This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe&#8217;s group at Northwestern University for providing helpful information.

注：倫理に関する声明：すべての手順は、実験動物の管理と使用に関する指針（全米アカデミープレス、第8版、2011）、およびウィスコンシン大学の施設内動物管理使用委員会によって承認されたプロトコルに準拠しています。すべての手術は（速度2ml /分流量、吸入を介して）、イソフルラン麻酔下で実施し、全ての努力が苦痛を最小限にするために行われた。動物を、徐々にCO 2で満たされたチャンバー内で安楽死させた。 全身麻酔と皮膚切開の1の誘導<ol><li> 23〜28グラムの12〜16週の年齢と体重と雄のC57BL / 6マウスを使用してください。閉回路の麻酔​​装置を通過イソフルランにより動物を麻酔。そのつま先をつまんで、適切な麻酔を確認してください。乾燥を防ぐために、目にいくつかの人工涙液軟膏を適用します。 </li><li>少し拉致左後肢膝関節のSLと仰臥位でマウスを置きますightly撓む。左大腿上部領域の周囲の毛を剃るとクロルヘキシジンで皮膚を消毒、滅菌した綿先端アプリケーターを用いて3回を適用した。手術中に滅菌楽器を保つためにホットビーズ滅菌器を使用してください。 </li><li>左足の上に滅菌ガーゼドレープを配置します。鼠径部（ 図1A）に膝関節からの左大腿内側に1.5センチメートルストレート、縦皮膚切開を行います。 </li></ol>大腿動脈と枝の2暴露メモ： 図1（b）は、大腿上部における総血管系の解剖学を示している。皮膚切開が適切に大腿内側の正中線に沿って行われたとき浅大腿動脈（SFA）は、解剖顕微鏡で観察することができる。 <ol><li>細かいピンセットでSFAの上にある薄い筋膜を開き、9-0ナイロン縫合糸とループSFA。 注：このプロセスでは、大腿神経が容易ように置かれたその壁は、それが簡単に穿孔と出血していることを非常に薄いので、IDEが、隣接する大腿静脈は細心の注意が必要です。キー技術は動脈自体に集中することです。限り切開が近い動脈に行われるように、任意の偶発的静脈の損傷が回避される。 </li><li> SFAがループされると、モスキート鉗子で縫合糸を保持し、やや下向きに引き出します。この操縦は簡単にさらに解剖になりますが、あまりにも強制的に引っ張らないでください動脈を損傷しないように。 </li><li> 9-0ナイロン縫合糸で総大腿動脈（CFA）を露出し、ループするように近位に動脈を解剖続行。 注：通常、上腹部動脈（ 図1B）に遠位の点でCFAのループは、電線を挿入するための十分です。同じようにモスキート鉗子でCFAの縫合糸ループを保持します。 </li><li>上向きのCFAの縫合糸ループを引いて、SFAの縫合糸ループを下方にSFAと深い大腿動脈（DFA）の分岐を明らかに。 DFA、CFAの主要な枝、大腿筋（ 図1B）に向かう。 注：ワイヤーは後でDFAから挿入されます。 </li><li>余分な慎重にループDFA隣接静脈を動脈の下に絡まっているため。静脈からの出血の場合には、ガーゼで30〜60秒程度の単純な圧縮は、止血をもたらすのに通常は十分である。 </li><li> DFAは ​​、9-0ナイロン縫合糸でループされると、すべての3つの縫合糸ループは、（ 図2A）が正しい位置にあることを確認してください。 </li><li>遠位に可能な限り分岐からポイントで動脈を結紮するDFAの解剖を続行します。余分な縫合糸をカットしますが、両端がDFAは、ワイヤの挿入を作るより簡単に安定させるモスキート鉗子で保持できるように一定の長さにしておきます。 </li></ol> 3.ワイヤー傷害を作成する<ol><li> DFAを経由して電線を挿入<ol><li>一時的な止血を確立するために、CFA、SFA、およびDFAの縫合糸をひずみ。 </li><li> DFに小さな穴を作るmicroscissorsとA。 0.015インチ径のワイヤーのために十分な大きさの穴を拡張するために湾曲したmicrotweezersの先端を挿入します（ 図2Bは、動脈とワイヤとの間のサイズの違いを示しています）。 <ol><li> DFAに穴を行う前に、CFAの縫合糸ループを緩め、静かにクランプ/鈍い先端がmicrotweezersでDFA数回絞る。この操縦ワイヤの挿入を容易にするために著しく動脈の直径を拡大する。 PBSで徹底的にワイヤーを加湿することもスムーズに挿入するために重要である。 </li><li> DFAに穴を作るときは、動脈壁をトリミングが、小さなV字型のニップを作成し、操作するための鋭い先端がmicrotweezersとフラップを挟まないでください。 </li></ol></li><li> DFAに適切に-拡張穴を通してワイヤーを挿入します。 CFAの周りに縫合糸ループを緩めながらゆっくりと腸骨動脈に向かって線を進める。 5〜8ミリメートルはほとんどの場合、すべてのCFAの長さをカバーするために十分である（ 図2C </strong&gt;）。動脈穿孔は通常、マウスの損失をもたらすように強制的にワイヤーを挿入しないでください。長さ約15ミリにワイヤを短くする処理に役立ちます。 </li></ol></li><li> 1分間の中にワイヤーを残し<ol><li>最適な長さにワイヤーを挿入した後、動脈壁を伸び過ぎ1分間放置する。乾燥を防ぐためにPBSワイヤおよび動脈を湿らせる。別の9-0ナイロン縫合糸は、この待ち時間の間にワイヤ挿入ポイントにDFA近位の周りに配置することができる。この縫合糸は、ワイヤの除去後DFAに穴を閉じるために後で使用される。 </li></ol></li><li>ワイヤーを取得し、血流を再開<ol><li>血管カニュレーション鉗子で線を持ち、ゆっくりと引き戻す。ワイヤーがスムーズに取得されるべきである。通常動脈壁へのワイヤの付着によって生じるいかなる抵抗を克服するために、穏やかに回転してワイヤーを引き戻すと便利です。ここでも、力強い操縦は、動物の損失につながる可能性があります。 </li><li> Aワイヤーを取り出すFTERは、9-0ナイロン縫合糸との分岐点でDFAを連結する。 SFAへの血液流出を危うく分岐、に近づきすぎ連結しないでください。 </li><li>正常な血流を再開するためにSFAとCFAの縫合糸ループを緩めます。 CFAは、ワイヤの挿入（ 図2D）で十分に展開されていることに注意してください。 </li></ol></li></ol> 4.皮膚閉鎖と術後ケア<ol><li>コントロールが出血。マイナーにじみ出るは、簡単な圧縮によって停止することができる。いくつかの主要な動脈採血を11-0ナイロン縫合糸でステッチが必要になります。これらの縫合動脈は、最終的には動脈閉塞になります。 </li><li>スキンステープラー（通常3-4ステープル）で皮膚創傷を閉じます。クロルヘキシジン外科ソリューションを適用することにより、滅菌創傷領域にしてください。 </li><li>術後の治療法を適用します。ブプレノルフィンの皮下注射でマウスの痛みを制御します。それはSUFを取り戻したまでは温暖化パッド上でマウスを保つ胸骨の横臥位を維持するために意識をficient。他の動物の会社のない清潔なケージ内のハウスには、完全に回復するまで。 </li></ol>

<ol>
	<li>Dzau, V. J., Braun-Dullaeus, R. C., Sedding, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+proliferation+and+atherosclerosis:+new+perspectives+and+therapeutic+strategies.">Vascular proliferation and atherosclerosis: new perspectives and therapeutic strategies.</a> Nat Med. 8 (11), 1249-1256 (2002).</li><li>Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+hyperplasia+in+murine+models.">Intimal hyperplasia in murine models.</a> Curr Drug Targets. 9 (3), 251-260 (2008).</li><li>Ferns, G. A., Avades, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanisms+of+coronary+restenosis:+insights+from+experimental+models.">The mechanisms of coronary restenosis: insights from experimental models.</a> Int J Exp Pathol. 81 (2), 63-88 (2000).</li><li>Sata, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+of+vascular+injury+that+induces+rapid+onset+of+medial+cell+apoptosis+followed+by+reproducible+neointimal+hyperplasia.">A mouse model of vascular injury that induces rapid onset of medial cell apoptosis followed by reproducible neointimal hyperplasia.</a> J Mol Cell Cardiol. 32 (11), 2097-2104 (2000).</li><li>Xu, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+arteriosclerosis:+from+arterial+injuries+to+vascular+grafts.">Mouse models of arteriosclerosis: from arterial injuries to vascular grafts.</a> Am J Pathol. 165 (1), 1-10 (2004).</li><li>Lv, L., Meng, Q., Ye, M., Wang, P., Xue, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=STAT4+deficiency+protects+against+neointima+formation+following+arterial+injury+in+mice.">STAT4 deficiency protects against neointima formation following arterial injury in mice.</a> J Mol Cell Cardiol. 74C, 284-294 (2014).</li><li>Zhu, B., Kuhel, D. G., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+E+inhibits+neointimal+hyperplasia+after+arterial+injury+in+mice.">Apolipoprotein E inhibits neointimal hyperplasia after arterial injury in mice.</a> Am J Pathol. 157 (6), 1839-1848 (2000).</li><li>Fu, Y., Duru, E. A., Davies, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+metabolic+syndrome+on+the+response+to+arterial+injury.">Effect of metabolic syndrome on the response to arterial injury.</a> J Surg Res. , (2014).</li><li>Kuhel, D. G., Zhu, B., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinction+in+genetic+determinants+for+injury-induced+neointimal+hyperplasia+and+diet-induced+atherosclerosis+in+inbred+mice.">Distinction in genetic determinants for injury-induced neointimal hyperplasia and diet-induced atherosclerosis in inbred mice.</a> Arterioscler Thromb Vasc Biol. 22 (6), 955-960 (2002).</li><li>Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differential+neointimal+response+in+vein+grafts+and+wire-injured+arteries.">Mouse strain differential neointimal response in vein grafts and wire-injured arteries.</a> Circ J. 71 (10), 1649-1652 (2007).</li><li>Ishigami, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficiency+of+CuZn+superoxide+dismutase+promotes+inflammation+and+alters+medial+structure+following+vascular+injury.">Deficiency of CuZn superoxide dismutase promotes inflammation and alters medial structure following vascular injury.</a> J Atheroscler Thromb. 18 (11), 1009-1017 (2011).</li></ol>

The authors declare no competing financial interests.

ワイヤ傷害手順は、我々は12-16週齢の雄C57BL / 6マウスを使用する限り、それらの解剖学的構造が類似しているように4本研究では、マウスのすべての株に適用可能である。私たちは私たちの予備的研究から学んだように、10週より若いマウスの大腿動脈は、多くの場合、線挿入は非常に困難であることが非常に小さい。一方、16週間以上経過したマウスにおける電線挿入は、技術的に容?...

In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata&#8217;s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.

<table border="0" cellpadding="0" cellspacing="0" width="644">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Reagent/ Equipment</td>
			<td style="width:101px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:208px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fixed Core Wire Guide</td>
			<td style="width:101px;">Cook</td>
			<td style="width:119px;">G02426</td>
			<td>Diameter 0.015 inch, Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dilation Forceps</td>
			<td style="width:101px;">Roboz</td>
			<td style="width:119px;">RS-4927</td>
			<td>Curved, blunt tips</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dumont Tweesers #5</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">14095</td>
			<td>Straight, sharp tips</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dumont Vessel Cannulation Forceps</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">503373</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">McPherson-Vannas Scissors</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">501234</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Mosquito Forceps</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">501291</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ethilon Nylon Suture 9-0</td>
			<td style="width:101px;">Ethicon</td>
			<td style="width:119px;">7717G</td>
			<td>9-0, Black Nylon Monofilament</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Micro AROSuture, Sterile 11-0, 70 Microns, MET Point</td>
			<td style="width:101px;">AROSurgical</td>
			<td style="width:119px;">VT4A00N07</td>
			<td>11-0, Black Nylon Monofilament</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3M Precise Multi-Shot DS Disposable Skin Stapler</td>
			<td style="width:101px;">3M</td>
			<td style="width:119px;">DS-25</td>
			<td></td>
		</tr>
	</tbody>
</table>

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

アテローム性動脈硬化症により引き起こされる心血管疾患は、先進国における死亡の主な原因である。動脈硬化性プラークの開発や確立プラークの破裂による血管内腔の狭窄は、このような心筋梗塞や脳卒中などの様々な病的状態につながる正常な血流を遮断する。診療所では、そのような血管形成術などの血管内の手順は、一般的に内腔を再度開くために行われている。しかし、これらの治療は、必然的に、過剰な治癒反応と血管狭窄（​​再狭窄）を引き起こすルーメン内に延びて、新生内膜プラークの発達を誘発する、血管壁、ならびに血管内皮を損傷する。再狭窄は、アテローム性動脈硬化症のための血管内治療の失敗の主要な原因である。したがって、再狭窄の前臨床動物モデルは、病態生理学的機序ならびに脈管介入翻訳アプローチを調査するために極めて重要である。いくつかのネズミexperimeうち、それは密接に内皮および血管壁の両方を傷つける血管形成術に似ているので、NTALモデルは、大腿動脈ワイヤ傷害は広く血管形成術後の再狭窄の研究のために最も適していると認められている。しかし、多くの研究者が、技術的難易度の高い程度に、このモデルを利用して困難である。金属ワイヤが顕著新生を誘導するのに十分な損傷を生成するために、ワイヤの約3倍薄い大腿動脈に挿入する必要があるため、これは主にある。ここでは、効果的に、このモデルの主要な技術的困難を克服するために不可欠の手術の詳細を説明します。 、提示された手順に従って実行することによって、マウスの大腿動脈ワイヤ傷害が容易になる。慣れた後、全体の手順は、20分以内に完了することができる。

手術後4週間、ワイヤー損傷大腿動脈を収穫し、新生内膜形成を分析するためにパラフィンブロック標本を作る。上記のように全身麻酔を誘導する。すぐに胸を開き、左心室に20ゲージの翼状針を挿入します。点滴ボトルにラインを接続します。 4％パラホルムアルデヒド（通常は10〜20ミリリットル、各）に続いて、PBSの点滴を経由して灌流固定を行います。動物は、ワイヤ損傷CFAを取?...

Watch this Scientific Journal Video about A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury at JoVE.com

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

再狭窄のマウス大腿動脈ワイヤ傷害モデルは、技術的に困難である。このプロトコルでは、我々は成功し、再狭窄の研究のための一貫性の新生内膜を誘導するために、ワイヤ傷害を実施するための不可欠なキー技術的な詳細を示している。

動脈再狭窄のマウスモデル：大腿ワイヤー傷害の技術的な側面

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

a-murine-model-arterial-restenosis-technical-aspects-femoral-wire

Research

JoVE Journal

Medicine

19.9K ビュー.  University of Wisconsin School of Medicine and Public Health. 再狭窄のマウス大腿動脈ワイヤ傷害モデルは、技術的に困難である。このプロトコルでは、我々は成功し、再狭窄の研究のための一貫性の新生内膜を誘導するために、ワイヤ傷害を実施するための不可欠なキー技術的な詳細を示している。 

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

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

ウサギモデルにおける顕微静脈ポーチ動脈 - 分岐瘤：技術的側面

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

医学

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

血液サンプリング、医薬品の管理と意識血圧および心拍数測定のための大腿動脈および静脈カテーテル

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

再狭窄の研究のための頸動脈内ステント留置のマウスモデル

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

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.

マウスAortocaval瘻の技術的な側面

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

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

くも膜下出血の急性および後期後遺症の研究のためのウサギの血液シャントモデル：技術的側面

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

マウスの簡単な重要なサイズの大腿欠陥モデル

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

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.

血管周囲ドラッグデリバリーパッチの移植に大腿動脈ワイヤ傷害のマウスモデル

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

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

腸管虚血 - 再灌流傷害のマウスモデル

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

マウスにおける加速アテローム性動脈硬化症の誘導:ワイヤー傷害モデル

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

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

最大急性非致死肺損傷における急性肺損傷の結果のオープン気管切開胃酸吸引マウスモデル

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...