この作品は、SNF [31003A-138528]、OctavとマルセラBotnar財団、ノバルティス財団とエママスチャンプ財団からの補助金によってサポートされていました。私たちは、その優れた技術支援のためにマルティーヌLambelet、ジャン=クリストフシュテーレに感謝します。

ローザンヌ大学の倫理委員会は、ヒト組織の使用のために1983年に改訂され、1975年のヘルシンキ宣言で概説した原理に従っている実験を、承認した。 1人間の大伏在静脈の収穫<ol><li>虚血下肢バイパス手術を受けた患者から非静脈瘤人間伏在静脈の余剰部分を取得します。手術室では、ヨウ素溶液で脚全体を消毒し、足に脚の付け根から足を露出するために患者をドレープ。 </li><li>膝への鼠径部から正中切開を行います（中断された皮膚の部分を残して）。 </li><li>周囲の組織（ノータッチ技術）の椎弓と大伏在静脈を収穫。 4-0シルクネクタイと静脈のセキュアな側枝。すぐにRPMI-1640、G、4℃で9センチメートル2.5〜4ミリメートルの外径と大伏在静脈の長い余剰セグメントの最小値を格納するlutamax培地、12.5％ウシ胎児血清を補充した、実験室に持ってきて。 </li></ol> 2 EVPSデザイン<ol><li> 図1に示されている一般的な機器を組み立てます。すべての機器をオートクレーブ滅菌条件下ですべてのコンポーネントを保つ。また、システムは、防水であり、培地中に化学物質を漏洩しないことを確認してください。表紙にポリメチルメタクリレート（PMMA-GS）を使用します。スチール（X5クロムニッケル18 10）と静脈のサポートなどポリオキシメチレンプラスチック（POM）。 </li><li>静脈との接続の配置を可能にするために所望の形状に灌流チャンバーを設計します。 （円筒構造を使用している場合、または半径）の深さを確認してください、それが培養培地により一定報道（ 図1）と一緒に容器の最小限の屈曲および拡張を可能にするように少なくとも2.5 cmです。シールは、主要な問題であり、長方形のPMMA-GS建設が使用されている理由である。 </li><li>希望geometrに静脈のサポートをデザインyを。静脈よじれや膨満の上を回避するには、（静脈がねじで一緒にねじられるようねじは、その目的のために使用することはできません）押したり引っ張って長さ調節が可能です。 NOTE：L字型（血管に適合するように直径5mm）2静脈シリンダをサポートピース及び静脈（ 図1Bおよび図2）スライド2によって接続されたフルスチールロッドは、ここで使用されている。 </li><li>のp流体柱の= 0-10 = hxをρ×gではp =圧力（N / m 2であり、ペンシルベニア州）は、h =高さ（m）ρ：システムに適用される「静止圧」であることなど、低圧塔を設計する=液体の密度（㎏/ m 3）であり、g =重力定数（9.81メートル/秒2）。上から下へのデザイン4つの接続ダクト、：（静脈へ）、（静脈）からの流出のために、流入圧力を加えると培地交換を可能にするために。 </li><li>メディアを準備します。これまでの研究5,11-14に基づき、12.5グルタマックスを補充した、RPMI-1640]を選択します％ウシ胎児血清および1％抗生物質 - 抗真菌溶液（10,000 U / mlのペニシリンG、プラス10 mg / mlの硫酸ストレプトマイシンを加えた25 mg / mlのアムホテリシンB、プラス0.5 / mlの：ゲンタマイシン）。剪断応力（SS）は3 Qは流速（ml /秒）であるSS = 4μQ/π* rで与えられる、静脈セグメントの半径（cm）のrは、μは灌流媒体の粘度である。 <ol><li> 70kDaのデキストランを添加して粘度を調整することでSSを調節する。粘度計粘度を測定します。ここでは、9〜15ダイン/ cm 2とSSを設定するために、8％の70kDaデキストランを追加します。 </li></ol></li><li> 60パルス/分、圧力システムに適用され、コンピュータによって制御から独立した150±15ml /分の一方向の流れを生成する一定振幅の拍カージオイド信号を誘導するために歯車ポンプを設定する。運転ソフトウェアは一定の取得と監視圧力、流速、脈​​拍数、及び信号を積分いることを確認してください。必要に応じて、第二のポンプを使用します（非同期hronized）非層流、乱流を生成する。 </li></ol> 3 EVPSアセンブリ（図1） <ol><li>開始する前に、すべての機器が滅菌されていることを確認してください。層流フード内で無菌状態の下で、以下のすべての手順を実行します。 </li><li>媒質で満たされたペトリ皿に静脈を置きます。外科用メスを使用して、3等分静脈を分割。 </li><li>すぐに、PBS中の1セグメントをすすぐ。 3部にセグメントを分割し、形態計測のためにホルマリンに1を修正。定量的な転写産物（RT-PCR）とタンパク質（ウェスタンブロット）分析のために他の二つをフリーズします。コントロールとして、これらのセグメントを考慮して、非灌流静脈。 </li><li>灌流2残りのセグメントを使用してください。 <ol><li>非常に穏やかに静脈内に媒体を注入し、通常の流れ方向を決定する;弁の存在下で、静脈が反転される。 </li><li>静脈をシーリング実験成功に最も重要である。担保を通して漏れがないか確認してください。 6-0絹縫合糸を持つ任意のリークを固定します。 </li> &lt;/オール&gt; </li><li>時間（2.3、 図1）での静脈2つの金属シリンダー間のセグメント、一方の端を接続します。くぼみの周りEthibon 3-0（ 図1AおよびB）でシリンダーを固定します。 <ol><li>以前媒質で満たされた灌流チャンバー内に全体を静脈セグメントを配置します。第二のセグメントに対して同じ手順を繰り返します。 注記：正しくシリンダに静脈をシールしないと、漏れの原因となる再介入を必要とし、大幅に感染し、実験失敗のリスクが増加します。 </li></ol></li><li> （メッシュ）第二のセグメントを強化するには、L字型のピース（2.3及び図1）から（添付静脈で）2シリンダーを離します。 <ol><li>やわらかに、任意の楽器と静脈を触れないでください。静脈へのその後のシリンダの最初のメッシュをスライドさせます。プッシュ/プル押し合いは、静脈上のメッシュを取得します。 </li><li>メッシュはJを確保静脈の全面を覆うたらEthibon 3-0とシリンダにacketed静脈。 </li><li> L字型の支持体に静脈/シリンダ化合物を再構成し、以前媒質で満たされた灌流チャンバーに移す。 </li></ol></li><li> 3.2ミリメートルの内径過酸化物で処理したシリコンチューブを使用して、Yスプリッタにそれぞれ金属製の円筒（インおよびアウトフロー）を接続します。 </li><li>チューブの同じタイプを使用して第2のYスプリッタに流出スプリッタを接続します。このYスプリッタからは、両方の血管を通る灌流圧を測定するために一本のチューブを使用しています。閉ループ系（ 図2）を形成するために戻って列に、もう一方を接続します。 </li><li>インキュベーターの内部では、ポンプヘッドに低圧塔を接続するために長い（1メートルの長さ）のチューブを使用しています。 </li><li>他の長尺管を流入Yスプリッタにポンプヘッドを接続することにより設定します（ 図1）完了します。 </li></ol> 4。静脈灌流<ol><li> EVPSアセンブリは、同時された後mpleted、（補充できるように、静脈流出ダクトの下に留まる）培地でカラムを埋める。システムが一杯になるまでカラムに多くの媒体を追加します。 pHが37±0.1℃に維持インキュベーターにすべてのシステムを移動（ヘンダーソン·ハッセルバッハ式に基づいてCO 2 / pHのアルゴリズムを使用して）7.40±0.01で一定に保った。 </li><li>インキュベーターの外歯車ポンプヘッドを持参し、ギアポンプドライブに接続します。アセンブリを固定する棒をねじ込み。 </li><li>ポンプパワーのスイッチを入れ、それが駆動ソフトウェア上で起動しても同様に、すべての区画内に分配される媒体5分を許可していることを確認してください。 </li><li>圧力を監視するには、動脈ラインの監視を使用しています。コンピュータにリンクされた圧力変換器にEVPS圧力出力を（それが動脈カテーテルに相当）を接続します。 <ol><li>チューブが完全に媒体で充填され、任意の気泡を含んでいないことを確認してください。 「動脈リチウムを通じて脱泡培養システムね&quot;チューブ（ 図2）。ディスプレイに注意を払い、60パルス/一定の振幅の分の拍動カーディオイド信号を探します。この時点で、平均圧力0〜10 mmHgでの間である。圧力が&lt;0で、列には、漸進的に（静脈とチューブとの間の静脈担保または不十分なシール）漏れを探して空になります。 </li></ol></li><li>静脈試験用または動脈試験のための90ミリメートルHgで6ミリメートルHgのに最小限の圧力を設定してください。これらの条件下で、空気インジェクタ、カラムおよびシステムに必要な圧力を加える。 </li><li>高圧塔に接続されたチューブを使用して、2日ごとに培地を変更します。圧力変化の損傷を防止するために、最初の列プラグを開く。 </li></ol>灌流の5。完了<ol><li>灌流の3または7日後：インキュベーターからEVPSを取ると静脈をディスマウント。捨てる5mmの近位および遠位静脈が機器に付着し終了します。中央、厚さ5mm厘をカット残りのセグメントからのGSとホルマリンに（形態計測）を固定。残りのフラグメントを凍​​結し、さらなる分子解析のための粉末に減らす。 </li></ol>

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著者らは、開示することは何もない。

この研究は、ヒトの静脈に広範囲血行動態の研究を実行するためのex vivoで静脈灌流システム（EVPS）をuncovers。このシステムは、 生体内で細胞を循環させることによって放出された炎症性悪化や成長因子の非存在下で定義された血行動態パラメータの下に伏在静脈灌流を可能にします。このように、人間の静脈グラフト5,11,12,15におけるIHの制御に関与根底にある経路のよ...

心血管疾患は、欧米諸国1における罹患率および死亡率の主要な原因である。血管内治療の進歩にもかかわらず、バイパス手術は、このように万人以上の静脈グラフトは、米国で毎年行われている、現代的な治療法の柱のまま。しかし、研究の数十年にもかかわらず、下肢静脈グラフト30〜60％が内膜肥厚（IH）2による第一年以内に失敗する。機械的な力、特に低せん断応力（SS）と高い壁張力が、この過形成応答3,4の開始と発展に極めて重要である。この問題に対処するために、ex vivoで静脈灌流システム（EVPS）は厳密に制御され、血行動態条件（圧力およびせん断応力）、ヒト伏在静脈の行動の下で、研究し作成した。本研究では、動脈のような循環への挿入に続いて、高圧（= 100mmHgの平均）prolifを刺激するのに十分であったや機能の内膜層（IH）5への平滑筋細胞の遊走。 哺乳類の研究は、「動脈血静脈」をサポートし、急性の血行動態は静脈が一度動脈環境に移植の顔に変化に対抗するための効率的な方法として、外部の補強を使用することを示唆している。過膨張を防止メッシュは、せん断応力が増加し、壁張力、その結果、IH 6-10を減少させた。しかし、根底にある機序およびバイパス開通性を向上させる上で、人間の静脈への適用は十分に特徴付けられていない。私たちのEVPS静脈を一旦動脈レジメン（高いせん断応力および圧力）、外部マクロ多孔質ポリエステル管状メッシュの非存在下および存在下でのヒト伏在静脈の挙動に挿入面する変化を模倣する条件において、比較した。病理学的リモデリングおよびIHを防止することにより、メッシュは、その潜在的な臨床効率11の証拠を提供</sup&gt;。 この研究は1）は、外部マクロ多孔質ポリエステルメッシュはIHを減少させ、その潜在的な臨床応用するために重要な情報を提供することを実証制御された圧力とせん断応力の2）の下でex vivoでのヒト伏在静脈灌流のモデルを紹介します。

<table><tbody><tr><td>RPMI 1640 - Glutamax</td><td>Life Technologies</td><td>61870-010</td><td></td></tr><tr><td>Penicilline/Streptomycine/Fungizone</td><td>Bioconcept</td><td>4-02F00-H</td><td></td></tr><tr><td>Dextran from Leuconostoc spp. 500 g</td><td>Sigma-Aldrich</td><td>31390</td><td></td></tr><tr><td>Tampon PBS CHUV pH 7.1-7.3 1 L</td><td>Laboratorium und Grosse Apotheke Dr. G. Bichsel AG</td><td>100 0 324 00</td><td></td></tr><tr><td>Cryosectionning embedding medium - Tissue-Tek OCT Compound</td><td>Fisher Scientific</td><td>14-373-65</td><td></td></tr><tr><td>Silicon Tubing (Peroxide) L/S 16 (96400-16 ) - 7.5 m</td><td>Idex Health &amp;amp; Science GMBH</td><td>MF0037ST</td><td></td></tr><tr><td>Y-splitter&amp;nbsp;</td><td>Idex Health &amp;amp; Science GMBH</td><td>Y-connector</td><td></td></tr><tr><td>35 mm Culture dish</td><td>Sigma-Aldrich</td><td>CLS430165-100EA</td><td></td></tr><tr><td>15 ml Falcon tube</td><td>BD Bioscence</td><td>352096</td><td></td></tr><tr><td>50 ml Falcon tube</td><td>BD Bioscence</td><td>352098</td><td></td></tr><tr><td>Gearing pump - Reglo-Z</td><td>Idex Health &amp;amp; Science GMBH</td><td>SM 895&amp;nbsp;</td><td>&amp;nbsp;App-Nr 03736-00194</td></tr><tr><td>Pump Head</td><td>Idex Health &amp;amp; Science GMBH</td><td>MI0008&amp;nbsp;</td><td></td></tr><tr><td>Monitoring Kit TRANSPAC IV</td><td>icumedical</td><td>011-0J736-01</td><td></td></tr><tr><td>20 ml Syringes</td><td>B. Braun Medical SA</td><td>4612041-02</td><td></td></tr><tr><td>Etibon 3-0 FS-2</td><td>Ethicon- Johnson&amp;amp;Johnson</td><td>EH7346H</td><td></td></tr><tr><td>Mesh ProVena 6-8mm</td><td>B. Braun Medical SA</td><td>1105012-14</td><td></td></tr><tr><td>NaCl: Sodium chloride solution perfusion 0.9% (100 ml)</td><td>B. Braun Medical SA</td><td>534534</td><td></td></tr><tr><td>Masterflex L/S Standard Drive</td><td>Cole-Parmer Instrument Co</td><td>7521-10</td><td></td></tr><tr><td>Acquisition card</td><td>National Instruments</td><td>PCI-6024 E</td><td></td></tr><tr><td>Flowmeter module</td><td>Transonic Systems Inc.</td><td>TS410 and T402</td><td></td></tr><tr><td>Stopcock with 3-ways</td><td>BD Connexta Luerlock</td><td>394600</td><td></td></tr><tr><td>Millex Filter</td><td>Milian</td><td>SE2M229I04</td><td></td></tr></tbody></table>

procedure for human saphenous veins ex vivo perfusion and external reinforcement

大規模な閉塞性動脈疾患のための現代的な治療の主力は、静脈バイパス移植である。しかし、その耐久性は、最終的には血管閉塞及び移植の失敗につながる内膜肥厚（IH）の脅威にさらされている。機械的な力、特に低せん断応力と高い壁の張力は、開始するために、これらの細胞および分子の変化を維持すると考えられているが、それらの正確な寄与は解明されていない。選択的にIHの生物学上の圧力とせん断応力の役割を評価するために、ex vivoで灌流システム（EVPS）は、動脈レジメン（高せん断応力および高圧）の下で人間の伏在静脈のセグメントを灌流するために作成されました。さらなる技術革新は、同じ静脈、外部メッシュで補強された1から2のセグメントの同時灌流を可能にした。静脈は、非接触技術を使用して採取し、直ちにEVPSアセンブリ実験室に移した。たてISOLの一つのセグメントated静脈（コントロール、0日目）に灌流されていませんでした。他の二つのセグメントが最大7日間灌流し、1は完全に4ミリメートル（直径）外部メッシュで保護されている。圧力、流速、および脈拍数を連続的にモニターし、大腿動脈内で優勢な血行力学的条件を模倣するように調整した。潅流が完了すると、静脈がディスマウントされ、組織学的および分子分析のために使用される。 エキソビボ条件下では、高圧灌流（動脈、平均値= 100 mmHgで）がIHおよびヒト静脈のリモデリングを生成するのに十分である。これらの変化は、外部ポリエステルメッシュの存在下で還元されている。

EVPSは、人間の伏在静脈の再構築およびIHをグラフト上で独立して血流力を評価するための貴重なツールを提供します。 図1は、灌流チャンバーと静脈のサポートを示しています。 図1AおよびBにおいて、前静脈サポート（ 図1A）および後（ 図1B）アセンブリは、それぞれ描かれている。これは、簡単に（左か...

Watch this Scientific Journal Video about Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement at JoVE.com

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

内膜過形成（IH）及び静脈グラフト不全の発症を導くメカニズムはまだよく理解されていない。本研究では、制御された流れと圧力の下で、人間の静脈を灌流するためにex vivoでシステムが記載されている。さらにIHの開発を制限する外部メッシュ補強の効率を評価した。

人間の伏在静脈のための手順<em&gt;生体外</em&gt;灌流および外部補強

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

procedure-for-human-saphenous-veins-ex-vivo-perfusion-external

Research

JoVE Journal

Medicine

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

11.2K ビュー.  Brigham and Women's Hospital/Harvard Medical School. 内膜過形成（IH）及び静脈グラフト不全の発症を導くメカニズムはまだよく理解されていない。本研究では、制御された流れと圧力の下で、人間の静脈を灌流するためにex vivoでシステムが記載されている。さらにIHの開発を制限する外部メッシュ補強の効率を評価した。 

ビデオ: Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

中に組み込み血管新生による組織工学<em&gt;インビボ</em&gt;ティッシュ・エンジニアリング商工会議所

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

生物工学

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in pathological situations such as cancer, vascularization plays a leading role in disease progression. Consequently, there is a strong need for a standardized and well-characterized in vivo model in order to elucidate the mechanisms of neovascularization and develop different vascularization approaches for tissue engineering and regenerative medicine.
We describe a microsurgical approach for a small animal model for induction of a vascular axis consisting of a vein and artery that are anastomosed to an arteriovenous (AV) loop. The AV loop is transferred to an enclosed implantation chamber to create an isolated microenvironment in vivo, which is connected to the living organism only by means of the vascular axis. Using 3D imaging (MRI, micro-CT) and immunohistology, the growing vasculature can be visualized over time. By implanting different cells, growth factors and matrices, their function in blood vessel network formation can be analyzed without any disturbing influences from the surroundings in a well controllable environment. 
In addition to angiogenesis and antiangiogenesis studies, the AV loop model is also perfectly suited for engineering vascularized tissues. After a certain prevascularization time, the generated tissues can be transplanted into the defect site and microsurgically connected to the local vessels, thereby ensuring immediate blood supply and integration of the engineered tissue. By varying the matrices, cells, growth factors and chamber architecture, it is possible to generate various tissues, which can then be tailored to the individual patient's needs.

The Arteriovenous AV Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

We describe a microsurgical approach for the generation of an arteriovenous (AV) loop as a model for analyzing vascularization in vivo in an isolated and well-characterized environment. This model is not only useful for the investigation of angiogenesis, but is also optimally suited for engineering axially vascularized and transplantable tissues.

血管新生および血管組織工学を研究するための小さな動物モデルにおける動静脈（AV）ループ

A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in ...

Decellularization and Recellularization Methodology for Human Saphenous Veins

Vascular conduits used during most vascular surgeries are allogeneic or synthetic grafts that often lead to complications caused by immunosuppression and poor patency. Tissue engineering offers a novel solution to generate personalized grafts with a natural extracellular matrix containing the recipient&#39;s cells using the method of decellularization and recellularization. We show a detailed method for performing decellularization of the human saphenous vein and recellularization by perfusion of peripheral blood. The vein was decellularized by perfusing 1% Triton X-100, 1% tri-n-butyl-phosphate (TnBP) and 2,000 Kunitz units of deoxyribonuclease (DNase). Triton X-100 and TnBP were perfused at 35 mL/min for 4 h while DNase was perfused at 10 mL/min at 37 &#176;C for 4 h. The vein was washed in ultrapure water and PBS and then sterilized in 0.1% peracetic acid. It was washed again in PBS and preconditioned in endothelial medium. The vein was connected to a bioreactor and perfused with endothelial medium containing 50 IU/mL heparin for 1 h. Recellularization was performed by filling the bioreactor with fresh blood, diluted 1:1 in Steen solution, and adding endocrine gland-derived vascular endothelial growth factors (80 ng/mL), basic fibroblast growth factors (4 &#181;L/mL), and acetyl salicylic acid (5 &#181;g/mL). The bioreactor was then moved into an incubator and perfused for 48 h at 2 mL/min while maintaining glucose between 3 - 9 mmol/L. Later, the vein was washed with PBS, filled with endothelial medium and perfused for 96 h in the incubator. Treatment with Triton X-100, TnBP and DNase decellularized the saphenous vein in 5 cycles. The decellularized vein looked white in contrast to normal and recellularized veins (light red). The hematoxylin &#38; eosin (H&#38;E) staining showed the presence of nuclei only in normal but not in decellularized veins. In the recellularized vein, H&#38;E-staining showed the presence of cells on the luminal surface of the vein.

Here, we describe a protocol for the saphenous vein decellularization using detergents and recellularization by perfusion of the peripheral blood and the endothelial medium.

Decellularization とひと伏在静脈の Recellularization 方法

Vascular conduits used during most vascular surgeries are allogeneic or synthetic grafts that often lead to complications caused by immunosuppression and ...

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. ...

医学

Synergizing Antegrade Endoscopic with Bridging Vein Harvesting for Improvement of Great Saphenous Vein Graft Quality from the Lower Leg

Antegrade endoscopic harvesting of autografts for bypass grafting may be an optimal strategy addressing excellent graft quality and reduced post-operative wound complications. This standardized protocol for antegrade endoscopic vein harvesting (EVH) from the lower leg has the potential to be introduced to routine coronary artery bypass grafting (CABG). Patients undergoing CABG surgery are positioned on a surgical table with two additional foam rollers below the extended legs, enabling antegrade EVH from the lower leg. Following minimally invasive surgical access through a bridging vein harvest technique, an endoscopic optical dissector is inserted antegrade into the wound. The main vessel and side branches are dissected under continuous optical control of vein quality status and the working channel. After, an endoscopic optical retractor is inserted with an internal bipolar electrocoagulation device for precise, safe, and tissue-protective interruption of side branches. After release of the vein, the vessel is cut off at the proximal and distal ends under optical control, retrieved from the wound, then cannulated and flushed with heparinized saline. Finally, all side branches of the vein graft are double-clipped. Vascular histology is analyzed in a randomized selection of vein samples. After applying this standardized EVH protocol, the learning curve was shown to be steep, and graft quality was sufficient for coronary artery bypass grafting in every case. There was no conversion to surgical harvesting and low risks for tissue damage and bleeding. Leg positioning and synergizing EVH with bridging vein harvesting improved procedural success and vein graft quality. In our hands, antegrade EVH from the lower leg was feasible, demonstrating straightforward graft dissection as well as adequate macroscopic and microscopic graft quality with preserved endothelial integrity. In conclusion, the introduced technique is safe, shows excellent vein autograft quality, and illustrates feasibility for elective and urgent isolated CABG and combined CABG scenarios.

Presented here is a protocol for antegrade endoscopic vein harvesting from the lower leg, which can safely be introduced in routine coronary artery bypass grafting. Vein grafts present excellent graft quality following this standardized protocol with positioning of the legs, minimally invasive access to the vein, and antegrade endoscopic vein harvesting.

下肢からの大きなサフェノス静脈移植品質の向上のための橋渡し静脈収穫と前立腺内視鏡を相乗的に

Antegrade endoscopic harvesting of autografts for bypass grafting may be an optimal strategy addressing excellent graft quality and reduced post-operative ...

A Training and Testing System for Performing Vascular Reconstruction In Vitro

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has yet to be developed. In this study, we introduce an in vitro training and testing system using a magnetic anchoring technique with which a trainee can practice manual vascular reconstruction individually. Additionally, this system can also be used to test the quality of the reconstruction. The described system includes a vascular reconstruction training machine, magnetic tractors, and a magnetic suture puller. In this manuscript, we detail an end-to-end vein anastomosis using porcine right and left iliac veins. To identify the potential damage caused by a magnetic suture puller on the suture, we created three groups with six segments of 4-0 polypropylene sutures each: a control group with no intervention on the polypropylene suture, a group in which the polypropylene suture is manually pulled with sterile gloves 20x, and a magnetic puller group in which the magnetic puller pulled the polypropylene suture 20x. These groups were tested by light microscopy and breaking strength tests, and the effect of reconstruction was assessed. In the light microscopy test, the control group was less likely to be damaged (p &#60; 0.05) and the number of damaged points of the manual group and magnetic puller group were similar (p &#62; 0.05). The results of the breaking strength test were compared across groups and no significant difference was observed (p &#62; 0.05). The end-to-end anastomosis of the porcine iliac veins was successfully performed using this training system, and the reconstructed veins could undergo 2.0 kPa perfusion pressure. Using this training and testing system the trainee can practice manual vascular reconstruction in vitro individually with the aid of magnetic tractors and a magnetic suture puller, and the quality of the reconstruction can be tested.

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

インビトロにおける血管再建を行うためのトレーニングと検査システム

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has ...

A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under Arterial Blood Pressure

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor because of the acceleration of intimal hyperplasia due to the exposure to arterial blood pressure. The present protocol is designed for the establishment of experimental venous intimal hyperplasia by interposing rabbit jugular veins to the ipsilateral carotid arteries. The protocol does not require surgical procedures deep in the body trunk and the extent of the incision is limited, which is less invasive for the animals, allowing long-term observation after implantation. This simple procedure enables researchers to investigate strategies to attenuate the progression of intimal hyperplasia of the implanted vein grafts. Using this protocol, we reported the effects transduction of microRNA-145 (miR-145), which is known to control the phenotype of vascular smooth muscle cells (VSMCs) from the proliferative to the contractile state, into harvested vein grafts. We confirmed the attenuation of intimal hyperplasia of vein grafts by transducing miR-145 before implantation surgery through the phenotype change of the VSMCs. Here we report a less invasive experimental platform to investigate the strategies that can be used to attenuate intimal hyperplasia of vein grafts in revascularization surgeries.

The present protocol aims to experimentally create venous intimal hyperplasia by subjecting veins to arterial blood pressure for developing strategies to attenuate venous intimal hyperplasia following revascularization surgery using vein grafts.

静脈移植を用いた血管形成術を模したウサギ静脈間位置モデルによる動脈血圧下の内膜過形成の評価

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor ...

Occlusion of the Great and Small Saphenous Vein Using Copolymeric Glue Based on N-Butyl Cyanoacrylate and Methacryloxy Sulfolane

We present the preliminary results of a longitudinal observational study aimed at evaluating the effectiveness and safety at different short- and long-term follow-ups of vascular occlusion of the great saphenous vein (GSV) and small saphenous vein (SSV) affected by severe pathological reflux, using an innovative modified cyanoacrylate surgical glue composed of N-butyl cyanoacrylate and methacryloxy sulfolane (NBCA+MS). Ninety patients, prospectively recruited for 1 year, underwent the study with EcoColor-Doppler (ECD) to evaluate the maximum diameters of the GSV and SSV in the orthostatic position and the reflux time (RT). An RT greater than 0.5 s was considered pathologic. Clinical, etiology, anatomy, and pathophysiology (CEAP) assessment was used for the complete evaluation of each patient in the study. All the patients were treated by NBCA+MS glue to obtain vein occlusion and observed before treatment (baseline; T0), within 6 h after treatment (T1), 1 month after treatment (T2), 3 months after treatment (T3), 6 months after treatment (T4), and 1 year after treatment (T5). Chi-square (&#967;) analysis was performed to evaluate the effectiveness and safety of the treatment. All the patients participated in the entire duration of the study. Complete occlusion was maintained in 100% of patients at T1, 98.9% at T2 and T3, and 97.8% at T4 and T5 (p &lt; 0.001). None of the patients suffered from post-surgical thrombosis. No blue hyperpigmentation, or paresthesia was observed during the entire observation period. Immediately after treatment, 7.7% of patients needed painkillers; 1 week after treatment, 100% of patients returned to normal life. Vascular occlusion of the great or small saphenous vein using NBCA+MS glue is a safe procedure with persistent benefits after a 1 year follow-up. This procedure can be performed with local anesthesia, allowing a quick return to normal life. Thanks to its low invasiveness, the treatment is not painful.

Here, we present a protocol to treat the great saphenous vein (GSV) and the small saphenous vein (SSV) affected by severe pathological reflux, using an original sclerosing and embolizing cyanoacrylate-based glue composed of N-butyl cyanoacrylate and methacryloxy sulfolane (NBCA+MS).

n-ブチルシアノアクリレートとメタクリロキシスルホランをベースとした共重合接着剤を用いた大小伏在静脈の閉塞

We present the preliminary results of a longitudinal observational study aimed at evaluating the effectiveness and safety at different short- and ...

Human Saphenous Vein Endothelial Cell Isolation and Exposure to Controlled Levels of Shear Stress and Stretch

Coronary artery bypass graft (CABG) surgery is a procedure to revascularize ischemic myocardium. Saphenous vein remains used as a CABG conduit despite the reduced long-term patency compared to arterial conduits. The abrupt increase of hemodynamic stress associated with the graft arterialization results in vascular damage, especially the endothelium, that may influence the low patency of the saphenous vein graft (SVG). Here, we describe the isolation, characterization, and expansion of human saphenous vein endothelial cells (hSVECs). Cells isolated by collagenase digestion display the typical cobblestone morphology and express endothelial cell markers CD31 and VE-cadherin. To assess the mechanical stress influence, protocols were used in this study to investigate the two main physical stimuli, shear stress and stretch, on arterialized SVGs. hSVECs are cultured in a parallel plate flow chamber to produce shear stress, showing alignment in the direction of the flow and increased expression of KLF2, KLF4, and NOS3. hSVECs can also be cultured in a silicon membrane that allows controlled cellular stretch mimicking venous (low) and arterial (high) stretch. Endothelial cells' F-actin pattern and nitric oxide (NO) secretion are modulated accordingly by the arterial stretch. In summary, we present a detailed method to isolate hSVECs to study the influence of hemodynamic mechanical stress on an endothelial phenotype.

We describe a protocol to isolate and culture human saphenous vein endothelial cells (hSVECs). We also provide detailed methods to produce shear stress and stretch to study mechanical stress in hSVECs.

ヒト伏在静脈内皮細胞の単離と制御されたレベルのせん断応力および伸張への曝露

Coronary artery bypass graft (CABG) surgery is a procedure to revascularize ischemic myocardium. Saphenous vein remains used as a CABG conduit despite the ...

Jugular Vein Graft Harvest From a Rabbit Model: A Surgical Procedure to Excise a Segment of Rabbit Jugular Vein for Ex Vivo Studies

Source: Nishio, H., et al. <a href="https://jove.unicartagenaproxy.elogim.com/v/60931">A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under Arterial Blood Pressure</a> J. Vis. Exp. (2020).
In this video, we demonstrate a procedure for surgically harvesting a segment of the jugular vein from a rabbit model. The resected segment can be used as an autologous vein graft to study intimal hyperplasia that develops due to arterial blood pressure.

ウサギモデルからの頸静脈移植片の収穫:ex vivo研究のためのウサギ頸静脈の一部を切除する外科的処置

Source: Nishio, H., et al. A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under ...

人間の伏在静脈のための手順

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中に組み込み血管新生による組織工学