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.

NOT: Etik Açıklama: Tüm işlemler Wisconsin Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanan Laboratuvar Hayvanları Bakım ve Kullanımı Kılavuzu (Ulusal Akademiler Press, 8. baskı, 2011), ve protokollere uygun. Tüm ameliyatlar (oran 2 ml / dak akış, teneffüs yoluyla) izofluran anestezi altında, ve tüm çabalar acı en aza indirmek için yapılmıştır. Hayvanlar kademeli CO2 ile doldurulmuş bir odacığm içine kurban edilmiştir. Genel Anestezi ve Cilt Kesiden 1. İndüksiyon <ol><li> 23-28 g bir 12-16 haftalık yaş ve vücut ağırlığı, erkek C57BL / 6 fareleri kullanın. Kapalı-devre anestezi aparatı ile izofluran ile hayvan anestezisi. Onun ayak kısma doğru anesthetization onaylayın. Kuruluğunu önlemek için gözlerin bazı yapay gözyaşı merhemi sürün. </li><li> Biraz kaçırılan sol arka bacak ve diz eklem sl ile yatar pozisyonda fare yerleştirinightly gerdi. Sol üst uyluk yöresinde saç tıraş ve klorheksidin ile cilt dezenfekte, steril pamuk uçlu aplikatörler kullanarak üç kez uygulandı. Ameliyat sırasında sterilize aletler tutmak için sıcak bir boncuk sterilizatör kullanın. </li><li> Sol bacak üzerinde bir sterilize gazlı bez örtü yerleştirin. Kasık (Şekil 1A) diz ekleminin sol medial uyluk üzerinde 1.5 cm düz, uzunlamasına deri kesi yapmak. </li></ol> Femoral Arter ve Şube 2. Pozlama NOT: Şekil 1B üst uyluk brüt damar anatomisini gösterir. cilt kesi düzgün medial uyluk orta hat boyunca yapıldığında yüzeyel femoral arter (SFA) bir mikroskop aracılığıyla görülebilir. <ol><li> Ince cımbız ve daha sonra döngü 9-0 naylon sütür ile SFA ile SFA örten ince fasya açın. NOT: Bu süreçte, femoral sinir kolayca konuronun duvara kolayca deler ve kanamaları çok ince olduğu için ide, ancak bitişik femoral ven yakın ilgi gerektirir. anahtar teknik arter kendisine odaklanmak; sürece diseksiyon yakın arterin yürütülen olarak, herhangi bir kaza sonucu damar hasarı önlenebilir. </li><li> SFA döngüye sonra, sivrisinek forseps ile dikiş tutun ve hafifçe aşağı doğru çekin. Bu manevra kolay daha diseksiyon yapar, ama çok zorla çekmeyin arter zarar vermemek için. </li><li> Proksimale 9-0 naylon sütür ile femoral arter (CFA) ortaya çıkarmak ve döngü arter diseksiyon devam edin. NOT: Genellikle, epigastrik arter (Şekil 1B) bir nokta distalinde CFA döngü tel yerleştirilmesi için yeterli. Aynı şekilde sivrisinek forseps ile CFA dikiş döngü tutun. </li><li> Yukarı CFA sütür döngü çekin ve SFA sütür döngü aşağı SFA ve derin femoral arter (DFA) çatallaşmayı ortaya çıkarmak. DFA, CFA büyük şube,uyluk kas (Şekil 1B) doğru gider. NOT: tel sonra DFA&#39;dan eklenecektir. </li><li> Ekstra dikkatli Döngü DFA bitişik ven arterin altında karışık çünkü. Venden kanama durumunda, gazlı bez ile bir 30- ila 60 sn, basit bir sıkıştırma genellikle hemostaz üretmek için yeterlidir. </li><li> DFA 9-0 naylon sütür ile döngüye sonra, her üç dikiş döngüler (Şekil 2A) onların doğru pozisyonlarda olduğundan emin olun. </li><li> Distale mümkün olduğunca bifurkasyonundan bir noktada arter bağlanması için DFA diseksiyonu ile devam edin. Ekstra dikiş kesin ama iki ucu DFA tel ekleme yapma kolay stabilize sivrisinek forseps ile tutulabilir, böylece belli bir uzunluğa bırakın. </li></ol> 3. Tel Yaralanması oluşturma <ol><li> DFA aracılığıyla Tel takın <ol><li> Geçici hemostat kurmak için CFA, SFA ve DFA sütür süzün. </li><li> DF küçük bir delik açınMicroscissors için uygun değildir. Bir 0.015 inç çaplı bir tel için yeterince büyük bir delik genişletmek için kavisli microtweezers ipuçlarını yerleştirin (Şekil 2B arter ve tel arasındaki boyut farkı gösterir). <ol><li> DFA&#39;da bir delik yapmadan önce, CFA sütür döngü gevşetin ve hafifçe kelepçe / donuk uçlu microtweezers ile DFA birkaç kez sıkın. Bu manevra tel ekleme kolaylaştırmak için son derece arteriyel çapını genişletir. PBS ile iyice tel Rutubet zamanda yumuşak sokulması için önemlidir. </li><li> DFA&#39;da bir delik yaparken, arter duvarı kırpmak, ama küçük bir v şekilli nip yapmak ve manipülasyon için keskin uçlu microtweezers ile flep çimdik yok. </li></ol></li><li> DFA üzerinde yeterince-genişlemiş delikten teli takın. CFA etrafında dikiş döngü gevşeterek ederken yavaşça iliak arter doğru tel ilerlemek. 5-8 mm çoğu durumda tüm CFA uzunluğu karşılamak için yeterli olduğunu (Şekil 2C </strong&gt;). Arteriyel perforasyon genellikle fare kaybına neden olduğu gibi zorla tel sokmayın. Uzunluğu yaklaşık 15 mm tel kısaltılması işleme yardımcı olur. </li></ol></li><li> Min 1 için içeride Tel bırakın <ol><li> Optimal uzunlukta tel taktıktan sonra, arter duvarı daraltabilir 1 dakika bırakın. Kurumasını önlemek için PBS ile tel ve arterler ıslatın. Başka bir 9-0 naylon dikiş bu bekleme süresi boyunca kablo ekleme noktasına DFA proksimal çevresinde yerleştirilebilir. Bu dikiş tel çıkarılmasını takiben DFA delik kapatmak için daha sonra kullanılacaktır. </li></ol></li><li> Tel Al ve Kan Akışı Devam <ol><li> Damar kanülasyon forseps ile tel tutun ve onu nazikçe geri çekin; Tel sorunsuz alınabilir olmalıdır. Nazik rotasyon ile tel geri çekerek, genellikle arter duvarına tel bağlılık kaynaklanan herhangi bir direnç, üstesinden gelmek için yardımcı olur. Yine, güçlü bir manevra hayvan kaybına neden olabilir. </li><li> Birtel almak fter, 9-0 naylon sütür ile çatallanma noktasında DFA Arter. SFA kan çıkışı tehlikeye atacağını bifürkasyonun, çok yakın Arter etmeyin. </li><li> Normal kan akışını sürdürmek için SFA ve CFA dikiş döngüler gevşetin. CFA tel yerleştirme (Şekil 2B) tarafından iyi genişletilmiş olduğunu unutmayın. </li></ol></li></ol> 4. Cilt Kapatma ve Ameliyat Sonrası Bakım <ol><li> Kontrol kanar. Küçük sızıntı basit sıkıştırma durdurulabilir; bazı büyük arter kanamaları bir 11-0 naylon sütür ile dikiş gerektirir. Bu dikildi arterler sonunda arter tıkanıklığına neden olur. </li><li> Cilt Zımbalar (genellikle 3-4 zımba) ile yara cilt kapatın. Klorheksidin cerrahi çözüm uygulayarak sterilize yara alan tutun. </li><li> Ameliyat sonrası tedavileri uygulayın. Buprenorfin subkutan enjeksiyonu ile fare ağrıyı kontrol. Bu suf kavuşana kadar ısınma pad üzerinde fare tutmaksternum yatma korumak için bilinç ficient. Diğer hayvanların şirket olmadan temiz bir kafes içinde Evi onu tamamen iyileşene kadar. </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.

tel yaralanma prosedürü, biz 12-16 haftalık bir yaş erkek C57BL / 6 fareler kullanıldı sürece onların anatomik yapıları benzer olarak. 4 Bu yazıda farelerin tüm suşları için geçerlidir. Bizim ön çalışmalardan öğrendiğimiz gibi, 10 haftadan küçük farelerin femoral arter genellikle tel ekleme oldukça zorlu olduğunu çok küçük. Diğer yandan, 16 haftadan daha yaşlı farelerin tel ekleme teknik olarak daha kolay olan, ancak çok büyük bir çeşitlilik neointimal hiperplazi neden ...

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.

<table><tbody><tr><td>Fixed Core Wire Guide</td><td>Cook</td><td>G02426</td><td>Diameter 0.015 inch, Straight</td></tr><tr><td>Dilation Forceps</td><td>Roboz</td><td>RS-4927</td><td>Curved, blunt tips</td></tr><tr><td>Dumont Tweesers #5</td><td>World Precision Instruments</td><td>14095</td><td>Straight, sharp tips</td></tr><tr><td>Dumont Vessel Cannulation Forceps</td><td>World Precision Instruments</td><td>503373</td><td></td></tr><tr><td>McPherson-Vannas Scissors</td><td>World Precision Instruments</td><td>501234</td><td></td></tr><tr><td>Mosquito Forceps</td><td>World Precision Instruments</td><td>501291</td><td></td></tr><tr><td>Ethilon Nylon Suture 9-0</td><td>Ethicon</td><td>7717G</td><td>9-0, Black Nylon Monofilament</td></tr><tr><td>Micro AROSuture, Sterile 11-0, 70 micrometers, MET Point</td><td>AROSurgical</td><td>VT4A00N07</td><td>11-0, Black Nylon Monofilament</td></tr><tr><td>3M Precise Multi-Shot DS Disposable Skin Stapler</td><td>3M</td><td>DS-25</td><td></td></tr></tbody></table>

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

Ateroskleroz neden Kardiyovasküler hastalık gelişmiş dünyada önde gelen ölüm nedenidir. Nedeniyle aterosklerotik plak gelişimi veya kurulmuş plakların çatlaması, damar lümeninin daralması gibi miyokard enfarktüsü ve inme gibi çeşitli hastalıklar giden normal kan akışını keser. Anjiyoplasti gibi klinik endovasküler işlemlerde yaygın lümen yeniden açmak için yapılır. Bununla birlikte, bu tedaviler, kaçınılmaz olarak bir fazla iyileşme yanıtı ve damar restenozu (yeniden daralma) neden olan lümen içine uzanan bir neointimal plak gelişimini tetikleme, damar duvarı hem de vasküler endotel hasar. Restenoz ateroskleroz için endovasküler tedavi başarısızlığı önemli bir nedenidir. Böylece, restenoz preklinik hayvan modelleri öteleme vasküler müdahalelere yaklaşımları yanı sıra patofizyolojik mekanizmaları araştırmak için hayati önem taşımaktadır. Bir çok murin experime arasındayakından endotel ve damar duvarını hem yaralı anjiyoplasti işlemi benzer çünkü ntal modelleri, femoral arter tel yaralanması yaygın anjiyoplasti sonrası restenoz çalışmaları için en uygun olarak kabul edilir. Ancak, birçok araştırmacı nedeniyle teknik zorluk yüksek derecede bu modeli kullanan zorluk. Bu, bir metal tel belirgin neointima indüklemek için yeterli darbe üretmek için, yaklaşık olarak tel üç kat daha incedir femoral arter içine takılması gerekir öncelikle çünkü. Burada, biz etkili bu modelin önemli teknik zorlukların üstesinden gelmek için gerekli cerrahi ayrıntıları açıklar. , Sunulan prosedürleri takip gerçekleştirerek fare femoral arter tel yaralanması kolaylaşır. Aşina sonra, bütün işlem 20 dakika içinde tamamlanabilir.

Ameliyatı takiben dört hafta, tel yaralı femoral arter hasat ve neointima oluşumunu analiz etmek için bir parafin blok örneği olun. Yukarıda tarif edilen genel anestezi neden. Hemen göğüs açın ve sol ventrikül bir 20-gauge kelebek iğne takın. Bir infüzyon şişe hattını bağlayın. % 4 paraformaldehid (genellikle 10-20 ml, her biri), ardından PBS damla infüzyon yoluyla perfüzyon fiksasyon gerçekleştirin. Hayvan daha sonra tel yaralı CFA çıkar ve 4 ° C&#39;de gece boyunca% 4 paraforma...

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

restenoz fare femoral arter yaralanması tel modeli teknik olarak zordur. Bu protokolde başarıyla restenoz çalışmaları için tutarlı neointima ikna etmek tel yaralanması gerçekleştirmek için anahtar teknik detaylar önemlidir göstermektedir.

Arter Restenozun bir fare modeli: Femur Tel Yaralanma Teknik Yönleri

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

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20.1K Görüntüleme.  University of Wisconsin School of Medicine and Public Health.  restenoz fare femoral arter yaralanması tel modeli teknik olarak zordur. Bu protokolde başarıyla restenoz çalışmaları için tutarlı neointima ikna etmek tel yaralanması gerçekleştirmek için anahtar teknik detaylar önemlidir göstermektedir. 

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

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Mikrovasküler Fonksiyon ve Tadilat üzerine Arteriolar Ligasyonu etkisini değerlendirmek için mürin Spinotrapezius Modeli

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

Tıp

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.

Restenoz Çalışmaları Karotis Arter Stent İmplantasyonu bir murin Modeli

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

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

Üretim ve Farelerde arteriyel Lezyonlarının 3-Boyutlu kantitasyonu Optik Projeksiyon Tomografi ile

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

Endovasküler Aort Anevrizma Onarımı Yeni Bir murin Modeli

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

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.

Bir Perivasküler Drug Delivery Patch İmplantasyonunda ile Femoral Arter Tel Yaralanma sıçan Modeli

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

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 ...

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 ...

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering from its most severe form, chronic limb-threatening ischemia (CLTI), face substantial impairments to daily living, including chronic pain, limited walking distance without pain, and nonhealing wounds. Preclinical models have been developed in various animals to study PAD, but mouse hindlimb ischemia remains the most widely used. There can be significant variation in response to ischemic insult in these models depending on the mouse strain used and the site, number, and means of arterial disruption. This protocol describes a unique method combining femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase (NOS) inhibitor to reliably induce footpad gangrene in Friend Virus B (FVB) mice that resembles the tissue loss of CLTI. While traditional means of assessing reperfusion such as laser Doppler perfusion imaging (LDPI) are still recommended, intracardiac perfusion of the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) is used to label the vasculature. Subsequent whole-mount confocal laser scanning microscopy allows for high-resolution, three-dimensional (3D) reconstruction of footpad vascular networks that complements traditional means of assessing reperfusion in hindlimb ischemia models.

The present protocol describes a unique, clinically relevant model of peripheral arterial disease that combines femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase inhibitor to induce hindlimb gangrene in FVB mice. Intracardiac DiI perfusion is then used for high-resolution, three-dimensional imaging of the footpad vasculature.

Murin Arka Ekstremite Kangren Modelinde Footpad Vaskülatürünün Yüksek Çözünürlüklü Üç Boyutlu Görüntülenmesi

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering ...

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell (VSMC) proliferation. Common murine models study small vessels such as the carotid and femoral arteries. Herein we describe an in vivo system in which both VSMC proliferation and endothelial barrier function can be simultaneously assessed in a large vessel. We studied the infrarenal aortic response to injury in C57BL/6 mice. The aorta was injured from the left renal vein to the aortic bifurcation by 30 transmural crushes of 5-seconds duration with a cotton-tipped applicator. Morphological changes were assessed with conventional histology. Aorta wall thickness was measured from the luminal surface to the adventitia. EdU integration and counter staining with DAPI and alpha-actin was used to demonstrate VSMC proliferation. Activation of ERK1/2, a known moderator of intimal hyperplasia formation, was determined by Western Blot analysis. The effect of inflammation was determined by immunohistochemistry for B-cells, T-cells, and macrophages. En face sections of endothelium were visualized with scanning electron microscopy (SEM). Endothelial barrier function was determined with Evans Blue staining. Transmural injury resulted in aortic wall thickening. This injury induced VSMC proliferation, most prominently at 3 days after injury, and early activation of ERK1/2 and decreased p27kip1 expression. Injury did not result in increased B-cells, T-cells, or macrophages infiltration in the vessel wall. Injury caused partial endothelial cell denudation and loss of cell-cell contact. Injury resulted in a significant loss of endothelial barrier function, which returned to baseline after seven days. The murine transmural blunt aortic injury model provides an efficient system to simultaneously study both VSMC proliferation and endothelial barrier function in a large vessel.

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Restenosis following cardiovascular procedures (bypass surgery, angioplasty, or stenting) is a significant problem reducing the durability of these procedures. An ideal therapy would inhibit smooth muscle cell (VSMC) proliferation while promoting regeneration of the endothelium. We describe a model for simultaneous assessment of VSMC proliferation and endothelial function in vivo.

Murin Aort Kırma Yaralanması: Verimli<emIn vivo</em&gt; Düz Kas Hücresi çoğalması ve Endotel Fonksiyon Modeli

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...