著者は、このモデルの開発における彼らの指導のために博士N. Banik博士とD.ミッチェルに感謝しています。

1。脊髄損傷前の準備この手順に必要な手術器具は、歯とないメス、ピックアップ、止血、自己保持リトラクター、細かい先端骨鉗子、針ドライバ、吸収性縫合糸、皮膚クリップアプリケーターです。必要な他の手術用品外科用ドレープ、手術野のための無菌シーツ、ガーゼスポンジ、綿チップアプリケーター、及び金属箔である。手術の前に手術器具や消耗品をオートクレーブ。一匹のための機器および消耗品の個々のセットを使用します。アルコールワイプで手術領域および装置（インパク、光源、定位フレーム、ガラスビーズ滅菌、加熱パッド）を清掃してください。 外科用ドレープを開き、慎重に汚染を回避手術野をドレープに滅菌手袋を使用しています。無菌個々の楽器を開き、慎重に手術野のものを配置します。番目のノブやハンドルカバー電子装置は、おそらく無菌の金属箔との手順の実行中に必要としていました。スイッチオンガラスビーズ滅菌手順中に使用可能な状態になるように。 2。動物の作製実際の手順の前に研究室エリア少ない時間にラットを持参。前もって予想手順（通常0.006 mg / mlの皮下のbuprinorphine 1.5ミリリットル）の少なくとも1時間鎮痛薬を管理します。術前の抗生物質（通常はバイトリル4 mg / kgを皮下）を管理。ケタミンの90 mg / kg体重を使ってラットを麻酔しないつま先ピンチ応答がなくなるまで待つ。胸椎で最も著名な棘突起を触診する。このレベルは、通常、T10棘突起に対応しています。 T10との関係で意図されたレベルの位置をマークします。私たちの研究室では、T10けがを実行します。次の物語は、T10 SCIの技術が記載されている。縦3センチメートル×6センチの長方形を剃るとT10のレベルを中心。清潔な肌三回のWi番目ベタジンソリューション。それぞれの目に眼科潤滑剤を適用します。慎重に汚染を回避手術野に楽な姿勢でラットを転送します。コア温度を監視し、正常に近い（〜37.5℃）できるだけのような動物の温度を保つためにそれに応じて加熱を調節するために直腸の温度プローブを挿入します。手術部位上記のウィンドウで外科用ドレープとラットをカバーしています。 3。外科手術 T10マークを中心に＃10のブレードを使用して近似横4cm切開で始まる。辛抱強く離れT9-T11棘突起と薄層から筋膜と筋肉層を解剖に進みます。骨から筋肉や筋膜を離れて撤回​​するトラクターを配置します。 T9の棘突起を安定しながら鋭く細かいハサミを使用してT9とT10の間に棘靭帯を分ける。同様にT10棘突起を安定させ、T10-T11の間に棘靭帯を分ける。補完するために、ループ倍率を使用黄色靱帯（ 図1）を介してTE靭帯の部門はすべての方法。黄色靱帯が切断された後莢膜嚢は容易に明らかである。慎重にT10で両側ピース食事椎弓切除を実行するために細い先端骨鉗子を使用してください。細心の注意は、骨鉗子先端から莢膜嚢と不注意傷害に下向きの圧力を避けるために取られる。ラミナとT10の棘突起が完全に削除されます。 安定化プラットフォーム上の位置に動物を移動します。 T9椎体の左右側面に続いT11椎体の横方向の側面をクランプして背骨を固定する安定クランプを使用してください。これは呼吸の動きのためのスペースを制限し、不必要なストレスを動物を追加しますと安定クランプでプラットフォームにラットを圧縮しないように注意してください。コンピュータ制御のインパクタで脊柱の設定を固定した後にチェックされます。我々は一般的に、SPで3.0ミリメートルのインパクチップを使用2mmの深さと0.3秒の滞留時間で4センチメートル/秒のEED。インパクチップを拡張し、それだけで脊髄表面に触れるまでそれを下げる。先端を撤回し、脊髄表面に向かってデバイス2ミリメートルを下げる。重度の挫傷、脊髄損傷を引き起こすために、4センチメートル/秒の速度でピストンを離します。コー​​ドの上に不要な圧力を作成しないように注意しながら、所定の位置に綿チップアプリケーターを維持するだけの十分な圧力を使って止血を達成。 8の字ステッチがきつすぎる使って吸収性縫合糸を引っ張らないように注意してとの縫合の筋肉と筋膜層。 2つの小さなステープルの最小値で皮膚を閉じ、切開部の一部が第2又は3のステープル後に開いたままにする場合、最大4または5のステープルを使用することができる。 3。術後ケア 24時間後の手術のため、約33-35°Cの暖かい環境で場所ラット。彼らが移動し始めたら、これがインキュベーター（彼らは無意識でありながら）、加熱されたケージのスペースを必要とする。かつてラットは完全にAWですAKEは、すべての皮下に生理食塩水5mlの、buprinorphine 1.5mlの（0.006 mg / ml）であり、バイトリルの0.1ミリリットルを投与。 1日1回7日間最初の24〜48時間とバイトリルため皮下一日二回buprinorphineに進みます。膀胱を手動膀胱機能のリターン（3日間連続早朝式で尿の&lt;2ミリリットル）まで一日三回を表明すべきである。動物も感染症（尿中の血液、白っぽい色、または悪臭）、この時間中にチェックされるべきであり、創傷治癒と身体活動や問題を減少させた。感染症の存在は地元の獣医師と相談の上、抗生物質の投与量が増加（または再起動）で対抗すべきである。毎日彼らの回復を評価するために、手術の翌日から始まるラットの重量を量る。

<ol>
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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+series+of+systematic+reviews+on+the+treatment+of+acute+spinal+cord+injury:+a+foundation+for+best+medical+practice.">A series of systematic reviews on the treatment of acute spinal cord injury: a foundation for best medical practice.</a> J. Neurotrauma. 28, 1329-1333 (2011).</li><li>Iseda, T., Okuda, T., Kane-Goldsmith, N., 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=Single,+high-dose+intraspinal+injection+of+chondroitinase+reduces+glycosaminoglycans+in+injured+spinal+cord+and+promotes+corticospinal+axonal+regrowth+after+hemisection+but+not+contusion.">Single, high-dose intraspinal injection of chondroitinase reduces glycosaminoglycans in injured spinal cord and promotes corticospinal axonal regrowth after hemisection but not contusion.</a> J. 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V., 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=Impact+speed+does+not+determine+severity+of+spinal+cord+injury+in+mice+with+fixed+impact+displacement.">Impact speed does not determine severity of spinal cord injury in mice with fixed impact displacement.</a> Journal of Neurotrauma. 26, 1395-1404 (2009).</li><li>Kwon, B. K., Hillyer, J., Tetzlaff, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+research+in+spinal+cord+injury:+a+survey+of+opinion+from+the+SCI+community.">Translational research in spinal cord injury: a survey of opinion from the SCI community.</a> J. Neurotrauma. 27, 21-33 (2010).</li><li>Kwon, B. K., Okon, E. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+treatment+trials+in+human+spinal+cord+injury:+issues,+difficulties,+and+recommendations.">Review of treatment trials in human spinal cord injury: issues, difficulties, and recommendations.</a> Neurosurgery. 59, 957-982 (2006).</li></ol>

財務情報の開示なし 資金調達の開示なし。

いくつかの新規治療法が最近SCI研究の分野における初期の約束を示している。これらの治療法の3は十分な評価が最大の翻訳可能性のある戦略を選択するためにSCIの臨床的に関連するモデルでは必須である。グレーディングのスキームは、最近、前臨床試験の強度を評価するために開発されました。9このスキームは厳しいSCIの挫傷モデルを活用することの重要性を強調した。?...

適切な損傷モデルの選択は、脊髄損傷のための新たな治療法（SCI）の前臨床評価のために重要です。医師や神経外傷挫傷モデルの分野における科学者の最近の調査では、1,2,13、片側切断または完全離断モデルとは対照的に、 、普遍的に臨床的に関連のあることが受け入れられた。8この意見は、ヒトの脊髄損傷の大部分が自然の中で打撲であるという観察に基づいている。10挫傷の生物学は、また片側切断または離断モデルとは異なるように見えます。11 Iseda らはらは片側切断及び挫傷モデルで別々に神経再生に脊髄コンドロイチナーゼABC注射の効果を比較した4軸索再生が片側切断におけるニューロンの橋ではなく、挫傷SCI群で観察された。片側切断または完全な横断モデルもCLのごく小さなサブセットに存在することが知られている条件を作成するinical状況。たとえば、複数の研究者が片側切断または再生を促進するための完全な離断後に病変空洞内に注入するための足場ベースの介入を採用しました。6負傷脊髄内の空洞の作成 ​​は非現実的で、おそらく非倫理的であるため、このアプローチは、臨床的には無関係になります。 機能回復にばらつきが挫傷モデルの主要な課題である。5,12このばらつきは、脊髄ボリューム全体で均一な力の配信のためのインパクトは特に腹側に位置する運動経路の前にコンピューター制御のインパクと背骨の安定化を使用することによって最小限に抑えることができます。それは生き残った軸索からの可塑性と担保貢献は脊髄損傷後の回復の主なメカニズムであることに注意しなければならない。挫傷技法で1したがって些細なバリエーションが大幅に異なる結果をもたらす可能性があります。この目的を達成するために、我々は開発してきた一貫挫傷量と離断モデルと同等の機能回復をもたらす深刻な脊髄損傷のモデル。このモデルは、治療効果の概念の証拠として、神経保護及び神経再生戦略の両方を調査するために利用され得る。

<table border="1">
<tbody>
 <tr>
 <td>Instrument/Drugs</td>
 <td>Company</td>
 <td>Cat #</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Computer controlled impactor</td>
 <td>Leica or the Infinite Horizons (formerly OSU) impactor</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Surgical instruments </td>
 </tr>
 <tr>
 <td>Scissors</td>
 <td>Fine Science Tools Inc
 </td>
 <td>14094-11 or 14060-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Forceps</td>
 <td>Fine Science Tools Inc </td>
 <td>11006-12 and 11027-12 or 			11506-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Hemostats</td>
 <td>Fine Science Tools Inc </td>
 <td>13009-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Retractors</td>
 <td>Fine Science Tools Inc </td>
 <td>17011-10</td>
 <td></td>
 </tr>
 <tr>
 <td>Rongeurs</td>
 <td>Fine Science Tools Inc </td>
 <td>16020-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle driver</td>
 <td>Fine Science Tools Inc </td>
 <td>12001-13</td>
 <td></td>
 </tr>
 <tr>
 <td>Stereotactic frame</td>
 <td> Leica or RWD Life Science Co. or
 TSE systems</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprinorphine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Baytril</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Ketamine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

a contusion model of severe spinal cord injury in rats

小説の治療の並進ポテンシャルは厳しい脊髄損傷（SCI）挫傷モデルで調査する必要があります。詳細な方法は重度のSCIの一貫したモデルを得ることが記載されている。定位フレームとコンピュータ制御のインパクタを使用することにより、再現性の傷害の作成が可能になります。低体温症と尿路感染症は、術後の期間に重要な課題を提起。毎日の体重記録と膀胱表情を持つ動物の注意深い監視が術後合併症の早期発見が可能になります。この挫傷モデルの機能的な結果は、離断モデルと同等です。挫傷モデルは、神経保護及び神経再生両方のアプローチの有効性を評価するために利用することができる。

病変ボリューム私たちは、技術は、上記に従うことによって大規模かつ一貫性のある病変のボリュームを取得しています。 2.04ミリメートル3（1.9から2.18 95％CI）（N = 5匹）の平均病変容積が得られたルクソールファストブルー染色を用いて、図2に示すと、病変震源を通してルクソールファストブルーを使用して代表的な染色で病変ボリュームを意味し?...

Watch this Scientific Journal Video about A Contusion Model of Severe Spinal Cord Injury in Rats at JoVE.com

A Contusion Model of Severe Spinal Cord Injury in Rats

重度の脊髄損傷の挫傷モデルが記載されている。詳細、術前手術と術後の手順は一貫したモデルを得るために説明されています。

ラットにおける重度の脊髄損傷挫傷モデル

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

a-contusion-model-of-severe-spinal-cord-injury-in-rats

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

30.3K ビュー.  Medical University of South Carolina. 重度の脊髄損傷の挫傷モデルが記載されている。詳細、術前手術と術後の手順は一貫したモデルを得るために説明されています。

ビデオ: A Contusion Model of Severe Spinal Cord Injury in Rats

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

ラット脊髄損傷後の急性および慢性の触覚官能検査

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

医学

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

慢性脊髄損傷患者で呼吸器モータ制御評価（RMCA）を使用して呼吸筋活動の評価

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

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, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

ラジオ遠隔測定システムは、脊髄離断および胚性神経細胞移植片幹ラットの心臓血管機能を監視する

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

脊髄損傷の実験的挫傷モデルにおける神経幹細胞移植

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

脊髄圧縮傷害の校正済み鉗子モデル

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

実験的脊髄損傷における脊髄血流の評価のための強化された超音波画像コントラスト

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

マウスの脊髄損傷のモデルとしての光血栓誘発性局所虚血

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

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 Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

マウスにおける組織変位ベースの脊柱挫傷モデルの検討

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...