Name: vascular balloon injury and intraluminal administration in rat carotid artery
Uploaded: 2014-12-23T13:00:00.000+00:00
Duration: 9 min 41 s
Description: Watch this Scientific Journal Video about Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery at JoVE.com

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

Not: Aşağıdaki deneyler için hayvanların kullanımı gözden ve Kurumsal Hayvan Bakım ve Kullanım Kurulu (IACUC) tarafından onaylanmıştır. 1. Ameliyat öncesi Prosedürler <ol style="margin-left: 40px;"><li> Kullanmadan önce cerrahi aletler sterilize. <ol><li> Tüm cerrahi enstrümanlarını ameliyattan önce 24 saat ya da daha az otoklavlayın. Birden ameliyatlar aynı gün yapılırsa, ameliyatlar arasında kuru boncuk sterilizatör tarafından aletleri sterilize. </li></ol></li><li> -Filtre sterilize kullanmadan önce tuzlu su çözeltisi. </li><li> Sıçan tartılır ve anestezi ilaçlarının (ketamin 80 mg / kg ketamin hidroklorid ve 7 mg / kg) dozu hesaplamak. </li><li> Intra-peritoneal (ip) anestezi ilaçları yönetme. <ol><li> 5-10 dakika içinde ayak tutam tarafından sedasyon yeterliliğini doğrulayın. Sedasyon tam değilse, ilaç (ketamin 7 mg / kg ve ksilazin, 0.6 mg / kg) ilave küçük doz uygulayın. </li><li> Emin olun iğne teslim ilaçlar içi başına yeterli derinitoneally periton boşluğuna tüm ilaç çözüm sunmak için başarısızlık yetersiz sedasyon neden olacaktır çünkü. NOT: Enjeksiyon yerinde deride Ülser ve saç dökülmesi nedeniyle yanlışlıkla subkutan (sc) ilaç solüsyon enjekte ameliyattan sonra görünür birkaç gün olacak. </li></ol></li><li> 3 ml steril tuzlu su çözeltisi deri altından (sc) enjekte edilir. Steril bir pamuklu çubuk kullanarak, kurumasını önlemek için kornealar hem gözlere oftalmik merhem küçük bir miktar uygulayın. </li><li> Isıtma cihazları hazırlayın. Mikrodalga veya su banyosu öncesi sıcak ısı pedleri. </li><li> Cerrahi platformda supinely hayvan koyun. <ol><li> Ventral boyun bölgesinde saç çıkarın. Bir pamuklu çubukla saç çıkarıcı uygulayın 30 saniye bekleyin ve gazlı bez ile tamamen silin. </li><li> Povidon-İyot fırçalayın ve% 70 etanol ile boyun çubukla. </li></ol></li><li> Cüppe, saç kapağı, cerrahi maske ve gözlük gibi kişisel koruyucu donanımlar, giyin. Ster koyunSteril cerrahi aletler ve malzemeleri dokunmadan önce sonunda cerrahi eldiven Ile. </li><li> Maruz kalan sadece boyun bölgesi ile steril bir cerrahi örtü ile sıçan örter. </li></ol> 2. Cerrahi İşlemleri <ol style="margin-left: 40px;"><li> Cerrahi işlem sırasında, her 15 dakikada ayak tutam tarafından hayvan sedasyon derinliğini kontrol edin. Hayvan ayak tutam yanıt verirse, ketamin ve ksilizin ek küçük doz (başlangıç ​​dozunun% 10) ekleyin. </li><li> Sol ortak karotid arteri teşrih (CCA) <ol><li> Boyun ortasında düz bir uzunlamasına bir kesi yapmak için bir neşter kullanın. cilt kesisi yaklaşık uzunluğu arterin 1.5-2 cm bölümünü izole amacıyla, 1.5-2 cm. değişebilir uzunluğu çalışmanın amacına bağlıdır. </li><li> Açık açık cilt bağ dokusunu incelemek. Forseps ipuçları tutun ve deri ya da altta yatan doku delinme değil emin olun. </li><li> Uzunlamasına alon kas katmanları teşrihg trakea sol tarafında. </li><li> Kas tabakası yukarı açarken, yakından bağlı vagus siniri ile sol CCA görselleştirmek. Çok dikkatli sol karotis arter az germe ile vagus siniri ayırmak için yanında açık açık teşrih. </li><li> Çatallanma kadar distalde CCA ayır. Internal karotis arter (ICA) ve dış karotid arter (ECA) - dikkatlice çatallanma ve iki şube teşrih. </li><li> Arterin 1.5-2 cm yaklaşık bir kısmı çevreleyen dokulardan izole kadar, CCA etrafında diseksiyon tutun. </li></ol></li><li> Balon yaralanması <ol><li> Kalıcı yaklaşık 5 mm uzaklıkta bifurkasyonundan ECA bir ligatürün yapmak. Kalıcı ECA ve ICA çatallanma yakın ECA oksipital dalı Arter. Ayrıca kalıcı çatallanma ve ECA dikilen arasındaki yerini, diğer dalları bağlayan varsa - örneğin, üstün tiroid dalı. Tüm ligatures için kullanılan dikiş 4-0 siyahipek. CCA proksimal ucu ve ICA uzak ucunda Clip. NOT: Şimdi, kan akımı durduruldu ya kalıcı ligasyon yoluyla (ECA on) veya geçici (CCA ve ICA üzerine) kırpma yoluyla. Çatallanma bölgesinde lümen içeriği sistemik dolaşımdan izole edilmiştir. </li><li> Küçük mikro makas ile ECA bir arteriyotomi kesi yapmak. Kesi uzak dikiş düğüm yakın olduğundan emin olun. Tuzlu ve pamuklu temiz kan. </li><li> ECA lümen içine Şişirilmemiş 2F balon kateteri yerleştirin. CCA lümen proksimal balon kateteri ilerletin. Klip kalır nerede ucu ulaşana kadar proksimal kateter ilerleyen tutun. </li><li> 3-yollu bir dişi luer kilit balon şişirme cihazı bağlayın ve balon kateter 3-yollu erkek luer kilit takın. </li><li> Yavaş yavaş çapının 1.5 kat karotid arteri gerilmek üzere, yaklaşık 1.5 atm basınçta balon şişirmek. </li&gt; <li> Yavaşça dönerek geri çatallanma Balonu çekin. </li><li> Balon havasını ve yakın ucuna geri ilerlemek. Tekrar şişirmek ve çekerek geri prosedür iki kez daha tekrarlayın. </li><li> Arter lumeninden balon kateteri. </li></ol></li><li> Reaktiflerin intralüminal uygulaması (örneğin, siRNA ilaçlar) Not: Burada kullanılan reaktif ya da shRNA stromal etkileşim Molekül 1 (STIM1) ya da olmayan hedefleme shRNA kontrol hedef kodlayan lentiviral parçacıkları ihtiva eden bir çözelti olmaktadır. STIM1 plazma zarı Ca 2 + kanal aktivasyonu kontrol eden ER Ca 2 + sensör ve bir proliferatif göçmen fenotip 12,30-33 içine damar düz kas dediferansiyasyon sırasında regüle kadar tek bir transmembran endoplazmik retikulum (ER) proteindir. <ol><li> Bir şırınga (24 g, 1.6 cm) bir damar aşırı iğne kateter takın. Test Reag aspire 30 ul çözümühastalar. ECA aynı kesi içine kateter yerleştirin. </li><li> CCA içine kateter ucu Advance ve kateter düzeltmek ve geçici kesi kapatmak için ECA tek düğüm ile sütür bir parça kravat. </li><li> CCA lümenine test ayıracı çözeltisi enjekte edilir. 30 dakika damar lümeninde çözüm tutun. Tuzlu su ile nemli maruz doku tutun ve ıslak bir gazlı bez parçası ile örtün. </li><li> İnkübasyondan sonra, kalan çözelti aspire. Bu tek düğüm gevşetin ve kateteri. </li><li> Arteriyotomi kesi proksimalde dikiş bir parça ile ECA Kravat. Çatallanma mümkün olduğunca yakın düğüm yapın. </li></ol></li><li> Yarayı kapatın. <ol><li> Balon yaralanma ve kateter giriş kan damarının bir sızıntı veya delinme neden olabileceğinden, ICA klibi kaldırmak ve herhangi bir sızıntı olup olmadığını kontrol edin. Kanama görülürse, gazlı bez parçası uygulamak ve kanamayı durdurmak için hafif bir baskı uygulayın. </li><li> KaldırCCA diğer klip. </li><li> Kanama belirtisi olmadığından emin olun ve daha sonra tüm kelepçeler ve diğer cerrahi aletler kaldırın. Aşırı sütür kes. </li><li> Kapat yara kullanarak cilt sütür (4-0 siyah ipek). Povidon-İyot veya diğer anti-septik / bakterisit / virüsit maddesi ile kapalı yara her tarafta bezi. Steril tuzlu sc 3 ml enjekte </li></ol></li></ol> 3. Ameliyat sonrası Prosedürler <ol style="margin-left: 40px;"><li> Ameliyat sonrası ısı yastığı sıçan tutun. Fareye intramüsküler enjeksiyon (im) ile 0.05 mg / kg buprenorfin bir dozunun tatbik edilmesi. Kurtarma işlemi sırasında, nemli hayvanların gözleri ve ağzı tutmak. Uyanık ve ayakta kadar hayvan izleyin. </li><li> Tamamen iyileşti kadar herhangi yatak olmadan temiz bir kafeste hayvan yerleştirin. İyileştikten sonra, hayvan, hayvan odasına döndü ve tek tek yer olacak. </li></ol>

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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=RNA+interference+targeting+STIM1+suppresses+vascular+smooth+muscle+cell+proliferation+and+neointima+formation+in+the+rat.+Molecular+Therapy.">RNA interference targeting STIM1 suppresses vascular smooth muscle cell proliferation and neointima formation in the rat. Molecular Therapy.</a> The Journal Of The American Society Of Gene Therapy. 17, 455-462 (2009).</li><li>Berra-Romani, R., Mazzocco-Spezzia, A., Pulina, M. V., Golovina, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ca2++handling+is+altered+when+arterial+myocytes+progress+from+a+contractile+to+a+proliferative+phenotype+in+culture.+American+journal+of+physiology.">Ca2+ handling is altered when arterial myocytes progress from a contractile to a proliferative phenotype in culture. American journal of physiology.</a> Cell Physiology. 295, C779-C790 (2008).</li><li>Bisaillon, J. 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=Essential+role+for+STIM1/Orai1-mediated+calcium+influx+in+PDGF-induced+smooth+muscle+migration.+American+journal+of+physiology.">Essential role for STIM1/Orai1-mediated calcium influx in PDGF-induced smooth muscle migration. American journal of physiology.</a> Cell Physiology. 298, C993-C1005 (2010).</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>Zhang, W., Trebak, M., Szallasi, A., B&#237;r&#243;, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon+Injury+in+Rats+as+a+Model+for+Studying+TRP+Channel+Contribution+to+Vascular+Smooth+Muscle+Remodeling.">Balloon Injury in Rats as a Model for Studying TRP Channel Contribution to Vascular Smooth Muscle Remodeling.</a> T TRP Channels in Drug DiscoveryMethods in Pharmacology and Toxicology. , 101-111 (2012).</li><li>Tulis, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+and+morphometric+analyses+for+rat+carotid+balloon+injury+model).">Histological and morphometric analyses for rat carotid balloon injury model).</a> Methods In Molecular Medicine. 139, 31-66 (2007).</li></ol>

The authors declare that they have no competing financial interests.

fare karotid balon zedelenmesi de. 2007 34 Tulis tarafından tarif edilmiştir Bu kapsamlı Dr Tulis bu işlemin tüm ayrıntılarını tartışılmıştır. Bu yordamı gerçekleştirmek isteyen okuyucular oldukça Tulis &#39;protokolü okumak için tavsiye edilir. Ancak, biz Dr Tulis ile kabul edilmez bir şey var: yerine tuzlu su veya herhangi bir sıvı ile balon şişirme, biz hava ile şişirmek için önerdi. Kişisel deneyim göre, sıvı ile şişirme pek hava kabarcıkları önleyebilirsiniz. Buna ek olarak, bu esnek ya...

Balon kateter bir kan damarı içinde aterom ya da trombozun tıkanma yer (ler) genişletilmesi amacıyla, anjiyoplasti prosedüründe kullanılan tıbbi cihazlardır. daralmış damar lümen şişirilmiş balon ile açmak zorunda kalır ve kan temini gibi anjin, miyokard infarktüsü ve bacak ağrısı gibi mansap iskemi belirtileri, rahatlatmak için sırayla restore edilecek. Bununla birlikte, anjiyoplasti büyük başarı gibi damar lümen (restenoz) 1 yeniden daralma vasküler barotravma (balon yaralanması), yani damar duvarı yenileme neden olan kuvvetin sonuçları gibi birçok durumda post-operatif komplikasyonlar azalmıştır. Hayvan modellerinde bir dizi araştırmacılar balon-yaralanma ilgili damar duvarı yenileme 2 altında yatan mekanizmaları anlamak için anjiyoplasti işlemi taklit geliştirilmiştir. Modelleme için kullanılan tüm hayvan türleri arasında, sıçan, en sık kullanılan biridir. Ctavşanlar, köpekler, domuzlar grubu ile karşılaştırıldığında, sıçan avantajları düşük maliyet kullanım görece kolaylığı ve sıçan fizyolojisi mevcut bilgi bulunmaktadır. Farenin Genetik olarak manipüle edilmiş suşların geniş bir aralıkta ek bir avantaja sahipse de, fareler damar balon kateter yerleştirmek üzere, çok küçüktür. Son üç yılda, deneysel sıçan araştırmacılar 3-6 remodeling neointima oluşumunu ve damar yatan moleküler ve hücresel mekanizmaların daha iyi anlayabilmek için izin vermiş. Balon yaralanma ötesinde, vasküler remodeling ayrıca ateroskleroz 7,8, hipertansiyon 9, ve anevrizma 10 gibi en büyük damar hastalıkları, katılmaktadırlar. Böylece, bilgi balon yaralanma modeli ile kazanılan genel olarak tüm damar duvar hastalığı çalışmaları için faydalıdır. Sıçan balon yaralanması modelinde genel amacı değil sadece daha fazla damar hastalıkları anlamak değil, aynı zamanda yeni ajanların gücünü test etmekhastalık kontrolü 11,12. Restenozla Güncel klinik ilaç tedavisi sağ anjiyoplasti sonrası damar lümen yoluyla yerleştirilen ilaç salınımlı stentler ile uygulanır. Hayvan modellerinde, yeni ajan test için verimli henüz daha ekonomik şekilde iyi gelişmiş yerel intraluminal perfüzyon yöntemidir. Bu yöntemle test edilmiştir Aday ajanlar küçük molekül uyuşturucuyu 13,14, sitokin içeren veya büyüme, 15,16 faktörleri gen manipüle ajanlar (cDNA klonları, siRNA&#39;nın, vb) 17-20 ve yeni farmasötik formülasyonlar 21,22. Şimdiye kadar, sıçan balon hasarı içeren model vasküler hastalıklar / bozuklukların araştırılması için en faydalı model biridir. Bu ilk adım in vivo in vitro hareket genellikle yatağa tezgah temel adım, ama sonuncusu olmamalıdır. fare deneylerinin sonuçları, kasıtlı ve ayrıca insan çevrilmesinden önce, özelliği gerekennedeniyle vasküler yatak ve damar anatomisi yanı sıra insan ve sıçan 23-26 arasındaki içsel tür farklar fark klinik kullanımı,. Yine, hala öteleme tıbbi araştırmalarda önemli bir araçtır. Böyle bir araştırma genetiği değiştirilmiş farelerde eksikliği ile sınırlı kullanılan iken, çinko-parmak gibi yeni genomik yaklaşımlar 29 nakavt fareler kolayca erişilebilir yaptık, 27 nükleazlara Talens 28 ve CRISPR-Cas yana, artık bir sorun olmuştur.

<table><tbody><tr><td>Fogarty balloon embolectomy catheters, 2 French&amp;nbsp;</td><td>Edwards Lifesciences, Germany</td><td>&amp;nbsp;120602F</td><td></td></tr><tr><td>Deltaphase Operating Board - Includes 2 Pads &amp;amp; 2 Insulators</td><td>Braintree Scientific, Inc.</td><td>39OP</td><td></td></tr><tr><td>LED light source</td><td>Fisher Scientific</td><td>12-563-501&amp;nbsp;</td><td></td></tr><tr><td>Hartmann Mosquito Forceps 4&amp;rdquo; curved</td><td>Apiary Medical, Inc. San Diego, CA</td><td>gS 22.1670</td><td></td></tr><tr><td>Crile Retractor 4&amp;rdquo; double ended</td><td>Apiary Medical, Inc.</td><td>gS 34.1934</td><td></td></tr><tr><td>Other surgical instruments</td><td>Roboz Surgical Instrument Company, Inc., Gaithersburg, MD</td><td></td><td></td></tr><tr><td>Peripheral Intravenous (I.V.) Cannula, 24 G</td><td>BD</td><td>381312</td><td></td></tr><tr><td>Ketamine HCl, 100 mg/ml, 10 ml</td><td>Ketaset- Patterson Vet</td><td>07-803-6637&amp;nbsp;</td><td></td></tr><tr><td>Xylazine (AnaSed), 20 mg/ml, 20 ml</td><td>Ketaset- Patterson Vet</td><td>07-808-1947</td><td></td></tr><tr><td>Buprenex, 0.3 mg/1 ml (5 Ampules/Box)</td><td>Ketaset- Patterson Vet</td><td>07-850-2280</td><td></td></tr><tr><td>Nair Baby Oil Hair Removal Lotion - 9 oz</td><td>Amazon/Walmart/CVS</td><td>N/A</td><td></td></tr><tr><td>Inflation Device</td><td>Demax Medical</td><td>DID30</td><td></td></tr><tr><td>D300 3-way Stopcock</td><td>B.Braun Medical Inc.</td><td>4599543</td><td></td></tr><tr><td>Artificial Tears Ointment&amp;nbsp;</td><td>Rugby Laboratories, Duluth, GA</td><td>N/A</td><td></td></tr></tbody></table>

vascular balloon injury and intraluminal administration in rat carotid artery

sıçanlarda karotis arter balon yaralanma modeli de iki yılı aşkın süredir kurulmuştur. Bu damar düz kas dediferansiyasyon neointima oluşumu ve vasküler yeniden katılan moleküler ve hücresel mekanizmaları incelemek için önemli bir yöntem olmaya devam etmektedir. Erkek Sprague-Dawley sıçanları bu model için en sık kullanılan hayvan bulunmaktadır. Kadın hormonları damar hastalıklarına karşı koruyucu ve dolayısıyla bu prosedür bir varyasyonu tanıtmak gibi dişi sıçanlar tercih edilmemektedir. Sol karotis tipik sağ karotid bir negatif kontrol olarak hizmet veren yaralandı. Sol karotis yaralanma endoteli denudes ve Cuz damar duvarını gerilmek şişirilmiş balon neden olur. Yaralanma sonrasında, bu tür farmakolojik bileşikler ve gen ya da shRNA transferi ya da kullanılması potansiyel terapötik stratejiler değerlendirilebilir. Tipik haliyle, gen ya da shRNA transferi için, damar lümeninin yaralı bölümü lokal olarak v 30 dakika içinde aktarılmasıyaralı damar duvarında teslimat ve ifade için bir protein veya shRNA ya kodlayan iral parçacıklar. Proliferatif vasküler düz kas hücrelerini temsil neointimal kalınlaşma genellikle yaralanma sonrası 2 hafta zirve. Gemiler, özellikle hücresel ve moleküler hücre sinyal yollarının analizi aynı zamanda gen ve protein ifadesi için bu zaman noktasında hasat edilir. Kaplar ayrıca amaçlanan deney amaçlarına bağlı olarak, özel bir protein ya da yolunun sentezlenmesi ve / veya aktivasyonu başlangıcını saptamak için, daha önceki zaman noktalarında hasat edilebilir. Kaplar histolojik lekeleme, immünohistokimya, bir protein / mRNA tahlilleri ve etkinlik tahlilleri kullanılarak karakterize edilmiş ve değerlendirilebilir. Aynı hayvandan sağlam sağ karotis arter ideal iç kontrol edilmesidir. Moleküler ve hücresel parametreleri Yaralanma kaynaklı değişimler iç Sağ kontrol artere yaralı arter karşılaştırarak değerlendirilebilir. Benzer şekilde, tedavi seçenekleri yaralayabilir karşılaştırarak değerlendirilebilirKontrol d ve tedavi arter tek arter yaralandı.

İki hafta yaralanma sonrası, karotid arteri hasat kesit ve morfolojik analizine tabi tutulurlar. Arterler çapraz kesit ve H &amp; E ile boyanmış olan (Şekil 1, 2b, C ve 3). Sıçan karotid arter duvar pembe çizgiler olarak görünür elastik lamina dört katmanları içerir. dış lamina, dış elastik lamina (EEL) ve içteki lamina arasındaki alan, internal elastik lamina (IEL) medya düz kas tabakası (Şekil 1). iel bir iç bölge intima, sağlam damarlarında endotel hücrel...

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

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Bu protokol, sıçan karotis arter üzerinde bir lümen içi yaralanmalara neden ve bundan sonra neointimal hiperplazi ortaya çıkarmak için bir balon kateter kullanır. Bu yaralanmaya yanıt olarak vasküler yeniden mekanizmalarını incelemek için iyi bilinen bir modeldir. Ayrıca yaygın potansiyel terapötik yaklaşımların geçerliliğini belirlemek için kullanılır.

Sıçan Karotis Arter Vasküler Balon Yaralanma ve intraluminal İdaresi

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

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Research

JoVE Journal

Medicine

16.3K Görüntüleme.  State University of New York College of Nanoscale Science and Engineering (SUNY CNSE).  Bu protokol, sıçan karotis arter üzerinde bir lümen içi yaralanmalara neden ve bundan sonra neointimal hiperplazi ortaya çıkarmak için bir balon kateter kullanır. Bu yaralanmaya yanıt olarak vasküler yeniden mekanizmalarını incelemek için iyi bilinen bir modeldir. Ayrıca yaygın potansiyel terapötik yaklaşımların geçerliliğini belirlemek için kullanılır. 

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

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

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

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

Sıçanlarda Common Karotid Arter interpozisyon Harici Juguler Ven Greft: Gelişmiş Mikrocerrahi Tekniği Eğitimi

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

Tıp

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ischemia produces damage to specific brain regions shown to be highly sensitive to ischemia 1. Hippocampal neurons have higher sensitivity to ischemic insults compared to other cell populations, and specifically, the CA1 region of the hippocampus is particularly vulnerable to ischemia/reperfusion 2.
The design of therapeutic interventions, or study of mechanisms involved in cerebral damage, requires a model that produces damage similar to the clinical condition and in a reproducible manner. Bilateral carotid vessel occlusion with hypotension (2VOH) is a model that produces reversible forebrain ischemia, emulating the cerebral events that can occur during cardiac arrest and resuscitation. We describe a model modified from Smith et al. (1984) 2, as first presented in its current form in Sanderson, et al. (2008) 3, which produces reproducible injury to selectively vulnerable brain regions 3-6. The reliability of this model is dictated by precise control of systemic blood pressure during applied hypotension, the duration of ischemia, close temperature control, a specific anesthesia regimen, and diligent post-operative care. An 8-minute ischemic insult produces cell death of CA1 hippocampal neurons that progresses over the course of 6 to 24 hr of reperfusion, while less vulnerable brain regions are spared. This progressive cell death is easily quantified after 7-14 days of reperfusion, as a near complete loss of CA1 neurons is evident at this time.
In addition to this brain injury model, we present a method for CA1 damage quantification using a simple, yet thorough, methodology. Importantly, quantification can be accomplished using a simple camera-mounted microscope, and a free ImageJ (NIH) software plugin, obviating the need for cost-prohibitive stereology software programs and a motorized microscopic stage for damage assessment.

Bilateral carotid occlusion coupled with systemic hypotension produces global brain ischemia in the rat, resulting in damage to the hippocampus with reproducible severity. Animal subjects are impaired with predictable patterns of brain damage, they recover expediently, and mortality rates are comparatively low.

2-Gemi Tıkanıklığı / Hipotansiyon: Küresel Beyin İskemi Rat Modeli

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ...

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.

Hidrolize Çapraz bağlayıcılar yoluyla Adenoviral vektörler gergin kullanma Metalik Stent Yüzeylerden Vasküler Gen Transferi

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

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.

Rat Karotis Balon Yaralanma Modeli Anti-vasküler Tadilat Therapeutics Test

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 ...

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.

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.

Farelerde Hızlandırılmış Ateroskleroz Indüksiyonu: "Tel-Yaralanma" Modeli

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

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.

Bir Fare Modelinde İnfrarenal Abdominal Aort Bağlama Ekipmanları tarafından tekrarlanabilir Arter denüdasyon Yaralanma

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

Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing and treating this complication using experimental models of spinal cord ischemia. The model described here demonstrates varying degrees of paraplegia that relate to the length of occlusion following thoracic aortic occlusion in a rat spinal cord ischemia model.
A 2-Fr. balloon-tipped catheter was advanced through the femoral artery into the descending thoracic aorta until the catheter tip was placed at the left subclavian artery in anesthetized male Sprague-Dawley rats. Spinal cord ischemia was induced by inflating the catheter balloon. After a set period of occlusion (9, 10, or 11 min), the balloon was deflated. Neurologic assessment was performed using the motor deficit index at 24 h after surgery, and the spinal cord was harvested for histopathological examination.
Rats that underwent 9 min of aortic occlusion showed mild and reversible motor impairment in the hind limb. Rats subjected to 10 min of aortic occlusion presented with moderate but reversible motor impairment. Rats subjected to 11 min of aortic occlusion displayed complete and persistent paralysis. The motor neurons in the spinal cord sections were more preserved in rats subjected to shorter duration of aortic occlusion.
Researchers can achieve a reproducible hind limb motor deficit following thoracic aortic occlusion using this spinal cord ischemia model.

This study demonstrates the technique to make a minimally invasive and easily reproducible model of spinal cord ischemia in rats. Various degrees of hind limb motor deficit can be produced by controlling the aortic occlusion time.

Spinal Kord İskemisinin Sıçan Modelinde Aortik Oklüzyon Sonrası Tekrarlanabilir Motor Bozukluğu

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

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

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Periadventitial Terapötik Uygulama ile Sıçan Karotis Arter Basınç Kontrollü Segmental Balon Yaralanması

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

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of coronary artery disease. In-stent restenosis (ISR) is determined by an excessive proliferation of neointimal tissue within the stent and limits the long-term success of stents. A variety of animal models have been used to elucidate pathophysiological processes underlying in-stent restenosis (ISR), with the porcine coronary and the rabbit iliac artery models being the most frequently used. Murine models provide the advantages of high throughput, ease of handling and housing, reproducibility, and a broad availability of molecular markers. The apolipoprotein E deficient (apoE-/- ) mouse model has been widely used to study cardiovascular diseases. However, stents must be miniaturized to be implanted into mice, involving important changes of their mechanical and (potentially) biological properties. The use of apoE-/- rats can overcome these shortcomings as apoE-/- rats allow for the evaluation of human-sized coronary stents while at the same time providing an atherogenic phenotype. This makes them an excellent and reliable model to investigate ISR after stent implantation. Here, we describe, in detail, the implantation of commercially available human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access.

This protocol describes the implantation of human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access. Compared with other animal models, murine models carry the advantages of high throughput, reproducibility, ease of handling and housing, and a broad availability of molecular markers.

Trans-Femoral Erişim Kullanılarak İnsan Büyüklüğünde Koroner Stentlerin Rat Abdominal Aort'a yerleştirilmesi

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of ...

Derin Ven Tedavisi Çalışmaları İçin Kemirgen Venoplasti Balon Modeli

Rodent Inferior Vena Cava Venoplasty Balloon Model

Balloon venoplasty is a commonly used clinical technique to treat deep vein stenosis and occlusion as a consequence of trauma, congenital anatomic abnormalities, acute deep vein thrombosis (DVT), or stenting. Chronic deep venous obstruction is histopathologically characterized by thrombosis, fibrosis, or both. Currently, no direct treatment is available to target these pathological processes. Therefore, a reliable in vivo animal model to test novel interventions is necessary. The rodent survival inferior vena cava (IVC) venoplasty balloon model (VBM) allows the study of balloon venoplasty in non-thrombotic and post-thrombotic conditions across multiple time points. The local and systemic effect of coated and uncoated venoplasty balloons can be quantified via tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent assay (ELISA), zymography, vein wall and thrombus cellular analysis, whole blood and plasma assays, and histological analysis. The VBM is reproducible, replicates surgical human interventions, can identify local vein wall-thrombi protein changes, and allows multiple analyses from the same sample, decreasing the number of animals required per group.

Yazar Spot Işığı: Derin Damarlar için Cerrahi Müdahaleleri Simüle Etmek İçin Kemirgenlerde Venoplasti Balon Modelinin Kullanılması

The protocol presents a survival rodent model to test venoplasty balloon (VB) interventions in non-thrombotic, thrombotic, and post-thrombotic deep veins.