This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe&#8217;s group at Northwestern University for providing helpful information.

Declaração de Ética: NOTA: Todos os procedimentos estão em conformidade com o Manual sobre Cuidados e Uso de Animais de Laboratório (National Academies Press, 8ª edição, 2011), e os protocolos aprovados pelo Comitê de Uso e Cuidado Animal Institucional da Universidade de Wisconsin. Todas as cirurgias foram realizadas sob anestesia isoflurano (através de inalação, taxa de fluxo de 2 ml / min), e foram feitos todos os esforços para minimizar o sofrimento. Os animais foram sacrificados numa câmara gradualmente cheio com CO 2. 1. A indução da anestesia geral e incisão na pele <ol><li> Use macho C57BL / 6 com idade de 12-16 semanas, com peso de 23-28 g. Anestesiar o animal por isoflurano através de um aparelho anestésico-circuito fechado. Confirme anesthetization adequada por beliscar seu dedo do pé. Aplique um pouco de pomada lágrimas artificiais nos olhos para evitar o ressecamento. </li><li> Posicione o mouse em decúbito dorsal com o membro posterior esquerdo ligeiramente sequestrado e do joelho sl conjuntaightly flexionados. Raspar o cabelo em torno da região da coxa esquerda e desinfectar a pele com clorexidina, aplicado três vezes usando algodão estéril aplicadores derrubado. Use um esterilizador talão quente para manter os instrumentos esterilizados durante a cirurgia. </li><li> Coloque uma cortina de gaze esterilizada sobre a perna esquerda. Faça, uma incisão na pele longitudinal 1,5 centímetros em linha reta na coxa medial esquerdo da articulação do joelho até a virilha (Figura 1A). </li></ol> 2. Exposição da artéria femoral e Ramos NOTA: A Figura 1B mostra a anatomia vascular grave na coxa superior. A artéria femoral superficial (SFA) pode ser observado através de um microscópio de dissecação, quando a incisão na pele é adequadamente feita ao longo da linha média da coxa medial. <ol><li> Abra-se a fáscia fina que recobre a SFA com uma pinça fina e, em seguida, enrole o SFA com 9-0 nylon sutura. NOTA: Neste processo, o nervo femoral é facilmente colocar comoide, mas a veia femoral adjacente requer muita atenção, pois sua parede é tão fina que facilmente perfura e sangra. A técnica chave é se concentrar na própria artéria; enquanto a dissecção é realizada perto da artéria, veia qualquer dano acidental é evitável. </li><li> Uma vez que a SFA é enrolado, segure o fio de sutura com uma pinça mosquito e puxe-o levemente para baixo. Essa manobra torna ainda mais dissecção mais fácil, mas para evitar danificar a artéria não puxar com muita força. </li><li> Continue dissecção da artéria proximal para expor e laço da artéria femoral comum (CFA) com 9-0 nylon sutura. NOTA: Normalmente, o looping CFA em um ponto distal à artéria epigástrica (Figura 1B) é suficiente para a inserção do fio. Segure o laço de sutura CFA com uma pinça mosquito da mesma forma. </li><li> Puxe o laço de sutura CFA para cima e para o laço de sutura SFA para baixo para descobrir a bifurcação da SFA e artéria femoral profunda (DFA). O DFA, o principal ramo do CFA,vai para o músculo da coxa (Figura 1B). NOTA: O fio será inserido a partir da DFA mais tarde. </li><li> Passe o DFA com cuidado extra porque a veia adjacente está embaraçada por baixo da artéria. Em caso de hemorragias a partir da veia, um de 30 a 60 segundos de compressão simples com gaze é geralmente suficiente para produzir a hemostasia. </li><li> Uma vez que o DFA é intercalado com 9-0 nylon sutura, certifique-se todos os três laços de sutura estão em suas posições corretas (Figura 2A). </li><li> Prossiga com a dissecção da DFA distal ao ligar a artéria em um ponto, na medida do possível a partir da bifurcação. Cortar sutura extra, mas deixe um determinado comprimento de modo que as duas extremidades pode ser realizada com uma pinça de mosquito, que estabilizam o DFA tornando mais fácil a inserção do fio. </li></ol> 3. Criação de Lesão Fio <ol><li> Insira fio via DFA <ol><li> Coe as suturas CFA, SFA, e DFA para estabelecer uma pinça hemostática temporária. </li><li> Faça um pequeno furo no DFUm com microtesoura. Inserir as pontas das micropinças curvas para dilatar o orifício suficientemente grande para um fio com um diâmetro de 0,015 polegadas (Figura 2B demonstra a diferença de tamanho entre a artéria e o fio). <ol><li> Antes de fazer um buraco na DFA, afrouxar o laço de sutura CFA e gentilmente apertar / apertar o DFA várias vezes com micropinças maçantes de ponta. Esta manobra aumenta o diâmetro arterial notavelmente para facilitar a inserção do fio. Humedecer o fio cuidadosamente com PBS também é importante para uma inserção suave. </li><li> Ao fazer um buraco na DFA, não colhe a parede arterial, mas fazer uma pequena alfinetada em forma de V e beliscar a aba com micropinças de ponta aguçada para a manipulação. </li></ol></li><li> Insira o fio através do furo adequadamente-dilatada no DFA. Gentilmente avançar o fio em direção à artéria ilíaca, enquanto soltando o laço de sutura em torno do CFA. 5-8 mm é suficiente para cobrir todo o comprimento CFA na maioria dos casos (Figura 2C </strong&gt;). Não insira o fio com força como uma perfuração arterial geralmente resulta em perda do mouse. Encurtar o arame para cerca de 15 mm de comprimento ajuda com a manipulação. </li></ol></li><li> Deixar fio dentro de um Min <ol><li> Depois de inserir o fio com o comprimento óptimo, deixá-lo durante 1 min para esticar a parede arterial. Umedeça o fio e artérias com PBS para evitar a secagem. Outra sutura de nylon 9-0 pode ser colocada em torno da DFA proximal ao ponto de inserção de fio durante este tempo de espera. Esta sutura será utilizado mais tarde para fechar o buraco no DFA após a remoção do fio. </li></ol></li><li> Recuperar fio e retomar o fluxo de sangue <ol><li> Segure o fio com uma pinça de punção embarcação, e puxe-o para trás suavemente; o fio deve ser recuperada sem problemas. Para ultrapassar qualquer resistência, geralmente causado pela adesão do arame para a parede arterial, puxando para trás o fio com uma rotação suave é útil. Mais uma vez, uma manobra forçada poderia resultar em perda de animal. </li><li> AD epois de recuperar o fio, ligar o DFA no ponto de bifurcação com sutura de nylon 9-0. Não ligar muito próximo à bifurcação, o que comprometeria a saída de sangue para o SFA. </li><li> Solte os laços de sutura SFA e CFA para retomar o fluxo de sangue normal. Note-se que o CFA é bem expandido pela inserção de fio (Figura 2D). </li></ol></li></ol> 4. Pele Encerramento e Cuidados Pós-Operatórios <ol><li> Controle sangra. Exsudação Minor pode ser interrompido por compressão simples; algumas grandes hemorragias arteriais vai precisar de pontos com uma sutura 11-0 nylon. Estas artérias suturadas, acabará por resultar em oclusão arterial. </li><li> Feche a pele ferida com grampeadores da pele (geralmente 3-4 grampos). Mantenha a área ferida esterilizado através da aplicação de solução cirúrgica clorexidina. </li><li> Aplicar tratamentos pós-cirúrgicos. Controlar a dor do mouse com injeções subcutâneas de buprenorfina. Mantenha o mouse sobre a almofada de aquecimento até que recuperou sufficiente de consciência para manter decúbito esternal. Casa-lo em uma gaiola limpa sem a companhia de outros animais até que esteja totalmente recuperado. </li></ol>

<ol>
	<li>Dzau, V. J., Braun-Dullaeus, R. C., Sedding, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+proliferation+and+atherosclerosis:+new+perspectives+and+therapeutic+strategies.">Vascular proliferation and atherosclerosis: new perspectives and therapeutic strategies.</a> Nat Med. 8 (11), 1249-1256 (2002).</li><li>Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+hyperplasia+in+murine+models.">Intimal hyperplasia in murine models.</a> Curr Drug Targets. 9 (3), 251-260 (2008).</li><li>Ferns, G. A., Avades, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanisms+of+coronary+restenosis:+insights+from+experimental+models.">The mechanisms of coronary restenosis: insights from experimental models.</a> Int J Exp Pathol. 81 (2), 63-88 (2000).</li><li>Sata, M., 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=A+mouse+model+of+vascular+injury+that+induces+rapid+onset+of+medial+cell+apoptosis+followed+by+reproducible+neointimal+hyperplasia.">A mouse model of vascular injury that induces rapid onset of medial cell apoptosis followed by reproducible neointimal hyperplasia.</a> J Mol Cell Cardiol. 32 (11), 2097-2104 (2000).</li><li>Xu, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+arteriosclerosis:+from+arterial+injuries+to+vascular+grafts.">Mouse models of arteriosclerosis: from arterial injuries to vascular grafts.</a> Am J Pathol. 165 (1), 1-10 (2004).</li><li>Lv, L., Meng, Q., Ye, M., Wang, P., Xue, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=STAT4+deficiency+protects+against+neointima+formation+following+arterial+injury+in+mice.">STAT4 deficiency protects against neointima formation following arterial injury in mice.</a> J Mol Cell Cardiol. 74C, 284-294 (2014).</li><li>Zhu, B., Kuhel, D. G., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+E+inhibits+neointimal+hyperplasia+after+arterial+injury+in+mice.">Apolipoprotein E inhibits neointimal hyperplasia after arterial injury in mice.</a> Am J Pathol. 157 (6), 1839-1848 (2000).</li><li>Fu, Y., Duru, E. A., Davies, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+metabolic+syndrome+on+the+response+to+arterial+injury.">Effect of metabolic syndrome on the response to arterial injury.</a> J Surg Res. , (2014).</li><li>Kuhel, D. G., Zhu, B., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinction+in+genetic+determinants+for+injury-induced+neointimal+hyperplasia+and+diet-induced+atherosclerosis+in+inbred+mice.">Distinction in genetic determinants for injury-induced neointimal hyperplasia and diet-induced atherosclerosis in inbred mice.</a> Arterioscler Thromb Vasc Biol. 22 (6), 955-960 (2002).</li><li>Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differential+neointimal+response+in+vein+grafts+and+wire-injured+arteries.">Mouse strain differential neointimal response in vein grafts and wire-injured arteries.</a> Circ J. 71 (10), 1649-1652 (2007).</li><li>Ishigami, N., 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=Deficiency+of+CuZn+superoxide+dismutase+promotes+inflammation+and+alters+medial+structure+following+vascular+injury.">Deficiency of CuZn superoxide dismutase promotes inflammation and alters medial structure following vascular injury.</a> J Atheroscler Thromb. 18 (11), 1009-1017 (2011).</li></ol>

The authors declare no competing financial interests.

O procedimento de lesão fio é aplicável a todas as linhagens de camundongos, desde que as suas estruturas anatômicas são semelhantes. 4 Neste trabalho, utilizou-se do sexo masculino camundongos C57BL / 6 com uma idade de 12-16 semanas. Como aprendemos a partir de nossos estudos preliminares, as artérias femorais de ratos mais jovens de 10 semanas são muitas vezes tão pequeno que a inserção do fio é bastante desafiador. Por outro lado, a inserção do fio nos ratos mais velhos que 16 semanas, é tec...

In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata&#8217;s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.

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a murine model of arterial restenosis: technical aspects of femoral wire injury

A doença cardiovascular causadas por aterosclerose é a principal causa de morte no mundo desenvolvido. O estreitamento da luz do vaso, devido ao desenvolvimento da placa aterosclerótica ou a ruptura de placas estabelecidos, interrompe o fluxo normal do sangue levando a várias morbidades, tais como infarto do miocárdio e acidente vascular cerebral. Na clínica procedimentos endovasculares, como a angioplastia são comumente realizados para reabrir o lúmen. No entanto, estes tratamentos inevitavelmente danificar a parede do vaso, bem como o endotélio vascular, desencadeando uma resposta de cicatrização excessiva e o desenvolvimento de uma placa que se estende da neointima no lúmen fazendo com que a restenose do vaso (re-estreitamento). A reestenose permanece uma das principais causas do fracasso dos tratamentos endovasculares para aterosclerose. Assim, os modelos animais pré-clínicos de reestenose são de vital importância para a investigação dos mecanismos fisiopatológicos, bem como translacional abordagens para intervenções vasculares. Entre vários experime murinomodelos ntal, lesão fio artéria femoral é amplamente aceito como o mais adequado para estudos de reestenose pós-angioplastia, pois se assemelha ao procedimento de angioplastia que prejudica a ambos endotélio e da parede do vaso. No entanto, muitos pesquisadores têm dificuldade em utilizar este modelo devido ao seu alto grau de dificuldade técnica. Isto é primeiramente porque um fio de metal tem de ser inserido dentro da artéria femoral, o que é cerca de três vezes mais fino do que o fio, para gerar suficiente para induzir lesão neoíntima proeminente. Aqui, descrevemos os detalhes cirúrgicos essenciais para efetivamente superar as maiores dificuldades técnicas deste modelo. Seguindo os procedimentos apresentados, realizando a lesão de arame artéria femoral de rato torna-se mais fácil. Uma vez familiarizado, todo o processo pode ser concluído no prazo de 20 min.

Quatro semanas após a cirurgia, dissecção da artéria femoral feridos fios e fazer espécime Bloco A parafina para analisar a formação neointima. Induzir anestesia geral, tal como descrito acima. Abra o peito imediatamente e insira uma de calibre 20 borboleta agulha para o ventrículo esquerdo. Ligue a linha para um frasco de infusão. Realizam a fixação de perfusão através de infusão de gotas de PBS, seguido por paraformaldeído a 4% (geralmente 10-20 mL, cada um). O frasco de infusão é colocado ce...

Watch this Scientific Journal Video about A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury at JoVE.com

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

O mouse modelo de lesão fio artéria femoral de reestenose é tecnicamente desafiador. Neste protocolo, mostrar os principais detalhes técnicos essencial para a realização com êxito lesão fio para induzir neoï¿½tima consistente para estudos de reestenose.

Um modelo murino de Arterial Restenosis: Aspectos Técnicos do Femoral Injury Fio

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

a-murine-model-arterial-restenosis-technical-aspects-femoral-wire

Research

JoVE Journal

Medicine

19.9K visualizações.  University of Wisconsin School of Medicine and Public Health.  O mouse modelo de lesão fio artéria femoral de reestenose é tecnicamente desafiador. Neste protocolo, mostrar os principais detalhes técnicos essencial para a realização com êxito lesão fio para induzir neoï¿½tima consistente para estudos de reestenose. 

Vídeo: A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

Microcirúrgica venoso Bolsa Bifurcação Arterial-aneurismas no Modelo de Coelho: Aspectos Técnicos

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

Medicina

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

Cateterismo arterial e venoso femoral para coleta de sangue, Drug Administration e Pressão Arterial Consciente e Medidas de freqüência cardíaca

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Um modelo murino de implante de stent na artéria carótida para o Estudo da reestenose

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspectos técnicos da artocava Fístula Rato

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

O coelho Modelo Blood-shunt para o Estudo da aguda e tardia Sequelas de hemorragia subaracnóidea: Aspectos Técnicos

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Um Defeito modelo simples Critical porte Femoral em Ratos

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

Modelo murino de artéria femoral Injury fio com a implantação de uma droga patch Entrega perivascular

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

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, ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Indução de Aterosclerose Acelerada em Camundongos: O Modelo de "Lesão por Fio"

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

Abrir traqueostomia ácido gástrico aspiração Modelo murino de lesão pulmonar aguda Resultados em Maximal aguda não-letal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...