この作品は、CKBのNIH（RO1 HL098228）からの助成金によって部分的にサポートされていました。

注：すべての動物の手続きは、全国の小児病院施設内動物管理使用委員会によって承認された。 1。グラフトづくり<ol><li>ヒュームフードの下に2ミリリットルのジオキサンに100mgのPは（LA / CL）を追加することにより、ε-カプロラクトンとL-ラクチド共重合体P（LA / CL）の溶液を作製。渦上に溶液を置き、完全に溶解する1〜1.5時間、連続して混ぜる。 </li><li>一方、ポリグリコール酸（PGA）のシートを削除冷凍庫から感じた、いくつかの5×8ミリメートルの部分を切り取る。また、単にフィルター上0.1〜10μlのピペットの先端を切り落とした。 </li><li>ピペットチップの先端部に19gの針（1.5長さ）を挿入し、PGAはマイクロピンセットを用いて、針の周りに感じ包む。 </li><li> 19 Gの針をその中に挿入されている間、慎重に、内腔が鈍い18 Gの針を使用して、近位部よりも直線的であるピペットチップの先端にフェルトを押してください。 </li><li>ピペット上からピペットチップに40μlのP（CL / LA）ソリューションを提供します。 PGAは、溶液と感じ飽和する。次いで、ピペットディスペンサーを用いて気泡を押し出す。 必要な場合には、このプロセスを繰り返す。 </li><li>場所の50ミリリットルチューブ内移植片および20分間-80℃の冷凍庫に入れてください。針の頭は下向きにしてください。 </li><li> 24時間凍結乾燥し、真空中にチューブを移す。空気の流れを持っているチューブの蓋を開けていることを確認してください。 </li><li>移植片を取り出し、針から削除します。 〜5ミリメ​​ートルのセクションを残して、移植片の両端をカットし、その形状を維持するために、針の上に戻ってそれらを置く。デシケーター中での移植片を保管してください。 </li><li>バイオセーフティーフードのO / Nの前に細胞播種のUV光の下で足場を置きます。 </li></ol> 2。骨髄単核細胞の回収および単離<ol><li>ケタミン/キシラジンの過剰摂取（ケタミン、200 mg / kgのキシラジンと、20 mg / kg）を持つマウスを安楽死させる。 </li><li>骨（大腿骨をAN削除する10 CCのRPMIでペトリ皿中10移植注入や場所のための3匹のマウスからDの脛骨）。 3のCC RPMIで新しいペトリ皿に25 G針で注射器を用いて骨とフラッシュ骨髄の両端をカットします。 15 CC管内に骨髄およびRPMI液を収集し、BMの残りの部分を収集するために、追加の2のCC RPMIでペトリ皿を洗う。 </li><li>サンプル（5〜10μl）を取り、 自動細胞カウンターまたは血球計数器を用いて細胞を数える。その結果を記録します。 </li><li> 15 CC遠心管に5のCCフィコールを入れて、骨髄およびRPMIソリューションを追加します。フィコールと混合するのを防ぐために非常に穏やかに溶液を入れる。 </li><li> 24℃で「ブレーキなし」と30分間528×gで遠心分離</li><li>上側のピンクの層を除去する。 MNC層であるミドルクリア層の図1を収集し、PBS 1時01分でそれを希釈する。 </li><li>遠心分離機24℃で10分間528×gで希薄MNCソリューション</li><li>上清およびジを削除5ミリリットルのPBSでペレットをリュート。 </li><li> 24℃で10分間、528×gでペレット溶液を遠心分離</li><li>上清を取り除きます。 RPMI中（〜200μL）の適切な量で、ペレットを希釈します。 </li><li> 5〜10μlのサンプルを取り、 自動細胞カウンターまたは血球計算板を用いて細胞を数える。その結果を記録します。 1より多くの時間をカウントし、細胞を繰り返して、平均セル数を計算する。 </li><li>のRPMIを使用してμlの1,000,000 cells/10に細胞濃度を希釈する。 </li></ol> 3。細胞播種<ol><li>のRPMIを削除して、5分間の管腔内5μLのRPMIを追加することで、予備湿潤足場。 </li><li>足場内腔へのステップ2.12からRPMI中で10μlの骨髄由来の単核細胞を加え、10分、細胞が足場に取り付けることができるようにするのを待ちます。 </li><li> 1cmの長さに19 G針をカットし、足場の形状を維持するために、足場の内腔に針を入れた。 24ウェルプレートに載せてください。&lt;/李&gt; <li>各ウェルに千μLのRPMIを加え、インキュベーター中で、O / Nインキュベートする。 </li></ol> 4。グラフト移植<ol><li>手術前に、すべての手術器具をオートクレーブ：細かいハサミ、3Xマイクロ鉗子、2倍のマイクロ血管クランプ、鉗子を適用する1Xクランプ、1Xマイクロニードルホルダー、1X春のはさみ、1Xリトラクターを1X。 </li><li> 6〜8週齢の雌C57BL / 6は、組織工学的血管移植片のレシピエントとして使用されている。そのケージからマウスを削除し、ケタミン/キシラジンカクテルで腹部の右下の象限（ケタミン、100 mg / kgのキシラジンと、10 mg / kg）を腹腔内注射により麻酔して、それを比較検討する。ケトプロフェン（5 mg / kgを、IPは）事前に麻酔鎮痛剤として使用されます。 </li><li>物語つまんで鎮静のレベルを確認し、腹部の毛をクリップ。無菌眼軟膏で目を潤滑し、パッド上の背側横臥位にマウスを置きます。ベタジンとアルコールパッドで腹部を消毒。 CO無菌ドレープでマウスをVERだけ切開領域を公開します。 </li><li>恥骨領域への剣状突起下から正中開腹切開を行い、自己保持リトラクターを挿入します。生理食塩水湿らせたガーゼで腸を包む。ぶっきらぼう大動脈大動脈と大静脈を定義します。 </li><li>近位大動脈と大静脈の遠位両側の2のマイクロ血管のクランプを配置し、ぶっきらぼうに大静脈トランセクト大静脈から大動脈を分離する。必要に応じて、テーパー針に10-0モノフィラメント縫合糸で腹部大動脈の枝を結紮。 </li><li>無菌10-0縫合糸を使用して吻合を終了するために、近位端および遠位端と下大静脈の介在移植片を移植する。マウスの解剖学に依存し、移植片、出荷までmmで、トリム。近位端および遠位端の両方で1ステッチで移植性を確保し、移植片の反対側から4-5 stichesで連続縫合を開始します。前面を終えた後、クランプおよび移植片を反転反対側に、グラフトの裏側を縫合。注入中に、急性血栓症を予防するために頻繁にヘパリン溶液で移植片をフラッシュします。 </li><li>近位クランプを外し、局所吸収性無菌止血剤を塗布して出血を制御します。出血が完全に停止すると、遠位クランプを取り外して、出血を同じように制御します。血液が移植片を流れて​​いることを確認してください。 </li><li>含まねじ切り針で6-0黒色ポリアミドモノフィラメント縫合糸を使用した2層に腹部の筋肉と皮膚を閉じます。 </li><li> 0.5ミリリットルの生理食塩水を皮下注射すると、マウスが完全にモバイルになるまで、温暖化パッド上で回復ケージにマウスを置きます。復旧時には、紙の寝具で新しいケージにマウスを返す。 48時間、鎮痛薬（イブプロフェン、30 mg / kgで、飲料水）を得た。 </li></ol>

<ol>
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著者らは、開示することは何もありません。

TEVGのマウスモデルは、新組織形成と狭窄の開発の細胞および分子メカニズムを研究するための貴重なツールです。シードされたBM-MNCが移植11に播種した細胞の組織学的およびSEM画像の両方で示された。細胞播種の効率はまた、DNAアッセイ7を用いて示された。このモデル系を用いて、我々は、細胞播種が私たち人間の臨床試験3の故障の主なモードだっTEVG狭窄の開発の...

先天性心疾患は、米国の出生の約8％に影響を与える深刻な状況である。先天性心疾患または2.4千あたりの出生とのそれらの乳児の約25％が、自分の人生1の最初の年に侵襲的な治療を必要とする。先天性心疾患のための最も有効な治療は、再建手術である。残念ながら、現在利用可能な血管導管の使用に起因する合併症は術後の罹患率と死亡率の最も重要な原因である。 この問題に対処するために、我々は、臨床使用のために第2組織工学血管移植片（TEVGs）を開発した。 TEVGsは、自家骨髄由来の、単核細胞（BM-MNCを）を播種し、先天性心臓手術のための静脈の導管として移植生分解性ポリエステルチューブから構築した。結果は、フォローアップの1〜3年で優れた開存率を示したが、狭窄の重要な発生率<s3,4&gt;まで。これは、血管新組織形成とTEVG狭窄の開発の基礎となるメカニズムのよりよい理解が必要であることは明らかであった。より良いTEVGsの開発や狭窄開発のメカニズムを理解するためには、ヒツジモデルは5,6を作成しました。このモデルでは、TEVGs正常生体血管に変換され、天然の静脈の形態と機能の両方において同様であった。大型動物モデルの使用はTEVGsの臨床使用量を助け、重要な前臨床情報を提供する上で良い最初のステップでした。しかし、大規模な動物モデルを用いTEVGsの血管新組織形成の細胞および分子メカニズムの完全な理解は原因種特異的分子ツールが不足しているため血管細胞の表現型の分子特性の限界に制限されています。これらの欠点を克服するために、TEVGsのマウスモデルは、マウス遺伝学の急速な進歩とその広範なmoleculaの理由によって開発されました短縮された時間スケールの付加的な利点を持つRの特性評価。 マウスのIVC介在モデルを忠実に大型動物やヒトで発生する新生血管形成の過程をで要約するが、はるかに短い時間経過6-9以上。ここで、生分解性の足場を用いた小規模な移植片の製造のための詳細なプロトコルは、BM-MNCの回収と単離、足場上のBM-MNCの播種、およびマウスモデルにおける移植片植え込みについて説明した。

<table border="0" cellpadding="0" cellspacing="0" style="width:616px;" width="615">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">Name</td>
			<td style="width:139px;">Company</td>
			<td style="width:133px;">Catalog Number</td>
			<td style="width:168px;">Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">polyglycolic acid (PGA) felt</td>
			<td>Biomedical Structures</td>
			<td></td>
			<td>Custome ordered</td>
		</tr>
		<tr height="45">
			<td height="45" style="height:45px;width:176px;">&#603;-caprolactone and L- 
			lactide copolymer P(LA/CL)&#160;</td>
			<td>Gunze Inc.</td>
			<td></td>
			<td>Custome ordered</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Pipet tip, 0.1-10 &#956;l&#160;</td>
			<td>Fisher Sientific</td>
			<td>02-707-456</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lyophilizer&#160;</td>
			<td>Labconco</td>
			<td>7070020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">RPMI medium 1604</td>
			<td>gibco</td>
			<td>11875-093</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">Petri dish</td>
			<td>BD</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:176px;">24 well plate</td>
			<td>Corning</td>
			<td>3526</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">15cc tube&#160;</td>
			<td>BD</td>
			<td>352096</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">Ficoll</td>
			<td>Sigma</td>
			<td>10831-100ml</td>
			<td>Also called &#39;Histopaque&#39;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">DPBS</td>
			<td>gibco</td>
			<td>14190-144</td>
			<td></td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:176px;">Littauer Bone Cutter 4.5&#34; Straight</td>
			<td>Roboz</td>
			<td>RS-8480</td>
			<td>For BM harvesting</td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:176px;">Forceps 4.5&#34;</td>
			<td>Roboz</td>
			<td>RS-8120</td>
			<td>For BM harvesting</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:176px;">Scissors 4.5&#34;</td>
			<td>Roboz</td>
			<td>RS-5912</td>
			<td>For BM harvesting</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:176px;">Microscope</td>
			<td style="width:139px;">Leica</td>
			<td style="width:133px;">M80</td>
			<td style="width:168px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:176px;">C57BL/6J (H-2b), Female</td>
			<td style="width:139px;">Jackson Laboratories&#160;</td>
			<td style="width:133px;">664</td>
			<td style="width:168px;">8-12 weeks</td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:176px;">Ketamine Hydrochloride Injection</td>
			<td style="width:139px;">Hospira Inc.</td>
			<td style="width:133px;">NDC 0409-2053</td>
			<td style="width:168px;"></td>
		</tr>
		<tr height="23">
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			<td height="24" style="height:24px;width:176px;">10 ml Syringe</td>
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			<td height="20" style="height:20px;width:176px;">18G 1 1/2 in, Needle</td>
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			<td height="20" style="height:20px;width:176px;">25G 1 in., Needle</td>
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			<td style="width:133px;">305125</td>
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			<td height="20" style="height:20px;width:176px;">30G 1 in., Needle</td>
			<td style="width:139px;">BD</td>
			<td style="width:133px;">305106</td>
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			<td height="23" style="height:23px;width:176px;">Warm Water Recircultor</td>
			<td style="width:139px;">Gaymar</td>
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implantation of inferior vena cava interposition graft in mouse model

骨髄単核細胞（のBMC）を播種した生分解性の足場は、しばしば、先天性心臓異常を治療するための再建手術のために使用される。長期臨床結果は、狭窄の有意な発生率が、優れた開存率を示した。血管新組織形成の細胞および分子機構を調査し、組織工学血管移植片（TEVGs）に狭窄発症を予防するために、我々は、約1mmの内径を有する移植片のマウスモデルを開発した。まず、ポリグリコール酸不織布から製造生分解性の管状の足場から組み立てたTEVGs、ε-カプロラクトンとL-ラクチド共重合体でコーティングされたメッシュと感じました。足場は、次いで、凍結乾燥機に入れ、24時間真空にし、細胞播種するまでデシケーター中に保存した。次に、骨髄を、ドナーマウスから採取し、単核細胞を、密度勾配遠心分離によって単離した。第三に、約100万個の細胞であった足場に播種し、O / Nインキュベート最後に、播種された足場は、その後、C57BL / 6マウスにおける大動脈大静脈介在移植片として移植した。注入された移植片は、血栓塞栓合併症や動脈瘤の形成の証拠がなく、優れた開存性（&gt; 90％）を示した。このマウスモデルは、理解で私たちを助け、TEVGに新組織形成の細胞および分子メカニズムを定量化します。

TEVG注入の概略図を図1に示されている。骨髄は、ドナーマウスから採取し、単核細胞を密度遠心分離を用いて単離し、その後、生分解性足場上に播種する。シードされた足場は、O / Nインキュベートし、下大静脈の介在移植片としてレシピエントマウスに移植した。 図2は、PGA-P（CL / LA）骨格の走査型電子顕微鏡を示す。内径は約1mmであり、壁の...

Watch this Scientific Journal Video about Implantation of Inferior Vena Cava Interposition Graft in Mouse Model at JoVE.com

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

細胞および分子新組織形成の我々の知識を向上させるために、TEVGのマウスモデルは、最近開発されました。グラフトは、C57BL / 6マウスにおける大動脈大静脈介在移植片として移植した。このモデルは、私たちの臨床研究で達成されるものと同様の結果を達成するが、はるかに短縮され、時間経過とともに。

マウスモデルにおける下大静脈介在移植の移植

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

implantation-of-inferior-vena-cava-interposition-graft-in-mouse-model

Research

JoVE Journal

Medicine

11.8K ビュー.  Nationwide Children's Hospital. 細胞および分子新組織形成の我々の知識を向上させるために、TEVGのマウスモデルは、最近開発されました。グラフトは、C57BL / 6マウスにおける大動脈大静脈介在移植片として移植した。このモデルは、私たちの臨床研究で達成されるものと同様の結果を達成するが、はるかに短縮され、時間経過とともに。 

ビデオ: Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential therapeutic compounds (1). New compounds to be utilized in the treatment and prevention of DVT are currently being developed. The delivery of potential therapeutic antagonist compounds to an affected thrombosed vein has been problematic. In the context of therapeutic applications, a model that uses partial stasis and consistently generates thrombi within a major vein has been recently established. The Electrolytic Inferior vena cava Model (EIM) is mouse model of DVT that permits thrombus formation in the presence of continuous blood flow. This model allows therapeutic agents to be in contact with the thrombus in a dynamic fashion, and is more sensitive than other models of DVT (1). In addition, this thrombosis model closely simulates clinical situations of thrombus formation and is ideal to study venous endothelial cell activation, leukocyte migration, venous thrombogenesis, and to test therapeutic applications (1). The EIM model is technically simple, easily reproducible, creates consistent thrombi sizes and allows for a large sample (i.e. thrombus and vein wall) which is required for analytical purposes.

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

The electrolytic induction of endothelial activation to the internal surface of the Inferior Vena Cava results in venous type thrombus formation due to endothelial activation and partial blood stasis, two components of Virchow's triad.

静脈血栓症の電解下大静脈モデル（EIM）

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

医学

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

下大静脈血栓症のマウス完全な停滞のモデル

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

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

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

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 ...

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

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

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

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. ...

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

ラット大動脈または下大静脈内のパッチ血管形成術

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

同所性同種のモデルラット Suprahepatic 上大静脈再建のため新規磁気吻合法を用いた肝移植

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

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

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

ワイヤー図法を使用してマウスの腸間膜動脈の等尺性収縮力測定

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

A Rodent Model of The Ross Operation: Syngeneic Pulmonary Artery Graft Implantation in A Systemic Position

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding root replacement, the possible dilation of the pulmonary autograft and subsequent aortic regurgitation is described. Several animal models have been proposed. However, these are usually limited to ex-vivo models or in-vivo experiments with relatively expensive large animal models. In this study, we sought to establish a rodent model of pulmonary artery graft (PAG) implantation in a systemic position. A total of 39 adult Lewis rats were included. Immediately after euthanasia, the pulmonary root was harvested from a donor animal (n=17). Syngeneic recipient (n=17) and sham-operated (n=5) rats were sedated and ventilated. In the recipient group, the PAG was implanted with an end-to-end anastomosis in infra-renal abdominal aortic position. Sham-operated rats underwent only transection and re-anastomosis of the aorta. Animals were followed with serial ultrasound studies for two months and post-mortem histological analysis. The median PAG diameter in the native position was 3.20 mm (IQR=3.18-3.23). At follow-up, the median diameter of the PAG was 4.03 mm (IQR=3.74-4.13) at 1 week, 4.07 mm (IQR=3.80-4.28) at 1 month, and 4.27 mm (IQR=3.90-4.35) at 2 months (p&lt;0.01). Peak systolic velocity was 220.07 mm/s (IQR=210.43-246.41) at 1 week, 430.88 mm/s (IQR=375.28-495.56) at 1 month, and 373.68 mm/s (IQR=305.78-429.81) at 2 months (p=0.02) and did not differ from the sham-operated group at the end of the experiment (p=0.5). Histological analysis did not show any sign of endothelial thrombosis. This study showed that rodent models may allow for the evaluation of the long-term adaptation of the pulmonary root to a high-pressure system. A systemically placed syngeneic PAG implantation represents a simple and feasible platform for the development and evaluation of novel surgical techniques and drug therapies to further improve the outcomes of the Ross operation.

We demonstrate how to establish a murine model of pulmonary root implantation into the descending aorta to simulate the Ross procedure. This model enables the medium/long-term evaluation of pulmonary autograft remodeling in a systemic position, representing the basis of developing therapeutic strategies to promote its adaptation.

ロス手術のげっ歯類モデル:全身位置における同系肺動脈移植移植

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding ...

Image Acquisition Method for the Sonographic Assessment of the Inferior Vena Cava

Over the past several decades, clinicians have incorporated several applications of diagnostic point-of-care ultrasound (POCUS) into medical decision-making. Among the applications of POCUS, imaging the inferior vena cava (IVC) is practiced by a wide variety of specialties, such as nephrology, emergency medicine, internal medicine, critical care, anesthesiology, pulmonology, and cardiology. Although each specialty uses IVC data in slightly different ways, most medical specialties, at minimum, attempt to use IVC data to make predictions about intravascular volume status. While the relationship between IVC sonographic data and intravascular volume status is complex and highly context-dependent, all clinicians should collect the sonographic data in standardized ways to ensure repeatability. This paper describes standardized IVC image acquisition including patient positioning, transducer selection, probe placement, image optimization, and the pitfalls and limitations of IVC sonographic imaging.&#160;This paper also describes the commonly performed anterior IVC long-axis view and three other views of the IVC that can each provide helpful diagnostic information when the anterior long-axis view is difficult to obtain or interpret.

Point-of-care ultrasound evaluation of the inferior vena cava (IVC) is commonly utilized to identify, among other things, the volume status. Imaging should be performed systematically to ensure repeatability. This manuscript reviews the methods and pitfalls of sonographic IVC examination.

下大静脈の超音波評価のための画像取得方法

Over the past several decades, clinicians have incorporated several applications of diagnostic point-of-care ultrasound (POCUS) into medical ...

マウス肝移植における効率的な血管吻合のための戦略

Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. However, the technical challenges pose a barrier to utilizing this valuable experimental model and passing on these skills to the next generation. The most challenging aspect of this procedure is vascular reconstruction, including the portal vein (PV), infrahepatic inferior vena cava (IHIVC), and suprahepatic inferior vena cava. The use of plastic cuffs, rather than sutures, allows for smoother PV and IHIVC reconstruction. Vessels are reconstructed by attaching a cuff made from an intravenous catheter to the tip of the graft vessel and interposing the cuff into the recipient vessel. The two most crucial aspects are properly visualizing the inner lumen of the vessel and avoiding the use of excessive force. Our aim is to provide a technical overview of vascular reconstructions using the cuff technique in recipient surgery. These technical tips for the cuff technique are expected to help microsurgeons facilitate vascular reconstruction and advance their research.

著者スポットライト:脂肪肝移植における冷虚血と温再灌流障害の探索

This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.

マウス同所性肝移植におけるカフ技術による血管再建

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. ...