This work was supported in part by the United States Department of Veterans Affairs Biomedical Laboratory Research and Development Program Merit Review Award I01-BX002336, the National Institute of Health grant R56-HL095498, as well as with the resources and the use of facilities at the VA Connecticut Healthcare System, West Haven, CT.

Uygun Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onay elde edilir. 1. Anestezi ve Ameliyat öncesi Prosedürleri <ol><li> akrilik indüksiyon odasına uygulanan buharlaşmış% 3 izofluran ve 0.8 L / dk oksijen ile erkek C57BL / 6 buz, yaşlı 8 hafta, uyutmak. </li><li> ayak tutam tepki eksikliği yeterli anestezi onaylayın. Sürekli solunum yoluyla% 3 izofluran - ameliyat masasında fare yatar pozisyonda ve bir silikon maske pozisyon 2 buharlaşmış verebilen. </li><li> kimyasal tüy dökücü krem ​​kullanarak karın alt boyun ventral tüylerden. </li><li> Ilgi alanları da arteriyel ve venöz akış ve damar çapının temel özelliklerini kaydetmek için önce AVF ameliyat Doppler ultrason muayenesi yapın. 1,2 </li><li> 25 G iğne bir 1 ml şırınga takın ve istenilen ilaç ile şırınga yükleyin. Viraj ihtiyaçtan 60 derecelik bir açıyla iğne yaklaşık 4 mmle ucu. kavisli iğne tutucu ile 25 G iğne kavrayın. </li></ol> 2. Operatif Prosedürler <ol><li> Topikal antiseptik ile kesi sitesi hazırlayın ve bir cerrahi örtü uygulayın. ameliyat boyunca aseptik teknik korumak için steril eldiven ve aletler kullanın. </li><li> Bir neşter sadece pubis üstünde alt karaciğer kenarının seviyesinden uzanan bir orta hat karın kesi yapmak. </li><li> retractor takın ve sağ tarafına doğru karın boşluğundan tüm bağırsakları eviscerate. serum fizyolojik ile ıslatılmış gazlı bez bağırsakları sarın. aorta ve IVC tam görünümünü elde etmek için retro periton ve alt kolon bağlantı membran teşrih. </li><li> proksimal ve distal sıkma için hazırlanıyor, çevre dokulardan kızıl-böbrek aorta ve IVC teşrih. </li><li> Proksimal aort ve sadece sol renal venin altında seviyesinde proksimal IVC hem genelinde tek bir mikrocerrahi klibi yerleştirin. d hem genelinde ikinci bir mikrocerrahi klip yerleştirinISTAL aort ve distal İVC. </li><li> Aorta çevreleyen bağ dokusu kavrayın ve aort dorsal yüzeyi hafif arteriyel ponksiyon maruz böylece daha önce açıklandığı gibi, medial döndürün. 1 </li><li> Hızla Delinen bölgeyi maruz kalmaktadır. delinme sitesi damarları, aort bifürkasyon sol renal ven mesafe yaklaşık dörtte üçü kaudal olacaktır. sağ elle ponksiyon izin için yeterli maruz kalmayacak şekilde sol el ile döndürülmüş bir pozisyonda aort tutulması, aortun sol yanal marjı teşrih. aorta ve IVC arasında teşrih için dikkatli olun. </li><li> Bir döndürülmüş pozisyonda aort korumak ve ekli 1 ml şırınga içeren ilaç solüsyonu ile 25 G iğne kullanılarak IVC içine doğru aort delinme. (Şekil 1A) </li><li> sol elini kullanarak - (200 ul 100) ilaç çözümü demlenmeye. İğne dilate ve ince boyunca görülebilirŞeffaf ilaç solüsyon IVC (Şekil 1B, C) ​​dışında venöz kan yerinden IVC duvar. Hala kalır ve 15 dakika boyunca bir konumda iğne korumak. </li><li> sadece distal aorta ve distal IVC-kelepçesi de distal mikrocerrahi klibi çıkarın. </li><li> İğneyi çıkarın ve sonra komşu retro periton dokuyu çekerek aort Delinen bölgeyi kapsamaktadır. </li><li> proksimal aorta ve proksimal IVC de-kelepçe proksimal mikrocerrahi klibi çıkarın. de-sıkma üzerine, arter kan IVC yerine koyu venöz kan akımı akan gözlenen edilir. 1 dakika boyunca delinme delik kapsayan tutun. </li><li> sıkıştırma olmadan 30 sn gözlem hemostaz onaylanmasından sonra, onların doğal pozisyona bağırsakları dönmek ve onaylanmış hayvan protokole göre çalışan bir sütür ile karın kapatın. </li></ol> 3. Post-operatif prosedürler <ol><li> Karnın kapanmasından sonra, Anest durdurmalerine. Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından tavsiye edilen talimatlara uygun olarak analjezi ve yara bakımı da dahil olmak üzere post-operatif bakım uygulayın. analjezi için biz / kg intrasmuscularly 24 saat her 12 saatte cerrahi prosedürler izlenerek 0.1 mg buprenorfin kullanın. </li><li> Ameliyattan sonra ilk gününde, AVF açıklığını onaylamak için Doppler ultrasonografi gerçekleştirin. Buna ek olarak, diğer damar ölçülmesi ve seri akış karakteristikleri ve ameliyat öncesi taban çizgisi değerlerine göre değişiklikler karşılaştırma. 1,2 </li></ol><img alt="Şekil 1" src="/files/ftp_upload/53905/53905fig1.jpg" /> AVF Cerrahi sırasında intraluminal Teslim Gösterilen Şekil 1. (A) Operatif Fotoğraf. yakın ve uzak aort, ayrıca, mikro klipleri uygulayarak IVC sıkıştırın. Ekli şırınga içeren ilaç solüsyonu ile 25 G iğne kullanılarak IVC içine doğru aort delinme. (B) higOnu Güç Resmi İnfüzyon önce Patlak IVC (4X büyütme). İğne ucu koyu renkli venöz kan altında kalmıştır. Sarı ok uçları İnfüzyon sonra Patlak IVC İVC. (C) Higher Power Resim (4X büyütme) duvar duvara çapı belirtir. İğne ucu (siyah ok) venöz kan yerinden şeffaf ilaç çözüm olarak hafifçe şişmiş ve ince İVC duvarının (sarı ok başları) aracılığıyla görülebilir.

<ol>
	<li>Yamamoto, K., Li, X., Shu, C., Miyata, T., Dardik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+Aspects+of+the+Mouse+Aortocaval+Fistula.">Technical Aspects of the Mouse Aortocaval Fistula.</a> J. Vis. Exp. (77), e50449 (2013).</li><li>Yamamoto, K., 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=The+mouse+aortocaval+fistula+recapitulates+human+arteriovenous+fistula+maturation.">The mouse aortocaval fistula recapitulates human arteriovenous fistula maturation.</a> Am. J. Physiol.: Heart Circ. Physiol. 305 (12), H1718-H1725 (2013).</li><li>Escobar-Ch&#225;vez, J. J., L&#242;pez-Cervantes, M., Na&#239;k, A., Kalia, Y. N., Quintanar-Guerrero, D., Ganem-Quintanar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+thermo-reversible+pluronic+F-127+gels+in+pharmaceutical+formulations.">Applications of thermo-reversible pluronic F-127 gels in pharmaceutical formulations.</a> J. Pharm. Pharm. Sci. 9 (3), 339-358 (2006).</li><li>Almeida, H., Amaral, M. H., Lob&#227;o, P., Lobo, J. M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pluronic&#174;+F-127+and+Pluronic+Lecithin+Organogel+(PLO):+main+features+and+their+applications+in+topical+and+transdermal+administration+of+drugs.">Pluronic&#174; F-127 and Pluronic Lecithin Organogel (PLO): main features and their applications in topical and transdermal administration of drugs.</a> J. Pharm. Pharm. Sci. 15 (4), 592-605 (2012).</li><li>Karper, J. 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=Toll-like+receptor+4+is+involved+in+human+and+mouse+vein+graft+remodeling,+and+local+gene+silencing+reduces+vein+graft+disease+in+hypercholesterolemic+APOE*3Leiden+mice.">Toll-like receptor 4 is involved in human and mouse vein graft remodeling, and local gene silencing reduces vein graft disease in hypercholesterolemic APOE*3Leiden mice.</a> Arterioscler., Thromb., Vasc. Biol. 31 (5), 1033-1040 (2011).</li><li>Redmond, E. M., Hamm, K., Cullen, J. P., Hatch, E., Cahill, P. A., Morrow, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+patched-1+prevents+injury-induced+neointimal+hyperplasia.">Inhibition of patched-1 prevents injury-induced neointimal hyperplasia.</a> Arterioscler., Thromb., Vasc. Biol. 33 (8), 1960-1964 (2013).</li><li>Wong, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ephrin+type-B+receptor+4+activation+reduces+neointimal+hyperplasia+in+human+saphenous+vein+in+vitro.">Ephrin type-B receptor 4 activation reduces neointimal hyperplasia in human saphenous vein in vitro.</a> J. Vasc. Surg. , (2014).</li><li>Matsumoto, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemagglutinating+virus+of+Japan-liposome-mediated+gene+transfer+of+endothelial+cell+nitric+oxide+synthase+inhibits+intimal+hyperplasia+of+canine+vein+grafts+under+conditions+of+poor+runoff.">Hemagglutinating virus of Japan-liposome-mediated gene transfer of endothelial cell nitric oxide synthase inhibits intimal hyperplasia of canine vein grafts under conditions of poor runoff.</a> J. Vasc. Surg. 27 (1), 135-144 (1998).</li><li>Petrofski, J. A., 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=Gene+delivery+to+aortocoronary+saphenous+vein+grafts+in+a+large+animal+model+of+intimal+hyperplasia.">Gene delivery to aortocoronary saphenous vein grafts in a large animal model of intimal hyperplasia.</a> J. Thorac. Cardiovasc. Surg. 127 (1), 27-33 (2004).</li><li>Hata, J. A., 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=Modulation+of+phosphatidylinositol+3-kinase+signaling+reduces+intimal+hyperplasia+in+aortocoronary+saphenous+vein+grafts.">Modulation of phosphatidylinositol 3-kinase signaling reduces intimal hyperplasia in aortocoronary saphenous vein grafts.</a> J. Thorac. Cardiovasc. Surg. 129 (6), 1405-1413 (2005).</li><li>Ohta, S., 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=Intraluminal+gene+transfer+of+endothelial+cell-nitric+oxide+synthase+suppresses+intimal+hyperplasia+of+vein+grafts+in+cholesterol-fed+rabbit:+a+limited+biological+effect+as+a+result+of+the+loss+of+medial+smooth+muscle+cells.">Intraluminal gene transfer of endothelial cell-nitric oxide synthase suppresses intimal hyperplasia of vein grafts in cholesterol-fed rabbit: a limited biological effect as a result of the loss of medial smooth muscle cells.</a> Surg. 131 (6), 644-653 (2002).</li><li>Baldwin, Z. K., 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=Modulation+of+vascular+remodeling+induced+by+a+brief+intraluminal+exposure+to+the+recombinant+R7020+strain+of+Herpes+simplex-1.">Modulation of vascular remodeling induced by a brief intraluminal exposure to the recombinant R7020 strain of Herpes simplex-1.</a> J. Vasc. Surg. 41 (1), 115-121 (2005).</li><li>Eslami, M. H., 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=Gene+delivery+to+in+situ+veins:+differential+effects+of+adenovirus+and+adeno-associated+viral+vectors.">Gene delivery to in situ veins: differential effects of adenovirus and adeno-associated viral vectors.</a> J. Vasc. Surg. 31 (6), 1149-1159 (2000).</li><li>Globerman, A. S., 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=Efficient+transgene+expression+from+naked+DNA+delivered+into+an+arterio-venous+fistula+model+for+kidney+dialysis.">Efficient transgene expression from naked DNA delivered into an arterio-venous fistula model for kidney dialysis.</a> J. Gene Med. 13 (11), 611-621 (2011).</li><li>Yang, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adventitial+transduction+of+lentivirus-shRNA-VEGF-A+in+arteriovenous+fistula+reduces+venous+stenosis+formation.">Adventitial transduction of lentivirus-shRNA-VEGF-A in arteriovenous fistula reduces venous stenosis formation.</a> Kidney Int. 85 (2), 289-306 (2014).</li><li>Brahmbhatt, A., 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=The+role+of+Iex-1+in+the+pathogenesis+of+venous+neointimal+hyperplasia+associated+with+hemodialysis+arteriovenous+fistula.">The role of Iex-1 in the pathogenesis of venous neointimal hyperplasia associated with hemodialysis arteriovenous fistula.</a> PLoS ONE. 9 (7), e102542 (2014).</li><li>Karram, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+cardiac+hypertrophy+by+a+controlled+reproducible+sutureless+aortocaval+shunt+in+the+mouse.">Induction of cardiac hypertrophy by a controlled reproducible sutureless aortocaval shunt in the mouse.</a> J. Investig. Surg. 18 (6), 325-334 (2005).</li><li>Perry, G. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+variation+in+angiotensin-converting+enzyme+does+not+prevent+development+of+cardiac+hypertrophy+or+upregulation+of+angiotensin+II+in+response+to+aortocaval+fistula.">Genetic variation in angiotensin-converting enzyme does not prevent development of cardiac hypertrophy or upregulation of angiotensin II in response to aortocaval fistula.</a> Circ. 103 (7), 1012-1016 (2001).</li><li>Guzman, R. J., Krystkowiak, A., Zarins, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+and+sustained+medial+cell+activation+after+aortocaval+fistula+creation+in+mice.">Early and sustained medial cell activation after aortocaval fistula creation in mice.</a> J. Surg. Res. 108 (1), 112-121 (2002).</li><li>Alexander, J. H., 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=Efficacy+and+safety+of+edifoligide,+an+E2F+transcription+factor+decoy,+for+prevention+of+vein+graft+failure+following+coronary+artery+bypass+graft+surgery:+PREVENT+IV:+a+randomized+controlled+trial.">Efficacy and safety of edifoligide, an E2F transcription factor decoy, for prevention of vein graft failure following coronary artery bypass graft surgery: PREVENT IV: a randomized controlled trial.</a> JAMA. 294 (19), 2446-2454 (2005).</li><li>Khaleel, M. S., 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=High-pressure+distention+of+the+saphenous+vein+during+preparation+results+in+increased+markers+of+inflammation:+a+potential+mechanism+for+graft+failure.">High-pressure distention of the saphenous vein during preparation results in increased markers of inflammation: a potential mechanism for graft failure.</a> Ann. thorac. surg. 93 (2), 552-558 (2012).</li></ol>

The authors have nothing to disclose.

fare AVF modelinin Bu değişiklik AVF oluşturulması sırasında venöz endotel intraluminal ilaç dağıtım içermektedir. AVF 25 G iğne ile alt böbrek aort delme, aynı iğne ile ilaç çözeltisi enjekte edilerek ve ardından IVC Karşıt aort duvarı ile delik uzanan oluşturuldu. Çözelti, intra-kaval korunur yani., AVF venöz ekstremite, de-sıkma kadar. Başka hangi fare AVF modellerinde 17-19 bu modeli ayıran moleküler analizi ile darlık, akut tromboz ve olası parazitlere neden olab...

Fare aortovenous fistül (AVF) aort ve inferior vena kava (IVC) arasında delinme modeli şimdi doğru çıkan bu modelde bir tekniktir. 1, 25 G iğne ile delinir ötesi böbrek aortun her iki duvarları olduğunu infra-böbrek vena kava bitişik; Anterior aort giriş deliği basit sıkıştırma ile tamir edilir, ve dikiş onarım gerektirmez. Yüksek çözünürlüklü Doppler ultrasonografi ve histolojik analiz seri izlem muayenesi olgunlaşma evresini ve sonra başarısız faz, insan AVF bilinen patofizyolojisi yansıtan olması AVF gösterir. 2 AVF olgunlaşmasını modüle eden mekanizmaları incelemek için, olgunlaşma AVF endotele terapötik maddelerin uygulanması için geliştirilmiş yöntemlere ihtiyaç vardır. kaplara terapötik maddelerin teslimi ya endovasküler iletim yoluyla lümen veya adventisya dış iletim yoluyla olabilir. Dış teslimat bir örneği, comkip eden bacak Pluronic jel adventisya uygulama kullanılabilir. vücut sıcaklığına ısıtıldı, bu kopolimer, termo-geri ve katı jel, sıvı dönüştürülür. Pluronic jel karışık madde in vivo topik olarak uygulandığında Önceki çalışmalar göstermiştir ilaç salınımı elde edilir. Viral vektörler ya Pluronik jel siRNA&#39;nın 3,4 Adventisyal uygulama perivasküler dağıtım sistemi olarak etkili olduğu rapor edilmiştir. 5,6 Biz Ayrıca, peptidler tarafından adventisya uyarım ile eksplante insan safen damarların tedavisi endotel reseptör protein fosforilasyonu ile sonuçlandığını rapor vardır. 7 Öte yandan, buluş sahipleri ayrıca belirtmişlerdir damar greft 8-10 köpek ve tavşan modellerinde 11,12 hem viral ve viral olmayan vektörler lümen içi teslim kullandık. Bu raporlarda, gen transferi hasat ven sonra ex vivo gerçekleştirilmiştir. Eslami ve ark., Endovasküler viral gen teslim rapory bypass yaratmadan yerinde damarları karotis için. 13 Gloverman ve ark. Sıçan femoral arter-yüzeyel epigastrik ven fistül çıplak DNA bildirdi lümen içi ve adventisyal teslim. 14 Mayo grup fare karotid arter-juguler ven fistül içinde adventisyal ilaç dağıtım bildirdi. 15,16 Ancak, bu daha önce bildirilen modeller oluşturmak için dikilmiş anastomoz gerekli AVF. Bu yazıda, farelerde aynı anda AVF oluşturulması ile lümen içi ilaç verme AVF oluşturma dikiş daha az model kullanılarak, tarif edilecektir. bu değiştirilmiş sıçangil AVF modeli kullanarak fistül venöz bacak için lümen içi ilaç verilmesi için basit bir yöntem gerçekleştirilebilir.

<table><tbody><tr><td>Pluronic F-127</td><td>Sigma-Aldrich</td><td>P2443-250G</td><td>Used as 30% solution in d-water</td></tr><tr><td>GFP antibody</td><td>NOVUS BIOLOGICALS INC</td><td>NB100-1770</td><td></td></tr><tr><td>Ad-CMV-GFP</td><td>VECTOR BIOLABS</td><td>1060</td><td></td></tr><tr><td>0.9% Sodium Chloride Irrigation, USP</td><td>Baxter</td><td>2F7122</td><td></td></tr><tr><td>BD PrecisionGlide Needle 25 G x 5/8</td><td>BD</td><td>305122</td><td></td></tr><tr><td>BD 1 ml Syringe Tuberculin Slip Tip</td><td>BD</td><td>309659</td><td></td></tr><tr><td>Scalpel</td><td>Surgical Design Inc</td><td>22079707</td><td></td></tr><tr><td>6-0 ETHILON P-1 11 mm 3/8c Reverse Cutting</td><td>ETHICON INC</td><td>697G</td><td></td></tr><tr><td>Vevo 770 ultrasound machine&amp;nbsp;</td><td>Visualsonics&amp;nbsp;</td><td>20 - 60 Mhz scan head; RMV-704</td><td></td></tr><tr><td>Vascular clamp&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5424</td><td></td></tr><tr><td>Clamp applying forceps&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5410</td><td></td></tr></tbody></table>

intraluminal drug delivery to the mouse arteriovenous fistula endothelium

Terapötik maddelerin teslimi arteryovenöz fistül (AVF) olgunlaşması ya lümen içi ya da dış yollar ile uygulanabilir geliştirmek için. Basit bir sıçangil AVF modeli fistül oluşturulması ile aynı anda venöz endotele ilaç solüsyonu intralüminal uygulaması ile bir araya getirilmiştir. Bu modelin teknik özellikleri tartışılmıştır. Genel anestezi altında, karın kesi yapılır ve aort ve vena kava inferior (VKİ) maruz kalmaktadır. infra-böbrek aort ve IVC sıkma için disseke edilir. proksimal ve distal sıkma sonra, delinme sitesi maruz kalır ve 25 G iğne aorta ve IVC içine hem duvarları delinerek için kullanılır. Hemen delme sonra, raportör geni ifade eden viral vektörü inkübasyon 15 dakika süre ile, aynı iğne yoluyla IVC infüze edildi. intraluminal uygulama yöntemi, dış yoldan uygulamaya kıyasla venöz endotel daha sağlam viral gen teslim sağladı. bu noveTeslimat l yöntemi AVF olgunlaşma endotel rolünü araştırmak ve cerrahi operasyon sırasında intraluminal ilaç dağıtımını sağlayacak çalışmalar kolaylaştıracaktır.

33 farelerin bir dizi, ilk post-operatif gün hayatta kalma% 97.0 idi; ultrason ile belirlenen AVF açıklık,% 84.9 idi. Geleneksel dış rota ile bu endovasküler teslim rotanın Gen transdüksiyon etkinliği karşılaştırıldı. Lümen içi doğum (ILD) hemen delme işlemi adenovirüs-GFP 200 ul (Ad-GFP) vektörü çözeltisi (1 x 10 9 PFU / ml) 15 dakika inkübasyon süresi ve ardından delme iğnesi ile IVC i...

Watch this Scientific Journal Video about Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium at JoVE.com

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Fare Arteriovenöz Fistül Endotelyumunun için intraluminal Drug Delivery

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

intraluminal-drug-delivery-to-mouse-arteriovenous-fistula

Research

JoVE Journal

Medicine

9.3K Görüntüleme.  Yale University. After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Video: Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

Mikro-Bilgisayarlı Tomografi Görüntüleme için Microfil ile İskemik arka ayaklarında ve damarları bir Perfüzyon için osmotik İlaç Taşıyıcı

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

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Technical Aspects of the Mouse Aortocaval Fistula

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

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

Fare Aortocaval Fistül Teknik Yönleri

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

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 ...

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high molecular weight compounds from the bloodstream and into brain tissue. Some direct delivery success in humans has been achieved via implantation of transcranial catheters; however this method is highly invasive and associated with numerous complications. A less invasive alternative would be to dose the brain through a surgically implanted, semipermeable membrane such as the nasal mucosa that is used to repair skull base defects following endoscopic transnasal tumor removal surgery in humans. Drug transfer though this membrane would effectively bypass the BBB and diffuse directly into the brain and cerebrospinal fluid. Inspired by this approach, a surgical approach in mice was developed that uses a donor septal mucosal membrane engrafted over an extracranial surgical BBB defect. This model has been shown to effectively allow the passage of high molecular weight compounds into the brain. Since numerous drug candidates are incapable of crossing the BBB, this model is valuable for performing preclinical testing of novel therapies for neurological and psychiatric diseases.

A mouse model of human endoscopic skull base reconstruction has been developed that creates a semipermeable interface between the brain and nose using nasal mucosal grafts. This method allows researchers to study delivery to the central nervous system of high molecular weight therapeutics which are otherwise excluded by the blood-brain barrier when administered systemically.

Farelerde Beyin Doğrudan İlaç Teslim heterotopik mukozal aşılanması Prosedürü

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high ...

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. ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

Arteriovenöz Fistül Yetersizliğinin A Novel Fare Modeli: Detay Cerrahi Prosedür

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

Bir outflow stenozu ile Aokaval fistül kullanarak Merkezi venöz stenozis murine modeli

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for analyzing collateral arteries and leukocytes in arteriogenesis is ex vivo (immuno-) histological methodology. However, this technique does not allow the measurement of dynamic processes such as blood flow, shear stress, cell-cell interactions, and particle velocity. This paper presents a protocol to monitor in vivo processes in growing collateral arteries during arteriogenesis utilizing intravital imaging. The method described here is a reliable tool for dynamics measurement and offers a high-contrast analysis with minimal photo-cytotoxicity, provided by multiphoton excitation microscopy. Prior to analyzing growing collateral arteries, arteriogenesis was induced in the adductor muscle of mice by unilateral ligation of the femoral artery.
After the ligation, the preexisting collateral arteries started to grow due to increased shear stress. Twenty-four hours after surgery, the skin and subcutaneous fat above the collateral arteries were removed, constructing a pocket for further analyses. To visualize blood flow and immune cells during in vivo imaging, CD41-fluorescein isothiocyanate (FITC) (platelets) and CD45-phycoerythrin (PE) (leukocytes) antibodies were injected intravenously (i.v.) via a catheter placed in the tail vein of a mouse. This article introduces intravital multiphoton imaging as an alternative or in vivo complementation to the commonly used static ex vivo (immuno-) histological analyses to study processes relevant for arteriogenesis. In summary, this paper describes a novel and dynamic in vivo method to investigate immune cell trafficking, blood flow, and shear stress in a hindlimb model of arteriogenesis, which enhances evaluation possibilities notably.

The recruitment of leukocytes and platelets constitutes an essential component necessary for the effective growth of collateral arteries during arteriogenesis. Multiphoton microscopy is an efficient tool for tracking cell dynamics with high spatio-temporal resolution in vivo and less photo-toxicity to study leukocyte recruitment and extravasation during arteriogenesis.

Bir Murin Arka Bacak Modelinde Arteriyogenez Sırasında Lökosit Alımını İzlemek için Multifoton İntravital Görüntüleme

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

Hemodiyaliz Erişimi ile İlişkili El Disfonksiyonunun Bir Murine Modeli

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

Drug Treatment by Central Venous Catheter in a Mouse Model of Angiotensin II Induced Abdominal Aortic Aneurysm and Monitoring by 3D Ultrasound

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse models, are applied to advance the understanding of the disease pathogenesis and to identify potential therapeutic targets. Testing novel drug candidates to block AAA growth in these models generally requires repeated drug administration during the time course of the experiment. Here, we describe a compiled protocol for AAA induction, insertion of an intravenous catheter to facilitate prolonged therapy, and serial AAA monitoring by 3D ultrasound. Aneurysms are induced in apolipoprotein E (ApoE) deficient mice by angiotensin II release over 28 days from osmotic mini-pumps implanted subcutaneously into the mouse back. Subsequently, the surgical procedure for external jugular vein catheterization is conducted to allow for daily intravenous drug treatment or repeated blood sampling via a subcutaneous vascular access button. Despite the two dorsal implants, the monitoring of AAA development is readily facilitated by sequential semi-automated 3D ultrasound analysis, which yields comprehensive information on the expansion of aortic diameter and volume and on aneurysm morphology, as illustrated by experimental examples.

This protocol describes the consecutive implantation of an osmotic pump to induce abdominal aortic aneurysm by angiotensin II release in apolipoprotein E (ApoE) deficient mice and of a vascular access port with a jugular vein catheter for repeated drug treatment. Monitoring of aneurysm development by 3D ultrasound is effectively conducted despite dorsal implants.

Anjiyotensin II İndüklü Abdominal Aort Anevrizmasının Bir Fare Modelinde Santral Venöz Kateter ile İlaç Tedavisi ve 3D Ultrason ile İzleme

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse ...