著者のこの作品の技術的な側面で彼女の援助ヴァレリー シュレーダーを感謝したいです。このモデルの開発に関連する作業は、クック バイオ テクノロジー社 (米国で、ウェスト ラファイエット) によって支えられました。

このプロトコルにおける動物の使用ノートルダム大聖堂機関動物ケアおよび使用委員会の大学によって承認された、すべての規制当局の要件とガイドライン、続いて、によってみなされている施設で実施された、(AAALAC)、国際実験動物管理認定協会。
 1 です動物の選択 <ol><li>取得病原体フリー スプレイグ ラット評判のベンダーまたは繁殖コロニーから。 。
	注: 多くの場合、施設の獣医またはマネージャーことができます直接適切な情報源を調査官。約 250 g の体重のラットはこの手順に適しています</li>。
	<li>1 日外科前に臨床的に健康であることを確認するための動物を確認します。この点で、動物の重量を量るし、身体の状態を評価呼吸、傷が開く、または大衆の存在と動物の一般的な活動レベルに異常に注意してください。潜在的な除外基準としてそのような観察をして手術記録に異常に注意してください</li>。
</ol> 2。麻酔と無菌手術のための動物の準備 <ol><li>鎮痛、ラット 2 時間前に手術します両方の目に眼軟膏を適用する徐放ブプレノルフィンの 1.1 1.2 mg/kg を皮下管理および管理。2 mL 加温滅菌生理食塩水を皮下</li>。
	<li>は、塩酸ケタミン (90 mg/kg) およびキシラジン (10 mg/kg の腹腔内投与でラットを麻酔します。角膜とした乾燥を防ぐためにそれぞれの目に眼潤滑剤の少量を適用します。一度動物 &#8217; s 呼吸が安定し、はやしっかりつま先ピンチへの応答、ラットの麻酔手術のため十分なレベルに達しています。麻酔の許容レベルは、深いの存在によってさらに確認できる通常の呼吸と目 (対 蒼白、不十分な心臓血管の灌流を示唆) の通常の色</li>。
	<li>は、胸骨に恥骨と、ventrum では、横余白から商業バリカンで腹側腹部を剃る。湿ったペーパー タオルを領域に適用することによって切り取られた髪の毛を削除します</li>。
	<li>は、70% エタノールで拭いて続いて、iodophor 防腐剤と剃毛エリアをスクラブします。切開用のウィンドウを潜在的な汚染から隔離できるように手術用ドレープを適用します</li>。
	<li>ドン滅菌手術用ガウン、マスク、髪ボンネット。徹底的に洗浄し、石鹸とドン滅菌手術用手袋で手をスクラブします</li>。
</ol> 3。作成および腹側腹部ヘルニアの修復 <ol><li>術前滅菌オートクレーブによる手術器具</li>。
	<li>メスを使用すると、腹部の筋肉のレベルまで腹側正中線に沿って約 3-4 cm 縦切開。鉗子で腹壁を慎重に持ち上げ、リネアを通じて小さな開口部を作成するアルバ。基になる内臓を保護するために穴に通し、筋肉壁の切開を作成すると保護のガイドとして機能する腹腔内に鉗子を挿入、約 2 cm × 2 cm 完全な厚さの欠陥、両端から約 1 cm 外側を切断リネアの両端の正中線上のアルバ切開します</li>。
	<li>欠陥の上機械ブリッジ材料、この場合は (図 1) で 4 層 SIS のカット済み 2 cm × 2 cm のセクションに置く 。
	注: また、アンダーレイのアプローチ使用できますある材料部が置かれる腹腔内にインテリアの観点から欠陥をカバーするためです</li>。
	<li>縫合材料簡単な縫合パターンで 4-0 絹またはナイロンを使用して腹壁の筋肉にブリッジの四隅 。
	注: 非吸収性縫合糸の使用により、吸収性縫合糸材料と収穫時に注入材料を簡単に識別します</li>。
	<li>は、等間隔で 3 つの追加縫合材料のそれぞれの端を固定します。腹部の壁を通して内臓の通過を許可する可能性があります、大きなギャップが存在しないことを確保するための全体の円周端を検査します。このようなギャップが存在する場合さらに余白を確保するため追加縫合糸を配置します</li>。
	<li>は、単純な中断パターンで 4-0 吸収性縫合糸で皮下組織を閉じます。縫合の単純な中断パターンの適用はまた使用するかもしれない外科ステープルで肌を終了します</li>。
	<li>放置しないでください動物まで胸骨の横臥を取り戻すために十分な意識を取り戻したが、それは完全に麻酔から回復するまで、他の動物の会社に手術を受けた動物を返さない。この時間の間に低体温症を防ぐために暖かい区域で動物を飼うです</li>。
	完全復旧し、正常に治癒できるようにこの手順を受けている毎日 <li>調査の動物。術後 7-10 日ステープル リムーバーで皮膚ステープルを削除します</li>。
</ol> 4。注入材料の収穫 <ol><li>一貫した研究のニーズ、安楽死、商工会議所で管理される二酸化炭素過剰摂取でフィル レート上昇 10 ~ 30%、事前に決められた時は多くの場合 21 - 動物の術後 28 日</li>。
	<li>使用標準的な剖検ツール、皮膚と腹壁を開き、インプラントの領域を表示します 。
	注: インプラントを確保する非吸収性縫合糸を使用した場合、素材によって識別できます元縫合糸 (図 2).</li>
	<li>副作用 (例えば 創傷の裂開、膿瘍、漿液腫、血腫) と接着程度基になる内臓に粘り強さを含む任意の総病理学的観察を観察</li>。
	<li>は、インプラントを含む組織の広いセクションを切断することによってインプラントのサイトを削除します。定着剤、組織学的検査および評価の処理に備えて、10% 中性緩衝ホルマリンなどでサンプルを配置します</li>。
</ol>

<ol>
	<li>Poussier, M., 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=A+review+of+available+prosthetic+material+for+abdominal+wall+repair.">A review of available prosthetic material for abdominal wall repair.</a> J. Visc. Surg. 150 (1), 52-59 (2013).</li><li>Hjort, H., Mathisen, T., Alves, A., Clermont, G., Boutrand, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-year+results+from+a+preclinical+implantation+study+of+a+long-term+resorbable+surgical+mesh+with+time-dependent+mechanical+characteristics.">Three-year results from a preclinical implantation study of a long-term resorbable surgical mesh with time-dependent mechanical characteristics.</a> Hernia. 16 (2), 191-197 (2012).</li><li>Ko, R., Kazacos, E. A., Snyder, S., Ernst, D. M. J., Lantz, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tensile+strength+comparison+of+small+intestinal+submucosa+body+wall+repair.">Tensile strength comparison of small intestinal submucosa body wall repair.</a> J. Surg. Res. 135 (1), 9-17 (2006).</li><li>Garcia-Moreno, F., 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=Comparing+the+host+tissue+response+and+peritoneal+behavior+of+composite+meshes+used+for+ventral+hernia+repair.">Comparing the host tissue response and peritoneal behavior of composite meshes used for ventral hernia repair.</a> J. Surg. Res. 193 (1), 470-482 (2015).</li><li>Soiderer, E. E., Lantz, G. C., Kazacos, E. A., Hodde, J. P., Wiegand, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphologic+study+of+three+collagen+materials+for+body+wall+repair.">Morphologic study of three collagen materials for body wall repair.</a> J. Surg. Res. 118 (2), 161-175 (2004).</li><li>Liu, Z., Tang, R., Zhou, Z., Song, Z., Wang, H., Gu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+two+porcine-derived+materials+for+repairing+abdominal+wall+defect+in+rats.">Comparison of two porcine-derived materials for repairing abdominal wall defect in rats.</a> PLOS One. 6 (5), e20520 (2011).</li><li>Zhang, J., Wang, G. Y., Xiao, Y. P., Fan, L. Y., Wang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biomechanical+behavior+and+host+response+to+porcine-derived+small+intestine+submucosa,+pericardium+and+dermal+matrix+acellular+grafts+in+a+rat+abdominal+defect+model.">The biomechanical behavior and host response to porcine-derived small intestine submucosa, pericardium and dermal matrix acellular grafts in a rat abdominal defect model.</a> Biomaterials. 32 (29), 7086-7095 (2011).</li><li>Ma, J., Sahoo, S., Baker, A. R., Derwin, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigating+muscle+regeneration+with+a+dermis/small+intestinal+submucosa+scaffold+in+a+rat+full-thickness+abdominal+wall+defect+model.">Investigating muscle regeneration with a dermis/small intestinal submucosa scaffold in a rat full-thickness abdominal wall defect model.</a> J.Biomed.Mater.Res.Part B. 103 (2), 355-364 (2015).</li><li>Greca, F. H., Souza-Filho, Z. A., Giovanni, A., Rubin, M. R., Kuenzer, R. F., Reese, F. B., Araujo, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+porosity+on+the+integration+histology+of+two+polypropylene+meshes+for+the+treatment+of+abdominal+wall+defects+in+dogs.">The influence of porosity on the integration histology of two polypropylene meshes for the treatment of abdominal wall defects in dogs.</a> Hernia. 12 (1), 45-49 (2008).</li><li>Hiles, M., Record-Ritchie, R. D., Altizer, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+biologic+grafts+effective+for+hernia+repair?+A+systematic+review+of+the+literature.">Are biologic grafts effective for hernia repair? A systematic review of the literature.</a> Surg. Innov. 16 (1), 26-37 (2009).</li><li>Helton, W. S., Fisichella, P. M., Berger, R., Horgan, S., Espat, N. J., Abcarian, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+outcomes+with+small+intestinal+submucosa+for+ventral+abdominal+hernia.">Short-term outcomes with small intestinal submucosa for ventral abdominal hernia.</a> Arch. Surg. 140 (6), 560-562 (2005).</li><li>Franklin, M. E., Trevino, J. M., Portillo, G., Vela, I., Glass, J. L., Gonzalez, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+porcine+small+intestinal+submucosa+as+a+prosthetic+material+for+laparascopic+hernia+repair+in+infected+and+potentially+contaminated+fields:+long-term+follow-up.">The use of porcine small intestinal submucosa as a prosthetic material for laparascopic hernia repair in infected and potentially contaminated fields: long-term follow-up.</a> Surg. Endosc. 22 (9), 1941-1946 (2008).</li><li>Clarke, K. M., Lantz, G. C., Salisbury, S. K., Badylak, S. F., Hiles, M. C., Voytik, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestine+submucosa+and+polypropylene+mesh+for+abdominal+wall+repair+in+dogs.">Intestine submucosa and polypropylene mesh for abdominal wall repair in dogs.</a> J. Surg. Res. 60 (1), 107-114 (1996).</li><li>Sarikava, A., Record, R., Wu, C. C., Tullius, B., Badylak, S. F., Ladisch, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+activity+associated+with+extracellular+matrices.">Antimicrobial activity associated with extracellular matrices.</a> Tissue Eng. 8 (1), 63-71 (2002).</li><li>Serra, R., Grande, R., Butrico, L., Rossi, A., Settimio, U. F., Caroleo, B., Amato, B., Gallelli, L., de Franciscis, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+wound+infections:+the+role+of+Pseudomonas+aeruginosa+and+Staphylococcus+aureus.">Chronic wound infections: the role of Pseudomonas aeruginosa and Staphylococcus aureus.</a> Expert Rev. Anti. Infect. Ther. 13 (5), 605-613 (2015).</li></ol>

クック バイオ テクノロジー株式会社、医療グレード SIS の製造元によって (CJ) の著者の 1 つを採用します。

腹側腹部ヘルニアの修復のための材料は大きな関心は、特にその初期の機械式ブリッジを提供され、ホスト組織に組み込むことができます。この点では、ブタの心膜やブタの無細胞真皮マトリックス SIS を含む様々 な材料が検討されてきた。6,7これらの材料はそれらのティッシュの細胞外マトリックスを表す、足場に細胞が移行でき、増殖組織設?...

腹壁ヘルニアは、先天性欠損、外傷、または失敗した閉鎖、腹壁を含む手術の傷の結果として発生する可能性があります一般的臨床問題です。修理が一般的で腹壁を縫合糸で閉じる単にそれらと比較してインプラント治療を受けた患者の再発率が減少で、腹壁を補強するためにインプラントを使用して含まれます。1
外科ヘルニア修復は、注入材料の欠陥のメカニカル ブリッジを必要があります。この点では、合成や天然素材は、ヘルニア修復のため使用されています。多くの合成物質は通常非吸収性、他合成インプラントは、吸収性と非吸収性材料の一つを結合しながら展開されたポリテトラフルオロ エチレン、ポリエチレン テレフタ レート ポリエステル、ポリプロピレンが含まれますpolygalactine などのポリマーです。1対照的に、自然材料インプラントが動物や人間の死体ソースから派生した一般的に。ヘルニア修復のため使用される自然な材料、通常コラーゲンの豊富な内部成長とホスト組織の統合のための足場として機能します。
欲望を開発し、腹側腹部ヘルニア修復のための新しい材料を最適化は、候補材料の評価のための適切な動物モデルを必要とします。研究は、様々 な種、羊、豚2 、3ウサギ、4ラット、5,6,7,8犬などで行われています。9このような研究のラットを使用しての利点は、さまざまな遺伝的変異性の制御を許可する近交系で使用可能な扱いやすいモデルを提供することです。また、上記の他の種と比較してラットの全体的な本体表面の小さい領域を許可可能性があります自然の実験材料の評価、したがって、豊富でないです。 ラット対他の種に関連付けられているさらに、削減のコストを相対的に増加させた能力画面候補材料腹側腹部ヘルニアの修復に使用するため許可します。 材料に関する考慮事項には通常外科医、強さ、感染に対する抵抗性、生体適合性、ホスト組織に組み込むことで扱いやすさが含まれます。
例として小さなブタ腸粘膜下層 (SIS) はさまざまな組織修復の徴候、腹部、鼠径部、腹部、横隔膜ヘルニアの修復を含むため使用されている天然生体材料です。10,11,12 SIS、急速にホスト組織に組み込まれて、一般的に感染症に耐性があることがわかったし、材料が汚染されたフィールドで特定のユーティリティを持ってそう。13,14ラットのモデルでは、SIS は、一軸引張破壊試験および組織のコラーゲンと血管新生の面で成長の促進によって決まる良い引張強度の処理の容易さを実証しています。6,7
欠陥の正確な手術ブリッジは、候補者の修復材料を評価するときに生産的なモデリングに不可欠な機能です。動物は厳密に続く十分な麻酔や無菌技術をする必要があります。さらに、標準サイズの腹壁欠損を慎重に作成し、十分に欠陥のマージンに材料を固定する方法で、ブリッジする必要があります。8このプロトコルは、ラットの材料を埋める例として SIS を使用して腹側腹部ヘルニア手術の作成、および、修理の後にする標準的な方法を提供します。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Isoflurane (Isoflo)</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">17579</td>
			<td style="width:167px;">gas anesthetic</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Optixcare Eye Lubricant</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td style="width:119px;">OPX4240</td>
			<td>Ocular lubricant</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Oster clippers</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">6092</td>
			<td>hair clippers</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Betadine surgical scrub</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">1618</td>
			<td>antiseptic iodophor</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Sterile scalpel</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">329</td>
			<td>#10 scalpel</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">9&#34; x 12&#34; disposable surgical drape</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">SP-RPS</td>
			<td>Surgical drape</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">4-0 nylon suture</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">SUT 812</td>
			<td>Suture material</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">4-0 Absorbable Suture</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">29242</td>
			<td>Synthetic absorbable suture</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">9 mm Autoclips and applier</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">ACS KIT</td>
			<td>Surgical staples for closure of skin incision</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Torbugesic/analgesic</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">12084</td>
			<td>Butorphanol tartarate for post-operative analgesia</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Small intestinal submucosa (SIS)</td>
			<td style="width:71px;">Cook Biotech, Inc.</td>
			<td style="width:119px;"></td>
			<td>Implant material for bridging experimental abdominal wall defect</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Rats</td>
			<td style="width:71px;">Harlan, Inc.</td>
			<td style="width:119px;">Sprague Dawley</td>
			<td>Animal for hernia modeling</td>
		</tr>
	</tbody>
</table>

use of a rat model to study ventral abdominal hernia repair

腹側腹部ヘルニアは時々 herniorraphy (手術) が必要な比較的一般的な臨床症状です。腹側腹部ヘルニアの修復術、通常腹部壁の欠陥でメカニカル ブリッジとして機能する材料の注入が必要です。小さなブタ腸粘膜下層 (SIS) のような生体もインプラントに細胞増殖の格子として役立つので、ホスト組織に組み込むことが自然。このような修復材料の開発動物モデル実験的腹部壁の欠陥が簡単に作成しは、再現可能な方法で修復するの使用からの利点します。ここで提供されるメソッドでは、外科的創造とラットモデルにおける腹側腹部ヘルニアの修復について説明します。このモデルでの実験的腹側腹部ヘルニアを修復する SIS を使用すると、注入後 28 日以内ホスト組織に組み込まれて急速に。組織学的に、ホスト組織への注入材料の取り込みは、堅牢な線応答によって特徴づけられます。将来の改良とラット腹部ヘルニア モデルの応用可能性が高い人間のもっと密接に模倣の一般的な併存する手段として糖尿病や肥満の動物を伴うことがあります。

注入後 28 日後、SIS は通常良いホスト組織 (図 2) 混入を示します。ほとんどの場合がでは残留 SIS、明白な組織は、しばしば注入材料を識別するために調査が必要があります。組織学的解析は、主線組織と非常に小さな残留 SIS と指摘した (図 3) と良い組織定款を示す総の病理学的観察を確認しました。これらの結果はラット腹側腹部ヘルニア ...

Watch this Scientific Journal Video about Use of a Rat Model to Study Ventral Abdominal Hernia Repair at JoVE.com

Use of a Rat Model to Study Ventral Abdominal Hernia Repair

本稿では、作成および腹側腹壁欠損 (ヘルニア) の修復に関連付けられたメソッドについて説明します。このモデルは、注入材料を使用するものなどの修復戦略を研究に使用できます。本稿では、小さなブタの腸粘膜と実験的ヘルニアの修復は、例として提示されます。

腹側腹部ヘルニア修復術を勉強するラットのモデルの使用

Ventral abdominal hernia is a relatively common clinical condition that sometimes requires herniorraphy (surgical repair). The repair of ventral abdominal ...

use-of-a-rat-model-to-study-ventral-abdominal-hernia-repair

Research

JoVE Journal

Bioengineering

9.1K ビュー.  University of Minnesota. 本稿では、作成および腹側腹壁欠損 (ヘルニア) の修復に関連付けられたメソッドについて説明します。このモデルは、注入材料を使用するものなどの修復戦略を研究に使用できます。本稿では、小さなブタの腸粘膜と実験的ヘルニアの修復は、例として提示されます。

ビデオ: Use of a Rat Model to Study Ventral Abdominal Hernia Repair

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells behave differently when they are grown within a 3D extra-cellular matrix and also interact with other cells (1-5). Mouse models have been broadly utilized to study tissue morphogenesis in vivo. However mouse and human skin have significant differences in cellular architecture and physiology, which makes it difficult to extrapolate mouse studies to humans. Since melanocytes in mouse skin are mostly localized in hair follicles, they have distinct biological properties from those of humans, which locate primarily at the basal layer of the epidermis. The recent development of 3D human skin reconstruct models has enabled the field to investigate cell-matrix and cell-cell interactions between different cell types. The reconstructs consist of a "dermis" with fibroblasts embedded in a collagen I matrix, an "epidermis", which is comprised of stratified, differentiated keratinocytes and a functional basement membrane, which separates epidermis from dermis. Collagen provides scaffolding, nutrient delivery, and potential for cell-to-cell interaction. The 3D skin models incorporating melanocytic cells recapitulate natural features of melanocyte homeostasis and melanoma progression in human skin. As in vivo, melanocytes in reconstructed skin are localized at the basement membrane interspersed with basal layer keratinocytes. Melanoma cells exhibit the same characteristics reflecting the original tumor stage (RGP, VGP and metastatic melanoma cells) in vivo. Recently, dermal stem cells have been identified in the human dermis (6). These multi-potent stem cells can migrate to the epidermis and differentiate to melanocytes.

In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.

三次元ヒト皮膚は、モデルを再構築：ノーマルスキンとメラノーマ進行を勉強するツールを

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

生物工学

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic valve leaflets are composed of interstitial cells suspended within an extracellular matrix (ECM) and are lined with an endothelial cell monolayer. The valve withstands a harsh, dynamic environment and is constantly exposed to shear, flexion, tension, and compression. Research has shown calcific lesions in diseased valves occur in areas of high mechanical stress as a result of endothelial disruption or interstitial matrix damage1-3. Hence, it is not surprising that epidemiological studies have shown high blood pressure to be a leading risk factor in the onset of aortic valve disease4. 
The only treatment option currently available for valve disease is surgical replacement of the diseased valve with a bioprosthetic or mechanical valve5. Improved understanding of valve biology in response to physical stresses would help elucidate the mechanisms of valve pathogenesis. In turn, this could help in the development of non-invasive therapies such as pharmaceutical intervention or prevention. Several bioreactors have been previously developed to study the mechanobiology of native or engineered heart valves6-9. Pulsatile bioreactors have also been developed to study a range of tissues including cartilage10, bone11 and bladder12. The aim of this work was to develop a cyclic pressure system that could be used to elucidate the biological response of aortic valve leaflets to increased pressure loads. 
The system consisted of an acrylic chamber in which to place samples and produce cyclic pressure, viton diaphragm solenoid valves to control the timing of the pressure cycle, and a computer to control electrical devices. The pressure was monitored using a pressure transducer, and the signal was conditioned using a load cell conditioner. A LabVIEW program regulated the pressure using an analog device to pump compressed air into the system at the appropriate rate. The system mimicked the dynamic transvalvular pressure levels associated with the aortic valve; a saw tooth wave produced a gradual increase in pressure, typical of the transvalvular pressure gradient that is present across the valve during diastole, followed by a sharp pressure drop depicting valve opening in systole. The LabVIEW program allowed users to control the magnitude and frequency of cyclic pressure. The system was able to subject tissue samples to physiological and pathological pressure conditions. This device can be used to increase our understanding of how heart valves respond to changes in the local mechanical environment.

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

A cyclic pressure bioreactor capable of subjecting heart valve tissue to physiological and pathological pressure conditions has been designed. A LabVIEW program allows users to control pressure magnitude, amplitude and frequency. This device can be used to study the mechanobiology of heart valve tissue or isolated cells.

のための周期的圧力のバイオリアクターの設計生体外で</em大動脈心臓弁の>研究

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges1,2. These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production3. In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (&lambda;=532 nm, Fluence~110 J/cm2, spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 &plusmn; 6 kPa, (n=30). The adhesion strength drops to 2 &plusmn; 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

レーザー組織修復にローズベンガル-キトサン膜の作製と応用

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two ...

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This technique allows for easier venous cuff-anatomoses after a reasonable learning curve. The device is composed of two handles, which are carved out from scalpel blades, one approximator, which is obtained by modifying Kocher's forceps, and cuffs designed from fine-bore polyethylene tubing. The whole process can be performed at a low-cost using common laboratory material. The present report provides a step-by-step protocol for the design of the required pieces and includes stencils.

We describe the design of the &#x201C;quick-linker&#x201D; device for easier orthotopic rat liver transplantation.

簡単ラット肝移植のための製造装置や計測器

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This ...

Athymic Rat Model for Evaluation of Engineered Anterior Cruciate Ligament Grafts

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without intervention. Current treatment strategies include ligament reconstruction with either autograft or allograft, which each have their associated limitations. Thus, there is interest in designing a tissue-engineered graft for use in ACL reconstruction. We describe the fabrication of an electrospun polymer graft for use in ACL tissue engineering. This polycaprolactone graft is biocompatible, biodegradable, porous, and is comprised of aligned fibers. Because an animal model is necessary to evaluate such a graft, this paper describes an intra-articular athymic rat model of ACL reconstruction that can be used to evaluate engineered grafts, including those seeded with xenogeneic cells. Representative histology and biomechanical testing results at 16 weeks postoperatively are presented, with grafts tested immediately post-implantation and contralateral native ACLs serving as controls. The present study provides a reproducible animal model with which to evaluate tissue engineered ACL grafts, and demonstrates the potential of a regenerative medicine approach to treatment of ACL rupture.

Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.

エンジニアード前十字靭帯移植片の評価のための無胸腺ラットモデル

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without ...

In Vitro Permeation of FITC-loaded Ferritins Across a Rat Blood-brain Barrier: a Model to Study the Delivery of Nanoformulated Molecules

Brain microvascular endothelial cells, supported by pericytes and astrocytes endfeet, are responsible for the low permeation of large hydrosoluble drugs through the blood-brain barrier (BBB), causing difficulties for effective pharmacological therapies. In recent years, different strategies for promoting brain targeting have aimed to improve drug delivery and activity at this site, including innovative nanosystems for drug delivery across the BBB. In this context, an in vitro approach based on a simplified cellular model of the BBB provides a useful tool to investigate the effect of nanoformulations on the trans-BBB permeation of molecules. This study describes the development of a double-layer BBB, consisting of co-cultured commercially available primary rat brain microvascular endothelial cells and astrocytes. A multiparametric approach for the validation of the model, based on the measurement of the transendothelial electrical resistance and the apparent permeability of a high molecular weight dextran, is also described. As proof of concept for the employment of this BBB model to study the effect of different nanoformulations on the translocation of fluorescent molecules across the barrier, we describe the use of fluorescein isothiocyanate (FITC), loaded into ferritin nanoparticles. The ability of ferritins to improve the trans-BBB permeation of FITC was demonstrated by flux measurements and confocal microscopy analyses. The results suggest this is a useful system for validating nanosystems for delivery of drugs across the BBB.

In Vitro Permeation of FITC-loaded Ferritins Across a Rat Blood-brain Barrier: a Model to Study the Delivery of Nanoformulated Molecules

A method to establish an in vitro model of blood-brain barrier based on a co-culture of rat brain microvascular endothelial cells and astrocytes is described and validated. This system proved to be a valid tool to study the effect of nanoformulation on the trans-barrier permeation of fluorescent molecules.

Nanoformulated分子の送達を研究するためのモデル：ラット血液脳関門を通過して、FITCロードフェリチンのin vitro浸透で

Brain microvascular endothelial cells, supported by pericytes and astrocytes endfeet, are responsible for the low permeation of large hydrosoluble drugs ...

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from traumatic injury, disease and various congenital, genetic and acquired conditions are quite common. Tissue engineering and regenerative medicine technologies have enormous potential to provide a therapeutic solution. However, utilization of biologically relevant animal models in combination with longitudinal assessments of pertinent functional measures are critical to the development of improved regenerative therapeutics for treatment of VML-like injuries. In that regard, a commercial muscle lever system can be used to measure length, tension, force and velocity parameters in skeletal muscle. We used this system, in conjunction with a high power, bi-phase stimulator, to measure in vivo force production in response to activation of the anterior crural compartment of the rat hindlimb. We have previously used this equipment to assess the functional impact of VML injury on the tibialis anterior (TA) muscle, as well as the extent of functional recovery following treatment of the injured TA muscle with our tissue engineered muscle repair (TEMR) technology. For such studies, the left foot of an anaesthetized rat is securely anchored to a footplate linked to a servomotor, and the common peroneal nerve is stimulated by two percutaneous needle electrodes to elicit muscle contraction and dorsiflexion of the foot. The peroneal nerve stimulation-induced muscle contraction is measured over a range of stimulation frequencies (1-200 Hz), to ensure an eventual plateau in force production that allows for an accurate determination of peak tetanic force. In addition to evaluation of the extent of VML injury as well as the degree of functional recovery following treatment, this methodology can be easily applied to study diverse aspects of muscle physiology and pathophysiology. Such an approach should assist with the more rational development of improved therapeutics for muscle repair and regeneration.

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

We describe an in vivo protocol to measure dorsiflexion of the foot following stimulation of the peroneal nerve and contraction of the anterior crural compartment of the rat hindlimb. Such measurements are an indispensable translational tool for evaluating skeletal muscle pathology and tissue engineering approaches to muscle repair and regeneration.

の応用<em&gt;インビボ</em組織工学骨格筋の修復を評価するためのラット前脛骨の&gt;機能テスト

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from ...

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 ...

The Use of a &#946;-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

Biosensors are becoming increasingly important and implemented in various fields such as pathogen detection, molecular diagnosis, environmental monitoring, and food safety control. In this context, we used &#946;-lactamases as efficient reporter enzymes in several protein-protein interaction studies. Furthermore, their ability to accept insertions of peptides or structured proteins/domains strongly encourages the use of these enzymes to generate chimeric proteins. In a recent study, we inserted a single-domain antibody fragment into the Bacillus licheniformis BlaP &#946;-lactamase. These small domains, also called nanobodies, are defined as the antigen-binding domains of single chain antibodies from camelids. Like common double chain antibodies, they show high affinities and specificities for their targets. The resulting chimeric protein exhibited a high affinity against its target while retaining the &#946;-lactamase activity. This suggests that the nanobody and &#946;-lactamase moieties remain functional. In the present work, we report a detailed protocol that combines our hybrid &#946;-lactamase system to the biosensor technology. The specific binding of the nanobody to its target can be detected thanks to a conductimetric measurement of the protons released by the catalytic activity of the enzyme.

In this work, we report a new method to study protein-protein interactions using a conductimetric biosensor based on the hybrid &#946;-lactamase technology. This method relies on release of protons upon hydrolysis of &#946;-lactams.

生体分子相互作用を検出する β-ラクタマーゼによる伝導度バイオ センサー法の使用

Biosensors are becoming increasingly important and implemented in various fields such as pathogen detection, molecular diagnosis, environmental ...

An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) is a surgical technique leading to bone regeneration after an osteotomy. The osteotomized extremities are moved away from each other by gradual distraction to reach the desired elongation. This procedure is widely used in humans for lower and upper limb lengthening, treatment after a bone nonunion, or the regeneration of a bone defect following surgery for bone tumor excision, as well as in maxillofacial reconstruction. Only a few studies clearly demonstrate the efficiency of their protocol in obtaining a functional regenerated bone, i.e., bone that will support physiological weight-bearing without fracture after removal of the external fixator. Moreover, protocols for DO vary and reproducibility is limited by lack of information, making comparison between studies difficult. The aim of this study was to develop a reproducible protocol comprising an appropriate external fixator design for rat limb lengthening, with a detailed surgical technique that permits physiological weight-bearing by the animal after removal of the external fixator.

This study describes a reproducible and detailed protocol using a newly developed external fixator for distraction osteogenesis (DO) in a femoral rat model which permits physiological weight-bearing by the animal after removal of the external fixator.

機能再生大腿骨につながるラットモデルにおける仮骨延長法のための効率的かつ再現可能なプロトコル

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) ...

腹側腹部ヘルニア修復術を勉強するラットのモデルの使用

要約

さらに動画を探す

この動画の章

三次元ヒト皮膚は、モデルを再構築：ノーマルスキンとメラノーマ進行を勉強するツールを

のための周期的圧力のバイオリアクターの設計生体外で研究

レーザー組織修復にローズベンガル-キトサン膜の作製と応用

簡単ラット肝移植のための製造装置や計測器

エンジニアード前十字靭帯移植片の評価のための無胸腺ラットモデル

Nanoformulated分子の送達を研究するためのモデル：ラット血液脳関門を通過して、FITCロードフェリチンのin vitro浸透で

の応用

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

生体分子相互作用を検出する β-ラクタマーゼによる伝導度バイオセンサー法の使用

機能再生大腿骨につながるラットモデルにおける仮骨延長法のための効率的かつ再現可能なプロトコル

腹側腹部ヘルニア修復術を勉強するラットのモデルの使用

要約

さらに動画を探す

この動画の章

三次元ヒト皮膚は、モデルを再構築：ノーマルスキンとメラノーマ進行を勉強するツールを

のための周期的圧力のバイオリアクターの設計生体外で研究

レーザー組織修復にローズベンガル-キトサン膜の作製と応用

簡単ラット肝移植のための製造装置や計測器

エンジニアード前十字靭帯移植片の評価のための無胸腺ラットモデル

Nanoformulated分子の送達を研究するためのモデル：ラット血液脳関門を通過して、FITCロードフェリチンのin vitro浸透で

の応用

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

生体分子相互作用を検出する β-ラクタマーゼによる伝導度バイオ センサー法の使用

機能再生大腿骨につながるラットモデルにおける仮骨延長法のための効率的かつ再現可能なプロトコル

生体分子相互作用を検出する β-ラクタマーゼによる伝導度バイオセンサー法の使用