguida, parafina mais fina-seccionados e corados com hematoxilina-eosina (H &amp; E). análises Histo-morfológicas são realizadas utilizando um sistema de análise de imagem digital. O...

Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

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

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.

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

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Uma lesão de balão segmental controlada por pressão da artéria de rato com aplicação terapêutica periadventicional

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion

Um modelo murino de lesão isquêmica da retina induzida pela oclusão da artéria da carótida comum bilateral transitória

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. ...