TWP was funded by the Irish Cardiac Society Brian McGovern Travelling Fellowship. WTP was funded by R01 HL095712 and an AHA Established Investigator Award.

すべての説明する手順は、ボストン小児病院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の下で実施した。 の調製<ol><li>心筋内注射のための最適な固定トランスデューサ位置が解剖学と機能の評価の輪郭を描くための最適な標準のビューを得られない可能性がある、以前の注入プロトコルを開始する心エコー検査によるベースライン心臓解剖学的および機能評価を行います。 </li><li>注入のセットアップは、 図1A〜1Cに示されている。シリンジクランプで、ベベル指向上向き、シース針を空のシリンジを置きます（ 図1Cを参照）、（ 図1Bを参照）スキャンヘッドクランプに超音波トランスデューサ·プローブを固定します。スキャンヘッドクランプボールロックジョイント（ 図1B）をゆるめ、それが針の軸に平行に位置合わせされるように、変換器の向きを操作する。スキャを修正スキャンヘッドクランプボールロックジョイントを締めてヘッド位置。 注：成体マウスでの注射のために、1 / 2.5cmの長さが30のGの針が最適である。気密シリンジより少量（5〜10μl）をより正確に制御するために使用することができながら、1mlシリンジは、より大きな体積のために使用することができる。 </li><li>その全てに沿って頭をカバーするためにスパチュラでトランスデューサ先端に自由に超音波ゲルを適用します。慎重に針を鞘から抜くと針を直接変換器の下と可視化のための超音波ゲル内に針を前進させるためにコントロールをマウントしてください。針、針が超音波画像上でその長さに沿って明確に可視化されるように制御をマウントを使用して微調整を行います。トランスデューサが適切にステップ1.4において、針に平行に整列された場合は、前進及び噴射制御ノブ（ 図1C）に引き出されるように、次いで、針は、撮像面内にとどまるべきである。 </li><li>その後の工程では、必要性を乱すことがありません横軸のいずれかに移動させることにより、LE /トランスデューサ水平方向の配置。むしろ、マウント針の垂直（y軸）の位置を変更することにより、動物のプラットフォームを移動させることにより、注射用心臓の特定の領域を標的とする。 </li><li>動物プラットフォームに、麻酔したマウスのその後の配置を可能にするためにスキャン高さ制御（ 図1B）を用いて動物プラットフォームから上方にトランスデューサを移動します。これは、針の長軸にトランスデューサのx軸の位置合わせは中断されません。 </li><li>シリンジクランプから位置合わせに使用した注射器を取り外し、慎重に捨てる。シリンジチップにデッドスペースを可能にし、最終的なターゲットボリュームへの注入物を使用して新しい針と注射器をロードします。気泡を除去するように注意してください。そのx軸アライメントを調整することなく、シリンジクランプに注射器を置きます。完全噴射制御を使用して注射器を後退させる。 注：初期トレーニングのために、エバンスブルーの使用染料（1％）、トリパンブルー染色（0.4％）、または注入液のような蛍光マイクロスフェアの懸濁液をコンピテンシーと目標と注射の成功を確認したオペレータを支援することができます。 </li></ol> 2インジェクション<ol><li>加熱プラットフォームの統合ウォーマーの電源を入れ、37℃のためにそれを設定してください。麻酔ホースクランプ、オペレータに最も近い動物の頭で、通常の画像形成の向きから、動物のプラットホームを180°に配置します。これは（胸の左側にある）の心は、シリンジクランプと針と同側にすることができます。注：動物プラットフォームに追加のマイナー時計回りの回転調整は注射（ 図2A）のために使用される胸骨傍短軸像面に対して正しく心を配向する必要があるかもしれない。 </li><li>以前に9を報告したように心エコー検査のためにマウスを準備します。 2％のイソフルランで誘導チャンバ内でマウスを麻酔。脱毛クリームのANと胸毛を削除する強膜の乾燥を防ぐために両眼にゲルを適用する潤滑dは。 </li><li>スキャンヘッド高さ制御（ 図1B）を使用して、変換器を持ち上げます。 1〜3％イソフルラン（ 図2A-2B）を配信ノーズコーン内の鼻で加熱動物プラットフォームの上に、麻酔したマウスの仰臥位を置きます。ゆっくりと直腸プローブを挿入し、電気的に接触するための電極ゲルを適用し、ECG電極に4本の足をテープで固定します。 注：麻酔の適切なレベルは、動物の人道的な扱いのために確保されなければならない。心拍数に変化し、胸壁を通して針の配置に無応答でなければなりません。低体温症は相対徐脈、心室拡張、および可能な不快感になりますように、動物のプラットフォームの統合された恒常性、温度コントロールは、正常体温（37±0.5℃）を維持するために使用されるべきである。 </li><li>マウスは、動物のプラットフォーム上で安全であると、depila上にトランスデューサを下げるテッド胸部スキャンヘッド高さ制御（ 図1B）を使用して。心臓が標準の心エコー手法に従って、胸骨傍短軸方向で可視化されるために、注射のための最適な超音波のセットアップがある。を20-30°動物プラットフォームを回転させるには短軸像面内の注入のための最適な音響窓（ 図2A-2B）を得るために時計回り。注：あるいは、注入は、動物のプラットフォーム（ 図2C）の反時計回りの回転によって胸骨傍長軸の向きから行うことができる。 </li><li>視野を調整するために、左心室の心筋内の任意の所望の注射部位を標的とする動物プラットフォーム調整コントロールを使用する。左心室の心筋（ 図3A-3C）内の所望の噴射位置を標的とするために、心臓のベース頂部から前後パン。注：midpapillary胸骨傍短軸（ 図3A）が reproduciblを提供しています注射部位のフォローイメージングを可能にする電子ランドマーク。注意：代替的に、胸骨傍長軸ビューは、事前定義された注入部位（ 図3D）を標的化するために使用することができる。 </li><li>完全に後退したシリンジクランプで注射器から始めて、ゆっくりと噴射制御を時計回り（ 図4A）を回して、動物の胸に向けて注射器を前進させる。それは胸に近づくにつれて、心臓と針の先端の両方の明確な超音波可視化を可能にするには、胸の左側の上に超音波ゲルをたっぷり使用し、心エコーコントロールで広い視野を設定することで、音響窓を最適化します。注射のための標的部位での焦点/ゾーンを設定します。針の微調整は、コントロールは、その長さに沿って針の画像を最適化することができますマウント。 注：一部の超音波装置は、デジタルターゲット心筋へ通る針の長軸に沿った線を延長するニードルガイドソフトウェア機能を有している（ 図4B）。このようなソフトウェアツールが役立ちますが、必須ではないことができます。 </li><li>動物は適切に鎮静させて、慎重に全くベベル針の先端の位置を観察し、マウスの胸壁を通って心筋に針を前進させる（1〜3％イソフルラン0.5〜0.8 L /分で100％酸素と混合）回。針の先端がターゲット心筋（ 図4C）内にあるときに前進を停止する。全体ベベル先端が心膜腔に注入液漏れを避けるために、しっかりと心筋内になければならない。 </li><li>先端が目的の位置にあるとき、シリンジプランジャを押すことにより、注入物をお届けします。 5-30秒かけて、ゆっくりと注入液を配信（音量に応じて配信されている）。注入液を50μlまでの心室機能を損なうことなく送達することができる。注入された心筋領域への過渡echobright外観が成功し、注射後明らかであろう。相対bradycardの簡単な（秒）の期間iaが時折心筋への注入に留意し、迅速に解決されている。 </li><li>注入液を投与された後、速やかに噴射制御ノブの反時計回りの回転によって針を引き抜く。マウスは保存心室機能なしpostprocedural合併症を確認するために、心エコー観察の数分間麻酔下で維持されるべきである。示された場合は、心筋の複数の領域を連続動物プラットフォームの調整によってアプローチの針の角度の再配置によって注入することができる。心筋内注射後、マウスは、それ自身のケージに入れ、観察下に麻酔から回復させる。 </li><li>それは胸骨横臥位を維持するのに十分な意識を取り戻したまでは無人の動物を放置しないでください。完全に回復するまで、他の動物の会社に心筋内注射を受けた動物を返さないでください。ケージは、再を恒温パッド上に配置する必要があります水とマウスの食事のADY規定。 Postprocedural不快感が期待されておらず、障害が適切に（説明を参照）介入は潜在的な合併症を示唆した直後に通常の動作を再開すること。 </li><li>慎重に作業者や近くに鋭利器材損傷のリスクを最小限に抑えるために、すぐに心筋内注入が完了した後に針を捨てる。針を再利用することをより困難な心筋を貫通するようになっ先端の鈍化や合併症のリスクが高い、その結果をもたらす可能性がある。 </li></ol>

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The authors have nothing to disclose.

生物製剤は、血流を介して、直接心筋内注射、心膜内注射、または間接的な投与により心筋に送達することができる。心筋梗塞モデルにおける最近の細胞ベースの治療試験は、注入物12〜14の配信に開放開胸アプローチを記載している。心筋治療的介入の成功の重要な要因は、配信経路の選択にかかっている。生物学の最も高い局所的投与量は、心筋内デリバリー15,16によって...

心臓病は、毎年60万死者1を占め、米国の男性と女性の両方のための主要な死亡原因である。心血管疾患のマウスモデルは、病態生理学的メカニズムを調査するため、潜在的な治療法を探索するのに非常に重要である。遺伝子治療ベクターの心筋送達、幹細胞は、修飾RNA、および他の治療剤は、心臓病2-7に対するそれらの治療可能性の調査を可能にする。現在、マウスモデル6における治療薬の心筋の送達のための限られた選択肢があります。直接可視化の下で心筋注射が一般的に使用されますが、胸骨切開または開胸術を必要とし、心臓の露出領域に限定されているされている。そのようなLAD結紮などの別の実験操作時に行う際に便利な一方で、心筋内送達のための侵襲的処置の必要性が潜在的な実験デザインを制限し、紹介する手順（開胸術によるもので、 例えば線維症）からdditional効果。ウイルスベクターの経皮的心膜送達が報告されているが、サイトおよび治療 ​​剤の分布は均一ではなく、コントロール8に困難である。注入された材料のより均一な分布における経皮的冠動脈注入の結果が、効率的で再現性の冠動脈配信マウスモデルにおいて困難である。 ここでは、超音波ガイド下の治療薬の低侵襲性、オペレータ制御されたターゲティングを可能にする、閉じた胸の心筋内注入技術について説明します。技術は習得が容易で、開胸や胸骨切開とそれらに付随する実験的な合併症を不要、タイミングや心筋内注射部位に大きな柔軟性を提供します。このように、心エコー検査補助心筋内注入は、操作する技術的に簡単かつ非常に効果的な方法を示し、マウス実験モデルにおいて心筋。

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			<td height="21" style="height:21px;width:313px;">Vevo 2100 ultrasound imaging system&#160;</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;width:313px;">Vevo Integrated Rail System III</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS400 transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS550D transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">PrecisionGlide needles</td>
			<td style="width:144px;">BD</td>
			<td style="width:119px;">305128</td>
			<td>30G x 1 inch</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">1cc slip tip syringe</td>
			<td style="width:144px;">Exel International&#160;</td>
			<td style="width:119px;">26048</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Gastight 50ul glass syringes</td>
			<td style="width:144px;">Hamilton&#160;</td>
			<td style="width:119px;">1705</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Trypan blue stain (0.4%)</td>
			<td style="width:144px;">Gibco&#160;</td>
			<td style="width:119px;">15250</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Isoflurane</td>
			<td style="width:144px;">Baxter</td>
			<td style="width:119px;">AHN3640</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Aquasonic 100</td>
			<td style="width:144px;">Parker Laboratories</td>
			<td style="width:119px;">(01-08)</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Polystyrene microspheres (red fluorescent)</td>
			<td style="width:144px;">Life Technologies</td>
			<td style="width:119px;">F-8842</td>
			<td>or equivalent</td>
		</tr>
	</tbody>
</table>

ultrasound-guided transthoracic intramyocardial injection in mice

心血管疾患のマウスモデルは、病態生理学的メカニズムを調査し、潜在的な再生治療を探索するのに重要である。心筋の注入を含む実験は、現在、開胸を通じて直接外科的アクセスによって実行されます。そのような冠動脈結紮などの別の実験操作時に行う際に便利な一方で、心筋内送達のための侵襲的処置の必要性が潜在的な実験デザインを制限します。これまでの超音波の解像度と、高度な非侵襲イメージングモダリティを改善すると、それは日常的に超音波ガイド下、経皮的心筋内注射を行うことが今や可能である。この様式は、効率的かつ確実に心筋の標的領域にエージェントを提供します。この技術の利点は、外科的罹患率の回避、選択的に超音波ガイド下心筋の領域を標的とする機能、およびmyocardiuに注入液を送達する機会を含む複数、所定の時間間隔でmである。練習技術では、心筋内注射による合併症はまれであり、マウスはすぐに麻酔からの回復に通常の活動に戻る。このプロトコルで説明されている手順に続いて、基本的な心エコー検査の経験を持つオペレータは、すぐにこの汎用性、低侵襲性技術で有能になることができます。

青色染料または蛍光マイクロスフェアとマウス心筋内注射エバンスブルー色素の注入は、トレーニング目的のために有用である。すぐに注射した後、注入された青色色素の位置を可視化するために、マウスを安楽死させると心臓を取り外します。 図5は、青色色素半ば乳頭筋レベル（ 図5A、囲まれた領域での心筋浸潤して、成功した注入の例を示し...

Watch this Scientific Journal Video about Ultrasound-guided Transthoracic Intramyocardial Injection in Mice at JoVE.com

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

心エコーガイド下経皮的心筋内注入は、マウスの心臓への遺伝子導入剤又は細胞の送達のための、効率的で信頼性が高く、標的可能なモダリティを表しています。このプロトコルで説明されている手順に続いて、オペレータは、すぐにこの汎用性、低侵襲性技術で有能になることができます。

マウスにおける超音波ガイド下経胸壁心筋内注射

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. ...

ultrasound-guided-transthoracic-intramyocardial-injection-in-mice

Research

JoVE Journal

Medicine

14.8K ビュー.  Boston Children's Hospital. 心エコーガイド下経皮的心筋内注入は、マウスの心臓への遺伝子導入剤又は細胞の送達のための、効率的で信頼性が高く、標的可能なモダリティを表しています。このプロトコルで説明されている手順に続いて、オペレータは、すぐにこの汎用性、低侵襲性技術で有能になることができます。 

ビデオ: Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

マウスにおける経胸壁心エコー

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

医学

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

心筋梗塞のマウスモデルにおける冠動脈結紮および心筋内注入

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

Delivery of Therapeutic Agents Through Intracerebroventricular (ICV) and Intravenous (IV) Injection in Mice

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most often results in failure of potential therapeutics designed for neurodegenerative disorders 1,2. Neurodegenerative diseases such as Spinal Muscular Atrophy (SMA), in which the lower motor neurons are affected, can benefit greatly from introducing the therapeutic agents into the CNS. The purpose of this video is to demonstrate two different injection paradigms to deliver therapeutic materials into neonatal mice soon after birth. One of these methods is injecting directly into cerebral lateral ventricles (Intracerebroventricular) which results in delivery of materials into the CNS through the cerebrospinal fluid 3,4. The second method is a temporal vein injection (intravenous) that can introduce different therapeutics into the circulatory system, leading to systemic delivery including the CNS 5. Widespread transduction of the CNS is achievable if an appropriate viral vector and viral serotype is utilized. Visualization and utilization of the temporal vein for injection is feasible up to postnatal day 6. However, if the delivered material is intended to reach the CNS, these injections should take place while the blood brain barrier is more permeable due to its immature status, preferably prior to postnatal day 2. The fully developed blood brain barrier greatly limits the effectiveness of intravenous delivery. Both delivery systems are simple and effective once the surgical aptitude is achieved. They do not require any extensive surgical devices and can be performed by a single person. However, these techniques are not without challenges. The small size of postnatal day 2 pups and the subsequent small target areas can make the injections difficult to perform and initially challenging to replicate.

Delivery of Therapeutic Agents Through Intracerebroventricular ICV and Intravenous IV Injection in Mice

This article demonstrates two very different methods of injection: 1) into the brain (intracerebroventricular) and 2) systemic (intravenous) to introduce the therapeutic agents into the central nervous system of neonatal mice.

マウスの脳室内（ICV）と静脈（IV）注入による治療薬のデリバリー

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most ...

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related hospitalizations and for which no effective treatments are available to date1,2. Because of the heterogeneity of ICH in humans, various preclinical models are needed to thoroughly explore prospective therapeutic strategies3. Experimental ICH is commonly induced in rodents by intraparenchymal injection of either autologous blood or bacterial collagenase4. The appropriate model is selected based on the pathophysiology of hemorrhage induction and injury progression. The blood injection model mimics a rapidly progressing hemorrhage. Alternatively, bacterial collagenase enzymatically disrupts the basal lamina of brain capillaries, causing an active bleed that generally evolves over several hours5. Resultant perihematomal edema and neurofunctional deficits can be quantified from both models. In this study, we described and evaluated a modified double injection model of autologous whole blood6 as well as an ICH injection model of bacterial collagenase7, both of which target the basal ganglia (corpus striatum) of male CD-1 mice. We assessed neurofunctional deficits and brain edema at 24 and 72 hr after ICH induction. Intrastriatal injection of autologous blood (30 &mu;l) or bacterial collagenase (0.075U) caused reproducible neurofunctional deficits in mice and significantly increased brain edema at 24 and 72 hr after surgery (p&lt;0.05). In conclusion, both models yield consistent hemorrhagic infarcts and represent basic methods for preclinical ICH research.

Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.

マウスにおけるモデリング脳出血：自己血または細菌性コラゲナーゼの注入

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related ...

Intramyocardial Cell Delivery: Observations in Murine Hearts

Previous studies showed that cell delivery promotes cardiac function amelioration by release of cytokines and factors that increase cardiac tissue revascularization and cell survival. In addition, further observations revealed that specific stem cells, such as cardiac stem cells, mesenchymal stem cells and cardiospheres have the ability to integrate within the surrounding myocardium by differentiating into cardiomyocytes, smooth muscle cells and endothelial cells.
Here, we present the materials and methods to reliably deliver noncontractile cells into the left ventricular wall of immunodepleted mice. The salient steps of this microsurgical procedure involve anesthesia and analgesia injection, intratracheal intubation, incision to open the chest and expose the heart and delivery of cells by a sterile 30-gauge needle and a precision microliter syringe.
Tissue processing consisting of heart harvesting, embedding, sectioning and histological staining showed that intramyocardial cell injection produced a small damage in the epicardial area, as well as in the ventricular wall. Noncontractile cells were retained into the myocardial wall of immunocompromised mice and were surrounded by a layer of fibrotic tissue, likely to protect from cardiac pressure and mechanical load.

Intramyocardial cell delivery in murine models of cardiovascular diseases, such as hypertension or myocardial infarction, is widely used to test the therapeutic potential of different cell types in regenerative studies. Therefore, a detailed description and a clear visualization of this surgical procedure will help to define the limits and advantages of cardiovascular cell therapeutic analyses in small rodents.

心筋内細胞送達：マウスハーツの観測

Previous studies showed that cell delivery promotes cardiac function amelioration by release of cytokines and factors that increase cardiac tissue ...

Substernal Thyroid Biopsy Using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the lack of a standardized definition for STG. Biopsy is often required to differentiate benign from malignant lesions. Unlike cervical thyroid, the overlying sternum precludes ultrasound-guided percutaneous fine needle aspiration of STG. Consequently, surgical mediastinoscopy is performed in the majority of cases, causing significant procedure related morbidity and cost to healthcare. Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA) is a frequently used procedure for diagnosis and staging of non-small cell lung cancer (NSCLC). Minimally invasive needle biopsy for lesions adjacent to the airways can be performed under real-time ultrasound guidance using EBUS. Its safety and efficacy is well established with over 90% sensitivity and specificity. The ability to perform EBUS as an outpatient procedure with same-day discharges offers distinct morbidity and financial advantages over surgery. As physicians performing EBUS gained procedural expertise, they have attempted to diversify its role in the diagnosis of non-lymph node thoracic pathologies. We propose here a role for EBUS-TBNA in the diagnosis of substernal thyroid lesions, along with a step-by-step protocol for the procedure.

Substernal thyroid lesions are common, and need to be differentiated from malignancy. Obtaining percutaneous fine needle biopsy is not possible due to its retrosternal location. This article proposes a protocol for biopsy of substernal thyroid lesions using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA).

気管支超音波ガイド下経気管支穿刺吸引を使用して、胸骨下甲状腺生検

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the ...

Ultrasound-guided Botulinum Toxin-A Injections: A Method of Treating Sialorrhea

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an impaired quality of life, often worsened by correlated complications such as aspiration pneumonia, oral infections, dental caries, and maceration of the skin. Diverse therapeutic approaches have been proposed for the treatment of sialorrhea, including surgery and the use of anticholinergic agents, with limited results and the possible occurrence of serious adverse events. Recently, botulinum toxin (BoNT) injection within the major salivary glands has been proposed in patients refractory to anticholinergic therapy, with the aim of inhibiting local acetylcholine release and gland activity. 
In order to obtain a better outcome in terms of reduction of saliva production, efficacy, duration, and avoidance of major adverse events, we developed an ultrasound-guided BoNT-type A injection technique accurately described in the text. Here we present a method of treating sialorrhea with bilateral parotid and submandibular gland BoNT-type A injections under ultrasound guidance. Four quadrants of the parotid gland and two quadrants of the submandibular gland are visualized and injected using two accesses and one access, respectively. 
The ultrasound-guided procedure provides a simple, non-invasive, real-time visualization of the muscular and glandular tissues and their surrounding structures, optimizing treatment efficacy and safety.

Botulinum toxin is injected into the salivary glands for the treatment of sialorrhea. The ultrasound-guided botulinum toxin injection in both the parotid and submandibular glands demonstrates a sustained efficacy and the absence of serious side effects.

ボツリヌス毒素Aの注射を超音波誘導：唾液分泌過多を治療する方法

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an ...

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness 1,3,4,5. Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn&#39;s disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNF&#916;RE6. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.

Murine studies in models of colonic inflammation have demonstrated that a small percentage (1 - 5%) of mesenchymal stem cells (MSC) injected intravenously or intraperitoneally home to the inflamed colon1,2. This study shows that ultrasound-guided intracardiac injections of MSCs result in increased localization to the intestine.

実験的炎症性腸病気のマウスモデルで使用するため、腸管へのホーミングを増加するヒト間葉系幹細胞の超音波ガイド下心臓内注射

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Percutaneous Contrast Echocardiography-guided Intramyocardial Injection and Cell Delivery in a Large Preclinical Model

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection fraction (HFrEF). Before they can be considered for use, and implemented in humans, extensive preclinical studies are required in large animal models to evaluate the safety, efficacy, and fate of the injectate (e.g., stem cells) once delivered into the myocardium. Small rodent models offer advantages (e.g., cost effectiveness, amenability for genetic manipulation); however, given inherent limitations of these models, the findings in these rarely translate into the clinic. Conversely, large animal models such as rabbits, have advantages (e.g., similar cardiac electrophysiology compared to humans and other large animals), whilst retaining a good cost-effective balance. Here, we demonstrate how to perform a percutaneous contrast echocardiography-guided intramyocardial injection (IMI) technique, which is minimally invasive, safe, well tolerated, and very effective in the targeted delivery of injectates, including cells, into several locations within the myocardium of a rabbit model. For the implementation of this technique, we also have taken advantage of a widely available clinical echocardiography system. After putting in practice the protocol described here, a researcher with basic ultrasound knowledge will become competent in the performance of this versatile and minimally invasive technique for routine use in experiments, aimed at hypothesis testing of the capabilities of cardiac regenerative therapeutics in the rabbit model. Once competency is achieved, the whole procedure can be performed within 25 min after anaesthetizing the rabbit.

Novel therapeutic strategies in cardiac regenerative medicine require extensive and detailed studies in large preclinical animal models before they can be considered for use in humans. Here, we demonstrate a percutaneous contrast echocardiography-guided intramyocardial injection technique in rabbits, which is valuable for hypothesis testing the efficacy of such novel therapies.

経皮的造影エコー ガイド心筋と大規模な前臨床モデルで携帯配信

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

ウサギモデルにおける経胸部心エコー検査

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...