Name: a rabbit model of durable transgene expression in jugular vein to common carotid artery interposition grafts
Uploaded: 2018-09-10T12:30:00
Description: Watch this Scientific Journal Video about A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts at JoVE.com

すべての動物のプロトコルと研究は、ワシントン事務所の動物福祉の大学によって承認されました。
1. 事前操作 (すべての手術)
<ol><li>棘筋の筋肉内注射 (IM) 30 mg/kg 1.5 mg/kg とケタミン ・ キシラジンとウサギを麻酔します。 注: 食料と水は、手術前に制限されません。</li>
	<li>十分な麻酔深度を待っている間、準備室と手術室 (OR) 内のテーブルを設定します。<ol>
<li>ウ?...

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	<li>Libby, P., Bornfeldt, K. E., Tall, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atherosclerosis:+Successes,+Surprises,+and+Future+Challenges.">Atherosclerosis: Successes, Surprises, and Future Challenges.</a> Circ Res. 118, 531-534 (2016).</li><li>Fowkes, F. G., 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=Comparison+of+global+estimates+of+prevalence+and+risk+factors+for+peripheral+artery+disease+in+2000+and+2010:+a+systematic+review+and+analysis.">Comparison of global estimates of prevalence and risk factors for peripheral artery disease in 2000 and 2010: a systematic review and analysis.</a> Lancet. 382, 1329-1340 (2013).</li><li>Mohr, F. W., 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=Coronary+artery+bypass+graft+surgery+versus+percutaneous+coronary+intervention+in+patients+with+three-vessel+disease+and+left+main+coronary+disease:+5-year+follow-up+of+the+randomised,+clinical+SYNTAX+trial.">Coronary artery bypass graft surgery versus percutaneous coronary intervention in patients with three-vessel disease and left main coronary disease: 5-year follow-up of the randomised, clinical SYNTAX trial.</a> Lancet. 381, 629-638 (2013).</li><li>de Vries, M. R., Simons, K. H., Jukema, J. W., Braun, J., Quax, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vein+graft+failure:+from+pathophysiology+to+clinical+outcomes.">Vein graft failure: from pathophysiology to clinical outcomes.</a> Nat Rev Cardiol. 13 (8), 451-470 (2016).</li><li>Harskamp, R. E., Lopes, R. D., Baisden, C. E., de Winter, R. J., Alexander, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saphenous+vein+graft+failure+after+coronary+artery+bypass+surgery:+pathophysiology,+management,+and+future+directions.">Saphenous vein graft failure after coronary artery bypass surgery: pathophysiology, management, and future directions.</a> Ann Surg. 257, 824-833 (2013).</li><li>Sabik, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+saphenous+vein+graft+patency.">Understanding saphenous vein graft patency.</a> Circulation. 124 (3), 273-275 (2011).</li><li>Owens, C. D., Ho, K. J., Conte, 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=Lower+extremity+vein+graft+failure:+a+translational+approach.">Lower extremity vein graft failure: a translational approach.</a> Vasc Med. 13, 63-74 (2008).</li><li>Robertson, K. E., McDonald, R. A., Oldroyd, K. G., Nicklin, S. A., Baker, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+of+coronary+in-stent+restenosis+and+vein+graft+failure:+does+vascular+gene+therapy+have+a+role.">Prevention of coronary in-stent restenosis and vein graft failure: does vascular gene therapy have a role.</a> Pharmacol Ther. 136, 23-34 (2012).</li><li>Yla-Herttuala, S., Baker, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+Gene+Therapy:+Past,+Present,+and+Future.">Cardiovascular Gene Therapy: Past, Present, and Future.</a> Mol Ther. 25, 1095-1106 (2017).</li><li>Schwartz, L. 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=Adenoviral-mediated+gene+transfer+of+a+constitutively+active+form+of+the+retinoblastoma+gene+product+attenuates+neointimal+thickening+in+experimental+vein+grafts.">Adenoviral-mediated gene transfer of a constitutively active form of the retinoblastoma gene product attenuates neointimal thickening in experimental vein grafts.</a> J Vasc Surg. (5), 874-881 (1999).</li><li>Eefting, D., 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=Local+lentiviral+short+hairpin+RNA+silencing+of+CCR2+inhibits+vein+graft+thickening+in+hypercholesterolemic+apolipoprotein+E3-Leiden+mice.">Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice.</a> J Vasc Surg. 50, 152-160 (2009).</li><li>Handa, 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=Adventitial+delivery+of+platelet-derived+endothelial+cell+growth+factor+gene+prevented+intimal+hyperplasia+of+vein+graft.">Adventitial delivery of platelet-derived endothelial cell growth factor gene prevented intimal hyperplasia of vein graft.</a> J Vasc Surg. 48 (6), 1566-1574 (2008).</li><li>Kloppenburg, G. T., Grauls, G. E., Bruggeman, C. A., Stassen, F. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenoviral+activin+A+expression+prevents+vein+graft+intimal+hyperplasia+in+a+rat+model.">Adenoviral activin A expression prevents vein graft intimal hyperplasia in a rat model.</a> Interact Cardiov Th. 8, 31-34 (2009).</li><li>Eefting, D., 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+novel+urokinase+receptor-targeted+inhibitor+for+plasmin+and+matrix+metalloproteinases+suppresses+vein+graft+disease.">A novel urokinase receptor-targeted inhibitor for plasmin and matrix metalloproteinases suppresses vein graft disease.</a> Cardiovasc Res. 88, 367-375 (2010).</li><li>Eichstaedt, H. 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=Gene+transfer+of+COX-1+improves+lumen+size+and+blood+flow+in+carotid+bypass+grafts.">Gene transfer of COX-1 improves lumen size and blood flow in carotid bypass grafts.</a> J Surg Res. 161, 162-167 (2010).</li><li>Kritz, A. 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=In+vivo+modulation+of+Nogo-B+attenuates+neointima+formation.">In vivo modulation of Nogo-B attenuates neointima formation.</a> Mol Ther. 16 (11), 1798-1804 (2008).</li><li>Peroulis, 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=The+role+of+ex-vivo+gene+therapy+of+vein+grafts+with+Egr-1+decoy+in+the+suppression+of+intimal+hyperplasia.">The role of ex-vivo gene therapy of vein grafts with Egr-1 decoy in the suppression of intimal hyperplasia.</a> Eur J Vasc Endovasc. 40, 216-223 (2010).</li><li>Kochanek, 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=A+new+adenoviral+vector:+Replacement+of+all+viral+coding+sequences+with+28+kb+of+DNA+independently+expressing+both+full-length+dystrophin+and+b-galactosidase.">A new adenoviral vector: Replacement of all viral coding sequences with 28 kb of DNA independently expressing both full-length dystrophin and b-galactosidase.</a> Proc Natl Acad Sci U S A. 93, 5731-5736 (1996).</li><li>Parks, R. 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=A+helper-dependent+adenovirus+vector+system:+Removal+of+helper+virus+by+Cre-mediated+excision+of+the+viral+packaging+signal.">A helper-dependent adenovirus vector system: Removal of helper virus by Cre-mediated excision of the viral packaging signal.</a> P Natl Acad Sci USA. 93, 13565-13570 (1996).</li><li>Chen, H. -. 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=Persistence+in+muscle+of+an+adenoviral+vector+that+lacks+all+viral+genes.">Persistence in muscle of an adenoviral vector that lacks all viral genes.</a> P Natl Acad Sci USA. 94, 1645-1650 (1997).</li><li>Wen, S., Graf, S., Massey, P. G., Dichek, D. 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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=Efficient+adenoviral+gene+transfer+to+early+venous+bypass+grafts:+comparison+with+native+vessels.">Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels.</a> Cardiovasc Res. 35, 505-513 (1997).</li><li>Flynn, R., 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=Expression+of+apolipoprotein+A-I+in+rabbit+carotid+endothelium+protects+against+atherosclerosis.">Expression of apolipoprotein A-I in rabbit carotid endothelium protects against atherosclerosis.</a> Mol Ther. 19, 1833-1841 (2011).</li><li>George, S. 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=Sustained+Reduction+of+Vein+Graft+Neointima+Formation+by+Ex+Vivo+TIMP-3+Gene+Therapy.">Sustained Reduction of Vein Graft Neointima Formation by Ex Vivo TIMP-3 Gene Therapy.</a> Circulation. 124, 135-142 (2011).</li><li>Chiu-Pinheiro, C. 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=Gene+transfer+to+coronary+artery+bypass+conduits.">Gene transfer to coronary artery bypass conduits.</a> Ann Thorac Surg. 74, 1161-1166 (2002).</li><li>Byrom, M. J., Bannon, P. G., White, G. H., Ng, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+the+assessment+of+novel+vascular+conduits.">Animal models for the assessment of novel vascular conduits.</a> J Vasc Surg. 52, 176-195 (2010).</li><li>Ribichini, F., 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=Effects+of+oral+prednisone+after+stenting+in+a+rabbit+model+of+established+atherosclerosis.">Effects of oral prednisone after stenting in a rabbit model of established atherosclerosis.</a> J Am Coll Cardiol. 50, 176-185 (2007).</li><li>Langheinrich, A. 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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atherosclerosis+in+rabbit+vein+grafts.">Atherosclerosis in rabbit vein grafts.</a> Arteriosclerosis. 9, 374-379 (1989).</li><li>Qiang, 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=Statin+therapy+prevents+expansive+remodeling+in+venous+bypass+grafts.">Statin therapy prevents expansive remodeling in venous bypass grafts.</a> Atherosclerosis. 223, 106-113 (2012).</li><li>Casa, L. D. C., Ku, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombus+Formation+at+High+Shear+Rates.">Thrombus Formation at High Shear Rates.</a> Annu Rev Biomed Eng. 19, 415-433 (2017).</li><li>Chen, C., Coyle, K. A., Hughes, J. D., Lumsden, A. B., Ku, D. 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このプロトコルの重要なステップには、麻酔の管理、動脈/グラフト静脈の抗凝固、手術操作、グラフト静脈の血行動態の測定が含まれます。2 つの比較的長い操作 (二国間静脈グラフトの通常 3 〜 3.5 h および二国間移植伝達の 1.5 〜 2.5 h) を含む複数この生存手術モデルの麻酔の適切な管理が欠かせません。両方、粗野で、気管内挿管による麻酔を投与し、挿管を考える肯定的な圧力換気、無...

動脈硬化は、脂質蓄積と血管壁の炎症で血管内腔、心臓発作、ストロークおよび手足1,2の損失の縮小につながる慢性炎症性疾患です。経皮的介入 (e.g、血管形成術とステント留置術) と薬物療法 (e.g。、スタチン系薬剤と抗血小板薬のエージェント); アテローム性動脈硬化症の治療は、。ただし、彼らはしばしば冠動脈および末梢循環に重度の閉塞性疾患の両方の治療に効果的です。バイパス術、自家静脈セグメントを使用して深刻なびまん性冠動脈および末梢血管疾患3,4の患者を治療するための一般的な手順のままです。ただし、両方の冠は、移植を静脈し、末梢循環悪い長期開存率があります。冠循環の静脈グラフトが 1 年間で 50% 遮蔽の約 10-20% は 10 年5,6に遮蔽されました。末梢循環の静脈グラフト故障率、5 年間7時 30-50% です。
病気のサイトに正確に治療遺伝子産物を配信できるため、遺伝子治療は静脈グラフト障害の防止のための魅力的なアプローチです。したがって、多くの前臨床研究は静脈グラフト遺伝子療法8,9をテストしています。ただし、本質的にこれらの研究のすべては、早い時間 (2-12 週) のポイント10、11,12,13,14,15,の有効性を調べられます16,17. 遺伝子療法の介入が耐久性を提供できるという証拠はないと意識しております (年) 通常起因する新生内膜過形成や動脈硬化4後半の静脈グラフト障害に対する保護。グラフト静脈で耐久性のある遺伝子発現をでき、後半と初期の時点で遺伝子治療介入のテストによりメソッドを開発しました。耐久性のある遺伝子発現を成し遂げるためには、メソッドには、HDAd ベクトルと遅延伝達戦略が組み込まれています。HDAd ベクトル免疫18,19,によって導入された細胞の認識 (と拒絶)20,を防止するウイルスの遺伝子がないために長期の遺伝子発現を提供する21. 遅延伝達 (グラフト留置後実行される 28 日) は22移植後早期に発生する大量の処理中に導入された細胞の損失を防ぐことができます。
移植配置10、11,12,15,16 時静脈グラフトの伝達に依存して静脈の血管壁における治療導入遺伝子発現を達成する他の方法 ,17。連続的に測定したとき遺伝子発現のこのアプローチを使用してすばやく伝達22,23後低下します。したがって、このアプローチを用いた研究がほとんどない 4 週間を超えて有効性を評価すると、グラフト静脈後 12 週を超えて有効性を検討していません。これに対し、本方式による静脈グラフトと-同じような研究に基づいて動脈可能性がありますで実行される、少なくとも 24 数週間安定持続する遺伝子発現のはるかに長い22,24を続けています。その他静脈グラフト遺伝子療法の介入なしこの期間の遺伝子発現の安定を実現すると意識しております。
家兎モデルを使用して、本手法を開発しました。他の齧歯動物、ウサギ、使用したまたは静脈をテストするより大きい動物移植遺伝子療法10、11,12,15,16,17,25、 26。齧歯動物モデルと比較して、ウサギはより高くより厳格な規制要件が適用されます。しかしより大きい動物に比較 (e.g。、ブタおよび犬)、ウサギを購入し家にはるかに高価な、はるかに扱いやすい。ウサギ血管生理人間の血管のようにまた、27、彼らが経皮的介入28,29をテストするため使用することができます、彼らは十分な組織を提供する十分に大きい複数のエンドポイント (e.g。、組織学、タンパク質、RNA) 単一の血管標本22,30を使用して調べることができます。さらに、静脈グラフトのウサギは、高脂肪食にうんざりした、彼ら開発静脈移植動脈硬化31,32, 冠動脈バイパス静脈グラフト障害4,5 の一般的な原因であります。.これらの動脈硬化家兎静脈グラフトは、遺伝子治療の介入は、この方法をテストするための基板として使用できます。指定されたプロトコルは、ウサギ静脈グラフトにおける遺伝子発現の耐久性を達成するために必要な技術的なスキルを習得する捜査官を助けることができます。

<table border="0" cellpadding="0" cellspacing="0" style="width:707px;" width="707">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Disposables</td>
			<td style="width:167px;"></td>
			<td style="width:119px;"></td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">3mL syringe with 24G needle</td>
			<td>Becton Dickinson</td>
			<td>309571</td>
			<td style="width:207px;">2x for vein graft surgery; 2x for gene transfer&#160; surgery</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">1 mL syringe with 27G needle</td>
			<td>Becton Dickinson</td>
			<td>309623</td>
			<td style="width:207px;">2x for vein graft surgery, 5x for gene transfer surgery</td>
		</tr>
		<tr height="20">
			<td height="40" rowspan="2" style="height:40px;width:215px;">19G needle</td>
			<td rowspan="2">Becton Dickinson</td>
			<td rowspan="2">305187</td>
			<td rowspan="2" style="width:207px;">Gene transfer surgery</td>
		</tr>
		<tr height="20">
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">20 mL syringe, luer lock</td>
			<td>Nipro Medical Corp</td>
			<td>JD+20L</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Catheters, 24Ga x 3/4&#8221;</td>
			<td>Terumo Medical Products</td>
			<td>SROX2419V</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">21G needle</td>
			<td>Becton Dickinson</td>
			<td>305165</td>
			<td style="width:207px;">Gene transfer surgery and for 20 mL syringe of saline</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Gauze 4&#8221; x 4&#8221;</td>
			<td>Dynarex</td>
			<td style="width:119px;">3242</td>
			<td style="width:207px;">~10-15 per surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">3-0 silk suture</td>
			<td>Covidien Ltd.</td>
			<td>S-244</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">5-0 silk suture</td>
			<td>Covidien Ltd.</td>
			<td>S-182</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">7-0 polypropylene suture</td>
			<td>CP Medical</td>
			<td>8703P</td>
			<td style="width:207px;">Vein graft surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">7-0 polypropylene suture</td>
			<td>CP Medical</td>
			<td>8648P</td>
			<td style="width:207px;">Gene transfer surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">5-0 polyglycolic acid suture</td>
			<td>CP Medical</td>
			<td>421A</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">3-0 polyglycolic acid suture</td>
			<td>CP Medical</td>
			<td>398A</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Alcohol swabs</td>
			<td>Covidien Ltd.</td>
			<td>6818</td>
			<td style="width:207px;">For the placement of I.V. line</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Catheter plug</td>
			<td>Vetoquinol</td>
			<td>411498</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Ketamine HCl, 100 mg/mL</td>
			<td>Vedco Inc.</td>
			<td>5098916106</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Xylazine, 100 mg/mL</td>
			<td>Akorn Inc.</td>
			<td>4821</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Lidocaine, 20 mg/mL</td>
			<td>Pfizer</td>
			<td>409427702</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Marcaine 0.5%</td>
			<td>Pfizer</td>
			<td>409161050</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Beuthanasia D-Special</td>
			<td>Intervet Inc.</td>
			<td>NDC 00061047305</td>
			<td style="width:207px;">Harvest surgery only</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Buprenorphine HCl, 0.3 mg/mL</td>
			<td>Patterson Veterinary</td>
			<td>12496075705</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Saline IV bag, 0.9% sodium chloride</td>
			<td>Baxter</td>
			<td>2B1309</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Heparin&#160; (5000 U/mL)</td>
			<td>APP Pharmaceuticals</td>
			<td>NDC 63323-047-10</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Papaverine (3.5 mg/ml)</td>
			<td>American Reagent Inc.</td>
			<td>NDC 0517-4002-25</td>
			<td style="width:207px;">Diluted from 30mg/mL stock; Use 1 mL maximum</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Fentanyl patch, 25 mcg/h</td>
			<td>Apotex Corp.</td>
			<td>NDC 60505-7006-2</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Isoflurane</td>
			<td>Multiple vendors</td>
			<td style="width:119px;">Catalog number not available</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">&#160;Viral vector</td>
			<td></td>
			<td style="width:119px;"></td>
			<td style="width:207px;">Gene transfer surgery only</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Surgical Instruments</td>
			<td></td>
			<td style="width:119px;"></td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Metzenbaum needle holder 7&#34; straight</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7900</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Operating scissors 6.5&#34; straight blunt/blunt</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-6828</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Needle holder /w suture scissors</td>
			<td>Miltex</td>
			<td style="width:119px;">8-14-IMC</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Castroviejo scissors</td>
			<td>Roboz</td>
			<td>RS-5658</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Castroviejo needle holder, 5.75&#34; straight with lock</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-6412</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Stevens scissors 4.25&#34; curved blunt/blunt</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-5943</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Alm retractor 4&#34; 4X4 5mm blunt prongs</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-6514</td>
			<td style="width:207px;">2x</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Backhaus towel clamp 3.5&#34;</td>
			<td>Roboz</td>
			<td style="width:119px;"></td>
			<td style="width:207px;">4x</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Micro clip setting forceps 4.75&#34;</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-6496</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Micro vascular clips, 11 mm</td>
			<td>Roboz</td>
			<td style="width:119px;"></td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Surg-I-Loop</td>
			<td>Scanlan International</td>
			<td style="width:119px;">1001-81M</td>
			<td style="width:207px;">5 cm length</td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:215px;">Bonaccolto forceps, 4&#8221; (10 cm) long longitudinal serrations, cross serrated tip, 1.2mm tip width</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-5210</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Dumont #3 forceps Inox tip size .17 &#215; .10 mm</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-5042</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Graefe forceps, 4&#8221; (10 cm) long serrated straight, 0.8 mm tip</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-5280</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Halstead mosquito forceps,&#160; 5&#34; straight, 1.3 mm tips</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7110</td>
			<td style="width:207px;">2x</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Halstead mosquito forceps,&#160; 5&#34; curved, 1.3mm tips</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7111</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:215px;">Jacobson mosquito forceps 5&#34; curved extra delicate, 0.9 mm tips</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7117</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Kantrowitz forceps, 7.25&#34; 90 degree delicate, 1.7 mm tips</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7305</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Tissue forceps 5&#34;, 1X2 teeth, 2 mm tip width</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-8162</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Allis-Baby forceps, 12 cm, 4x5 teeth, 3 mm tip width</td>
			<td>Fine Science Tools</td>
			<td style="width:119px;">11092-12</td>
			<td style="width:207px;">2x</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Adson forceps, 12 cm, serrated, straight</td>
			<td>Fine Science Tools</td>
			<td style="width:119px;">11006-12</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Veterinary electrosurgery handpiece and electrode</td>
			<td>MACAN Manufacturing</td>
			<td style="width:119px;">HPAC-1; R-F11</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Surgical Suite Equipment</td>
			<td></td>
			<td style="width:119px;"></td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Circulating warm water blanket and pump</td>
			<td>Multiple vendors</td>
			<td style="width:119px;">Catalog number not available</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Bair hugger warming unit</td>
			<td>3M</td>
			<td style="width:119px;">Model 505</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">IV infusion pump</td>
			<td>Heska</td>
			<td style="width:119px;">Vet IV 2.2</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Isoflurane vaporizer and scavenger</td>
			<td>Multiple vendors</td>
			<td style="width:119px;">Catalog number not available</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:215px;">Veterinary multi-parameter monitor</td>
			<td>Surgivet</td>
			<td style="width:119px;">Surgivet Advisor</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Veterinary electrosurgery unit</td>
			<td>MACAN Manufacturing</td>
			<td style="width:119px;">MV-9</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:215px;">Surgical microscope</td>
			<td style="width:167px;">D.F. Vasconcellos</td>
			<td style="width:119px;">M900</td>
			<td style="width:207px;">25X magnification for vein graft surgery; 16X magnification for gene transfer surgery</td>
		</tr>
	</tbody>
</table>

a rabbit model of durable transgene expression in jugular vein to common carotid artery interposition grafts

静脈グラフト バイパス手術は、閉塞性動脈疾患の一般的な治療法ただし、長期的な成功は、血栓症、肥厚、動脈硬化など血管障害によって制限されます。この記事の目標は、ウサギの二国間静脈移植を配置し、遺伝子発現の耐久性を実現する遺伝子の転送ベクトルと移植を伝達する方法を示すことです。メソッドは、通常静脈グラフトの恒常性の遺伝子とその蛋白質製品の生物学的役割の調査をことができます。例えば静脈グラフト障害を防ぐことができる活動のための遺伝子のテストを実行できます、transgene の式に新生内膜増殖を防ぐかどうかは、血管の炎症を軽減メソッドまたは家兎動脈硬化を軽減供給。高脂肪食。初期生残手術中に右と左の外頚静脈のセグメントは摘出、逆側の端に一般的な頚動脈移植として両側に配置されます。28 日後を実行、2 番目の生存手術中にグラフトのそれぞれは血管クリップで循環から隔離され、ルーメンがヘルパー依存 (HDAd) アデノウイルスベクターを含むソリューション (すなわち) 経由で満ちています。20 分インキュベーション後ベクトル ソリューションを吸気、切開を修理し、流れを復元します。静脈は、個々 の実験的プロトコルによって決まります時点で収穫されます。移植配置と情報伝達系の 28 日間の遅延は動脈循環の静脈グラフトの適応を確保するため必要です。この適応は、静脈グラフト導入前に、または直後移植で発生する遺伝子発現の急速な損失を回避できます。メソッドは、耐久性、安定した遺伝子発現によりグラフト静脈を達成するためにその能力でユニークです。ウサギ他動物大静脈血管モデルと比較して、低コストと簡単な処理の利点があります。ウサギ齧歯動物の静脈グラフトのモデルと比較して、解析のための豊富な組織を提供する大きく、操作が簡単の血管があります。

オペレーターは、実験データを生成するこのメソッドを使用できます前に、new 演算子の技術的な習熟度を検証する必要があります。New 演算子を実現する必要があります最初のマイルス トーンは、一貫性のある静脈グラフトの開存を初回血管移植手術とその後の遅延伝達手術後です。以上手術のそれぞれが望ましいと達成した後 90% 開存性。開存は、我々 は通常術後 ...

Watch this Scientific Journal Video about A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts at JoVE.com

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

耐久性のある遺伝子発現により一般的な頚動脈移植に頸静脈の家兎モデル

このメソッドでは、ウサギの介在静脈グラフトの配置、移植の伝達と遺伝子発現の耐久性の達成をについて説明します。これは遺伝子とグラフト静脈でタンパク質製品の生理学的および病理学的役割と静脈グラフト病に対する遺伝子治療のテストの調査をことができます。

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

Research

JoVE Journal

Medicine

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

ラットの総頚動脈で外頚静脈グラフトの介在：洗練された顕微鏡下手術の訓練

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

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

局所脳虚血への早期虚血耐性を誘導するために適切な前処理刺激として二国間の総頚動脈閉塞

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

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

成体マウス静脈性高血圧モデル：外頸静脈吻合部の総頸動脈。

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

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

配慮ラットモデルにおける血行動態の措置、煎茶や血液サンプリングを実行するために頸動脈と頸静脈のカテーテル

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

マウスにおけるタキサンの薬物動態学的および薬力学的分析のための頸動脈煎茶

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

持続的な障害と皮質虚血を研究するために高齢ラットにおける一般的な頸動脈閉塞と永久遠位中大脳の実行

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

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

静脈介在モデル：バイパスグラフト開存を研究するための適切なモデル

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

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

生体内でウサギの一般的な頚動脈内皮細胞への遺伝子導入

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

ラットの総頸動脈で位相コントラスト磁気共鳴イメージング

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

塩化第二鉄によるイヌ総頚動脈血栓症: 血管損傷の大動物モデル

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...