クリス・デイリー博士は、リチャード・ジープソン研究奨学金の手術のための基金の受領者です。著者たちは、Anne Gibbon博士、ドン・ジャン医師、モナッシュ・アニマル・サービス（Monash Animal Services、Monash University）のスタッフに、動物の手術とケアの支援について感謝したいと考えています。

この原稿で詳述されているプロトコルは、Monash University Animal Ethicsの動物保護ガイドラインに従っています。このプロトコルに関する動物倫理の承認は、Monash University動物倫理によって認められています。倫理承認番号：MMCA / 2014/55 1.羊の準備注：2〜4歳のEwesを使用した。 <ol><li>麻酔前18時間の高速羊。操作19の前に6-12時間まで動物に水を供給する。 </li><li>手術室への移送を容易にするために、塩酸メデトミジン（0.015-0.020mg / kg）の静脈注射により動物を鎮静させる。 注：メデトミジン塩酸塩は、手術室への移送のために、他の動物との分離に伴う動物のストレスおよびかき混ぜを軽減するのに役立つ。 </li><li>手術室への到着時に麻酔を誘導するためにチオペンオン（10〜13mL / kg）を注入する。 </li><li>予防的イントラを管理する静脈内抗生物質（アモキシシリン1g IV）をチオペンタール注射直後に投与する。 </li><li>気管内チューブ20内径7.5〜9mmの挿管羊。 </li><li> 2L /分の流速で100％酸素中で吸入イソフルラン（2〜3％）を用いて麻酔を維持する。羊の耳にパルスオキシメータを取り付けます。 </li><li>羊のバイタルサイン（心拍数、呼吸数、酸素飽和度、パルスオキシメーター、観察）と意識レベルを密接に監視します。 注：自発的な咀嚼、活動的な逆流、および自発的な動きなどの軽い麻酔の指標は、麻酔のレベルの上昇を促すはずです。緊急の麻酔の軽減を示す赤旗の徴候には、呼吸障害および重度の徐脈が含まれる。目の回転は、羊19の麻酔深度の一貫した指標ではありません。 </li></ol>ディスクレベルと切開<ol><li> sを集める獣医クリッパー、20mLルアーロックシリンジ、21G IV針、＃4メスハンドル、＃22メスブレード、Gillies組織鉗子、メッツェンバム湾曲解剖はさみ、Deaverリトラクター、Hohmannリトラクターブレード、3.5mm Bradドリルビットストップ、ドリル、オートクレーブ可能な獣医ドリルバッグ、針ホルダー、2-0吸収性合成編組縫合糸、3-0吸収性合成編組縫合糸およびMayo縫合はさみを含む。 </li><li>操作スイートを準備します。 70％エタノールで手術台と計器スタンドを掃除してください。手術前にすべての手術器具をオートクレーブする。術前麻酔検査を行う。 </li><li>羊を右の横の位置で手術台の上に置きます。 </li><li>電子クリッパを使用して、下側肋骨によって上方に定義された領域を、腸骨によって下側に、対側腰部横断プロセスによって内側に、同側腰部横断プロセスに対して約10cm横に剃る。 </li><li>外科切開部位のランドマークのための腸骨稜、腰部横断プロセス（L1〜6）および肋骨 - 脊柱角を触診する。これらのランドマークを無菌ペンでマークしてください。 </li><li>クロルヘキシジンとアルコール性ヨウ素消毒洗浄液で消毒することにより、側方腹部を準備する。 </li><li> 手術中の標準的な手術無菌技術を観察する。手術チームは手術前にスクラブを行う。外科手術部位の上に滅菌有孔の四角いドレープを置き、オーバーヘッドテーブルに大きな滅菌四角いドレープを置きます。 <ol><li>操作の前に手術部位内で使用されるすべての項目を滅菌する。手術中の手術部位の無菌性を監視し、維持する。滅菌現場に導入されたすべての品目が滅菌され、滅菌された状態で移送されるようにする。 </li></ol></li><li>外科的処置中に視覚化を容易にするために、手術用ルーペの倍率およびヘッドライトを使用する。 </li><li>縦切開する＃4のメスに取り付けられた＃22のメスブレードを、椎骨間の関心対象レベルの上下に1cmから2cmの腰の横方向のプロセスと平行に使用する。 注：切開計画に関する詳細は、ディスカッションに記載されています。 </li><li>下にある皮下組織と腹部の筋肉組織の側面を分けるために、単極ジアテルミーを使います。関心のある椎間板の上下の腰部横断プロセスの先端に向けて解剖を指示する。 </li><li>胸腰部筋膜を横方向のプロセスへの取り付け時に縦に分割する。 </li><li>基礎となる四肢腰部 、腰筋、および横行神経血管束を視覚化し、保存する。 </li><li>ジアテルミーを使用して止血を維持する。 </li><li>露出した椎間diで腹膜の平面と後部腹壁の筋肉の間で指を掃引するscレベルを用いてデジタル鈍的切開を行う。 </li><li>椎間円板をさらに露出させるために、Deaverリトラクターを使用して四分円腰部および腰筋を後部後退させる。 </li><li>凹型椎体および凸状椎間板のための触角。 </li><li>リトラクタをディスクのすぐ上に置き、腰部血管が損傷しないように注意してください。 </li><li>ヘッドライト付きの手術用ルーペ倍率を使用して、下側の終板に対して約1cmの尾側に位置する腰部血管を特定する。 </li><li>術中X線を行い、椎間板のレベルを確認します。 21 注：ラジオグラフ設定：47kV; 4mAs 21 </li><li> 必要なディスクレベルに応じて、周囲の構造と添付ファイルを以下のように分離して椎間板を露出させます。 <ol><li>レベルL3 / 4以上では、筋肉のアタッチメントをディscを使用した。 </li><li>レベルL4 / 5以下の場合は、双極性のジアテルミーとはさみを使用して、厚い筋肉のアタッチメントをディスク上に鋭く分割します。 注：L6 / S1ディスクは、腸骨稜の閉塞のためにアクセスするのが難しい場合があります。側方アプローチを介してアクセスが達成できない場合、前方アプローチを利用することができる。 </li></ol></li></ol> 3.ドリルビットの傷害注：術前計画には、傷害/治療レベルおよびコントロールレベルの割り当てが含まれます。レベル配分に関するさらなる情報は、ディスカッションで見つけることができます。 <ol><li>椎間円板の左の側方および前方の四肢を観察することによって、ドリルビットの進入点を定義する。 注：エントリーポイントは、この左前側四分円（ディスクの前側および側端で定義される）の中間点に位置しています。ドリルビットは、この進入点に、目的の軌跡で挿入されます椎間板の中心に向けて、頭側から鉛直方向にわずかに向けられる。 </li><li> ブラッドポイントのドリルビットをパワードリルに合わせます。ドリルビットの直径が椎間板の高さよりわずかに小さいこと、 すなわち 60-70kgのヒツジの腰椎椎間板の場合〜3.5mmであることを確認する。 <ol><li>ドリルビットストップを適用して、腰椎の直径の約半分の保護されていないドリルビット長、 すなわち 60-70kgのヒツジの腰椎椎間板の〜12mmを提供する。 </li></ol></li><li>エントリーポイントにドリルビットを適用し、椎間板の中心にわずかに頭蓋骨のある軌跡で指示します。わずかな頭蓋の角度は、終板損傷のリスクを最小限にすることである。 </li><li>ドリルビットを1秒間低出力でドリルで椎間板にゆっくりと進めます。過度の抵抗に遭遇した場合、わずかな頭蓋または尾の様式で軌道を調整する 注：そのような過度の抵抗はエンドプレートとの接触を示している可能性があります。 </li></ol> 4.閉鎖<ol><li>止血が達成されたら、Ringersの溶液で傷口を洗浄します。 </li><li>外側の腹壁組織に2-0の吸収性合成編組縫合糸を使用し、皮膚に連続した3-0個の未染色吸収性合成編組皮下縫合糸を使用して、層状閉鎖を行うのが好ましい。 </li></ol> 5.術後管理<ol><li>術後鎮痛のために鼠径部にフェンタニル経皮パッチ（75μg/ h）を3日間置く。 </li><li>さらに、必要に応じて、静脈内投与のブプレノルフィン（0.005-0.01 mg / kg）を使用してください。 </li><li>吸入麻酔薬を止める。自発呼吸が起こったら、気管内チューブを外す。 </li><li>一定の観察の下で動物を保持ケージ内で回復させる。 注：動物は十分な注意を回復するまで無人で放置しないでください胸骨臥位を保持するためのusness。 </li><li>動物が完全に警戒し立ったら、食物と水を再導入する。完全に回復したら、動物を他の動物とペンを持っている手術施設に戻してください。 </li><li> 24時間綿密に監視し、1週間の観察を続ける。術後の痛みや苦痛の証拠を監視する。 注：術後経皮フェンタニルパッチを3日間適用すると、十分な鎮痛作用が得られるはずです。追加の鎮痛薬の要求は、動物のレビューを促すはずである。 </li><li>正常に羊を供給し、羊が制限なく通常の活動を行うことを許可する。跛行のような神経学的欠損の兆候がないか羊を観察する。 注記：ドリルビット損傷法による椎間板欠損は、椎間板の前外側面にあり、損傷深さはドリルビットストップによって中核に制限されます。神経要素は、椎間板の後方/後方側に位置するので、症候性髄核に続発する神経妥協のリスクは遠隔である。モデルのこの解剖学的特徴は、髄核ヘルニアの有無にかかわらず、椎間板変性を区別するための神経学的検査の使用を排除する。 </li><li>実験期間の終わりに安楽死と剖検を待つために、羊を大学の農場に戻してください。 </li></ol> 6.安楽死<ol><li>ドリルビット椎間板損傷後の適切な時間間隔でヒツジ安楽死を実施する。 </li><li>安楽死のために静脈内ペントバルビトンナトリウム（&gt; 100 mg / kg）を注入する。 </li></ol>

<ol>
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W., Metzdorf, A., Zanetti, M., Hodler, J., Boos, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+classification+of+lumbar+intervertebral+disc+degeneration.">Magnetic resonance classification of lumbar intervertebral disc degeneration.</a> Spine. 26 (17), 1873-1878 (2001).</li></ol>

著者らは、開示すべき競合する金銭的利益がないと宣言している。

この最小侵襲性の側方アクセスアプローチは、このシリーズで観察された術後ヘルニア、腹部裂開または感染症がなく、有効かつ安全である。深さ止めを有するドリルビット椎間板損傷モデルの使用は、既知の寸法（ すなわち 、本研究では直径3.5mm×深さ12mmの損傷）の一貫した椎間板損傷を誘発する再現可能な方法を提供する。我々の経験では、この方法は、従来の卵形メス刃の腰椎...

腰痛は世界中の障害の主要原因です1 。腰椎椎間板変性に関連する椎間板形成性疼痛は、腰痛の重大な原因と考えられている2 。変性プロセスの理解を広げ、潜在的な治療法の研究のために、椎間板疾患の信頼できる動物モデルに対する需要が高まっている。 椎間板変性の数多くの動物モデルが存在する3 。変性椎間板疾患の研究に用いられる動物モデルは、マウス4から 、イヌ5 、ヒツジ6 、および非ヒト霊長類7などのより大きな哺乳動物までの大きさの範囲にある。椎間板変性を誘発するために使用される方法は、機械的（ 例えば、椎間板の圧迫<html> （ 例えば、化学的核分解5 ）、またはそれほど一般的ではない自発的変性（ 例えば 、砂ラット9 ）を含むが、これらに限定されない。 人間の椎間板変性の複雑さを考えると、完璧な動物モデルは存在しない。しかし、この状態を模倣するために適切な動物モデルを選択する際の重要な考慮事項が確認されている3 。このような考察には、脊索細胞（ヒト、羊、ヤギおよび軟骨異形成犬の成体核膿瘍には存在しないが、ほとんどの哺乳類には存在する可能性のある前駆細胞機能10を有する原始細胞）、ヒトおよび動物に対する椎間板および椎間板の類似性、臨床的状態と同等の生体力学的力、機械的および倫理的考察3 。 <html> jove_content &quot;&gt;ヒト以外の霊長類は、上記の基準の多くを満たしています。自然発生した椎間板変性症のBaboonとmacaqueモデルは、11,12,13に記載されています。どちらの種も、直立姿勢または半直立姿勢しかし、倫理的かつ実践的な考慮（ 例えば、費用、住宅、自発的変性の発症の遅延）は、多くの機関でその使用を制限している。 ヒツジ背骨は、ヒトの背骨10,14,15との細胞、生体力学的および解剖学的類似性を含む利点を有する、椎間板変性の確立されたモデルである。ヒツジの4倍の体格にもかかわらず、羊の腰椎椎間板はヒトの椎間板と同様のストレスを受けるs = &quot;xref&quot;&gt; 14。ヒツジモデルは、非ヒト霊長類モデルよりも、倫理的観点からより広く受け入れられている。変性プロセスを開始するための様々な方法が記載されており、その多くは椎間板への直接アクセスを必要とする。仙骨領域における脊髄の終結およびヒツジ腰椎における後縦靭帯の骨化のために、椎間板への後方アプローチは技術的に困難であり、ヒツジ16ではそれほど一般的ではない。羊の腰椎への伝統的な接近ルート、 すなわち 、前部または前外側アプローチを介して、大きな腹部の切開が必要であり、ヘルニアのリスクを伴い、内臓の内臓および神経血管構造に損傷を与える16 。従属腹部領域から離れた比較的小さい側方切開部の使用は、そのようなリスクを減少させる可能性がある17 。 私たちは羊毛を提示する最小侵襲性側方アプローチを介して実施されたドリルビット損傷を使用した変性腰椎椎間板疾患の1例を経験し、Zhang らの研究によって触発された。 18 。このプロトコールの目標は、容易に再現性があり、一貫した傷害を生じ、安全で耐容性が高い信頼性の高い腰椎椎間板損傷モデルを可能にすることである。このアプローチは、椎間板変性または再生療法の研究のための伝統的な外科的切除法（未発表データ）で観察されたものより軽度の腰椎椎間板変性を誘導しようとする研究者に適している。これらの知見は、今後の刊行物に記載される。

<table border="0" cellpadding="0" cellspacing="0" width="947">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:296px;">Medetomidine Hydrochloride (10 mL Injection)</td>
			<td style="width:132px;">Therapon/Zoetis</td>
			<td style="width:119px;">PFIDOM10</td>
			<td style="width:400px;">Multiple suppliers: Zoetis/Ilium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Thiopentone</td>
			<td style="width:132px;">Troy</td>
			<td style="width:119px;">Triothiopentone</td>
			<td>Multiple suppliers: Neon Laboratories, Jagsonphal Pharmaceuticals</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Isoflurane (2-3 % in oxygen)</td>
			<td style="width:132px;">Baxter</td>
			<td style="width:119px;">AHN3636</td>
			<td>Multiple suppliers: Baxter/VetOne</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Amoxicillin parenteral</td>
			<td style="width:132px;">GlaxoSmithKline</td>
			<td style="width:119px;">JO1CA04</td>
			<td>Multiple suppliers: GlaxoSmithKline/Merck</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Bupivacaine (0.5% Injection 20 mL)</td>
			<td style="width:132px;">Pfizer</td>
			<td style="width:119px;">005BUP001</td>
			<td>Multiple suppliers: Pfizer/AstraZeneca</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">PVD Iodine Solution</td>
			<td style="width:132px;">Jurox</td>
			<td style="width:119px;">61330</td>
			<td>Multiple suppliers: Jurox/Orion</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:296px;">Chlorhexidine 5%w/v</td>
			<td style="width:132px;">Jurox</td>
			<td style="width:119px;">Chlorhex C 5L (SCRUB)</td>
			<td>Multiple suppliers: Jurox/Pfizer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Transdermal Fental Patch (75 &#956;g/h)</td>
			<td style="width:132px;">Janssen-Cilag</td>
			<td style="width:119px;">S8-Dur7.5</td>
			<td>Multiple suppliers: Sandoz</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Buprenorphine iv</td>
			<td style="width:132px;">Jurox</td>
			<td style="width:119px;">504410</td>
			<td>Multiple suppliers: LGM Pharma</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:296px;">Atipamezole (Antisedan 0.06 mg/kg - 0.08 mg/kg)</td>
			<td style="width:132px;">Zoetis</td>
			<td style="width:119px;">PFIANT10</td>
			<td>Multiple suppliers: Ilium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Oster Golden A5 2-Speed Clippers</td>
			<td style="width:132px;">Oster</td>
			<td style="width:119px;">078005-140-003</td>
			<td>Oster</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">20 ml luer lock syringe</td>
			<td style="width:132px;">Terumo</td>
			<td style="width:119px;">6SS+20L</td>
			<td>Multiple suppliers: Medshop Australia/Terumo</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">21 G IV needle</td>
			<td style="width:132px;">Terumo</td>
			<td style="width:119px;">SG3-1225</td>
			<td>Multiple suppliers:Medshop Australia/Terumo</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">#4 scalpel handle</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;">AD010/04</td>
			<td>Multiple suppliers: Austvet/SurgicalInstruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">#22 scalpel baldes</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Gillies tissue forceps</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;">AB430/15</td>
			<td>Multiple suppliers: Austvet/SurgicalInstruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Metzenbaum curved dissecting scissors</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;">AC101/14</td>
			<td>Multiple suppliers: Austvet/SurgicalInstruments</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:296px;">Deaver retractor</td>
			<td style="width:132px;">Surgical Instruments</td>
			<td style="width:119px;">23.75.03</td>
			<td>Multiple suppliers: Surgical Instrument/Austvet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Hohmann retractor</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;">KA173/35</td>
			<td>Multiple suppliers: Austvet/SurgicalInstruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mayo suture scissors</td>
			<td style="width:132px;">Austvet</td>
			<td style="width:119px;">AC911/14</td>
			<td>Multiple suppliers: Austvet/SurgicalInstruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Needleholder 14 cm&#160;</td>
			<td style="width:132px;">EliteMedical</td>
			<td style="width:119px;">18-1030</td>
			<td>Multiple suppliers: EliteMedical/Austvet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">CMT 3.5 mm Brad-Point Drill</td>
			<td style="width:132px;">Carbatec</td>
			<td style="width:119px;">516-035-51</td>
			<td>Multiple suppliers: Southeast Tool/Carbatec</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Drill Bit Stop 4 mm</td>
			<td style="width:132px;">Drill Warehouse</td>
			<td style="width:119px;">20121600</td>
			<td>Multiple suppliers: Amazon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Bosch 10.8 V Cordless Angle Drill</td>
			<td style="width:132px;">Get Tools Direct</td>
			<td style="width:119px;">GWB10.8V-LIBB</td>
			<td>Multiple suppliers:Bunnings/Get Tools Direct</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">Autoclavable veterinary drill bag</td>
			<td style="width:132px;">AustVet</td>
			<td style="width:119px;">DRA043-AV</td>
			<td>AustVet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">2-0 absorbable synthetic braided sutures</td>
			<td style="width:132px;">Ethicon</td>
			<td style="width:119px;">VCP335H</td>
			<td>Ethicon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:296px;">3-0 absorbable synthetic braided sutures</td>
			<td style="width:132px;">Ethicon</td>
			<td style="width:119px;">VCP232H</td>
			<td>Ethicon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Siemens 3 Tesla Skyra Widebore MRI</td>
			<td style="width:132px;">Siemens</td>
			<td style="width:119px;">N/A</td>
			<td>Siemens</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">9.4 Tesla Agilent (Varian) MRI</td>
			<td style="width:132px;">Agilent Technologies</td>
			<td style="width:119px;">N/A</td>
			<td>Agilent Technologies</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Atomscope HF 200 A Radiogaph</td>
			<td style="width:132px;">Radlink</td>
			<td style="width:119px;">330003A</td>
			<td>Multiple Suppliers: Radlink/DLC Australia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Veterinary Pulse Oximiter</td>
			<td style="width:132px;">DLC&#160;</td>
			<td style="width:119px;">192500A</td>
			<td>Multiple suppliers: DLC Australi Pty Ltd/AustVet</td>
		</tr>
	</tbody>
</table>

ovine lumbar intervertebral disc degeneration model utilizing a lateral retroperitoneal drill bit injury

椎間板の変性は、背痛の発症および世界的な障害の主要原因に大きく寄与する。椎間板変性の数多くの動物モデルが開発されている。理想的な動物モデルは、形態、生体力学的性質、および脊索細胞の欠如に関して、ヒト椎間板を酷似して模倣すべきである。ヒツジ腰椎椎間板モデルは、これらの基準を満たす。本発明者らは、横方向の後腹膜アプローチによるドリルビット損傷を利用した椎間板変性のヒツジモデルを提示する。側方アプローチは、ヒツジ背骨への伝統的な前方アプローチに伴う切開および潜在的な罹患率を有意に減少させる。ドリルビットの損傷方法を利用することにより、一定の程度の椎間板変性を開始する、正確で寸法の一貫した再現性のある損傷を生成する能力が得られる。環状の焦点の性質核脊髄欠損は、焦点椎間板ヘルニアの臨床状態をより密接に模倣する。羊はこの手順に従って急速に回復し、通常は移動して1時間以内に食べています。椎間板変性が起こり、羊は8週間の期間で検死およびその後の分析を受ける。我々は、椎間板変性のドリルビット損傷モデルが、従来の環状損傷モデルよりも利点があると考えている。

術前に、ヒツジは根底にある椎間板の形態および変性の評価のためのベースライン3T磁気共鳴画像法（MRI）を受けた。羊は、椎間板レベルの確認および椎間板高さ指数の計算のために、追加の術中側方X線撮影を受けた。 図1に 、3T MRIの術前矢状面スライスと術中X線写真を示します。 <img alt="図1" sr...

Watch this Scientific Journal Video about Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury at JoVE.com

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

椎間円板の変性は、背痛および世界的な障害の主要な原因に大きく寄与する。椎間板変性の多くの動物モデルが存在する。一貫した椎間板損傷および再現性のある椎間板変性を達成するドリルビットを利用して、椎間板変性のヒツジモデルを実証する。

側方腹腔ドリルビット傷害を利用した羊の腰椎椎間板変性モデル

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous ...

ovine-lumbar-intervertebral-disc-degeneration-model-utilizing-lateral

Research

JoVE Journal

Medicine

11.5K ビュー.  Monash University. 椎間円板の変性は、背痛および世界的な障害の主要な原因に大きく寄与する。椎間板変性の多くの動物モデルが存在する。一貫した椎間板損傷および再現性のある椎間板変性を達成するドリルビットを利用して、椎間板変性のヒツジモデルを実証する。 

ビデオ: Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

腎臓虚血再灌流障害：障害のマウスモデルおよび再生

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

医学

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

組織の変性/再生と癌の研究におけるマウス誘導テロメラーゼ対立遺伝子を利用

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生児マウス脊髄圧縮損傷モデル

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

マウスにおける反復震とう頭部損傷モデル

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

Transvaginal Mesh Insertion in the Ovine Model

This protocol describes mesh insertion into the rectovaginal septum in sheep using a single vaginal incision technique, with and without the trocar-guided insertion of anchoring arms. Parous sheep underwent the dissection of the rectovaginal septum, followed by the insertion of an implant with or without four anchoring arms, both designed to fit the ovine anatomy. The anchoring arms were put in place using a trocar and an "outside-in" technique. The cranial arms were passed through the obturator, gracilis, and adductor magnus muscles. The caudal arms were fixed near the sacrotuberous ligament, through the coccygeus muscles. This technique allows for the mimicking of surgical procedures performed in women suffering from pelvic organ prolapse. The anatomical spaces and elements are easily identified. The most critical part of the procedure is the insertion of the cranial trocar, which can easily penetrate the peritoneal cavity or the surrounding pelvic organs. This can be avoided by a more extensive retroperitoneal dissection and by guiding the trocar more laterally. This approach is designed only for experimental testing of novel implants in large animal models, as trocar-guided insertion is currently not used clinically.

This protocol describes mesh implantation in the ovine rectovaginal septum using a single vaginal incision technique, with and without the trocar-guided insertion of anchoring arms.

ヒツジモデルにおける経膣メッシュの挿入

This protocol describes mesh insertion into the rectovaginal septum in sheep using a single vaginal incision technique, with and without the trocar-guided ...

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

片性および反復性軽度外傷性脳損傷のマウスモデル

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human Brown-S&#233;quard syndrome&#160;is a common type of incomplete SCI caused by a lesion to one half of the spinal cord which results in paralysis and loss of&#160;proprioception&#160;on the same (or ipsilesional) side as the injury, and loss of pain and temperature sensation on the opposite (or contralesional) side. Adequate methodologies for producing a spinal cord lateral hemisection (HX) and assessing neurological impairments are essential to establish a reliable animal model of Brown-S&#233;quard syndrome. Although lateral hemisection model plays a pivotal role in basic and translational research, standardized protocols for creating such a hemisection and assessing unilateralized function are lacking. The goal of this study is to describe step-by-step procedures to produce a rat spinal lateral HX at the 9th thoracic (T9) vertebral level. We, then, describe a combined behavior scale for HX (CBS-HX) that provides a simple and sensitive assessment of asymmetric neurological performance for unilateral SCI. The CBS-HX, ranging from 0 to 18, is composed of 4 individual assessments which include unilateral hindlimb stepping (UHS), coupling, contact placing, and grid walking. For CBS-HX, the ipsilateral and contralateral hindlimbs are assessed separately. We found that, after a T9 HX, the ipsilateral hindlimb showed impaired behavior function whereas the contralateral hindlimb showed substantial recovery. The CBS-HX effectively discriminated behavioral functions between ipsilateral and contralateral hindlimbs and detected temporal progression of recovery of the ipsilateral hindlimb. The CBS-HX components can be analyzed separately or in combination with other measures when needed. Although we only provided visual descriptions of the surgical procedures and behavioral assessments of a thoracic HX, the principle may be applied to other incomplete SCIs and at other levels of the injury.

Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury.

成人ラットにおける脊髄横片切と非対称行動評価

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human ...

A Mobile Outside-in Technique of Transforaminal Lumbar Endoscopy for Lumbar Disc Herniations

Percutaneous endoscopic transforaminal lumbar discectomy (PETLD) has now become a standard of care for the management of lumbar disc disease. There are two techniques for the introduction of a working cannula with respect to disc&#8212;outside-in and inside-out. The aim of this prospective study is to describe the technical aspects of a novel mobile outside-in method in dealing with different types of disc prolapse. A total of 184 consecutive patients with unilateral lower limb radiculopathy due to lumbar disc prolapse were operated on with the mobile outside-in technique of PETLD. Their clinical outcomes were evaluated based on the type of disc prolapse they had, a visual analog scale (VAS) leg pain score, the Oswestry Disability Index (ODI), and the Macnab criteria. The completeness of the decompression was documented with a postoperative magnetic resonance imaging. The mean age of the patients was 50 &#177; 16 years and the male/female ratio was 2:1. The mean follow-up was 19 &#177; 6 months. A total of 190 lumbar levels were operated on (L1&#8211;L2: n = 4, L2&#8211;L3: n = 17, L3&#8211;L4: n = 27, L4&#8211;5: n = 123, and L5&#8211;S1: n = 19). Divided into types, the patient distribution was central: n = 14, paracentral: n = 74, foraminal: n = 28, far lateral: n = 13, superior-migrated: n = 8, inferior migrated: n = 38, and high canal compromise: n = 9. The mean operative time was 35 &#177; 12 (25 - 56) min and the mean hospital stay was 1.2 &#177; 0.5 (1&#8211;3) days. The VAS score for leg pain improved from 7.5 &#177; 1 to 1.7 &#177; 0.9. The ODI improved from 70 &#177; 8.3 to 23 &#177; 5. According to the Macnab criteria, 75 patients (40.8%) had excellent results, 104 patients (56.5%) had good results, and 5 patients (2.7%) had fair results. Recurrence (including early and late) was seen in 15 out of the 190 levels that were operated on (7.89%). This article presents a novel outside-in approach that relies on a precise landing within the foramen in a mobile manner and does not solely depend upon the enlargement of the foramen. It is more versatile in application and useful in the management of all types of disc prolapse, even in severe canal compromise and high migration.

Here we present a protocol of a novel outside-in technique of transforaminal endoscopic discectomy for lumbar disc herniations. The technical aspects of the technique, the wide indications of use, and the results of the treatment in 184 patients are described in detail.

腰椎椎間板ヘルニアの切開腰椎内視鏡外でモバイルの技術

Percutaneous endoscopic transforaminal lumbar discectomy (PETLD) has now become a standard of care for the management of lumbar disc disease. There are ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

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

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

ワイヤー図法を使用してマウスの腸間膜動脈の等尺性収縮力測定

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

A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease.

Symptomatic intervertebral disc (IVD) degeneration (IDD) is a major socioeconomic burden and is characterized by inflammation and tissue degradation. Due to the lack of causative therapies, there is an urgent need for innovative experimental organ culture models to study the mechanisms involved in the progression of the disease, find therapeutic targets, and reduce the need for animal models. We here present a novel, three-dimensional organ culture model protocol mimicking the proinflammatory and catabolic microenvironment, which is present during IDD. 
Initially, bovine caudal IVDs were dissected, cleaned, and cultured in the tissue culture medium. Dynamic physiologic or pathologic loading was applied in a custom-made bioreactor for 2 hours per day. IVDs were assigned to a control group (high glucose medium, physiological loading, phosphate-buffered saline injection) and a pathological group (low glucose medium, pathological loading, tumor necrosis factor-alpha injection) for four days. Gene expression analysis from collected nucleus pulposus cells of the IVDs and enzyme-linked immunosorbent assay of the conditioned organ culture media was performed. 
Our data revealed a higher expression of inflammatory markers and reduced disc heights after loading in the pathological group compared to the control group. This protocol is reliable to simulate IVD inflammation and degeneration and can be further expanded to broaden its application scope.

This protocol presents a novel experimental model of proinflammatory, degenerative bovine organ culture to simulate early-stage intervertebral disc degeneration.

初期段階の椎間板疾患をシミュレートする炎症性の退行性臓器培養モデル。

Symptomatic intervertebral disc (IVD) degeneration (IDD) is a major socioeconomic burden and is characterized by inflammation and tissue degradation. Due ...