Yazarlar onun teknik yardım için Christiane Pahrmann teşekkür ederiz. Bu çalışma, Deutsche Stiftung für Herzforschung tarafından finanse edildi (F / 28/14). DW Kalp ve Akciğer Transplantasyonu Uluslararası Derneği ödülü ile desteklenmiştir. TD Else-Kröner-Fresenius-Stiftung (2012_EKES.04) den Else Kröner Mükemmellik maaş aldı. SS Deutsche Forschungsgemeinschaft araştırma hibe almıştır (DFG; DE2133 / 2-1, TD ve SCHR992 / 3- 1, SCHR992 / 4-1, SS).

Hayvanlar Laboratuvar Hayvanları İlkeleri, Laboratuar Hayvan Kaynakları Enstitüsü tarafından hazırlanan ve Ulusal Sağlık Enstitüleri tarafından yayınlanan Kılavuzu&#39;nda uygun insancıl bakıma aldı. Tüm hayvan protokolleri sorumlu yerel otorite ( &quot;Amt für Gesundheit und Verbraucherschutz, Hansestadt (Sağlık ve Tüketiciyi Koruma Dairesi) Hamburg&quot;) tarafından onaylanmıştır. 1. Hayvan Bakımı <ol><li> Laboratuvar Hayvanları Enstitüsü 300-350 g ağırlığında Lewis sıçanları (LEW / Crl) sıçan ve ROSA / lusiferaz LEW transgenik fareler elde edin. </li><li> havalandırmalı dolaplar geleneksel koşullar altında sıçan tutmak ve onlara standart sıçan yemi ve otoklava su ad libitum beslenirler. </li><li> donör ve alıcı olarak sinjenik LEW / Crl sıçanlar olarak ROSA / lusiferaz LEW transgenik fareler kullanarak bir greft nakli gerçekleştirmek. </li></ol> Donör Sıçan 2. Hazırlık <ol><li> Bir askere kullanıniyon odası izofluran (2.5-3%) ile bir sıçan uyuşturan için. </li><li> Sırtında sıçan yerleştirin ve ağız ve burun kapsayan bir yüz maskesi ile anestezi korumak. arka ayakları sıkışmasını ve reflekslerin yokluğu doğrulayarak anestezi yeterli derinliğe kontrol edin. anestezi altında iken kuruluğunu önlemek için gözlere bazı veteriner merhem sürün. </li><li> arka bacakları yaymak ve bant kullanarak kendi konumunu düzeltmek. </li><li> saç düzeltici ile inguinal saçlarını tıraş ve% 80 etanol ile takip povidon-iyot kullanarak tüm alanı dezenfekte edin. İki kez dezenfeksiyon işlemi tekrarlayın. NOT: Cerrahi alan, gazlı bez ve cerrahi aletler sterilize edilmelidir. süreç boyunca steril bir alan korumak ve tek kullanımlık, steril cerrahi eldiven, maske ve kep giymek. </li><li> Mikroskop altında, linea Inguinalis boyunca dikey bir kesi yapmak. nazikçe cilt altı dokuları ayırmak için iki forseps kullanın ve ori yüzeysel epigastrik ven maruzFemoral ven cin. Dikkatle çevre dokulardan yüzeyel epigastrik ven izole. </li><li> İki mikro kelepçeler kullanarak yüzeyel epigastrik ven kan akışını durdurun. </li><li> dikkatlice forseps ile izole ven kaldırma ve microscissors damar yoluyla keserek damar yaklaşık 0.5 ila 1 cm segmenti hasat. kan kaybını önlemek için damar güdük mikro kelepçeler bırakın. steril gazlı bez üzerine ven kaldırıldı parça yerleştirin. Dikkatle toplandı damarın bir ucunda içinde 30 G iğne yerleştirin ve heparin ile kap (50 birim / ml) yıkayın. NOT: Dikkatli damar tutun ve kaldırma, kesme ve yıkama sırasında zarar görmemesi. heparin uygun miktarda greft temizlemek için emin olun. </li><li> Alıcı sıçan içine nakli, bir gemi spazmı önlemek için kadar buz üzerinde% 1 lidokain damar segmentini tutun. </li><li> % 5 izofluran anestezi artırarak donör sıçan Euthanize. 2-3 dk, op sonralinea alba boyunca karın tr, diyafram üzerinden kesilmiş ve kan dolaşımını durdurmak için kalbi kaldırın. </li></ol> Alıcı Sıçan 3. hazırlanması <ol><li> Uyutmak ve donör sıçan aynı şekilde alıcı sıçan düzeltin. </li><li> saç düzeltici ile bacakların medial tarafını Tıraş ve povidon-iyot ve% 80 etanol kullanılarak üç kez dezenfekte edin. </li><li> anestezi derinliği izlemek ve arka ayakları kısma zaman refleksleri bulunmadığını teyit ederek yeterli olduğundan emin olun. </li><li> kasık kat diz medyan femur kesi gerçekleştirin. Mikroskop altında, çevresinden femoral arter ayırmak için 2 forseps kullanabilir. </li><li> kan akışını durdurmak için mikro kelepçeleri kullanın. Distal kelepçe ile takip ilk proksimal kelepçe, yerleştirin. </li><li> microscissors kenetlenmiş femoral arter kısa bir kesimi kesip atın. bir boşluk yaratarak microscissors kalan arter güdük kısaltılmasıven grefti daha büyük 1-2 mm. 30 G iğne kullanılarak heparin ile arteriyel güdük yıkayın. NOT: adventisya hafifçe damar güdük ötesinde çıkıntı halinde, geminin ucuna hafifçe çekin ve bir parça çıkarmak için forseps kullanabilir. </li><li> arteriyel kütükleri arasındaki adım 2.8 hasat ven yerleştirin ve boşluğa uygun sığacak şekilde uzunluğunu ayarlayın. ven yönüne dikkat edin. </li><li> ilk 10-0 prolen sütür kullanılarak proksimal anastomoz gerçekleştirin. (Şekil 1D) gösterilen sırayla tek dikiş yürütün. ventral tarafta üç sütür eklemeden önce her yan tarafında bir dikiş ile başlayın. Daha sonra, anastomoz tamamlamak için teknenin sırt tarafında üç dikiş yerleştirin. </li><li> Adım 3.8 açıklanan proksimal anastomoz için aynı tekniği kullanarak greft distal damarları bağlayın. Yine, her yan tarafında bir sütür ile başlamak ve daha sonra ventral sid üç sütür koyunE ve dorsal tarafı. </li><li> fibrin yapıştırıcı bileşen 1 ve 2 ile yük iki adet 1 mL&#39;lik şırınga dikkatlice forseps ile greft kaldırın ve bileşen 2, ardından greft altında fibrin yapıştırıcı bileşeninin 1 yaklaşık 100 ul bırakın. NOT:: 1 oranında parçalar 1 ve 2, bir 1 uygulanır emin olun. </li><li> geri pozisyonda greft yerleştirin ve greft üzerine bileşenleri 1 ve 2&#39;nin ek bir 100 ul bırakın. tutkal greft ve anastomoz yetmezliği ve damar grefti aşırı distansiyonu önlemek amacıyla anastomoz kapsar emin olun. </li><li> Dikkatle proksimal ardından uzak kelepçe açmak. </li><li> Nakledilen damar ve greft distal arter gözle görülür bir darbe kontrol ederek başarılı bir ameliyat onaylayın. </li><li> cilt kapatma engellemektedir aşırı tutkal, çıkarın. tedavi tutkal kaldırın ve microscissors aşırı çıkarmak için forseps kullanın. 5-0 prolen sütür ile cilt katmanları kapatın. </li><li> 4-5 m enjekteg / kg Carprofen subkutan sıçan uyanmak izin vermeden önce. sternal yatma korumak için yeterli bilinci yerine kadar sahipsiz hayvan bırakmayın. tamamen iyileşti kadar tek bir kafeste hayvan tutun. </li><li> Aşağıdaki 3 gün boyunca ağrı kesici ilaç olarak içme suyu (100 ml başına 50 mg metamizol) Metamizol ekleyin ve günlük hayvan izleyin. </li></ol> 4. Dubleks Sonografi NOT: duxplex sonografi sıçanlarda 14 non-invaziv kan akışını görselleştirmek için. <ol><li> indüksiyon odasında (izofluran% 2) bir sıçan Anesteziyoloji. Sırtında sıçan yerleştirin ve burnu kaplayan bir yüz maskesi ile anestezi korumak. </li><li> uyluk çevresi saç kaldırmak için saç makasları ve epilasyon kremi kullanın. </li><li> uyluk ultrason jeli uygulayın. hiçbir hava kabarcığı olmadığından emin olun. MS 400 transdüser kullanılarak dubleks sonografi görüntü elde (merkez frekansı: 30 M/ Sn 230-400 kare hızı ile Hz). </li></ol> 5. histopatolojisi NOT: Hasat morfometrik analiz 15 Masson trikrom boyama ile gemi leke ve. <ol><li> gece boyunca% 4 paraformaldehid içinde hasat kap saptamak etanol artan konsantrasyonlarda kurutmak. parafin örnek embed ve bir mikrotom kullanılarak 5 mikron kalınlığında dilimler halinde kesin. NOT: Paraformaldehyde toksik ve özel dikkatle ele alınmalıdır. </li><li> trikrom boyama solüsyonu ile onları boyamadan önce slaytlar Deparaffinize. Lekeli slaytlar dehydrate ksilen ile bunları temizleyin ve orta montaj takar. slaytlar kurutulduktan sonra, bir mikroskop ile örnekleri görmek. </li></ol> 6. Biyoparlaklık Görüntüleme (BLI) NOT: Ameliyat sonrası greft biyolüminesens sinyali 1 ölçerek in vivo zamanla takip edildi6. <ol><li> PBS 22 ml D-luciferase potasyum tuzu, 1 g çözülür ve intraperitoneal olarak sıçan (375 mg / kg vücut ağırlığı) enjekte. luciferin hayvan dönmesi için 15 dakika bekleyin. </li><li> gerçek zamanlı biyolüminesens miktar sistemi içine sıçan yerleştirin ve biyoparlaklık sinyali erişin. </li></ol>

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The authors have nothing to disclose.

Bu video ven greft kaybı araştırmak ve altta yatan patolojik süreçlerin araştırılması ve yeni ilaçlar ya da tedavi seçeneklerinin test etmek için izin veren bir fare ven interpozisyonu modelini göstermektedir. Anestezi cerrahi prosedürler çok önemli bir yönüdür. Bu özellikle uzun süreli operasyonlar sırasında, güvenli ve kolay bir yöntem olduğu için sürekli inhalasyon anestezi sistemi tavsiye edilir. Bu işlem en fazla 1 saat sürer eğitim safhasında büyük ön...

Koroner arter hastalıkları ve bunların komplikasyonları dünyada önde gelen ölüm nedenleri arasındadır. Güncel tedavi stratejileri yeniden kurulması kan akımı üzerine ya daralmış damar genişletici veya bir bypass oluşturarak odaklanmak. damar otogrefti kullanarak koroner arter bypass cerrahisi (CABG) ilk kez 1968 yılında tanımlanmıştır ve yılda rafine edilmiştir. Dışında sol ön revaskülarizasyon inen koroner arter gelen, safen ven kanallar en sık 1 kullanılmaktadır. Ancak, greft açıklık safen ven grefti (SVG) Aşil topuğu kalır. Cerrahiden bir yıl sonra, greft açıklık on yıl 2,3 sonra% 61 düşüyor,% 85&#39;dir. SVG açıklık kaybı patofizyolojik mekanizmaları ve nedenlerini açıklanması dolayısıyla önemli bir görevdir. Bu video ven greft kaybı araştırmak için bir sıçan ven interpozisyonu modelini göstermektedir. Bu yöntemin genel amaçları altında yatan biyopatolojik keşfetmek için vardırve ve hastalığın ilerlemesi sırasında -physiological süreçler ilaç veya tedavi seçeneği testi için uygun bir model geliştirmektir. arteriyel sisteme yüzeysel epigastrik ven nakli, bu model yakından koroner bypass klinik ayarını taklit eder. Cerrahi travma, iskemi ve duvar basıncı patolojik damar değişikliklerinin önemli tetikler ve tarif edilen modelinde taklit edilir. Farklı modeller ve türlerin ven greft açıklığı kaybı araştırmak için kullanılabilir. Bu domuzlar 4, koyun 5, köpeklerde 6 ve maymun 7 gibi büyük hayvan modelleri, insan damar ve anatomik yapıları benzer ve bu nedenle 8 test edilecek Bypass stent veya yeni cerrahi teknikler gibi kompleks terapötik stratejiler, sağlar. Ancak özel konut, ekipman ve personel gereklidir. Buna ek olarak, yüksek maliyetler ve ameliyat sırasında ek bir anestezist ihtiyacı onların geniş bir uygulama engel. Smsıçanlarda da dahil olmak üzere tüm hayvanlar, özel konut gerekmez, kullanımı kolay, ve yönetilebilir maliyetleri. Fare modellerinde 9,10 ile karşılaştırıldığında, sıçan modelleri sonuca daha iyi çalışabilirlik avantajı ve bu nedenle daha az değişkenliğe sahip. Sıçanlar fizyolojik ve genetik fareler 11,12 den insanlara daha benzer. Buna ek olarak, çoğu vahşi tip fareler sadece II hataları tip eğilimli fare modelleri yapmak sınırlı myointima 13, gelişir. Böyle inferior vena kava olarak ana fare damarlar, histolojisi, sadece birkaç hücre tabakalarından oluşur ve erken değerlendirme zor 13 vermektedir. Bir başka dezavantaj, greft kurtarma sonra bir sonraki analiz için uygun bir doku küçük bir miktardır. Bu video açıklanan model, gerçekleştirmek için, yeniden üretilebilir, ucuz ve kolay ve hızlı ve güvenilir bir tespit edilebilir. Bu viral vektörler gibi pahalı deneysel terapötik maddeleri değerlendirmek için uygundurekonomik bir şekilde gen terapisi için.

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			<td height="17" style="height:17px;width:236px;">Rat LEW/Crl</td>
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			<td style="width:156px;">http://www.anim.med.kyoto-u.ac.jp/NBR/</td>
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			<td align="right">27618</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Povidone-Iodine</td>
			<td>Betadine Purdue Pharma</td>
			<td>NDC:67618-152</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 Ethilon suture</td>
			<td>Ethicon</td>
			<td>2814G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer</td>
			<td style="width:145px;">400684.00.00</td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Novaminsulfon</td>
			<td>Ratiopharm</td>
			<td>PZN 03530402</td>
			<td>Metamizole</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Heparin</td>
			<td>Rotexmedica</td>
			<td>PZN: 3862340</td>
			<td>25.000 I.E./mL</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Xylocain 1%</td>
			<td>AstraZeneca</td>
			<td>PZN: 1137907</td>
			<td>Lidocain</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;">EVICEL</td>
			<td>J&#38;J Med.Ethicon Biosur</td>
			<td style="width:145px;">PZN 7349697 Art. Nr.:EVK01DE</td>
			<td>fibrin glue</td>
		</tr>
		<tr height="37">
			<td height="37" style="height:37px;">NaCl 0,9%</td>
			<td>B.Braun</td>
			<td style="width:145px;">PZN 06063042 Art. Nr.: 3570160</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:236px;">Vevo 770 high-resolution in vivo micro-imaging system</td>
			<td>VisualSonics</td>
			<td></td>
			<td>duplex sonography</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ecogel 100 ultrasound gel</td>
			<td>Eco-med</td>
			<td>30GB</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">D-Luciferin Firefly, potassium salt</td>
			<td>Biosynth</td>
			<td>L-8220</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PBS pH 7,4</td>
			<td>Gibco</td>
			<td>10010023</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Xenogen Ivis 200</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td>bioluminescence imaging</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Weigerts iron hematoxylin Kit</td>
			<td>Merck</td>
			<td>1.15973.0002</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Resorcine-Fuchsine Weigert</td>
			<td>Waldeck</td>
			<td>2.00E-30</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Acid Fuchsin</td>
			<td>Sigma-Aldrich</td>
			<td>F8129-25G</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ponceau S solution</td>
			<td>Serva Electrophoresis</td>
			<td>33427</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Azophloxin</td>
			<td>Waldeck</td>
			<td>1B-103</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:236px;">Molybdatophosphoric acid hydrate</td>
			<td>Merck</td>
			<td>1.00532.0100</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Orange G</td>
			<td>Waldeck</td>
			<td>1B-221</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Light Green SF</td>
			<td>Waldeck</td>
			<td>1B-211</td>
			<td>Trichrome staining</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vitro-Clud</td>
			<td>Langenbrinck</td>
			<td>04-0001</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Glacial Acetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td align="right">537020</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">37% HCl</td>
			<td>Sigma-Aldrich</td>
			<td>H1758</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Xylene</td>
			<td>Th. Geyer</td>
			<td align="right">3410</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Paraffin</td>
			<td>Leica biosystems</td>
			<td>REF 39602004</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ethanol absolute</td>
			<td>Th. Geyer</td>
			<td align="right">2246</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ethanol 96%</td>
			<td>Th. Geyer</td>
			<td align="right">2295</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Ethanol 70%</td>
			<td>Th. Geyer</td>
			<td align="right">2270</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Slide Rack</td>
			<td>Ted Pella</td>
			<td align="right">21057</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Staining dish</td>
			<td>Ted Pella</td>
			<td align="right">21075</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Bepanthen Eye and Nose ointment</td>
			<td>Bayer</td>
			<td align="right">1578675</td>
			<td>Eye ointment</td>
		</tr>
	</tbody>
</table>

vein interposition model: a suitable model to study bypass graft patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

sıçan ven interpozisyonu modeli myointima hiperplazi ve ven greft yetmezliği gelişimini incelemek için uygundur. Hayvanlar ameliyat sonrası iyi kurtarmak ve mükemmel fiziksel durumu sonrası operasyon göstermektedir. Şekil 1 anahtar cerrahi adımları gösterir. Linea Inguinalis boyunca cilt kesisi sonra, epigastrik yüzeysel ven ve femoral arter (Şekil 1A) tespit edilir. Bu erken greft yetmezliği ve açıklık kaybına yol açabilir olarak greft Hasat, greft (Şekil ...

Watch this Scientific Journal Video about Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency at JoVE.com

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

Ven interpozisyonu Model: baypas açıklığının Eğitim için uygun bir model

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

vein-interposition-model-suitable-model-to-study-bypass-graft

Research

JoVE Journal

Medicine

8.7K Görüntüleme.  University Heart Center Hamburg. This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats. 

Video: Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

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

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

Yetişkin Fare Venöz Hipertansiyon Modeli: Dış Boyun Ven Anastomoza Ortak Karotis Arter.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. In addition, measuring levels of a molecule of interest in human blood helps to further investigate its clinical relevance, but this represents systemic and not local inflammation. Therefore, we here describe a plaque culture model to study human atherosclerotic lesion biology ex vivo. In short, fresh plaques are obtained from patients undergoing endarterectomy or coronary artery bypass grafting and stored in RPMI medium on ice until usage. The specimens are cut into small pieces followed by random distribution into a 48-well plate, containing RPMI medium in addition to a substance of interest such as cytokines or chemokines alone or in combination for defined periods of time. After incubation, the plaque pieces can be shock frozen for mRNA isolation, embedded in Paraffin or OCT for immunohistochemistry staining or smashed and lysed for western blotting. Furthermore, cells may be isolated from the plaque for flow cytometry analysis. In addition, supernatants can be collected for protein measurement by ELISA. In conclusion, the presented ex vivo model opens the possibility to further study inflammatory lesional biology, which may result in identification of novel disease mechanisms and therapeutic targets.

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory process. This manuscript illustrates an easy to use ex vivo model to investigate fresh carotid or coronary artery plaques. The ex vivo model allows for the investigation of potential substances on the inflammatory milieu in human atherosclerotic lesions and results can be analyzed by various methods.

Bir İnsan<em&gt; Ex Vivo</em&gt; Aterosklerotik Plak Model Lezyon Biyoloji Eğitim için

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

Fare Modeli Inferior Vena Cava İnterpozisyon Greft İmplantasyonu

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

Kardiyopulmoner baypas gerektirmeyen bir fare modeli: yeni bir yaklaşım

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

Surgical Swine Model of Chronic Cardiac Ischemia Treated by Off-Pump Coronary Artery Bypass Graft Surgery

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of coronary artery disease (CAD) patients have HM, which in addition to causing impaired function, puts them at higher risk for arrhythmia and future cardiac events. The standard treatment for this condition is revascularization, but this has been shown to be an imperfect therapy. The majority of pre-clinical cardiac research focuses on infarct models of cardiac ischemia, leaving this subset of chronic ischemia patients largely underserved. To address this gap in research, we have developed a well-characterized and highly reproducible model of hibernating myocardium in swine, as swine are ideal translational models for human heart disease. In addition to creating this unique disease model, we have optimized a clinically relevant treatment model of coronary artery bypass surgery in swine. This allows us to accurately study the effects of bypass surgery on heart disease, as well as investigate additional or alternate therapies. This model surgically induces single vessel stenosis by implanting a constrictor on the left anterior descending (LAD) artery in a young pig. As the pig grows, the constrictor creates a gradual stenosis, resulting in chronic ischemia with impaired regional function, but preserving tissue viability. Following the establishment of the hibernating myocardium phenotype, we perform off-pump coronary artery bypass graft surgery to revascularize the ischemic region, mimicking the gold-standard treatment for patients in the clinic.

This protocol presents a surgical large animal model of chronic, single vessel ischemia that results in regional abnormalities but does not create infarct, known as hibernating myocardium. Following establishment of chronic ischemia, animals are treated with off-pump LIMA-LAD coronary artery bypass graft surgery to revascularize the ischemic tissue.

Kronik kalp iskemi "Off-Pump" koroner arter Bypass greft cerrahisi ile tedavi cerrahi domuz modeli

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

Tavşan manken dayanıklı Transgene ifade juguler ven ortak karotid arter interpozisyonu Greftler için

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

Farede Miyokard Enfarktüsü Çalışması İçin Kriyoyaralanma Modeli

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under Arterial Blood Pressure

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor because of the acceleration of intimal hyperplasia due to the exposure to arterial blood pressure. The present protocol is designed for the establishment of experimental venous intimal hyperplasia by interposing rabbit jugular veins to the ipsilateral carotid arteries. The protocol does not require surgical procedures deep in the body trunk and the extent of the incision is limited, which is less invasive for the animals, allowing long-term observation after implantation. This simple procedure enables researchers to investigate strategies to attenuate the progression of intimal hyperplasia of the implanted vein grafts. Using this protocol, we reported the effects transduction of microRNA-145 (miR-145), which is known to control the phenotype of vascular smooth muscle cells (VSMCs) from the proliferative to the contractile state, into harvested vein grafts. We confirmed the attenuation of intimal hyperplasia of vein grafts by transducing miR-145 before implantation surgery through the phenotype change of the VSMCs. Here we report a less invasive experimental platform to investigate the strategies that can be used to attenuate intimal hyperplasia of vein grafts in revascularization surgeries.

The present protocol aims to experimentally create venous intimal hyperplasia by subjecting veins to arterial blood pressure for developing strategies to attenuate venous intimal hyperplasia following revascularization surgery using vein grafts.

Arteriyel Kan Basıncı Altında İntimal Hiperplazi değerlendirmek için Ven Greftleri kullanarak Revaskülarizasyon Cerrahisi Taklit Bir Tavşan Venöz Interposition Modeli

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor ...