この作品はMLIAに再生医療のイーライとエディスブロードセンターからの助成金によってサポートされており、AIMにASSとAIM、フィリップ・J. WhitcomeフェローシップにUCLAトレーニングプログラムで幹細胞研究、および国立衛生研究所（HL130290）しました。

注：このプロトコルは、カリフォルニア大学ロサンゼルス校で動物研究委員会によって承認されています。 1.術前の準備と麻酔<ol><li>作業中の滅菌技術を必ず守ってください。 </li><li>蒸気オートクレーブを使用して、すべての手術用消耗品を滅菌します。 </li><li>それは、外科手術中に可視化するために実体顕微鏡下で適切な温度（37℃）の場所に温めることができるように前に麻酔導入に加熱された齧歯類の外科のプラットフォームをオンにします。 </li><li>麻酔誘導チャンバ内にマウスを置き、1リットル/分の流速で4％イソフルランで誘導します。 <ol><li>慎重に誘導時のマウスの呼吸数およびフットパッド肌の色を監視します。 </li><li>呼吸速度の減速した後、誘導チャンバーからマウスを削除し、2.5％の独立したノーズコーンとメンテナンスイソフルラン濃度の顔をした外科手術の準備のために温めたパッドの上に置きます。 </li><li>麻酔薬の妥当性を評価するためにつま先のピンチを実行します。 </li></ol></li><li>角膜に眼軟膏を適用し、（5ミリグラム/ kg）を皮下術前カルプロフェンを投与します。 </li><li>腹部から毛を除去するために、マウス仰臥位および使用バリカンを置きます。 </li><li>滅菌ガーゼパッドで適用される3ポビドンヨードの交互スクラブと70％イソプロピルアルコールで手術領域を準備します。 </li><li>ノーズコーン内の顔をして加熱齧歯類の外科のプラットフォームの上に置いてマウスの仰臥位とすべての四肢を固定し、慎重に腹部を実体顕微鏡で見えるようにマウスを置きます。 </li><li>準備された手術領域の各辺に沿って、滅菌、接着剤包帯を置きます。 </li><li>自発呼吸を維持しながら、手術中に十分な麻酔を維持するために、イソフルラン濃度を滴定します。 </li></ol> 2.腎臓下の大動脈クランプ<ol><li> approximatelを開始、メスを用いて腹部の正中線の下3センチメートル切開を作ります剣状突起に劣るyの0.5センチメートル。 </li><li>優しく鉗子で皮膚を撤回し、微細な結合組織を切断するために微細なハサミを使用して腹壁から肌を分析。 </li><li>筋肉壁に0.5％ブピバカイン0.05〜0.1 mlで適用し、腹部臓器を露出するために腹壁に2cmの切開を行います。 <ol><li>出血が腹壁に沿って発生した場合は、綿棒で穏やかな圧力を適用します。 </li></ol></li><li>静かに生理食塩水に浸した綿棒のアプリケーターを使用して腸を持ち上げます。 注：腹腔外に暖かい、滅菌生理食塩水に浸したガーゼスポンジで空腸と回腸動脈と場所への鈍的外傷を避けるように注意してください。 <ol><li>水分損失を回避するために別の暖かい、滅菌生理食塩水に浸したガーゼスポンジで腸をカバーしています。 </li></ol></li><li>直腸をlateralizeし、後腹膜を露出させるために開創器を置きます。 </li><li> retroperitoneの可視化のため、必要に応じて小さなガーゼパッドを配置umの使用数を追跡します。 </li><li>右腎の下極のレベルでは、大動脈への横retroperitonotomyを作るために、鋭い解剖鉗子を使用しています。 注：下大静脈や周囲の血管を傷つけないように注意してください。 <ol><li>ぶっきらぼうに大動脈から後腹膜組織を解剖 下大静脈や周囲の血管系を穿孔しないように注意してください。 </li></ol></li><li>少なくとも1分間、大動脈、または他の指定された時間をかけて血管クランプを配置し、視覚的に遠位大動脈における拍動性の欠如を観察することにより閉塞を確認します。 </li><li>血管クランプを外し、止血を確認します。止血を達成し、血液のアクティブな血管外漏出が見られないときに確保されています。 <ol><li>腹腔内に0.5ミリリットルの生理食塩水を添加し、生理食塩水がますます血を帯びたになると評価することによって、血管外漏出を確認します。このような場合は、保証するために、1分間生理食塩水に浸したアプリケーターを用いて穏やかな圧力をかけます止血。 </li></ol></li><li>後腹膜の可視化に役立つと腹部にその場で腸を置き換えるために置かれたガーゼパッドを取り外します。 </li><li>予め温めた滅菌生理食塩水を使用して腹腔を灌漑。 </li></ol>開腹し、皮膚の3閉鎖<ol><li> 5-0編組、吸収性単ランニング縫合糸を使用して腹壁筋層を閉じます。 </li><li> 1-3ポリマー接着剤の滴、その後創傷クリップを有する接着剤がセットされた後との緊密な肌。 </li></ol> 4.回復と術後評価<ol><li>ケージの床に食料と水を加熱パッド上で回復ケージにマウスを転送します。 </li><li>密接に呼吸困難の徴候についてマウスを監視します。機関のガイドラインあたり術後48時間毎日カルプロフェン（5 mgの/キログラム）を管理します。 <ol><li>それは胸骨recumを維持するのに十分な意識を取り戻したまで無人の動物を放置しないでくださいbency。 </li></ol></li><li>食料と水と通常のケージにマウスを返します。 <ol><li>完全に回復するまで、他の動物の会社に手術を受けた動物を返さないでください。 </li></ol></li><li>裂開のための日常的に傷を評価し、術後14日目に、クリップを取り外してください。 </li></ol> 5.大動脈解離と染色<ol><li>裸出損傷後の関心のある特定の時点に基づいて、犠牲のためのマウスを選択します。 </li><li>イソフルラン吸入によりマウスを屠殺し、直ちに血管平滑筋弛緩を引き起こすことは5mgのメタコリン塩化注入。 </li><li>呼吸停止、角膜反射および運動の欠如の欠如により死亡が確認されると、10分、左心室を経由して100 mmHgでの灌流圧でリン酸緩衝生理食塩水で4％パラホルムアルデヒドで灌流。 </li><li>注意深くから無傷の腹部大動脈を分離するために解剖実体顕微鏡を使用して、周囲の組織。 </li><li>腸骨分岐に腎動脈と尾側に大動脈吻側を横断し、背側表面に沿って縦切開を介して開かれました。 </li><li>無未満2時間までの固定のための管腔側と35ミリメートルシリコーンコートディッシュ以上は必要ない12時間に平らに大動脈をピン。その後、組織はどちらの切片用に包埋または顔全体マウント免疫細胞化学専用に使用することができます。 </li><li>共焦点顕微鏡用ガラススライド上のカバーガラスに面した腔側で染色した大動脈をマウントします。 </li></ol>

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著者らは、開示することは何もありません。

このようなバルーン血管形成術および血管ステント留置などの経皮的介入への動脈削剥損傷は、初期および後期の血管血栓症および再狭窄を生じ、再発性虚血性イベント3,12に貢献しています。興味深いことに、外科的血管クランプはまた、13経皮的またはオープンかどうか、任意の血管手術を受ける患者に問題の範囲を拡大し、動脈侵食の原因として関与しています。減損再...

血栓症および再狭窄は、血管内バルーン血管形成術及び1,2-ステント留置などの経皮的血管介入を受ける患者に深刻な初期および後期合併症です。いくつかの戦略は、特に、二重抗血小板療法と薬物溶出ステントをこれらの合併症に対処するために使用されています。しかし、少しの焦点は、血栓症および再狭窄、内皮細胞のカバレッジ（削剥）の、すなわち損失の根本的な原因に置かれています。裸出傷害は、血管壁への機械的外傷による介入手順の必然的な結果です。この機械的外傷、血液3循環の損傷および基底膜および血管平滑筋の保護内皮層と露光の除去をもたらすことができます。これらの分野での内皮細胞の損失は、血小板接着およびその後の血栓症を促進するだけでなく、プロ血栓性および前炎症性環境を作成しますLSOは、新生内膜肥厚及び再狭窄4で得られた血管平滑筋細胞の遊走および増殖を刺激します。これらの合併症、およびそれらに関連する治療法は、人間の健康に影響を与える重要な罹患率、最も顕著な再発虚血性疾患や出血事象につながります。 再内皮創傷縁から剥皮損傷の血栓症および再狭窄5を予防する上で極めて重要です。ステントのカバレッジが6,7支柱と剖検結果および動物モデルは、効果的に血栓症の縮小率を示しました。薬剤溶出ステントは、動脈の再内皮化および後期血栓3の増加率に有意な障害をもたらし、平滑筋の増殖および新生内膜過形成を抑制することにより再狭窄率を減少させるために考案。残念ながら、再内皮ためのメカニズムを理解することは、主にAPの不足によって制限され、ゆっくりとしたプロセスとなっていますpropriate動物モデル8。 動脈損傷後の内皮細胞および血管平滑筋細胞の役割を理解するためのいくつかの動物モデルは、7,9,10が作成されています。ラット頸動脈バルーン傷害モデルは、最高の特徴であると、細胞および分子レベル11、肉眼で裸出傷害の影響を研究するために使用されています。それにもかかわらず、優れた生存率と動脈裸出損傷の再現性の高いマウスモデルは、不足しているとはるかに優れた複数の設定の血管再生を解明するために利用可能な複数のトランスジェニック系統を利用するために必要とされています。 この原稿は、再現性があり、実行するのは簡単である動脈裸出損傷のマウスモデルを提示します。アプローチは、いくつかのトランスジェニック系統において最小の罹患率および死亡率を示しました。なぜなら遺伝的に改変されたマウス系統の広範な数の、このモデルに使用することができます削剥傷害後の再内皮化の根底にある分子メカニズムを解明。

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			<td height="20" style="height:20px;width:289px;">Adjustable Wire Retractor</td>
			<td style="width:187px;">Fine Science Tools</td>
			<td style="width:119px;">17004-05</td>
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			<td height="20" style="height:20px;">Schwartz Micro Serrefines - Sharp Bend</td>
			<td>Fine&#160;Science Tools</td>
			<td>18052-03</td>
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			<td height="20" style="height:20px;">Surgical Instruments</td>
			<td>Fine Science Tools</td>
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			<td>sharp dissecting forceps, blunt forceps, fine scissors, spring scissors, hemostat</td>
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			<td height="20" style="height:20px;width:289px;">0.5% Marcaine</td>
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			<td height="20" style="height:20px;width:289px;">5-0 Suture, Vicryl</td>
			<td style="width:187px;">Fisher Scientific</td>
			<td style="width:119px;">NC0189890</td>
			<td>tapered needle</td>
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			<td height="20" style="height:20px;width:289px;">Vetbond</td>
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			<td style="width:119px;">NC0304169</td>
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		<tr height="40">
			<td height="40" style="height:40px;width:289px;">Falcon&#174; 35 mm Not TC-Treated Easy-Grip Style Bacteriological Petri Dish</td>
			<td style="width:187px;">Corning</td>
			<td align="right">351008</td>
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			<td height="20" style="height:20px;width:289px;">Sylgard 184 Silicone Elastomer Kit</td>
			<td style="width:187px;">Dow Corning</td>
			<td style="width:119px;">3097358-1004</td>
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			<td height="20" style="height:20px;width:289px;">Dissecting pins</td>
			<td style="width:187px;">Fisher Scientific</td>
			<td>NC9681411</td>
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reproducible arterial denudation injury by infrarenal abdominal aortic clamping in a murine model

経皮的血管インターベンションを均一に続いて血栓症および再狭窄につながる動脈削剥傷害につながります。これらの合併症は、創傷縁内に再内皮に障害に起因することができます。しかし、再内皮の細胞および分子メカニズムが定義されていません。動脈裸出後の再内皮化を研究するためのいくつかの動物モデルが利用可能であるが、いくつかはあるため、外科的制限のマウスで行われています。これは、トランスジェニックマウス系統を利用し、再内皮の処理に特定の遺伝子の寄与を調査する機会を損ないます。ここでは、外部血管クランプを使用して、下腹部大動脈に動脈裸出損傷の再現性の高いマウスモデルを作成するためのステップバイステップのプロトコルを提示します。フィブリノゲンおよびβカテニンのため負傷した大動脈の免疫細胞化学染色はプロ血栓性表面ANの暴露を実証しますそれぞれ、無傷の内皮の境界線をdは。ここに提示された方法は、トランスジェニックマウスモデルにおける動脈裸出傷害を課すための一意の実用的なツールを作成する、速度、優れた全体的な生存率、および相対技術的容易さの利点を有します。この方法を使用して、研究者は、正常または病的な条件下での再内皮化のメカニズムを解明することができます。

八十-5マウスは、下腹部大動脈クランプのために、このレポートに記載され生存手術法を受けています。全体的な生存率は85.9％でした。手術合併症は、腸の出血および大血管穿孔、5.9％および3.5％の死亡率が得られた（ 表1）に含まれます。麻酔から回復した後、マウスは正常に歩くと下肢に虚血損傷の兆候を示しません。体重減少または食欲の欠如は認...

Watch this Scientific Journal Video about Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model at JoVE.com

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

動脈裸出損傷後の再内皮の細胞および分子メカニズムを理解することは、動脈の血栓症および再狭窄を予防する上で極めて重要です。ここでは、下腹部大動脈の再現性動脈裸出損傷のためのプロトコルについて説明します。手順は、マウスモデルを使用して、内皮の再生を制御する機序を研究するために開発されました。

マウスモデルにおける下腹部大動脈クランプすることにより、再現性動脈侵食に起因けが

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

reproducible-arterial-denudation-injury-infrarenal-abdominal-aortic

Research

JoVE Journal

Medicine

9.2K ビュー.  University of California, Los Angeles. 動脈裸出損傷後の再内皮の細胞および分子メカニズムを理解することは、動脈の血栓症および再狭窄を予防する上で極めて重要です。ここでは、下腹部大動脈の再現性動脈裸出損傷のためのプロトコルについて説明します。手順は、マウスモデルを使用して、内皮の再生を制御する機序を研究するために開発されました。 

ビデオ: Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

A New Murine Model of Endovascular Aortic Aneurysm Repair

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

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

血管内大動脈瘤の修復の新しいマウスモデル

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

医学

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

脊髄虚血再灌流のマウスモデルにおいて胸部大動脈閉塞によって再現可能対麻痺

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

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

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

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 ...

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, ...

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 ...

Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic aneurysms (AAAs). This article introduces and describes a standard protocol used to induce AAAs through adventitial VV hypoperfusion created with a combination of polyurethane catheter insertion into the aortic lumen and suture ligation of the infrarenal abdominal aorta.
The protocol involves the use of male rats weighing 300-400 g, which are provided food and water ad libitum. After laparotomy with a ventral midline abdominal incision, exfoliation of the aorta is performed, which blocks blood flow from the perivascular tissue. Aortotomy involving a small incision adjacent to the renal artery branches is performed, and a polyurethane catheter is inserted using an 18-gauge indwelling needle. After repairing the incision, tight ligation of the aorta over the catheter blocks VV blood flow from the proximal direction through the aortic wall without disturbing the aortic blood flow. This technique can induce an AAA with progressive aortic dilatation.
The greatest benefit of this model is that VV hypoperfusion causes tissue hypoxia and the development of an infrarenal AAA, which has morphological and pathological characteristics similar to those of a human AAA.

Polyurethane catheter insertion into the aortic lumen and suture ligation of the aorta induce chronic hypoxia due to hypoperfusion of the adventitial vasa vasorum. This article describes a novel animal model of abdominal aortic aneurysm (AAA) with characteristics similar to those of AAA in humans.

外ヴァーサ栄養血管の血流をブロックすることにより腹部の大動脈瘤の齧歯動物モデルの作成

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic ...

Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing and treating this complication using experimental models of spinal cord ischemia. The model described here demonstrates varying degrees of paraplegia that relate to the length of occlusion following thoracic aortic occlusion in a rat spinal cord ischemia model.
A 2-Fr. balloon-tipped catheter was advanced through the femoral artery into the descending thoracic aorta until the catheter tip was placed at the left subclavian artery in anesthetized male Sprague-Dawley rats. Spinal cord ischemia was induced by inflating the catheter balloon. After a set period of occlusion (9, 10, or 11 min), the balloon was deflated. Neurologic assessment was performed using the motor deficit index at 24 h after surgery, and the spinal cord was harvested for histopathological examination.
Rats that underwent 9 min of aortic occlusion showed mild and reversible motor impairment in the hind limb. Rats subjected to 10 min of aortic occlusion presented with moderate but reversible motor impairment. Rats subjected to 11 min of aortic occlusion displayed complete and persistent paralysis. The motor neurons in the spinal cord sections were more preserved in rats subjected to shorter duration of aortic occlusion.
Researchers can achieve a reproducible hind limb motor deficit following thoracic aortic occlusion using this spinal cord ischemia model.

This study demonstrates the technique to make a minimally invasive and easily reproducible model of spinal cord ischemia in rats. Various degrees of hind limb motor deficit can be produced by controlling the aortic occlusion time.

脊髄虚血ラットモデルにおける大動脈閉塞後の再現性のある運動障害

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

Intravital Microscopy of Monocyte Homing and Tumor-Related Angiogenesis in a Murine Model of Peripheral Arterial Disease

The therapeutic goal for peripheral arterial disease and ischemic heart disease is to increase blood flow to ischemic areas caused by hemodynamic stenosis. Vascular surgery is a viable option in selected cases, but for patients without indications for surgery such as progression to rest pain, critical limb ischemia, or major disruptions to life or work, there are few possibilities for mitigating their disease. Cell therapy via monocyte-enhanced perfusion through the stimulation of collateral formation is one of a few non-invasive options.
Our group examines arteriogenesis after monocyte transplantation into mice using the hindlimb ischemia model. Previously, we have demonstrated improvement in hindlimb perfusion using tetanus-stimulated syngeneic monocyte transplantation. In addition to the effects on the collateral formation, tumor growth could be affected by this therapy as well. To investigate these effects, we use a basement membrane-like matrix mouse model by injecting the extracellular matrix of the Engelbreth-Holm-Swarm sarcoma into the flank of the mouse, after occlusion of the femoral artery.
After the artificial tumor studies, we use intravital microscopy to study in vivo tumor-angiogenesis and monocyte homing within collateral arteries. Previous studies have described the histological examination of animal models, which presupposes subsequent analysis to post-mortem artifacts. Our approach visualizes monocyte homing to areas of collateralization in real time sequences, is easy to perform, and investigates the process of arteriogenesis and tumor angiogenesis in vivo.

Monocytes are important mediators of arteriogenesis in the context of peripheral arterial disease. Using a basement membrane-like matrix and intravital microscopy, this protocol investigates monocyte homing and tumor-related angiogenesis after monocyte injection in the femoral artery ligation murine model.

単球のホーミングと末梢動脈疾患のマウスモデルにおける腫瘍関連血管新生の生体顕微鏡

The therapeutic goal for peripheral arterial disease and ischemic heart disease is to increase blood flow to ischemic areas caused by hemodynamic ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

改変非縫合カフ技術を用いたマウス子宮頸動脈大動脈移植モデル

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...

Porcine Model of Infrarenal Abdominal Aortic Aneurysm

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal abdominal aortic aneurysms (AAA) in swine. To achieve this, we use a combination of balloon angioplasty, elastase and collagenase, and a lysyl oxidase inhibitor, called &#946;-aminopropionitrile (BAPN), to create clinically significant infrarenal aortic aneurysms, analogous to human disease.
Noncastrated male swine are fed BAPN for 7 days prior to surgery to achieve a steady state in the blood. A midline laparotomy is performed and the infrarenal aorta is circumferentially dissected. An initial measurement is recorded prior to aneurysm induction with a combination of balloon angioplasty, elastase (500 units)/collagenase (8000 units) perfusion, and topical elastase application. Swine are fed BAPN daily until terminal procedure on either postoperative day 7, 14, or 28, at which time the aneurysm is measured, and tissue procured. BAPN + surgery pigs are compared to pigs that underwent surgery alone.
Swine treated with BAPN and surgery had a mean aortic dilation of 89.9% &#177; 47.4% at day 7, 105.4% &#177; 58.1% at day 14, and 113.5% &#177; 30.2% at day 28. Pigs treated with surgery alone had significantly smaller aneurysms compared to BAPN + surgery animals at day 28 (p &#60; 0.0003). The BAPN + surgery group had macroscopic and immunohistochemical evidence of end stage aneurysmal disease.
Clinically significant infrarenal AAA can be induced using balloon angioplasty, elastase/collagenase perfusion and topical application, supplemented with oral BAPN. This model creates large, clinically significant AAA with hallmarks of human disease. This has important implications for the elucidation of AAA pathogenesis and testing of novel therapies and devices for the treatment of AAA. Limitations of the model include variation in BAPN ingested by swine, quality of elastase perfusion, and cost of BAPN.&#160;

This novel model creates robust infrarenal abdominal aortic aneurysms in swine using a combination of balloon angioplasty, elastase/collagenase perfusion, topical elastase application, and oral compound &#946;-aminopropionitrile administration, which interferes with collagen cross-linking.

尿毛腹部大動脈瘤の豚モデル

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal ...