Name: a rat carotid artery pressure-controlled segmental balloon injury with periadventitial therapeutic application
Uploaded: 2020-07-09T12:30:00.000+00:00
Duration: 6 min 53 s
Description: Watch this Scientific Journal Video about A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application at JoVE.com

A N.E.B. foi apoiada por uma bolsa de treinamento do Instituto Nacional de Ciências da Saúde Ambiental (5T32ES007126-35, 2018), e uma bolsa de pré-doutorado da American Heart Association (20PRE35120321). E.S.M.B. foi um estudioso da KL2 parcialmente apoiado pelo Programa de Acadêmicos do UnC Clinical and Translational Science Award-K12 (KL2TR002490, 2018), e pelo National Heart, Lung, and Blood Institute (K01HL145354). Os autores agradecem ao Dr. Pablo Ariel, do Laboratório de Serviços de Microscopia da UNC, por ajudar na LSFM. A Microscopia de Fluorescência de Folha de Luz foi realizada no Laboratório de Serviços de Microscopia. O Laboratório de Serviços de Microscopia, Departamento de Patologia e Medicina Laboratorial, é apoiado em parte pela bolsa de apoio do Centro de Câncer P30 CA016086 ao Centro de Câncer Integral UNC Lineberger.

Todos os métodos descritos aqui foram aprovados pelo Comitê Institucional de Cuidados e Uso de Animais (IACUC) da Universidade da Carolina do Norte em Chapel Hill.
1. Procedimentos pré-operatórios
<ol><li>Esterilizar instrumentos cirúrgicos. Autoclave todos os instrumentos cirúrgicos antes da cirurgia. Se realizar múltiplas cirurgias no mesmo dia, esterilize os instrumentos entre as cirurgias utilizando um esterilizador de contas secas.</li>
	<li>Prepare o terapêutico em 25% de gel plurônico-127 (diluído em água destilada estéril).</li>
	<li>Configure um cateter de balão Fogarty 2F no insuflê e coloque a extremidade do balão do cateter em uma seringa de 1 mL cheia de solução salina.</li>
	<li>Induzir anestesia colocando o rato em uma câmara com 5% de isoflurano.<ol>
<li>Retire o rato da câmara e grave o peso do rato. Use cortadores de cabelo para raspar peles na região do pescoço ventral.</li>
		<li>Coloque o rato de volta na câmara com 5% de isoflurano para garantir a indução da anestesia.</li>
	</ol>
</li>
	<li>Coloque o supino do rato em uma plataforma cirúrgica, inserindo a face no cone do nariz para que a face do rato esteja em direção ao cirurgião.<ol>
<li>Reduza a anestesia inalatória para 1,5% de isoflurane. Verifique a profundidade da anestesia por um reflexo de dedo do pé nos quatro pés.</li>
		<li>Fita as quatro pernas até a plataforma cirúrgica.</li>
	</ol>
</li>
	<li>Ligue a lâmpada de calor.</li>
	<li>Injete atropina (0,01 mg/kg) subcutânea para reduzir as secreções das vias aéreas.</li>
	<li>Injete Carprofen (5 mg/kg) subcutânea para o tratamento da dor profilática. Se os anti-inflamatórios não forem aceitáveis para o experimento, consulte as etapas 3.2.2 e 3.2.3.</li>
	<li>Aplique pomada lubrificante nos olhos em ambos os olhos usando um cotonete estéril para evitar que as córneas sequem durante a cirurgia.</li>
	<li>Cotonete o pescoço três vezes em um movimento circular alternando entre 70% de álcool etílico seguido por Betadine do centro da região raspada para fora para esterilizar o local de incisão.</li>
	<li>Infiltrar-se na linha de incisão s.c. com 0,25% bupivacaína.</li>
	<li>Coloque luvas cirúrgicas estéreis antes de manusear instrumentos cirúrgicos estéreis e suprimentos.</li>
	<li>Coloque todos os instrumentos cirúrgicos autoclavados em uma folha cirúrgica estéril.</li>
	<li>Corte três fios independentes de 1 polegada de sutura estéril 7-0 Prolene.</li>
	<li>Coloque cotonetes e gaze na folha cirúrgica.</li>
	<li>Drape o rato com uma folha cirúrgica estéril que só expõe a região do pescoço esterilizada.</li>
	<li>Corte uma abertura pequena adicional na folha que exponha parte do cone do nariz. Este será o local para gravar o cateter de balão durante a lesão.</li>
</ol>2. Procedimentos operacionais
<ol><li>Durante todo o procedimento cirúrgico, avalie a profundidade da anestesia monitorando a taxa respiratória (a taxa deve ser consistente e considerada normal) bem como por beliscar o dedo do pé a cada 15 minutos. Se a taxa respiratória aumentar ou houver uma resposta ao aperto do dedo do pé, então pausa a manipulação cirúrgica e aumente o isoflurane até 2,5%.</li>
	<li>Expor a artéria carótida comum (CCA).<ol>
<li>Faça uma incisão superficial, reta e longitudinal entre os ossos da mandíbula do rato. A incisão terá aproximadamente 1,5-2 cm de comprimento.</li>
		<li>Faça uma incisão através do tecido conjuntivo sob a pele até que a camada muscular seja exposta. Deslocar as glândulas salivares sob a pele para acessar o tecido muscular.</li>
		<li>Separe sem rodeios o tecido conjuntivo do músculo inserindo uma tesoura fechada entre a camada muscular e o tecido conjuntivo e abrindo suavemente a tesoura enquanto puxa a pele para cima.</li>
	</ol></li>
	<li>Disseque os dois músculos visíveis (estehisidóide e esternostoide) longitudinalmente ao longo do lado esquerdo da traqueia até que um terceiro músculo (omohyide) que corre perpendicular aos dois músculos superficiais seja observado.</li>
	<li>Use fórceps para criar uma janela que separe este músculo perpendicular (omohyide) do músculo longitudinal (estehiidaide) correndo sobre a traqueia. Realize suavemente esta separação para evitar traumas contundentes na traqueia.</li>
	<li>Atinja fórceps sob o músculo perpendicular e corte para separar os dois músculos longitudinais e expor a CCA.</li>
	<li>Disseca a CCA.<ol>
<li>Disseque a CCA perto da bifurcação até que a artéria carótida interna (ICA) e a artéria carótida externa (ECA) sejam expostas.</li>
		<li>Dissecar a artéria tireoide superior (STA), que se ramifica do TCE.</li>
		<li>Utilizando as suturas prolene pré-cortadas, ligar o STA e o ECA perto de sua respectiva bifurcação. Deixe a maior parte da sutura para um lado do nó e pegue cada sutura com um hemosta curvo.</li>
		<li>Finalize a dissecação ao redor do ICA, alcance fórceps por baixo e ao redor do ICA, e use um grampo vascular não esmagador para obter controle distal. Aperte a artéria occipital junto com o ICA.</li>
		<li>Dissecar o CCA proximal à bifurcação, garantindo separar o nervo vago da CCA.</li>
		<li>Alcance fórceps por baixo e ao redor da CCA e use um grampo vascular não esmagante para obter controle proximal. Coloque o grampo de pelo menos 5 mm da bifurcação.</li>
	</ol></li>
	<li>Faça lesão no balão.<ol>
<li>Manobrar os hemostatos curvos que seguram cada ramo da artéria ligada para expor a bifurcação entre o TCE e o ramo superior.</li>
		<li>Dissecar suavemente o tecido na bifurcação e, em seguida, fazer uma incisão de arteriotomia entre o ECA e o ramo superior usando tesouras de microdisseção.</li>
		<li>Use um cotonete para empurrar todo o sangue para fora da CCA e limpar o local da arteriotomia.</li>
		<li>Insira o cateter de balão não inflado através da arteriotomia e avance para dentro da CCA até que a extremidade proximal do balão passe da bifurcação.</li>
		<li>Cateter de fita no cone do nariz para que o balão não saia da artéria durante a inflação.</li>
		<li>Infle lentamente o balão a 5 atmosferas de pressão e deixe na CCA por 5 minutos para induzir lesões arteriais. Certifique-se de que a pressão permaneça constante durante todo o 5 min.</li>
		<li>Depois de 5 min, esvazie o balão e retire suavemente da CCA através da arteriotomia.</li>
		<li>Lave a CCA apertando suavemente o grampo na CCA. Não remova o grampo.</li>
		<li>Ligate o ECA proximal à arteriotomia e, em seguida, remova os grampos da CCA e ica para restaurar o fluxo sanguíneo através da CCA para o ICA. Certifique-se de que não há sangramento visível ao redor da arteriotomia e que a CCA está pulsando.</li>
	</ol></li>
	<li>Aplique 100 μL de veículo de gel terapêutico ou plurônico sozinho periadventitamente ao longo da CCA ferida. Faça isso aplicando 50 μL no lado esquerdo da CCA e, em seguida, 50 μL no lado direito da CCA para garantir o revestimento uniforme da artéria ferida.</li>
	<li>Feche o local da ferida.<ol>
<li>Corte suturas prolene em excesso.</li>
		<li>Feche a ferida usando camadas vicríl interrompidas 4-0 ou 6-0 ao longo do tecido conjuntivo.</li>
		<li>Finalize o fechamento da ferida usando sutura de nylon 4-0 ao longo da pele.</li>
	</ol></li>
</ol>3. Procedimentos pós-operatórios
<ol><li>Coloque o rato sozinho em uma gaiola limpa com metade da gaiola sob uma lâmpada de aquecimento e monitore até que o rato recupere a consciência suficiente para manter a recumbência severa. Mantenha o rato em uma gaiola separada até que o animal esteja totalmente alerta e móvel antes de ser transferido de volta para sua gaiola original.</li>
	<li>Monitore o rato diariamente pelos próximos três dias e depois três vezes por semana até a eutanásia. Eutanize usando overdose de isoflurane seguido de toracotomia bilateral como descrito abaixo.<ol>
<li>Utilize o Centro Nacional de Requinte e Redução de Animais em Pesquisa (NC3Rs) para identificar os níveis de dor pós-operatório. Se algum animal parece estar sentindo dor ou desenvolvendo qualquer compromisso neurológico, sacrifique imediatamente.</li>
		<li>Para animais que não recebem carprofeno, administre acetaminofeno 6 mg/mL em sua água potável 24h antes da cirurgia através de 48h após a cirurgia. O acetaminofeno fornece analgesia com efeitos anti-inflamatórios mínimos.</li>
		<li>Alternativamente, outras estratégias de analgesia com efeitos anti-inflamatórios mínimos podem ser utilizadas, como buprenorfina ou liberação estendida de buprenorfina. Consulte a equipe veterinária de sua instituição.</li>
	</ol>
</li>
</ol>4. Colheita e imagem de tecido
<ol><li>Duas semanas após a cirurgia, eutanize o rato por overdose de anestesia (5% isoflurane). Alternativamente, os ratos eutanizam em um momento anterior para analisar os vários aspectos da resposta à lesão arterial.<ol>
<li>Uma vez que a respiração pára, faça a toracotomia bilateral como um método secundário de eutanásia.</li>
	</ol></li>
	<li>Faça uma incisão lateral através do abdômen, e depois corte para cima, através do diafragma e costelas, expondo a cavidade torácica.</li>
	<li>Peruuse e conserte as artérias.<ol>
<li>Insira uma cânula de 18 G presa a um sistema de fixação gravitacional de perfusão através do ventrículo esquerdo. Mantenha pressão equivalente entre ratos marcando a altura do sistema de perfusão em relação à bancada (elevação de 120 cm, equivalente a 91 ± 3 mmHg).</li>
		<li>Aperte a cânula junto com o ventrículo usando um hemostato curvo.</li>
		<li>Faça um corte no átrio direito, abrindo o circuito vascular, e comece a perfusão com PBS seguido de 2-4% de paraformaldeído (cerca de 250 mL cada).</li>
		<li>Prepare paraformaldeído diluído na PBS no dia do sacrifício, ou no máximo na noite anterior ao sacrifício. Se se preparar no dia do sacrifício, certifique-se de que o paraformaldeído esfriou à temperatura ambiente antes de iniciar a perfusão. Armazenar paraformaldeído a 4 °C.</li>
	</ol></li>
	<li>Após a fixação, extraia as artérias carótidas esquerda e direita e armazene a 4 °C por 2h em 2-4% de paraformaldeído.</li>
	<li>Transfira as artérias para 30% de sacarose e armazene durante a noite a 4 °C.</li>
	<li>Após 16-24 h, incorpore as artérias no composto de temperatura de corte ideal (OCT) e congele os blocos de artérias embutidas por OCT.<ol>
<li>Condicionam artérias em OUTUBRO à temperatura ambiente por 10 minutos. Coloque a artéria paralela ao plano da criomolda cheia de OCT, marcando o lado da criomold para a qual a bifurcação arterial está voltada. Congelamento de snap em nitrogênio líquido.</li>
		<li>Armazene blocos congelados a longo prazo a -80 °C.</li>
	</ol></li>
	<li>Seção blocos congelados usando um criostat.<ol>
<li>Colete seis seções transversais arterials de 5 μm de espessura por slide, com slide 1 começando na bifurcação.</li>
		<li>Seção blocos congelados até que a hiperplasia não seja mais visível (cerca de 100 slides).</li>
	</ol></li>
	<li>Hematoxylin & eosin (H&E) slides de manchas18<ol>
<li>Encontre a área do ferimento manchando um em cada dez slides ao longo de toda a artéria a partir da bifurcação (por exemplo, slides 1, 10, 20, 30, 40, 50, 60, 70, 80, 90 e 100).</li>
		<li>Manchas adicionais desliza ao redor do local da lesão para encontrar o slide com oclusão de pico (por exemplo, slides 20, 30 e 40 tinham hiperplasia visível, assim manchas desliza 15, 25, 35 e 45).</li>
		<li>Manche e quantifique o slide com oclusão máxima e deslizamentos equidistantes antes e depois do deslizamento de oclusão de pico (por exemplo, oclusão máxima encontrada no slide 35, depois manche e quantifique slides 25, 45, etc.) para um total de 3-10 slides por rato.</li>
	</ol></li>
	<li>Para imagens de microscopia de fluorescência de folha de luz, armazene as artérias durante a noite a 4 °C após a fixação na etapa 4.4.<ol>
<li>Artéria da sonda com diluição de 1:500 de anticorpo primário anti-CD31 de coelho em diluente (pH 7.4) por 3 dias. Em seguida, a artéria de contra-mancha com diluição de 1:500 do anticorpo secundário Alexa Fluor 647 por 2 dias19.</li>
		<li>Limpe a artéria usando iDISCO+20.</li>
		<li>Imagem da artéria usando um microscópio de fluorescência de folha de luz21. Renderizar imagens usando software (por exemplo, Imaris)19.</li>
	</ol></li>
	<li>Quantifique a hiperplasia neointimal. Realize a quantificação de forma cega, se possível.<ol>
<li>Use o software ImageJ para traçar o perímetro da lâmina elástica intima, interna elástica (IEL) e lamina elástica externa (EEL) de uma artéria em cada um dos 3-10 slides determinados acima (passo 4.8.3).</li>
		<li>Quantifique a área de cada região traçada em ImageJ e exporte esses valores. O traço intima produz a área de lúmen, o traço IEL produz a área IEL, e o traço EEL produz a área da ENGUIA.</li>
		<li>Média dos valores obtidos dos slides 3-10 para obter a lesão média (% de oclusão, proporção intima:media (I:M), hiperplasia neointimal) por artéria carótida de rato. Hiperplasia Neointimal = Área IEL - Área de Lumen <img alt="Equation 1" src="/files/ftp_upload/60473/60473eq1.jpg"> <img alt="Equation 2" src="/files/ftp_upload/60473/60473eq2.jpg">
</li>
	</ol></li>
</ol>

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+restenosis+models:+challenges+and+successes.">Preclinical restenosis models: challenges and successes.</a> Toxicologic Pathology. 34 (1), 11-18 (2006).</li><li>Xiangdong, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+the+atherosclerosis+research:+a+review.">Animal models for the atherosclerosis research: a review.</a> Protein Cell. 2 (3), 189-201 (2011).</li><li>Chen, H., Li, D., Liu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Rat+Models+for+Atherosclerosis.">Novel Rat Models for Atherosclerosis.</a> Journal of Cardiology and Cardiovascular Sceinces. 2 (2), 29-33 (2018).</li><li>Xing, D., Nozell, S., Chen, Y. F., Hage, F., Oparil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrogen+and+mechanisms+of+vascular+protection.">Estrogen and mechanisms of vascular protection.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 29 (3), 289-295 (2009).</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=Rat+carotid+artery+balloon+injury+model.">Rat carotid artery balloon injury model.</a> Methods in Molecular Medicine. 139, 1-30 (2007).</li><li>Pellet-Many, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropilins+1+and+2+mediate+neointimal+hyperplasia+and+re-endothelialization+following+arterial+injury.">Neuropilins 1 and 2 mediate neointimal hyperplasia and re-endothelialization following arterial injury.</a> Cardiovascular Research. 108 (2), 288-298 (2015).</li><li>Wu, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perivascular+delivery+of+resolvin+D1+inhibits+neointimal+hyperplasia+in+a+rat+model+of+arterial+injury.">Perivascular delivery of resolvin D1 inhibits neointimal hyperplasia in a rat model of arterial injury.</a> Journal of Vascular Surgery. 65 (1), 207-217 (2017).</li><li>Tan, J., Yang, L., Liu, C., Yan, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MicroRNA-26a+targets+MAPK6+to+inhibit+smooth+muscle+cell+proliferation+and+vein+graft+neointimal+hyperplasia.">MicroRNA-26a targets MAPK6 to inhibit smooth muscle cell proliferation and vein graft neointimal hyperplasia.</a> Scientific Reports. 7, 46602 (2017).</li><li>Pearce, C. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beneficial+effect+of+a+short-acting+NO+donor+for+the+prevention+of+neointimal+hyperplasia.">Beneficial effect of a short-acting NO donor for the prevention of neointimal hyperplasia.</a> Free Radical Biology and Medicine. 44 (1), 73-81 (2008).</li><li>Cao, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=S100B+promotes+injury-induced+vascular+remodeling+through+modulating+smooth+muscle+phenotype.">S100B promotes injury-induced vascular remodeling through modulating smooth muscle phenotype.</a> Biochimica et Biophysica Acta - Molecular Basis of Disease. 1863 (11), 2772-2782 (2017).</li><li>Madigan, M., Entabi, F., Zuckerbraun, B., Loughran, P., Tzeng, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+inhaled+carbon+monoxide+mediates+the+regression+of+established+neointimal+lesions.">Delayed inhaled carbon monoxide mediates the regression of established neointimal lesions.</a> Journal of Vascular Surgery. 61 (4), 1026-1033 (2015).</li><li>Khurana, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiogenesis-dependent+and+independent+phases+of+intimal+hyperplasia.">Angiogenesis-dependent and independent phases of intimal hyperplasia.</a> Circulation. 110 (16), 2436-2443 (2004).</li><li>Tsihlis, N. D., Vavra, A. K., Martinez, J., Lee, V. R., Kibbe, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitric+oxide+is+less+effective+at+inhibiting+neointimal+hyperplasia+in+spontaneously+hypertensive+rats.">Nitric oxide is less effective at inhibiting neointimal hyperplasia in spontaneously hypertensive rats.</a> Nitric Oxide. 35, 165-174 (2013).</li><li>Chen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+neointimal+hyperplasia+in+the+rat+carotid+artery+injury+model+by+a+HMGB1+inhibitor.">Inhibition of neointimal hyperplasia in the rat carotid artery injury model by a HMGB1 inhibitor.</a> Atherosclerosis. 224 (2), 332-339 (2012).</li><li>Mano, T., Luo, Z., Malendowicz, S. L., Evans, T., Walsh, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversal+of+GATA-6+downregulation+promotes+smooth+muscle+differentiation+and+inhibits+intimal+hyperplasia+in+balloon-injured+rat+carotid+artery.">Reversal of GATA-6 downregulation promotes smooth muscle differentiation and inhibits intimal hyperplasia in balloon-injured rat carotid artery.</a> Circulation Research. 84 (6), 647-654 (1999).</li><li>Becher, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Imaging+Provides+Detailed+Atherosclerotic+Plaque+Morphology+and+Reveals+Angiogenesis+after+Carotid+Artery+Ligation.">Three-Dimensional Imaging Provides Detailed Atherosclerotic Plaque Morphology and Reveals Angiogenesis after Carotid Artery Ligation.</a> Circulation Research. 126 (5), 619-632 (2020).</li></ol>

Os autores declaram que não há conflitos de interesse em relação à publicação deste manuscrito.

A lesão do balão da artéria carótida de rato é um dos modelos animais de restenose mais utilizados e estudados. Tanto o modelo de lesão do balãooriginal 3 quanto a variação de lesão segmental controlada pela pressão modificada10 informaram muitos aspectos da resposta à lesão arterial que também ocorre em humanos, com as poucas limitações sendo que o trombo rico em fibrina raramente se desenvolve e a inflamação local é mínima em comparação com outros mo...

A doença cardiovascular (DCV) continua sendo a principal causa de morte em todo o mundo1. A aterosclerose é a causa básica da maior morbidade e mortalidade relacionadas à DCV. Aterosclerose é o acúmulo de placa dentro das artérias que resulta em um lúmen estreito, dificultando a perfusão sanguínea adequada aos órgãos e tecidos distais2. As intervenções clínicas para o tratamento da aterosclerose grave incluem angioplastia de balão com ou sem colocação de stent. Esta intervenção envolve o avanço de um cateter de balão para o local da placa, e a inflação do balão para comprimir a placa até a parede arterial, ampliando a área luminal. Este procedimento danifica a artéria, no entanto, iniciando a resposta da lesão arterial3. A ativação prolongada desta resposta à lesão leva à restenose arterial, ou re-estreitamento, secundária à hiperplasia neointimal e à remodelação do vaso. Durante a angioplastia, a camada intimal é desnudada de células endoteliais que levam ao recrutamento imediato de plaquetas e inflamação local. A sinalização local induz alterações fenotípicas nas células musculares lisas vasculares (VSMC) e nos fibroblastos adventitas. Isso leva à migração e proliferação de VSMC e fibroblastos para dentro do lúmen, levando à hiperplasia neointimal4,5. As células progenitoras circulantes e as células imunes também contribuem para o volume global de restenose6. Quando aplicável, os stents que eluem drogas (DES) são o padrão atual para inibir a restenose7. O DES inibe a re-endotelialização arterial, no entanto, criando assim um ambiente pró-trombótico que pode resultar em trombose tardia do stent8. Portanto, os modelos animais são fundamentais tanto para a compreensão da fisiopatologia da restenose, quanto para o desenvolvimento de melhores estratégias terapêuticas para prolongar a eficácia dos procedimentos de revascularização.
Vários modelos animais grandes e pequenos9 são utilizados para estudar essa patologia. Estes incluem lesão de balão3,10 ou lesão de fio11 do lado luminal de uma artéria, bem como ligadura parcial12 ou colocação de manguito13 ao redor da artéria. A lesão do balão e do arame desnude a camada endotelial da artéria, imitando o que ocorre clinicamente após a angioplastia. Em particular, os modelos de lesão de balão utilizam ferramentas semelhantes às do cenário clínico (ou seja, cateter de balão). A lesão do balão é melhor realizada em modelos de ratos, uma vez que as artérias de rato são um tamanho apropriado para cateteres de balão comercialmente disponíveis. Aqui descrevemos uma lesão arterial segmental controlada por pressão, uma versão bem estabelecida e modificada da lesão do balão da artéria carótida do rato. Esta abordagem controlada por pressão imita de perto o procedimento de angioplastia clínica, e permite a formação de hiperplasia neointimal reprodutível duas semanas após a lesão14,15. Além disso, esta lesão arterial controlada por pressão resulta em restauração completa da camada endotelial por 2 semanas após a cirurgia16. Isso contrasta diretamente com o modelo original de lesão do balão, descrito por Clowes, onde a camada endotelial nunca retorna à cobertura completa3.
Após a cirurgia, a terapêutica pode ser aplicada ou direcionada para a artéria ferida através de várias abordagens. O método descrito aqui usa a aplicação periadventitial de uma pequena molécula embutida em uma solução de gel plurônico. Especificamente, aplicamos uma solução de aldeído cinnâmico de 100 μM em gel plurônico-F127 na artéria imediatamente após lesão para inibir a formação de hiperplasia neointimal15. Pluronic-F127 é um gel termo-reversível não tóxico capaz de entregar drogas localmente de forma controlada17. Enquanto isso, a lesão arterial é local, portanto a administração local permite testar um princípio ativo, minimizando os efeitos fora do alvo. No entanto, a entrega efetiva de um uso terapêutico deste método dependerá da química da pequena molécula ou do uso biológico.

<table><tbody><tr><td>1 mL Syringe</td><td>Fisher</td><td>14955450</td><td></td></tr><tr><td>1 mL Syringe with needle</td><td>BD</td><td>309626</td><td></td></tr><tr><td>2 French Fogarty Balloon Embolectomy Catheter</td><td>Edwards LifeSciences</td><td>120602F</td><td></td></tr><tr><td>4-0 Ethilon (Nylon) Suture</td><td>Ethicon Inc</td><td>662H</td><td></td></tr><tr><td>4-0 Vicryl Suture</td><td>Ethicon Inc</td><td>J214H</td><td></td></tr><tr><td>7-0 Prolene Suture</td><td>Ethicon Inc</td><td>8800H</td><td></td></tr><tr><td>70% ethyl alcohol</td><td></td><td></td><td></td></tr><tr><td>Anti-Rabbit Alexa Fluor 647</td><td>Thermo Fisher Scientific</td><td>A21245</td><td></td></tr><tr><td>Atropine Sulfate</td><td>Vedco Inc</td><td></td><td>for veterinary use</td></tr><tr><td>Cotton Swabs</td><td>Puritan</td><td>806-WC</td><td></td></tr><tr><td>Curved Hemostats</td><td>Fine Science Tools</td><td>13009-12</td><td></td></tr><tr><td>Fine Curved Forceps</td><td>Fine Science Tools</td><td>11203-25</td><td></td></tr><tr><td>Fine Scissors</td><td>Fine Science Tools</td><td>14090-11</td><td></td></tr><tr><td>Gauze</td><td>Covidien</td><td>2252</td><td></td></tr><tr><td>IHC-Tek Diluent (pH 7.4)</td><td>IHC World</td><td>IW-1000</td><td></td></tr><tr><td>Insufflator</td><td>Merit Medical</td><td>IN4130</td><td></td></tr><tr><td>Iodine solution</td><td></td><td></td><td></td></tr><tr><td>Lubricating Eye Ointment</td><td>Dechra</td><td></td><td>for veterinary use</td></tr><tr><td>Mayo Scissors</td><td>Fine Science Tools</td><td>14010-15</td><td></td></tr><tr><td>Micro Serrefines</td><td>Fine Science Tools</td><td>18055-05</td><td></td></tr><tr><td>Microdissection Scissors</td><td>Fine Science Tools</td><td>15004-08</td><td></td></tr><tr><td>Micro-Serrefine Clamp Applying Forceps</td><td>Fine Science Tools</td><td>18057-14</td><td></td></tr><tr><td>Needle Holder</td><td>Fine Science Tools</td><td>12003-15</td><td></td></tr><tr><td>Pluronic-127 (diluted in sterile water)</td><td>Sigma-Aldrich</td><td>P2443</td><td>25% prepared</td></tr><tr><td>Rabbit Anti-CD31</td><td>Abcam</td><td>ab28364</td><td></td></tr><tr><td>Retractor</td><td></td><td></td><td>Bent paper clips work well</td></tr><tr><td>Rimadyl (Carprofen)</td><td>Zoetis Inc</td><td></td><td>for veterinary use</td></tr><tr><td>Saline solution</td><td></td><td></td><td></td></tr><tr><td>Standard Forceps</td><td>Fine Science Tools</td><td>11006-12</td><td></td></tr><tr><td>Sterile Drape</td><td>Dynarex</td><td>4410</td><td></td></tr><tr><td>T-Pins</td><td></td><td></td><td></td></tr></tbody></table>

a rat carotid artery pressure-controlled segmental balloon injury with periadventitial therapeutic application

As doenças cardiovasculares continuam sendo a principal causa de morte e incapacidade em todo o mundo, em parte devido à aterosclerose. A placa aterosclerótica reduz a área luminal da superfície nas artérias, reduzindo assim o fluxo sanguíneo adequado para órgãos e tecidos distais. Clinicamente, procedimentos de revascularização, como angioplastia de balão com ou sem colocação de stent, visam restaurar o fluxo sanguíneo. Embora esses procedimentos restabeleçam o fluxo sanguíneo reduzindo a carga da placa, eles danificam a parede do vaso, que inicia a resposta de cura arterial. A resposta de cura prolongada causa restenose arterial, ou re-estreitamento, limitando o sucesso a longo prazo desses procedimentos de revascularização. Portanto, modelos de animais pré-clínicos são fundamentais para analisar os mecanismos fisiodológicos que conduzem a restenose, e proporcionam a oportunidade de testar novas estratégias terapêuticas. Os modelos murinos são mais baratos e fáceis de operar do que os grandes modelos animais. Lesões de balão ou arame são as duas modalidades de lesão comumente aceitas usadas em modelos murinos. Os modelos de lesão de balão, em particular, imitam o procedimento de angioplastia clínica e causam danos adequados à artéria para o desenvolvimento da restenose. Aqui descrevemos os detalhes cirúrgicos para a realização e análise histologicamente do modelo de lesão do balão carótida de rato modificado e controlado por pressão. Além disso, este protocolo destaca como a aplicação periadventicial local da terapêutica pode ser usada para inibir a hiperplasia neointimal. Por fim, apresentamos a microscopia de fluorescência da folha de luz como uma nova abordagem para a imagem e visualização da lesão arterial em três dimensões.

A Figura 1 mostra todos os materiais e ferramentas cirúrgicas utilizados para a realização desta cirurgia. A coloração de hematoxilina e eosina (H&E) de seções transversais arterials feridas de duas semanas permite uma visualização clara da hiperplasia neointimal. A Figura 2 mostra imagens representativas de seções arterials manchadas de H&E de uma artéria saudável, ferida e tratada. A Figura 2 também descreve como qua...

Watch this Scientific Journal Video about A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application at JoVE.com

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

A lesão do balão carótida do rato imita o procedimento clínico de angioplastia realizado para restaurar o fluxo sanguíneo em vasos ateroscleróticos. Este modelo induz a resposta à lesão arterial distending da parede arterial, e desnuding a camada intimal de células endoteliais, causando, em última análise, remodelação e uma resposta hiperplástica intimal.

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 ...

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Research

JoVE Journal

Medicine

4.9K visualizações.  University of North Carolina at Chapel Hill. A lesão do balão carótida do rato imita o procedimento clínico de angioplastia realizado para restaurar o fluxo sanguíneo em vasos ateroscleróticos. Este modelo induz a resposta à lesão arterial distending da parede arterial, e desnuding a camada intimal de células endoteliais, causando, em última análise, remodelação e uma resposta hiperplástica intimal.

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

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

Implantação de um Cuff Carotid para Triggering Shear-estresse aterosclerose induzida em camundongos

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

Medicina

Vascular Gene Transfer from Metallic Stent Surfaces Using Adenoviral Vectors Tethered through Hydrolysable Cross-linkers

In-stent restenosis presents a major complication of stent-based revascularization procedures widely used to re-establish blood flow through critically narrowed segments of coronary and peripheral arteries. Endovascular stents capable of tunable release of genes with anti-restenotic activity may present an alternative strategy to presently used drug-eluting stents. In order to attain clinical translation, gene-eluting stents must exhibit predictable kinetics of stent-immobilized gene vector release and site-specific transduction of vasculature, while avoiding an excessive inflammatory response typically associated with the polymer coatings used for physical entrapment of the vector. This paper describes a detailed methodology for coatless tethering of adenoviral gene vectors to stents based on a reversible binding of the adenoviral particles to polyallylamine bisphosphonate (PABT)-modified stainless steel surface via hydrolysable cross-linkers (HC). A family of bifunctional (amine- and thiol-reactive) HC with an average t1/2 of the in-chain ester hydrolysis ranging between 5 and 50 days were used to link the vector with the stent. The vector immobilization procedure is typically carried out within 9 hr&#160;and consists of several steps: 1) incubation of the metal samples in an aqueous solution of PABT (4 hr); 2) deprotection of thiol groups installed in PABT with tris(2-carboxyethyl) phosphine (20 min); 3) expansion of thiol reactive capacity of the metal surface by reacting the samples with polyethyleneimine derivatized with pyridyldithio (PDT) groups (2 hr); 4) conversion of PDT groups to thiols with dithiothreitol (10 min); 5) modification of adenoviruses with HC (1 hr); 6) purification of modified adenoviral particles by size-exclusion column chromatography (15 min) and 7) immobilization of thiol-reactive adenoviral particles on the thiolated steel surface (1 hr). This technique has wide potential applicability beyond stents, by facilitating surface engineering of bioprosthetic devices to enhance their biocompatibility through the substrate-mediated gene delivery to the cells interfacing the implanted foreign material.

These studies report on reversible attachment of adenoviral gene vectors to coatless metal surfaces of stents and model mesh disks. Sustained release of transduction-competent viral particles contingent upon hydrolysis of cross-linkers used for vector immobilization results in a durable site-specific transgene expression in vascular cells and in stented arteries.

Vascular transferência de genes de Metallic Stent superfícies usando Adenoviral Vetores Tethered através hidrolisável reticulantes

In-stent restenosis presents a major complication of stent-based revascularization procedures widely used to re-establish blood flow through critically ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

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 cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

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 ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

Quantificação de Proteção Neurovascular sequência repetitiva Hypoxic Pré-condicionamento e Transient Oriente oclusão da artéria cerebral em ratos

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

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 ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Reprodutível Arterial Desnudamento Lesão por aorta abdominal infrarrenal fixação num modelo murino

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

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 ...

Transvenous Embolization of Carotid Cavernous Fistula through Inferior Petrosal Sinus with Detachable Coils and Ethylene Vinyl Alcohol Copolymer

Carotid cavernous fistula (CCF) is a rare disease caused by abnormal communications between the internal carotid artery (direct fistula) or meningeal branches of the external carotid artery (indirect fistula) and the cavernous sinus (CS). Trauma is the most common cause of CCF. The clinical presentation of CCF is closely related to the venous drainage pattern. Orbital and neuro-ophthalmological symptoms are the most common clinical presentation of CCF with drainage through the superior ophthalmic vein (SOV). Endovascular embolization by arterial or venous approaches is the most common management of CCF. Transvenous embolization using detachable coils and ethylene-vinyl alcohol copolymer (EVOH) is an alternative method for the treatment of CCF. Endovascular embolization offers different options to treat CCF by minimally invasive approach decreasing morbidity and residual fistulas. The purpose of this article is to report our treatment experiences via the inferior petrosal sinus (IPS), and immediate-term outcomes of endovascular embolization of CCF by using detachable coils and EVOH.

The purpose of the article is to present our experience in endovascular treatment via the inferior petrosal sinus of the carotid cavernous fistula with detachable coils and ethylene-vinyl alcohol copolymer.

Carotid cavernous fistula (CCF) is a rare disease caused by abnormal communications between the internal carotid artery (direct fistula) or meningeal ...

Complete and Partial Resuscitative Endovascular Balloon Occlusion of the Aorta for Hemorrhagic Shock

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for hemorrhage control. With the advent of purpose-built devices, REBOA has become increasingly common in civilian trauma and acute care settings. Currently available REBOA catheters were designed as complete aortic occlusion devices. However, the therapeutic window for complete aortic occlusion is time-limited due to ischemia-reperfusion injury. The partial procedure allows blood flow past the level of occlusion while maintaining targeted proximal pressure, which has been shown to reduce distal ischemia and adjunctive resuscitation requirements in preclinical studies with prolonged occlusion times as compared to traditional complete occlusion.
pREBOA-PRO is the first catheter designed to enable partial and complete aortic occlusion and is currently in limited market release at seven Level I trauma centers in North America. This paper will focus on procedural considerations for REBOA, including patient selection criteria and a comparison of complete and partial aortic occlusion in a simulator, along with highlighting critical steps to improve clinical outcomes. Additionally, this paper reviews a contrast-enhanced CT scan from a trauma patient that shows distal perfusion after 2 h of partial aortic occlusion using this newly designed catheter and discusses representative results from the limited market release to highlight the profound effect of technological innovation on outcomes in vascular emergencies.

A commercial catheter was designed to facilitate true partial resuscitative endovascular balloon occlusion of the aorta (REBOA) and address the complications associated with complete aortic occlusion. Initial clinical reports indicate that partial REBOA improves transition to reperfusion, reduction of distal ischemia, and extension of safe occlusion time compared to complete occlusion.

Balão Endovascular Completo e Parcial Ressuscitante Oclusão da Aorta para Choque Hemorrágico

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for ...

Real-Time Monitoring and Modulation of Blood Pressure in a Rabbit Model of Ischemic Stroke

Control of blood pressure, in terms of both absolute values and its variability, affects outcomes in ischemic stroke patients. However, it remains challenging to identify the mechanisms that lead to poor outcomes or evaluate measures by which these effects can be mitigated because of the prohibitive limitations inherent to human data. In such cases, animal models can be utilized to conduct rigorous and reproducible evaluations of diseases. Here we report refinement of a previously described model of ischemic stroke in rabbits that is augmented with continuous blood pressure recording to assess the impacts of modulation on blood pressure. Under general anesthesia, femoral arteries are exposed through surgical cutdowns to place arterial sheaths bilaterally. Under fluoroscopic visualization and roadmap guidance, a microcatheter is advanced into an artery of the posterior circulation of the brain. An angiogram is performed by injecting the contralateral vertebral artery to confirm occlusion of the target artery. With the occlusive catheter remaining in position for a fixed duration, blood pressure is continuously recorded to allow for tight titration of blood pressure manipulations, whether through mechanical or pharmacological means. At the completion of the occlusion interval, the microcatheter is removed, and the animal is maintained under general anesthesia for a prescribed length of reperfusion. For acute studies, the animal is then euthanized and decapitated. The brain is harvested and processed to measure the infarct volume under light microscopy and further assessed with various histopathological stains or spatial transcriptomic analysis. This protocol provides a reproducible model that can be utilized for more thorough preclinical studies on the effects of blood pressure parameters during ischemic stroke. It also facilitates effective preclinical evaluation of novel neuroprotective interventions that might improve care for ischemic stroke patients.

Continuous arterial blood pressure recording allows the investigation of impacts of various hemodynamic parameters. This report demonstrates the application of continuous arterial blood pressure monitoring in a large animal model of ischemic stroke for determination of stroke pathophysiology, impact of different hemodynamic factors, and the assessment of novel treatment approaches.

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

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