The authors have no acknowledgements.

これらのプロトコルは、当院の人間研究倫理委員会のガイドラインに従ってください。 1.従来の角膜コラーゲンCXL（C-CXL） 患者の調製<ol><li>手術前の5日は、ピロカルピンは、処置した眼で一日二回低下1％を置きます。 </li><li>手術室では、無菌条件下で、彼/彼女の背中に患者を横になります。 </li><li>このようなオキシブプロカイン0.4％などの局所麻酔を管理します。 </li><li>目と二回消毒ヨウ素と目の周りの皮膚をきれいにしてください。 </li><li>開いている目を保つために蓋鏡を使用してください。 </li></ol> 2.上皮の除去<ol><li>マーク・サークル角膜マーカーで角膜の中央9.0ミリメートル。 </li><li>鈍いスパチュラを使用して機械的デブリドマンによって角膜上皮の中央7.0〜9.0ミリメートルを削除します。 </li></ol> 3.リボフラビンアプリケーション<ol><li>目に20％デキストランでリボフラビン0.1％を適用します<html> Eの角膜20分間毎分。 </li></ol> 4. UVA照射<ol><li> 3ミリワット/ cm 2の（5.4 J / cm 2の表面線量）の照度で30分間5cmの作動距離で370 nmの波長のUVA光を角膜に照射。 </li></ol><img alt="図1" src="/files/ftp_upload/53119/53119fig1.jpg" /> 図1：C-CXL中でUVA照射角膜は、3ミリワット/ cm 2の（5.4 J / cm 2の表面線量）の照度で30分間、5 cmの作動距離で370 nmの波長のUVA光を照射します。 <a href="https://www.jove.com/files/ftp_upload/53119/53119fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <ol start="2"><li>照射中、角膜に5分ごとリボフラビンの滴を適用します。 </li><li>照射中、局所麻酔（オキシブプロカイン0.4％）は必要に応じて追加します。 </li></ol>外科 &quot;&gt; 5。終了<ol><li>入れ抗生物質滴（トブラマイシン0.3％）、運営の目に人工涙液（ヒアルロン酸0.18％低下）。 </li><li>再上皮が完了するまで、手術の終了時に、ソフト包帯コンタクトレンズを配置します。再上皮化は、通常は3日かかります。 </li><li>そのようなパラセタモール（500 mg）を加えたコデイン（30 mg）を、6丸薬日と鎮痛剤を処方します。 </li><li>角膜上皮再形成した後、ステロイドと局所療法（局所デキサメタゾン1 mg / mlの）を開始し、3〜4週間継続します。さらに、1月の人工涙液を1日4回を使用しています。 </li></ol> 2.加速角膜コラーゲンCXL（ - CXL） 患者の調製<ol><li>手術前の5日は、ピロカルピンは、処置した眼で一日二回低下1％を置きます。 </li><li>手術室では、無菌条件下で、彼/彼女の背中に患者を横になります。 </li><li>このようなオキシブプロカイン0.4％などの局所麻酔を管理します。 </li><li>電子メールをきれいにあなたがた二回消毒ヨウ素と目の周りの皮膚。 </li><li>開いている目を保つために蓋鏡を使用してください。 </li></ol> 2.上皮の除去<ol><li>マーク・サークル角膜マーカーで角膜の中央9.0ミリメートル</li><li>鈍いスパチュラを使用して機械的デブリドマンによって角膜上皮の中央7.0〜9.0ミリメートルを削除します。 </li></ol> 3.リボフラビンアプリケーション<ol><li> 10分毎に2分角膜上デキストランなしリボフラビン0.1％を適用します。 </li></ol> 4. UVA照射<ol><li> 30 mWの/ cm 2の（5.4 J / cm 2の表面線量）の照度で3分間5cmの作動距離で370 nmの波長のUVA光を角膜に照射。 </li><li> （oxybuproca＆＃239; 0.4％NE）照射中、局所麻酔を追加し、必要に応じて。 </li></ol>外科5.終了<ol><li>抗生物質滴（0.3％トブラマイシン）と人工涙液を配置（ヒアルロン酸が0をドロップ<html>操作の目に0.18パーセント）。 </li><li>再上皮が完了するまで、手術の終了時に、ソフト包帯コンタクトレンズを配置します。再上皮化は、通常は3日かかります。 </li><li>そのようなパラセタモール（500 mg）を加えたコデイン（30 mg）を、6丸薬日と鎮痛剤を処方します。 </li><li>角膜上皮再形成した後、ステロイドと局所療法（局所デキサメタゾン1 mg / mlの）を開始し、3〜4週間継続します。さらに、1月の人工涙液を1日4回を使用しています。 </li></ol> 3.イオントフォレーシス（I-CXL） 患者の調製<ol><li>手術前の5日は、ピロカルピンは、処置した眼で一日二回低下1％を置きます。 </li><li>手術室では、無菌条件下で、彼/彼女の背中に患者を横になります。 </li><li>このようなオキシブプロカイン0.4％などの局所麻酔を管理します。 </li><li>目と二回消毒ヨウ素と目の周りの皮膚をきれいにしてください。 </li><li>開いている目を保つために蓋鏡を使用してください。 </li></ol><html> _step &quot;&gt; 2。イオントフォレーシス装置を配置します。 <ol><li>術野の下で額に粘着性のパッシブ電極を適用します。 </li><li>開いた目に、活性電極、吸引リングを適用します。吸引をリリースする前に、角膜の周囲に吸引リングを中央に配置します。 </li></ol><img alt="図2" src="/files/ftp_upload/53119/53119fig2.jpg" /> 図2.イオントフォレーシス装置。受動電極は、術野の下で額と活性電極、吸引リング、開いた目に適用されるに適用されます。 <a href="https://www.jove.com/files/ftp_upload/53119/53119fig2large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> 3.リボフラビンアプリケーション<ol><li>デキストランのない低浸透圧0.1％のリボフラビンと吸引リングを入力します。 </li></ol><html> JPG &quot;/&gt; I-CXL図3.リボフラビンアプリケーション 。吸引リングは、デキストランずに低浸透圧0.1％のリボフラビンが充填されている。 <a href="https://www.jove.com/files/ftp_upload/53119/53119fig3large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <ol start="2"><li> 0.2ミリアンペアでの電流をスタートし、徐々に5分（ 図4）の総イオン導入時間を1.0ミリアンペアに増加します。 </li></ol><img alt="図4" src="/files/ftp_upload/53119/53119fig4.jpg" /> 浸透リボフラビン図4.イオントフォレーシス装置。電流は、最初は0.2ミリアンペアで、徐々に1.0ミリアンペアに増加しました。総イオン導入時間は5分である。 <a href="https://www.jove.com/files/ftp_upload/53119/53119fig4large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> EP &quot;&gt; 4。UVA照射<ol><li> 10ミリワット/ cm 2の（5.4 J / cm 2の表面線量）と9分間5cmの作動距離での放射照度で370 nmの波長のUVA光で角膜を照射。 </li><li> （oxybuproca＆＃239; 0.4％NE）照射中、局所麻酔を追加し、必要に応じて。 </li></ol>外科5.終了<ol><li>抗生物質滴（トブラマイシン0.3％）、運営の目に人工涙液を（ヒアルロン酸0.18％低下）を配置します。 </li><li>そのようなパラセタモール（500 mg）を加えたコデイン（30 mg）を、6丸薬日と鎮痛剤を処方します。 </li><li>手術後、ステロイドと局所療法（局所デキサメタゾン1mgの/ ml）を開始し、3〜4週間継続します。さらに、1月の人工涙液を1日4回を使用しています。 </li></ol>

<ol>
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K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+corneal+endothelial+changes+following+accelerated+collagen+cross-linking+for+the+treatment+of+progressive+keratoconus.">Transient corneal endothelial changes following accelerated collagen cross-linking for the treatment of progressive keratoconus.</a> Cutan Ocul Toxicol. 33 (2), 127-131 (2013).</li><li>Spoerl, E., Mrochen, M., Sliney, D., Trokel, S., Seiler, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+UVA-riboflavin+cross-linking+of+the+cornea.">Safety of UVA-riboflavin cross-linking of the cornea.</a> Cornea. 26 (4), 385-389 (2007).</li><li>Gokhale, N. 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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacokinetics+and+safety+of+transcorneal+iontophoresis+of+tobramycin+in+the+rabbit.">Pharmacokinetics and safety of transcorneal iontophoresis of tobramycin in the rabbit.</a> Invest Ophthalmol Vis Sci. 29 (9), 1397-1401 (1998).</li><li>Vinciguerra, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transepithelial+iontophoresis+corneal+collagen+cross-linking+for+progressive+keratoconus:+initial+clinical+outcomes.">Transepithelial iontophoresis corneal collagen cross-linking for progressive keratoconus: initial clinical outcomes.</a> J Refract Surg. 30 (11), 746-753 (2014).</li><li>Caporossi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Riboflavin-UVA-induced+corneal+collagen+cross-linking+in+pediatric+patients.">Riboflavin-UVA-induced corneal collagen cross-linking in pediatric patients.</a> Cornea. 31 (3), 227-231 (2012).</li><li>Buzzonetti, L., Petrocelli, G., Valente, P., Larossi, G., Ardia, R., Petroni, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iontophoretic+transepithelial+corneal+cross-linking+to+halt+keratoconus+in+pediatric+cases:+15-month+follow-up.">Iontophoretic transepithelial corneal cross-linking to halt keratoconus in pediatric cases: 15-month follow-up.</a> Cornea. 34 (5), 512-515 (2015).</li><li>Baiocchi, S., Mazzotta, C., Cerretani, D., Caporossi, T., Caporossi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+crosslinking:+riboflavin+concentration+in+corneal+stroma+exposed+with+and+without+epithelium.">Corneal crosslinking: riboflavin concentration in corneal stroma exposed with and without epithelium.</a> J Cataract Refract Surg. 35 (5), 893-899 (2009).</li><li>Wollensak, G., Iomdina, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanical+and+histological+changes+after+corneal+crosslinking+with+and+without+epithelial+debridement.">Biomechanical and histological changes after corneal crosslinking with and without epithelial debridement.</a> J Cataract Refract Surg. 35 (3), 540-546 (2009).</li><li>Soeters, N., Wisse, R. P., Godefrooij, D. A., Imhof, S. M., Tahzib, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transepithelial+versus+epithelium-off+corneal+cross-linking+for+the+treatment+of+progressive+keratoconus:+a+randomized+controlled+trial.">Transepithelial versus epithelium-off corneal cross-linking for the treatment of progressive keratoconus: a randomized controlled trial.</a> Am J Ophthalmol. 159 (5), 821-828 (2015).</li></ol>

The authors have nothing to disclose.

UVA照射及びリボフラビンを用いCXLは、円錐角膜の進行を阻止するための標準的な治療です。リボフラビンは、UVA 3を照射したときの化学共有結合（架橋）を誘導光増感剤です。角膜では、この現象は、角膜の剛性を高めるコラーゲン原線維間の架橋を生成します。この現象はよく記載されているが、今まで角膜内架橋の直接的な証拠はなかったです。それにもかかわらず、いくつか?...

円錐角膜は通常、角膜形状の変更の結果、一般集団1に2000で1に報告二国間及びプログレッシブ角膜拡張であるため、ビジョン2を減少させました 。円錐角膜は通常、早期思春期に存在し、進行は、患者の生涯にわたって可変とすることができるが、この疾患は、通常、安定化する傾向があるとき、人生の第四十年に第三まで進行します。円錐角膜の進行を停止させることにより、架橋は、延期または角膜移植を回避することを目的としています。 現在まで、臨床試験で証明された進歩的な円錐角膜の唯一の効率的かつ安全な治療は、剛性を高めるので、円錐角膜の進行3-8を停止することを目指し、従来の角膜コラーゲン架橋（C-CXL）プロトコルです。動作時間などの感染性角膜炎または間質ヘイズ9と C-CXLの他の可能な危険因子を減らすために、いくつかの改良されたプロトコルを持っています記載されて。まず、加速CXL（A-CXL）は、UVAのより高い放射照度は減少し、時間10にわたって角膜に送達されます。第二に、上皮の創面切除の必要性を回避するために、経上皮のアプローチが採用されています。従来のプロトコル11と比較した場合、残念ながら、それらは成功が限られています。 CXL中角膜リボフラビン送達のための最新の方法は、経上皮イオントフォレーシス（I-CXL）であるが、この治療法の厳密な評価はまだ12に行われていません。イオントフォレーシスは、小電流が組織を通ってイオン化された薬物の浸透を改善するために適用された非侵襲的技術です。イオントフォレーシスによりCXLでは、リボフラビンは上皮を介して角膜を貫通するようにイオン化されます。 生体内では、共焦点顕微鏡（IVCM）は、円錐角膜13のような疾患における異常な角膜の細胞の変化を強調することができ、角膜を画像化する方法です。実際、IVCMサブ基底神経叢および間質角膜実質13〜15の密度の特定の減少と円錐角膜で角膜のすべての層に変化を実証してきました。さらに、IVCMは、C-CXL 16後の角膜の微細構造解析のために非常に便利であることが証明されました。 角膜境界線は300ミクロン17,18の深さで、C-CXL後1ヶ月（10月AS）前眼部光干渉断層撮影で見hyperreflectiveラインとして記載されています。 IVCM次のC-CXLは、300ミクロンの深さに角膜実質細胞の不在を含む角膜の構造変化に関する情報を提供します。この無細胞ゾーンの深さだけでなく、角膜実質内の境界線の深さは、10月1日としてCXL処理19、及び角膜境界線の深さの測定の有効深さに関連すると思われる10月AS上で明らかにしましたCXLは、効率的な臨床として提案された後月CXL効果18の評価のための方法。 本研究では、10月、共焦点顕微鏡ASによって角膜実質境界線の測定を使用して、3つの異なる（従来、加速され、イオン導入）角膜コラーゲン架橋のプロトコルの効率を調査します。我々は、さらに定量的に3治療後の角膜の微細構造の変化を分析するためにIVCMを使用していました。

<table>
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td>Riboflavin&#160;&#160;&#160;&#160;&#160;&#160;&#160; Product number</td>
			<td></td>
		</tr>
		<tr>
			<td>C-CXL</td>
			<td>Sooft SPA, Montegiorgio, Italy</td>
			<td>Ricrolin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 468465-6</td>
			<td></td>
		</tr>
		<tr>
			<td>A-CXL</td>
			<td>Avedro Inc, Waltham, Massachusetts</td>
			<td>VibeX&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 520-01863-006</td>
			<td></td>
		</tr>
		<tr>
			<td>I-CXL</td>
			<td>Sooft SPA, Montegiorgio, Italy</td>
			<td>Ricrolin+&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 975481-6</td>
			<td>Passive electrode: PROTENS ELITE 4848LE/ Active electrode: IONTOFOR CXL</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td>UVA Machine</td>
			<td></td>
		</tr>
		<tr>
			<td>X-Vega</td>
			<td></td>
			<td></td>
			<td>UVA: 3 mW/cm2 30 min</td>
		</tr>
		<tr>
			<td>KXL System</td>
			<td></td>
			<td></td>
			<td>UVA: 30 mW/cm2 10 min</td>
		</tr>
		<tr>
			<td>X-Vega</td>
			<td></td>
			<td></td>
			<td>UVA: 10 mW/cm2 9 min</td>
		</tr>
	</tbody>
</table>

three different protocols of corneal collagen crosslinking in keratoconus: conventional, accelerated and iontophoresis

円錐角膜は、二国間及び進歩的な角膜拡張症です。その進行を遅くするために、角膜コラーゲン架橋（CXL）は最近、効率的な治療オプションとして導入されました。生物学的および化学的科学では、架橋は、反応性分子との間に形成された新たな化学結合を指します。したがって、角膜コラーゲンCXLの目的は、合成角膜基質中のコラーゲン原線維間の架橋の形成を増加させることです。従来のCXL（C-CXL）プロトコルの効率は、既にいくつかの臨床研究で示されているという事実にもかかわらず、それは、プロシージャと角膜上皮の除去の持続時間の改善から利益を得るかもしれません。したがって、二つの新しい及び最適化されたCXLプロトコルのコヒーレントな評価を提供するために、我々三CXL治療の受けていた円錐角膜の患者検討：イオン導入（I-CXL）を、CXL（-CXL）を加速し、そして従来のCXL（ C-CXL）。 -CXLは6時間より早くCXL手順Uです10時間、より高いUVA放射照度を歌うが、それでも上皮の除去を含みます。イオントフォレーシスは、小電流が角膜を通じてリボフラビン浸透を改善するために適用された上皮の非侵襲的技術です。 （OCT）およびインビボ共焦点顕微鏡（IVCM） に前部光コヒーレンストモグラフィを用いて、治療の浸透の深さについては、従来のCXLプロトコルはプログレッシブ円錐角膜を治療するための標準的なままであることを結論付けています。加速CXLは薄い角膜を治療するために、迅速、効果的かつ安全な代替手段であると思われます。イオントフォレーシスの使用はまだ調査されている、より大きな慎重に検討する必要があります。

<html>角膜境界線は301.6ミクロン（SD、73.6）の平均深さで例92％で10月ASに見えました<img alt="図5" src="/files/ftp_upload/53119/53119fig5.jpg" /> 358ミクロン（白矢印）の平均深さで角膜実質境界線を可視化（10月AS） のC-CXL。高解像度角膜前眼部光干渉断層撮影スキャン後の図5.境界線 、従来の角膜コラーゲンクロス後1ヶ月リン?...

Watch this Scientific Journal Video about Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis at JoVE.com

Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis

Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL).

円錐角膜で角膜コラーゲン架橋の三つの異なるプロトコル：従来は、加速してイオントフォレーシス

Keratoconus is a bilateral and progressive corneal ectasia. In order to slow down its progression, corneal collagen cross-linking (CXL) has recently been ...

three-different-protocols-corneal-collagen-crosslinking-keratoconus

Research

JoVE Journal

Medicine

19.8K ビュー.  Quinze-Vingts National Ophthalmology Hospital. Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL).

ビデオ: Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis

Staining Protocols for Human Pancreatic Islets

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. 
Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region.
The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&amp;E and IHC stains. 
The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections.

This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&amp;E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.

ヒト膵島の染色プロトコル

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and ...

医学

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the cornea, which can be caused by trauma, keratoplasty or infectious disease, breaks down the so called &lsquo;angiogenic privilege' of the cornea and forms the basis of multiple visual pathologies that may even lead to blindness. Although there are several treatment options available, the fundamental medical need presented by corneal neovascular pathologies remains unmet. In order to develop safe, effective, and targeted therapies, a reliable model of corneal NV and pharmacological intervention is required. Here, we describe an alkali-burn injury corneal neovascularization model in the mouse. This protocol provides a method for the application of a controlled alkali-burn injury to the cornea, administration of a pharmacological compound of interest, and visualization of the result. This method could prove instrumental for studying the mechanisms and opportunities for intervention in corneal NV and other neovascular disorders.

Neovascularization (NV) of the cornea can complicate multiple visual pathologies. Utilizing a controlled, alkali-burn injury model, a quantifiable level of corneal NV can be produced for mechanistic study of corneal NV and evaluation of potential therapies for neovascular disorders.

マウス角膜血管新生のアルカリ熱傷損傷モデル

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the ...

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes &#8211; but is not limited to &#8211; hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

従来のクローズド·チェスト法による心室細動と蘇生のラットモデル

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in rodents, with detailed descriptions of the appropriate methods for anesthesia, the echocardiographic windows used, the imaging planes and probe orientations for image acquisition, the methods for data analysis, and the limitations of emerging technologies for the evaluation of cardiac and valvular function. Importantly, we also highlight several future areas of research in cardiac and heart valve imaging that may be leveraged to gain insights into the pathogenesis of valve disease in preclinical animal models. We propose that using a systematic approach to evaluating cardiac and heart valve function in mice can result in more robust and reproducible data, as well as facilitate the discovery of previously underappreciated phenotypes in genetically-altered and/or physiologically-stressed mice.

This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice.

マウスの弁膜症の包括的表現型の特徴のための心エコーアプローチとプロトコル

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in ...

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

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

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

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

マウスにおける加速アテローム性動脈硬化症の誘導:ワイヤー傷害モデル

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

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

角膜上皮摩耗眼はりのモデルとして角膜創傷治癒

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The appearance of a mass on certain magnetic resonance (MR) sequences can help differentiate a thrombus from competing diagnoses such as a tumor. T1 and T2 signal characteristics of a thrombus are related to the evolution of hemoglobin properties. A thrombus typically does not enhance following contrast administration, which also helps differentiation from a tumor. We also highlight the emerging role of T1 mapping in the evaluation of a thrombus, which can add another level of support in diagnosis. Prior to any CMR exam, patient screening and interviews are critical to ensure safety and to optimize patient comfort. Effective communication during the exam between the technologist and the patient promotes proper breath holding technique and higher quality images. Volumetric post processing and structured reporting are helpful to ensure that the radiologist answers the ordering services&#39; question and communicates these results effectively. Optimal pre-MR safety evaluation, CMR exam execution, and post exam processing and reporting allow for delivery of high quality radiological service in the evaluation of a suspected cardiac thrombus.

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

疑わしい心臓血栓の評価のための心臓磁気共鳴:従来技術と新興技術

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...

Simultaneous Laryngopharyngeal and Conventional Esophageal pH Monitoring

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, hoarseness or asthma, which can be caused by laryngopharyngeal reflux (LPR). Due to their multifactorial origin, those symptoms can be a great diagnostic and therapeutic challenge. Esophageal pH-monitoring is commonly used to determine abnormal esophageal acid exposure and confirm the diagnosis of GERD. However, for better evaluation of acid exposure above the upper esophageal sphincter, a new laryngopharyngeal pH measurement system is now available and may lead to more reliable results in patients with predominantly extraesophageal symptoms. This article aims to present a standardized protocol for simultaneous pH measurement using esophageal and laryngopharyngeal pH probes in order to obtain acid exposure scores from both measurements.

Laryngopharyngeal pH monitoring has been specifically designed to measure acid exposure above the upper esophageal sphincter and complements diagnostic evaluation in patients that present with mainly extraesophageal reflux symptoms. Patients with suspected laryngopharyngeal reflux (LPR) were evaluated using distal esophageal and laryngopharyngeal pH testing simultaneously.

喉頭咽頭と従来の食道pHモニタリングの同時

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, ...

Three-Dimensional Collagen Matrix Scaffold Implantation as a Liver Regeneration Strategy

Liver diseases are the leading cause of death worldwide. Excessive alcohol consumption, a high-fat diet, and hepatitis C virus infection promote fibrosis, cirrhosis, and/or hepatocellular carcinoma. Liver transplantation is the clinically recommended procedure to improve and extend the life span of patients in advanced disease stages. However, only 10% of transplants are successful, with organ availability, presurgical and postsurgical procedures, and elevated costs directly correlated with that result. Extracellular matrix (ECM) scaffolds have emerged as an alternative for tissue restoration. Biocompatibility and graft acceptance are the main beneficial characteristics of those biomaterials. Although the capacity to restore the size and correct function of the liver has been evaluated in liver hepatectomy models, the use of scaffolds or some kind of support to replace the volume of the extirpated liver mass has not been assessed.
Partial hepatectomy was performed in a rat liver with the xenoimplantation of a collagen matrix scaffold (CMS) from a bovine condyle. Left liver lobe tissue was removed (approximately 40%), and an equal proportion of CMS was surgically implanted. Liver function tests were evaluated before and after the surgical procedure. After days 3, 14, and 21, the animals were euthanized, and macroscopic and histologic evaluations were performed. On days 3 and 14, adipose tissue was observed surrounding the CMS, with no clinical evidence of rejection or infection, as was vessel neoformation and CMS reabsorption at day 21. There was histologic evidence of an insignificant inflammation process and migration of adjacent cells to the CMS, observed with the hematoxylin and eosin (H&#38;E) and Masson&#39;s trichrome staining. The CMS was shown to perform well in liver tissue and could be a useful alternative for studying tissue regeneration and repair in chronic liver diseases.

Liver diseases are induced by many causes that promote fibrosis or cirrhosis. Transplantation is the only option for recovering health. However, given the scarcity of transplantable organs, alternatives must be explored. Our research proposes the implantation of collagen scaffolds in liver tissue from an animal model.

肝再生戦略としての三次元コラーゲンマトリックス足場移植

Liver diseases are the leading cause of death worldwide. Excessive alcohol consumption, a high-fat diet, and hepatitis C virus infection promote fibrosis, ...

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, and result in a large, persistent corneal epithelial defect. Epithelial defect with the following corneal opacity and peripheral neovascularization result in irreversible visual impairment and hinder future management, especially keratoplasty. Since the animal model can be used as an effective drug development platform, models of corneal injury to the mouse and alkali burn to rabbit corneal epithelium are developed here. New Zealand white rabbit is used in the alkali burn model. Different concentrations of sodium hydroxide can be applied onto the central circular area of the cornea for 30 s under intramuscular and topical anesthesia. After copious isotonic normal saline irrigation, residual loose corneal epithelium was removed with corneal burr deep down to the Bowman's layer within this circular area. Wound healing was documented by fluorescein staining under Cobalt blue light. C57BL/6 mice were used in the traumatic model of murine corneal epithelium. The murine central cornea was marked using a skin punch, 2 mm in diameter, and then debrided by a corneal rust ring remover with a 0.5 mm burr under a stereomicroscope. These models can be prospectively used to validate the therapeutic effect of eye drops or mixed agents such as stem cells, which potentially facilitate corneal epithelial regeneration. By observing corneal opacity, peripheral neovascularization, and conjunctival congestion with stereomicroscope and imaging software, therapeutic effects in these animal models can be monitored.

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Here, animal models based on mouse and rabbit are developed for mechanical and chemical injury of corneal epithelium to screen new therapeutics and the underlying mechanism.

角膜上皮の機械的および化学的損傷のためのEx Vivoおよびin vivo動物モデル

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, ...