このデバイスの開発はLISA㈱、ルイビル、ケンタッキー州によってサポートされていました。またXMXにCBS、およびNIH NS050243、NS052290、そしてNS059622に、ノートンのヘルスケアの継続的なサポートを認めるルイビル、ケンタッキー州。

1。背骨スタビライザーの動物の準備とアプリケーションマウス頸椎は、側面から見た腹凹である。 C3からT1への棘突起は小さく砕けており、一般的に3,4に記載されているようしたがって、脊椎の安定化には ​​適していません。私たちは、脊椎安定化横ファセット固定によって実行されることをお勧めします。固定装置は、横方向に各ファセットにクランプマウスと2つの調整ステンレス製のアームを支持するU字状の金属チャンネルから構成される。これは、ターゲットの椎骨の優れた固定化を提供します。脊椎固定した後、脊椎が若干脊髄のより良好な露光を提供するために、頸椎湾曲を平坦化するために上昇する。 <ol><li>鉗子、microscissorsの2組、30 G針、縫合糸と針ホルダー、皮膚クリップ、クリップアプリケーターの2-3ペア次手術器具を滅菌する。脊椎安定剤を消毒。 Anesケタミン/キシラジン（100ミリグラム/ 10 mg / kg）を腹腔内のカクテルを使用して、マウスをthetize。マウスの首から毛を剃る。 </li><li>ポビドンヨード液を、70％アルコールで皮膚洗浄した後、加熱パッドを温め手術台の上にマウスを移動します。角膜の乾燥を防ぐために、眼軟膏と動物の目をカバーしています。 </li><li>麻酔導入（マウステールピンチに応答しない場合に到達）した後、後頭部から下頚椎の皮下脂肪·パッドに後頸部正中皮膚切開を行います。倍率で、C2で僧帽筋の間に正中切開を行い、頭半棘筋の筋肉を分割。筋肉下の脂肪パッドの識別は正しい層に解剖を容易にします。 </li><li>信頼性のあるランドマークとして機能T2棘突起に尾側正中筋肉解剖を拡張します。 T2の椎骨に付着筋肉をカットし、T2の軟骨部分を除去棘突起。 </li><li>マイクロはさみのペアを使用してT2薄層を通してC2から傍脊柱筋群を解剖。筋肉解剖は棘突起に隣接始まり、椎間関節に両側​​及ぶ。出血を最小限に抑えるために棘突起とラミナ（骨膜層内）に直接隣接する筋肉を分離する。ラテラルファセットが露出された後、LISA段のU字型チャネル上にマウスを置きます。 </li><li>両側の面の下にさらさステンレスアームを取り付けます。腕が所定の位置にしたら、背骨を固定する鋼アームのつまみネジを締めます。これは、ターゲットの椎骨をしっかり固定を維持し、優れた露出を提供しています。アームは、脊椎の正確な水平配向を提供するように調整することができる。 </li><li>硬膜の基礎となる公開するC5とC6の間に黄色靱帯を切開。層間空間との間、microscissorsがに配置されるときに通過する小durotomyを作成するために30 G針を使用durotomyを拡張します。脊髄は、現在制御された病変裂傷を受けるように調製される。 </li></ol> 2。 LISAデバイスを使用した子宮頸脊髄裂傷<ol><li>頸髄拡大の幅が異なるレベルに変化する。 2.3ミリメートルフラットブレードを使用してC5-6で背片側切断病変を作り、脊髄の幅全体をカバーするために振動の振幅を設定します。ブレードはファイン科学ツール社（フォスターシティ、カリフォルニア州）から入手し、脊髄の裂傷のために変更されています。低い振幅レベルがコード裂傷のしやすさを減少するように、≥0.5ミリメートルでブレード振動の振幅を維持します。 </li><li> LISAの舞台で背骨の安定やマウスを置きます。ブレードは運動の3つの範囲が可能なマイクロドライバにより制御される地位にLISAに取り付けられている。 LISAとその機能の構成要素は、 図1に記載されている。 </li><li>電源にあるブレード·振動スイッチを。のmagnif下icationは、露出した脊髄を振動刃の直下に配置されるようにマウスを移動します。 </li><li>振動ブレードに向かってマウスをサポートしている段階に昇格。ブレードはかろうじて脊髄の背静脈に触れたときに &quot;0&quot;の位置が記録される。 &quot;0&quot;の位置に相対的な脊髄裂傷の深さを測定します。 </li><li>マイクロドライバ制御によるステージ位置を高める：マイクロドライバーノブの360°ターンは0.25ミリメートルでのステージを高めます。従って、0.75ミリメートル背側片側切断損傷部は、マイクロ·ドライバー·ノブを3回回すことによって作成される。病変の精度は±0.01 mmである。ブレードが脊髄を裂くし始めると、生理食塩水灌漑で手術野に注油してください。脊髄の切り込み深さは、縦マイクロドライバによって制御され、視覚的な指導とは無関係です。 </li><li>所定の深さに達すると、振動スイッチをオフにしてください。理想的には、揺動刃リットルに配置されている組織変形の証拠なしesionギャップ。切刃から段階を下げ、綿綿棒を使用して術野からの血液や生理食塩水を削除します。止血には、&lt;1分で自然に発生します。 </li><li>脊椎スタビライザーからマウスを離します。おおよそ6-0絹縫合糸を使用した傍脊柱筋とステンレスミシェルクリップを使用して皮膚創傷を閉じます。 </li></ol> 3。アニマルケア<ol><li>皮下に十分な水分補給を維持し、意識を取り戻しつつ加熱パッドに回復ケージにマウスを置いて1〜2ミリリットルの生理食塩水の合計を注入。 </li><li>水や柔らかい食品アドリブを提供し、術後48時間鎮痛薬を投与する。脊髄の背側片側切断後の膀胱心配する必要はない。 </li></ol>

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+spinal+cord+research+-+a+refined+strategy+for+the+International+Spinal+Research+Trust.">Progress in spinal cord research - a refined strategy for the International Spinal Research Trust.</a> Spinal Cord. 38 (8), 449 (2000).</li><li>Seitz, A., Aglow, E., Heber-Katz, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+from+spinal+cord+injury:+a+new+transection+model+in+the+C57Bl/6+mouse.">Recovery from spinal cord injury: a new transection model in the C57Bl/6 mouse.</a> J. Neurosci. 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<ol><li>いくつかの著者は（YPZ、XMX、CBS）ルイビルインパクシステム株式会社の金融関心を持っている</li><li>著者、李平張、リサBEシールズ、そしてクリストファーB.シールズは、ノートンヘルスケア、ルイビル、ケンタッキー州の従業員です。他の著者は、IN、インディアナ大学、インディアナ州で採用されています。 </li><li>著者は、この記事で利用試薬や楽器を生産する会社からの資金を受け取りませんでした。 </li></ol>

脊髄への裂傷の傷害の前に椎安定化が棘突起の固定によって得られた。マウスのT1を通してC3から砕けやすい短い頚椎棘突起へのクランプの頸椎前湾カーブと添付ファイルの両方が効果的な脊椎安定化を防ぐ。さらに、手動制御の下で利用カミソリの刃やmicroscissorsの使用は6病変の深さの変動を作成する重要な組織の変形を引き起こす。これは、特定の経路の軸索再生が研究され、特にデータの誤った解釈につながる可能性があります。背側皮質脊髄路が完全lesioning時に離断されていない場合、例えば、免れる背側皮質軸索が再生軸索として誤解されることがあります。これらの課題は、単一レベルおよび脊髄の正確なlesioningにおけるファセットに固定するとともに脊椎安定化装置を使用することによって克服することができる。また、ああを使用IGHの周波数振動ブレードは、隣接脊​​髄を粉砕またはcontusingなし鋭い裂傷を生成します。この方法は、マウス6の胸部脊髄裂傷を生成するための後続の修正を、ラット9,12,14における脊髄裂傷の傷害を生成するために使用されている。本通信では、マウスにおける信頼性子宮頸部病変裂傷を作成する方法を説明する。 脊髄の前後径は、マウスで&lt;2mmである限り、裂傷病変の正確な深さは信頼性のある実験モデルを作成する上で不可欠です。病変の深さの最小の変動性が大きく軸索再生と同様に、体積や行動の研究を評価する実験の結果を変更します。我々は、切断ブレードの位置を制御する高精度のマイクロドライバを使用したため、この方法を用いて病変部の深さの精度は±0.01 mmである。この方法はで矛盾を減少している裂傷SCIを作成する他のモデルでエラン。この方法は、このような皮質脊髄路、赤核脊髄路、および背側大動脈路として脊髄の背側半分に位置し、長い脊髄経路の軸索再生の研究に特に便利です。この方法では、これらの繊維路が完全かつ確実に切開することができる。この点において、データの解釈の誤差することにより、SCIに関する実験的研究の報告の信頼性を向上させる、最小限に抑えられる。 要約すると、我々は、マウスにおける頸髄裂傷の傷害のin vivoモデルにおいて再現を作成するための新規な技術を記載している。この手法は、振動ブレードを使用して脊髄の子宮頸部の側面と裂傷の固定により、脊椎安定化に基づいています。マウス6の背側胸部脊髄裂傷モデルでは、このメソッドを使用して、我々は裂傷の深さ、組織型との間に緊密な相関関係を示した、と行動回復。そのような技術はまた、いくつかの他の研究室2,12によって信頼性があることが見出されている。

遺伝子改変マウスの可用性は、SCIのメカニズムの役割を果たす特定の遺伝子の影響を識別するための強力なツールです。裂傷SCIこれは損傷8以下の効果的な治療を提供することができる治療薬または分子を調べるために使用される重要なモデルである。マウスで裂傷の傷害の作成 ​​時に棘突起の固定が原因で脊椎固定5,11の維持に関与して薄くて壊れやすい棘突起を把握するのが困難に不正確である。わずか0.2ミリメートル（マウス脊髄の直径の10％）の裂傷の深さのばらつきは、データの解釈を誤解を招く原因となる。脊髄裂傷病変の性質と程度を正確に10を定義する必要があります。この課題に対処するため、我々は、椎安定からなる新規技術を開発し、ルイビル傷害システム装置に接続されている製作のブレードを使用している（LISA）が裂傷SCI 7,14を生成する。この損傷は、裂傷プロセスの間、組織の変形を回避鋭い振動ブレードを使用して作成された。裂傷の深さは、裂傷の深さを制御するマイクロドライバを使用して0.01ミリメートルの精度に精密だった。カッティングブレードは希望裂傷輪郭9を作成するには 、特定の形状や幅に、カスタムメイドされています。我々は、1）頚椎露光、2の方法）両側性ファセット固定装置を用いて椎安定化する技術、及び振動ブレードを用いて子宮頸部裂傷損傷の3）の作成を示す。

<table border="1">
 <tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalogue Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Mice vertebral stabilizer </td>
 <td>Louisville Impactor System</td>
 <td></td>
 <td>Stabilize and expose the cervical vertebra</td>
 </tr>
 <tr>
 <td>LISA vibraknife </td>
 <td>Louisville Impactor System</td>
 <td></td>
 <td>Produce the laceration injury of the cervical spinal cord</td>
 </tr>
 <tr>
 <td>Spring Scissors</td>
 <td>Fine Science Tools (USA) </td>
 <td>15013-12</td>
 <td>Skin and trapezius muscle incision</td>
 </tr>
 <tr>
 <td>Spring Scissors</td>
 <td>Fine Science Tools (USA) </td>
 <td>15023-10</td>
 <td>Separate muscles from the laminae</td>
 </tr>
 <tr>
 <td>Spring Scissors</td>
 <td>Fine Science Tools (USA) </td>
 <td>15002-08</td>
 <td>Incision of dura</td>
 </tr>
 <tr>
 <td>Graefe forceps </td>
 <td>Fine Science Tools (USA) </td>
 <td>11154-10</td>
 <td>Retract skin</td>
 </tr>
 <tr>
 <td>Dumont #7 forceps </td>
 <td>Fine Science Tools (USA) </td>
 <td>11274-20</td>
 <td>Muscle retraction (tip modified)(Fig. A)</td>
 </tr>
 <tr>
 <td>Dumont SS forceps </td>
 <td>Fine Science Tools (USA) </td>
 <td>11203-25</td>
 <td>Fixation of vertebra (tip modified )(Fig.B)</td>
 </tr>
 <tr>
 <td>30G needle </td>
 <td>Becton Dickenson</td>
 <td>305106</td>
 <td>Create a dural opening </td>
 </tr>
 <tr>
 <td>6-0 suture </td>
 <td>Ethicon </td>
 <td>8806H</td>
 <td>Close muscle and fascial layers </td>
 </tr>
 <tr>
 <td>wound clip </td>
 <td>Fine Science Tools (USA) </td>
 <td>12031-07</td>
 <td>Skin closure</td>
 </tr>
 <tr>
 <td>Tribromoethanol (Avertin)</td>
 <td>Sigma-Aldrich</td>
 <td>90710-10G</td>
 <td>Anesthetic agent</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Louisville Impactor System, Inc, 210 E. Gray St., Suite 1102, Louisville, KY 40202, (502) 629-5510, E-mail: <a href="mailto:cbshields1@gmail.com">cbshields1@gmail.com</a>
 Fine Science Tools (USA), Inc, 373-G Vintage Park Drive, Foster City, CA 94404-1139, (800) 521-2109, E-mail: <a href="mailto:info@finescience.com">info@finescience.com</a> 
 Becton Dickenson, 1 Becton Drive, Franklin Lakes, NJ USA 07417, (201) 847-6800 Ethicon, Route 22 West,
 Somerville, NJ 08876 1-877-ETHICON 
 Sigma-Aldrich Corp. 
 St. Louis, MO, USA, 63178 (314) 771-5765, E-mail: cssorders@sial.com</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Figure A is the modified Dumont #7 forceps; B is the modified Dumont SS forceps.</td>
 </tr>
 </tbody>
 </table>

controlled cervical laceration injury in mice

遺伝子改変マウスの使用は、脊髄損傷（SCI）など、いくつかの神経疾患の根底にある分子機構の理解を向上させます。 SCIの裂傷モデルを生成するために使用されるフ​​リーハンド手動制御は、多くの場合、クラッシュまたは挫傷コンポーネントと、それゆえ、新しい技術が開発されたものに関連付けられている一貫性のない怪我を作成します。我々の子宮頸部裂傷SCIのモデル1を組み込むことにより、フリーハンド方式に固有​​の困難を解決した）椎骨ファセット固定によって頚椎安定化、2）強化された脊髄露出、と振動ブレードを使用して脊髄の再現可能な裂傷の3）の作成関連する挫傷せず、深さ±0.01ミリメートルの精度。このようなメスやはさみのフリーハンドの使用などのSCI裂傷を作成する標準的な方法に比べて、我々の手法は、一貫した病変を生産している。このメソッドはcorticospの軸索再生の研究に有用ですinal、赤核脊髄、そして背昇順広大。

ターゲット椎骨の固定化は、マウス脊髄の正確な病変を生成する際に非常に重要である。我々の脊椎安定化装置は、短い棘突起とマウス頸椎の腹側前弯の解剖学的な問題を克服する。頸椎は、我々頸椎安定します（ 図2）を使用して公開されます。我々のマウスの脊椎安定化装置は、頸髄手順については、背骨を製造するための信頼できる技術である。 LISAを使用して病変の深さは0.01ミリメートル6,13に正確です。正確な裂傷は、病変/組織インターフェース（ 図3）全く挫傷を引き起こしません。背片側切断病変の精度が0.9ミリメートル深い裂傷がちょうど脊髄1の病理セクションによって確認各標本における中央運河を超えて拡張する軸索再生に関する研究にC57BL / 6マウスで実証された。これらの動物のすべての歩行を回復この脊髄裂傷損傷後のED。 <img alt="図1" src="/files/ftp_upload/50030/50030fig1.jpg" /> 図1（A）LISAステージに載置された脊椎安定化剤でマウス。振動ブレードが引き裂かれる脊髄に向けられている。微小駆動制御は、ステージの下に配置され、適切な部位でマウスを位置決めするように設計されている。垂直マイクロドライバが病変の深さを制御し、チルトノブが裂傷の角度を防ぐために脊髄の水平面を制御する。オン·オフスイッチは、振動モータを制御し、別のノブは、その振幅を調整します（B）0.75ミリメートル背片側切断の裂傷病変が完全に層のアーチの下にカット。 <img alt="図2" src="/files/ftp_upload/50030/50030fig2.jpg" /> 図2。 </st栄&gt;（A）U字型チャネルと両腕とコネクタで構成され、マウスの脊椎スタビライザー。マウスは子宮頸部SCIに使用Cトラフや胸部SCIのためのTトラフに配置されます（B）頸椎がラテラルファセットの下に腕を置いた後、親指のネジをロックすることによって固定されています。硬膜、骨のいずれかの取り外しなしC5-6のラミナ、C6-7、およびC7-T1の間に露出されています。 <img alt="図3" src="/files/ftp_upload/50030/50030fig3.jpg" /> 図3 0.5の深さ、0.8、1.1、および矢状ビュー（シミクレ紫とエオシン）で観察された1.4ミリメートルで4背側脊髄裂傷は、この手法を用いた高精度を描いた。

Watch this Scientific Journal Video about Controlled Cervical Laceration Injury in Mice at JoVE.com

Controlled Cervical Laceration Injury in Mice

再現性を作成するための新たな手法<em生体内で&gt;</emマウスにおける頸髄損傷の裂傷&gt;モデルが記載されている。この技術は、±0.01程度の精度で振動ブレードを用いて脊髄の頸部のファセット及び裂傷の固定により、脊椎安定化に基づいている。

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

controlled-cervical-laceration-injury-in-mice

Research

JoVE Journal

Medicine

16.4K ビュー.  Norton Healthcare. 再現性を作成するための新たな手法モデルが記載されている。この技術は、±0.01程度の精度で振動ブレードを用いて脊髄の頸部のファセット及び裂傷の固定により、脊椎安定化に基づいている。

ビデオ: Controlled Cervical Laceration Injury in Mice

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged mechanical ventilation, contributes to morbidity and mortality of critical illness, and studies on novel genetic or pharmacological targets are areas of intense investigation (1-3, 5, 8, 26, 30, 33-36). ALI is defined by the acute onset of the disease, which leads to non-cardiac pulmonary edema and subsequent impairment of pulmonary gas exchange (36). We have developed a murine model of ALI by using a pressure-controlled ventilation to induce ventilator-induced lung injury (2). For this purpose, C57BL/6 mice are anesthetized and a tracheotomy is performed followed by induction of ALI via mechanical ventilation. Mice are ventilated in a pressure-controlled setting with an inspiratory peak pressure of 45 mbar over 1 - 3 hours. As outcome parameters, pulmonary edema (wet-to-dry ratio), bronchoalveolar fluid albumin content, bronchoalveolar fluid and pulmonary tissue myeloperoxidase content and pulmonary gas exchange are assessed (2). Using this technique we could show that it sufficiently induces acute lung inflammation and can distinguish between different treatment groups or genotypes (1-3, 5). Therefore this technique may be helpful for researchers who pursue molecular mechanisms involved in ALI using a genetic approach in mice with gene-targeted deletion.

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

マウスにおける急性肺障害を誘発するために圧力制御された換気

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

医学

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

筋萎縮性側索硬化症や外傷性脊髄損傷では頚椎前角をターゲットとするための髄腔内細胞移植

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

無限の地平線インパクターを用いた片側頚髄損傷の打撲のモデル

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

マウスにおける急性腎障害とポスト傷害線維の虚血再灌流モデル

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

ポスト病変呼吸神経可塑性を研究するための子宮頸脊髄損傷のマウスモデル

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

外傷性脳損傷のための制御皮質衝撃モデル

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

実験的子宮頸脊髄損傷における神経前駆細胞と自己組織化ペプチドの相乗使用

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

マウスにおける異子宮頸心臓移植

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

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 ...

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. ...