SN und MDD erhielt Unterstützung von NIH K08 NS083740 und der UCLA Klinik für Neurologie. AJG dankt für die Unterstützung durch die Dr. Miriam und Sheldon G. Adelson Medical Research Foundation und der Larry L. Hillblom Foundation. KLN dankt Unterstützung von der American Heart Association 14BFSC17760005 ASA-Bugher Stroke Center. ILL, EGS und STC wurden von NIH R01 NS071481 unterstützt. JDH dankt für die Unterstützung von NIH K08 NS083740.

Die Verwendung von Tieren in diesem Protokoll wurde in Übereinstimmung mit Verfahren, die von der University of California Los Angeles Animal Care und Use Committee genehmigt durchgeführt wurde. Hinweis: Beginnen Sie mit der Ziel murine Bevölkerung zu identifizieren. In früheren Studien, nur männliche Wildtyp-C57 / BL6-Mäusen verwendet wurden, jedoch verschiedene transgenen oder Knockout-Mäuse auch verwendet werden können. Beachten Sie, dass sind stereotaktischen Koordinaten basierend auf C57 / Bl6 Anatomie. Es wird empfohlen, dass jeder Benutzer zunächst die Lokalisierung des Hubs zu der weißen Substanz zu überprüfen. 1. Weiß Matter Stroke Induktion - Medial mit schräger Ansatz <ol><li> Beginnen Sie mit einer gezogenen Glaspipette unter Verwendung von 0,5 mm Kapillarrohr vorbereitet , so dass der distale Durchmesser zwischen 15 bis 25 &amp; mgr; m 11 ist. </li><li> Bereiten einer sterilen 10-ul-Aliquot von L-Nio (N (5) - (1) -iminoethyl-L-Ornithin HCl) bei 27,4 mg / ml (130 uM) in steriler 0,9% normale Kochsalzlösung. </li><li> Vorfüllen die gezogener Glaspipettemit einem kleinen Volumen von L-Nio (2-5 &amp; mgr; l) durch die Glaspipette Befestigung mit einer Vakuumleitung an die Rohrleitung. Legen Sie die Pipette flach auf der Tischplatte und setzen Sie das zog Ende in die L-Nio Lösung. <ol><li> Übernehmen des Vakuums, bis mindestens 2 mm von der 0,5 mm Abschnitt der Pipetten gefüllt ist. Schalten Sie das Vakuum aus, und die Pipette zurückziehen. Legen Sie sie beiseite, bis Schritt 1.12. </li></ol></li><li> Platzieren Sie die Maus in einer Induktionskammer und induzieren Anästhesie der Maus unter Verwendung von Standard 32% Isofluran floss durch einen Verdampfer (5 l / min inhaliert mit 5 l / min Sauerstoff und 0,5 l / min N 2) für 1 min oder bis tief anästhesiert. Übertragen Sie die Maus an einen stereotaktischen Apparat mit einem stereotaktischen Mikroskop ausgestattet. Geben Sie Isofluran Aufrechterhaltung der Narkose mit 32% floss durch einen Verdampfer (2 l / min inhaliert mit 5 l / min Sauerstoff und 0,5 l / min N 2) und einem Nasenkegel. Prüfen Tiefe der Anästhesie mit einer Zehe Prise. </li><li> Stellen Sie den Injektionsarm bis 36 °. </li><li> Affixa gezogen Glaspipette Halter an das distale Ende eines geringen Volumendruck-Einspritzsystem und befestigen Sie es an der Injektions Arm des Stereotaxie-Setup. </li><li> Mantel des narkotisierten Tieres Whiskers mit Vaseline und Ort künstliche Tränen Salbe über beide Augen. Bereiten Sie eine sterile OP-Feld durch einen sterilen Tuch über den Kopf des Tieres mit einer 5-10 cm Öffnung über den Kopf stellen. Bereiten Sie eine aseptischen OP-Oberfläche, die durch das Fell rasiert den Schädel überlagert. Reinigen Sie die Kopfhaut mit abwechselnden betadine und 70% Alkoholtupfer. </li><li> Machen Sie eine 1,5 cm Mittellinie Kopfhauteinschnitt mit sterilen feinen Schere, um die Schädeloberfläche zu belichten. Trocknen Sie den Schädel mit einem sterilen Wattestäbchen und unter Verwendung eines stereotaktischen Mikroskop bei 1-3x Vergrößerung, entfernen Sie alle darüber liegenden periostaler Gewebe eine sterile Mikropunkt-Werkzeug. </li><li> Markieren Sie die Bregma als Referenzpunkt einen feinen Punkt Marker. </li><li> Bohren Sie ein 2 mm ellipitical Craniotomie einer sterilen feinen stip chirurgische Bohrer mit0; posterior am Bregma beginnen und nach vorne gerade links von der Mittellinie erstrecken. Entfernen von Knochenfragmenten und Weichgewebe überlagert, so dass der Großhirnrinde sichtbar gemacht werden kann. </li><li> Halten Sie das OP-Feld und kortikalen Oberfläche feucht durch intermittierend Tropfen steriler Kochsalzlösung anwenden. </li><li> Affix eine gezogen Glaspipette mit dem Injektor Arm des stereotaktischen Apparat. Richten Sie das distale Ende der Pipette mit dem Bregma und Null, um die stereotaktischen Koordinaten. </li><li> Vorzurücken die Pipette mit dem ersten anterioren / posterioren (A / P) und medial / lateral (M / L) -Koordinaten in Tabelle 1 bereitgestellt. </li><li> Schieben Sie die Pipette auf die kortikalen Oberfläche und Null die dorsale / ventralen (D / V) Messung. </li><li> Passieren langsam die Pipette in das Gehirn bis zum Erreichen der ersten D / V in Tabelle 1 zu koordinieren. </li><li> Mit einem geringen Volumendruck-Einspritzsystem bei 20 psi für 20 ms Impulse gesetzt, injizieren 100 nl von L-Nio in das Gehirn ein und warten 5 Minuten Reflux zu verhindernup der Pipette Spur. <ol><li> Verwenden eines kalibrierten Retikel in das Okular des Stereotaxie-Mikroskop und einer Vergrößerung von 3X. </li><li> Dementsprechend insgesamt 0,100 mm 3 (0,5 mm Länge in einer 0,5 mm Durchmesser Pipette bis 100 nl entspricht) verdrängen aus der ausgezogenen Glaspipette für jeden Satz von Koordinaten. Mit der Einrichtung eines reticule mit, zu messen und zu standardisieren, da jeder einrichten variiert in Abhängigkeit von der Vergrößerung und Skalen verwendet. </li><li> Für eine genaue Volumenmessung während jedes Einspritz nähern sich die abgewinkelten Pipette mit dem Mikroskop von der Seite, so dass der Flüssigkeitsmeniskus eine Sagittalansicht aufweist. Der Meniskus sollte in der gleichen Brennebene erscheinen sowohl der Innen- und Außenwand der Pipette. </li></ol></li><li> Langsam die Pipette zurückziehen und die Schritte wiederholen 1.13-1.16.3 bei der zweiten und dritten Satz von Koordinaten in Tabelle 1 zur Verfügung gestellt. </li><li> Nach der letzten Injektion, entfernen Sie die Pipette und legen genug Knochenwachs die Kraniotomiestelle zu füllen. EINpproximate die Kanten der Kopfhaut Wunde und binden mit dermal Klebstoff. </li><li> Injizieren von 0,1 ml 0,5% Marcain in die Wundränder eine sterile 30 G-Nadel zu verhindern, lokale Schmerzen zu verhindern, mit der Kopfhaut Einschnitt verbunden. </li><li> Bringen Sie das Tier mit dem Gehäuse und Versorgung postoperativer Antibiotika (0,48 mg / ml Trimethoprim-Sulfamethoxazol oder 0,5 mg / ml Levofloxacin) im Trinkwasser für 5 Tage. </li></ol> 2. Weiß Matter Stroke Induktion - posterior gewinkelt Ansatz <ol><li> Führen Sie die Schritte 1,1-1,12 wie im medialen abgewinkelte Ansatz Protokoll, mit Ausnahme der Injektionsarm des stereotaktischen Setup anpassen, um 45 Grad ausgerichtet von vorne nach hinten. </li><li> Vorzurücken die Pipette mit dem ersten A / P und M / L - Koordinaten in Tabelle 2 bereitgestellt. </li><li> Vollständige verbleibenden Schritte 1,14-1,20 wie in der seitlichen abgewinkelten Ansatz Protokoll. </li></ol> 3. Rückläufige Neuronale Labeling <ol><li> Bereiten Sie eine sterile 10&amp; mgr; l Aliquot von L-Nio bei 54,8 mg / ml in 0,9% physiologischer Kochsalzlösung. </li><li> Bereiten Sie eine sterile aliquote Menge von 20% Fluororuby (oder 20% biotinyliertes Dextran Amin oder 2% Fluorogold) in 0,9% physiologischer Kochsalzlösung. </li><li> Verdünnte zusammen 1: 1 für Endkonzentrationen von 27,4 mg / ml L-NiO und 10% Fluororuby. </li><li> Führen Sie den Hub-Protokoll, wie oben in den Schritten 1,3-1,23. </li><li> Visualisieren Sie die nativ fluoreszierende Tracer in Gewebeschnitt durch Perfusionsfixierung, Kryoschneiden und Mikroskopie wie zuvor 8 beschrieben. </li></ol> 4. Gewebeverarbeitung für Immunofluoreszenz <ol><li> Zu einem geeigneten post-Takt-Intervall im Bereich von 3 Stunden bis 14 Tage nach Schlaganfall, einschläfern Mäuse über Isofluran Überdosierung oder lokalen IACUC Verfahren genehmigt. </li><li> Öffnen Sie die Brusthöhle mit abgewinkelten Schere und legen Sie eine 23 G Butterfly-Nadel in den linken Ventrikel. </li><li> Setzen Sie einen kleinen Schnitt im rechten Vorhof mit einer feinen Schere einen Abfluss Spur für die Perfusionsflüssigkeit zu ermöglichen. </li><li> Transkardial mit 30-40 ml kaltem Phosphat-gepufferter Kochsalzlösung perfundiert von 30-40 ml kaltem 4% Paraformaldehyd bei einer Rate von 10 ml / min bei RT verfolgt. </li><li> Enthaupten die Maus und entfernen Sie das Gehirn mit einer sterilen Schere den Schädel posterior zu öffnen und entfernen Sie dann vorsichtig den darüber liegenden Schädel mit einem Spatel, legen Sie das Gehirn in kalte 4% PFA für 24 h und dann übertragen auf 30% Saccharose in PBS für 48 Stunden . </li><li> Bereiten Sie vierzig Mikron schwebenden Abschnitte unter Verwendung eines Kryostats und Antikörper - Verarbeitung durchführen , wie zuvor 6-8 beschrieben. In dieser Studie wurden die folgenden Antikörper: Kaninchen-Anti-Neurofilament 200 (1: 500 Verdünnung); Kaninchen-anti-Vimentin (1: 500); Ziege-Anti-GFAP (1: 500); Kaninchen-anti-Iba-1 (1: 1000). </li></ol> 5. Gewebeverarbeitung für Protein oder RNA-Analyse <ol><li> Zu einem geeigneten post-Takt-Intervall im Bereich von 3 Stunden bis 14 Tagen nach dem Schlaganfall, einschläfern über Isofluran Überdosierung oder lokalen IACUC Verfahren genehmigt. </li><li> enthaupten dieMaus und das Gehirn mit einer sterilen Schere entfernen Sie den Schädel posterior zu öffnen und dann mit einem Spachtel die darüber liegende Schädel sanft entfernen. </li><li> Legen Sie eine sterile 4 mm Spachtel an der Vorderseite des Gehirns Riechkolben und Sehnerven zu trennen. Heben Sie vorsichtig das Gehirn aus dem Schädelkalotte aufnehmen und in eiskalten Dissektion Puffer (1x Hanks Balanced Salt Solution, 25 mM HEPES-KOH, pH 7,4, 35 mM Glucose, 4 mM Natriumbicarbonat und 0,01 mg / ml Cycloheximid). </li><li> Mit einem Gehirn blockieren und sterile neue Rasierklingen, vorbereiten 2-3 mm Platten den Hub und Ort in kalte Dissektion Puffer enthält. </li><li> Unter einem Binokular, identifizieren die weiße Substanz zugrunde liegenden motorischen Kortex in der injizierten Hemisphäre. Bei längeren post stroke Intervallen kann die Region visuell durch fokale Nekrosen und Myelin Blässe identifiziert werden. Hinweis: Bei früheren Post-Takt-Intervalle, die Injektion von L-Nio gemischt mit 1 ul 10% Fast Green kann eine visuelle Identifikation des Hubs ermöglichen ( <strong&gt; 4A). </li><li> Unter Anleitung eines Binokular und ein frisches Skalpell vorsichtig die Region der weißen Substanz sezieren den Hub enthält, identifiziert durch entweder Fast Green Kennzeichnung oder Gewebeverlust. Entfernen Sie darüber liegenden Kortex und der zugrunde liegenden Striatum nach Wunsch. </li></ol>

<ol>
	<li>Go, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2014+update:+a+report+from+the+American+Heart+Association.">Heart disease and stroke statistics--2014 update: a report from the American Heart Association.</a> Circulation. 129, 28-292 (2014).</li><li>Saini, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Silent+stroke:+not+listened+to+rather+than+silent.">Silent stroke: not listened to rather than silent.</a> Stroke. 43, 3102-3104 (2012).</li><li>Koton, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burden+and+outcome+of+prevalent+ischemic+brain+disease+in+a+national+acute+stroke+registry.">Burden and outcome of prevalent ischemic brain disease in a national acute stroke registry.</a> Stroke. 44, 3293-3297 (2013).</li><li>Jiwa, N. S., Garrard, P., Hainsworth, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+vascular+dementia+and+vascular+cognitive+impairment:+a+systematic+review.">Experimental models of vascular dementia and vascular cognitive impairment: a systematic review.</a> J Neurochem. 115, 814-828 (2010).</li><li>Sozmen, E. G., Hinman, J. D., Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+that+matter:+white+matter+stroke+models.">Models that matter: white matter stroke models.</a> Neurotherapeutics. 9, 349-358 (2012).</li><li>Sozmen, E. G., Kolekar, A., Havton, L. A., Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+white+matter+stroke+model+in+the+mouse:+axonal+damage,+progenitor+responses+and+MRI+correlates.">A white matter stroke model in the mouse: axonal damage, progenitor responses and MRI correlates.</a> J Neurosci Methods. 180, 261-272 (2009).</li><li>Rosenzweig, S., Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-dependent+exacerbation+of+white+matter+stroke+outcomes:+a+role+for+oxidative+damage+and+inflammatory+mediators.">Age-dependent exacerbation of white matter stroke outcomes: a role for oxidative damage and inflammatory mediators.</a> Stroke. 44, 2579-2586 (2013).</li><li>Hinman, J. D., Rasband, M. N., Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remodeling+of+the+axon+initial+segment+after+focal+cortical+and+white+matter+stroke.">Remodeling of the axon initial segment after focal cortical and white matter stroke.</a> Stroke. 44, 182-189 (2013).</li><li>McCall, T. B., Feelisch, M., Palmer, R. M., Moncada, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+N-iminoethyl-L-ornithine+as+an+irreversible+inhibitor+of+nitric+oxide+synthase+in+phagocytic+cells.">Identification of N-iminoethyl-L-ornithine as an irreversible inhibitor of nitric oxide synthase in phagocytic cells.</a> Brit j pharmacol. 102, 234-238 (1991).</li><li>Gadea, A., Aguirre, A., Haydar, T. F., Gallo, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelin-1+regulates+oligodendrocyte+development.">Endothelin-1 regulates oligodendrocyte development.</a> J Neurosci. 29, 10047-10062 (2009).</li><li>Dean, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+(pulling)+of+needles+for+gene+delivery+by+microinjection.">Preparation (pulling) of needles for gene delivery by microinjection.</a> CSH prot. , (2006).</li><li>Hughes, P. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+lesions+in+the+rat+central+nervous+system+induced+by+endothelin-1.">Focal lesions in the rat central nervous system induced by endothelin-1.</a> J. Neuropathol. Exp. Neurol. 62, 1276-1286 (2003).</li><li>Whitehead, S. N., Hachinski, V. C., Cechetto, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+between+a+rat+model+of+cerebral+ischemia+and+beta-amyloid+toxicity:+inflammatory+responses.">Interaction between a rat model of cerebral ischemia and beta-amyloid toxicity: inflammatory responses.</a> Stroke. 36, 107-112 (2005).</li><li>Frost, S. B., Barbay, S., Mumert, M. L., Stowe, A. M., Nudo, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+animal+model+of+capsular+infarct:+endothelin-1+injections+in+the+rat.">An animal model of capsular infarct: endothelin-1 injections in the rat.</a> Behav Brain Res. 169, 206-211 (2006).</li><li>Horie, 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=Mouse+model+of+focal+cerebral+ischemia+using+endothelin-1.">Mouse model of focal cerebral ischemia using endothelin-1.</a> J Neurosci Methods. 173, 286-290 (2008).</li><li>Wang, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cognitive+deficits+and+delayed+neuronal+loss+in+a+mouse+model+of+multiple+microinfarcts.">Cognitive deficits and delayed neuronal loss in a mouse model of multiple microinfarcts.</a> Neuroscience. 32, 17948-17960 (2012).</li><li>Shih, A. Y., 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=The+smallest+stroke:+occlusion+of+one+penetrating+vessel+leads+to+infarction+and+a+cognitive+deficit.">The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit.</a> Nat Neurosci. 16, 55-63 (2013).</li><li>Jung, K. J., 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=The+role+of+endothelin+receptor+A+during+myelination+of+developing+oligodendrocytes.">The role of endothelin receptor A during myelination of developing oligodendrocytes.</a> J Korean Med Sci. 26, 92-99 (2011).</li></ol>

Eine Anzahl von früheren Modellen von subkortikalen Schlaganfall einschließlich Brenn Injektionen von Endothelin-1 in die Capsula interna, subkortikalen weißen Substanz und Striatum in der Ratte und Maus - 12-14 6,15 beschrieben. Neuere Modelle von kleinen Brenn Hübe haben Cholesterin Mikroemboli Injektion in die Arterie carotis 16 verwendet und photothrombotic Okklusion eines einzelnen eindringenden arteriole 17. Jedes dieser Modelle hat sowohl Vorteile als auch Nachteile...

Stroke affecting the subcortical white matter is a common clinical entity, accounting for up to 25% of clinical strokes annually in the US 1. Ischemic damage to white matter also occurs silently at a significantly higher rate and contributes to the development of vascular dementia 2,3. Presently, patients with this form of cerebral ischemia have few, if any treatment choices. Despite the clinical importance of this disease, few clinically relevant animal models exist 4,5. 
The goal of this protocol is to produce a focal ischemic lesion within the murine white matter. This murine model of human disease allows the specific study of axonal injury response to stroke and how the cellular elements of white matter, namely oligodendrocytes and astrocytes along with axons, respond to and repair after stroke.
Previous reports have described a model of subcortical white matter stroke using endothelin-1 (ET-1) 6 that is similar to the one described here. Several key changes to the experimental protocol have been made thereby the potential uses of this model have expanded 7,8. This protocol provides a reliable and modifiable strategy to produce a focal stroke within mouse brain white matter. 
The major advantages of this model are the use of a chemical endothelial nitric oxide synthase (eNOS) inhibitor N(5)-(1)-iminoethyl-L-ornithine HCl (L-Nio) 9 with no known paracrine effects on cellular elements of white matter which had been a complication of models using endothelin-1 10. In addition, the stereotactic targeting of white matter in the mouse allows the use of any variety of transgenic or knockout strains, greatly expanding the available tools to determine the effect of stroke on brain white matter. Here, two variations on this technique are described and demonstrate some of the additional variations that can be utilized to enhance the understanding of axonal and white matter damage and repair after stroke.

<table border="0" cellpadding="0" cellspacing="0" width="725">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">L-N5-(1-Iminoethyl)ornithine, Dihydrochloride</td>
			<td style="width:204px;">Calbiochem</td>
			<td style="width:133px;">400600-20MG</td>
			<td style="width:81px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Phoenix Pharmaceutical, Inc.</td>
			<td>NDC 57319-559-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Capillary tubes</td>
			<td>World Precision Instruments</td>
			<td>50-821-807</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Picospritzer</td>
			<td>Parker Instrumentation</td>
			<td>Picospritzer II</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereotactic setup</td>
			<td>Kent Scientific</td>
			<td>KSC51725</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pipette puller</td>
			<td>KOPF</td>
			<td>Model 720</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereomicroscope SZ51</td>
			<td>Olympus</td>
			<td>88-124</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fine scissors</td>
			<td>Fine Scientific Tools</td>
			<td>14084-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps</td>
			<td>Harvard Apparatus</td>
			<td>PY2 72-8547</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Curved Forceps</td>
			<td>Harvard Apparatus</td>
			<td>PY2 72-8598</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blunt dissection tool</td>
			<td>Fine Scientific Tools</td>
			<td>10066-15</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Drill</td>
			<td>Dremel</td>
			<td>8220-1/28</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Drill bits</td>
			<td>Fine Scientific Tools</td>
			<td>19007-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vetbond</td>
			<td>3M</td>
			<td>1469SB&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Marcaine</td>
			<td>HOSPIRA</td>
			<td>NDC 0409-1610-50</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trimethoprim-Sulfamethaxole</td>
			<td>STI Pharmacy</td>
			<td>NDC 54879-007-16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluororuby</td>
			<td>Fluorochrome Inc</td>
			<td>30mg</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>Fisher</td>
			<td>O4042-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sucrose</td>
			<td>Fisher</td>
			<td>BP220-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cryostat</td>
			<td>Leica CM3050 S</td>
			<td>14047033518</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass slides</td>
			<td>Fisher</td>
			<td>12-544-7</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fast Green&#160;</td>
			<td>Sigma</td>
			<td>F7252-5G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissection microscope</td>
			<td>Nikon</td>
			<td>SMZ1500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">23 gauge butterfly needle</td>
			<td>Fisher</td>
			<td>14-840-35</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10X Hank&#39;s Balanced Salt Solution</td>
			<td>Life Technologies</td>
			<td>14065056</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1M HEPES-KOH, pH 7.4</td>
			<td>Affymetrix</td>
			<td>16924</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D-Glucose</td>
			<td>Sigma</td>
			<td>G8270</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium bicarbonate</td>
			<td>Sigma</td>
			<td>S5761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cyclohexamide</td>
			<td>Sigma</td>
			<td>01810</td>
			<td></td>
		</tr>
	</tbody>
</table>

a versatile murine model of subcortical white matter stroke for the study of axonal degeneration and white matter neurobiology

Schlaganfall beeinflussen weißen Substanz macht bis zu 25% der klinischen Schlaganfall Präsentationen, tritt leise auf Raten, die 5-10-fach größer, und trägt wesentlich zur Entwicklung von vaskulärer Demenz sein kann. Nur wenige Modelle von fokalen weißen Substanz Schlaganfall existieren und dieser Mangel an geeigneten Modellen hat Verständnis der neurobiologischen Mechanismen zu Verletzungen Antwort und Reparatur nach dieser Art von Schlaganfall beteiligt behindert. Die Hauptbeschränkung von anderen subkortikalen Schlaganfallmodellen ist, dass sie nicht fokal Infarkt zu der weißen Substanz zu beschränken oder haben in nicht-murine Arten validiert primär sind. Dies begrenzt die Fähigkeit, die Vielzahl von murinen Forschungswerkzeuge anzuwenden, um die Neurobiologie der weißen Substanz Schlaganfall zu studieren. Hier präsentieren wir eine Methode für die zuverlässige Herstellung eines fokalen Schlaganfalls in murine weißen Substanz eine lokale Injektion eines irreversiblen eNOS-Inhibitor verwendet wird. Wir präsentieren auch mehrere Varianten auf dem allgemeinen Protokoll einschließlich zwei einzigartige StereotaxieVariationen, retrograden neuronale Tracing sowie frisches Gewebe Kennzeichnung und Dissektion, die stark die möglichen Anwendungen dieser Technik erweitern. Diese Variationen erlauben für mehrere Ansätze, um die neurobiologischen Auswirkungen dieser gemeinsamen und erforschten Form von Schlaganfall zu analysieren.

Mit dem Modell dargestellt wird, kann die weiße Substanz zugrunde liegenden forelimb sensomotorischen Kortex zuverlässig ausgerichtet sein. Diese chemisch induzierte Hubmodell produziert Brenn axonalen und Myelinverlust, Astrozytose und Mikrogliose (Abbildung 1), wie sie typischerweise in menschlichen lacunar Infarkte zu sehen ist. Durch die Verwendung von drei Injektionen, ein klinisch nützliches Modell mit frühen Beeinträchtigung auf forelimb motorischen Aufgaben ...

Watch this Scientific Journal Video about A Versatile Murine Model of Subcortical White Matter Stroke for the Study of Axonal Degeneration and White Matter Neurobiology at JoVE.com

A Versatile Murine Model of Subcortical White Matter Stroke for the Study of Axonal Degeneration and White Matter Neurobiology

Here we present methodology for the production of a focal stroke in murine white matter by local injection of an irreversible endothelial nitric oxide synthase (eNOS) inhibitor (L-Nio). Presented are two stereotactic variations, retrograde neuronal tracing, and fresh tissue labeling and dissection that expand the potential applications of this technique.

Ein vielseitiges Mausmodell von Subcortical Weiß Matter Stroke für das Studium der axonalen Degeneration and White Matter Neurobiologie

Stroke affecting white matter accounts for up to 25% of clinical stroke presentations, occurs silently at rates that may be 5-10 fold greater, and ...

a-versatile-murine-model-subcortical-white-matter-stroke-for-study

Research

JoVE Journal

Medicine

8.0K Aufrufe.  University of California, Los Angeles. Here we present methodology for the production of a focal stroke in murine white matter by local injection of an irreversible endothelial nitric oxide synthase (eNOS) inhibitor (L-Nio). Presented are two stereotactic variations, retrograde neuronal tracing, and fresh tissue labeling and dissection that expand the potential applications of this technique.

Serie: A Versatile Murine Model of Subcortical White Matter Stroke for the Study of Axonal Degeneration and White Matter Neurobiology

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize their blood glucose levels, islet transplantation has been proposed to be a potential treatment for type 1 diabetes 1,2. More recently, advances in human islet transplantation have further strengthened this view 1,3. However, two major limitations prevent islet transplantation from being a widespread clinical reality: (a) the requirement for large numbers of islets per patient, which severely reduces the number of potential recipients, and (b) the need for heavy immunosuppression, which significantly affects the pediatric population of patients due to their vulnerability to long-term immunosuppression. Strategies that can overcome these limitations have the potential to enhance the therapeutic utility of islet transplantation.
Islet transplantation under the mouse kidney capsule is a widely accepted model to investigate various strategies to improve islet transplantation. This experiment requires the isolation of high quality islets and implantation of islets to the diabetic recipients. Both procedures require surgical steps that can be better demonstrated by video than by text. Here, we document the detailed steps for these procedures by both video and written protocol. We also briefly discuss different transplantation models: syngeneic, allogeneic, syngeneic autoimmune, and allogeneic autoimmune.

Transplantation of isolated islets has been proposed to be a potential treatment for type 1 diabetes. Here we describe a method to isolate islets from mouse pancreata and transplant them to the subcapsular space of the kidney.

Eine Methode zur Murine Islet Isolation und subkapsulären Nierentransplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

Forschung

Medizin

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarker in einem Tiermodell für die Aufdeckung Neural, hämatologischen und Behavioral Korrelate der PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Ein Mausmodell der Stentimplantation in der Arteria carotis für das Studium der Restenose

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Einem Mausmodell der Subarachnoidalblutung

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

Exzitotoxische Stimulation der Hirn Microslices als<em&gt; In-vitro-</em&gt; Modell der Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

DTI of the Visual Pathway - White Matter Tracts and Cerebral Lesions

DTI is a technique that identifies white matter tracts (WMT) non-invasively in healthy and non-healthy patients using diffusion measurements. Similar to visual pathways (VP), WMT are not visible with classical MRI or intra-operatively with microscope. DTI will help neurosurgeons to prevent destruction of the VP while removing lesions adjacent to this WMT. We have performed DTI on fifty patients before and after surgery between March 2012 to January 2014. To navigate we used a 3DT1-weighted sequence. Additionally, we performed a T2-weighted and DTI-sequences. The parameters used were, FOV: 200 x 200 mm, slice thickness: 2 mm, and acquisition matrix: 96 x 96 yielding nearly isotropic voxels of 2 x 2 x 2 mm. Axial MRI was carried out using a 32 gradient direction and one b0-image. We used Echo-Planar-Imaging (EPI) and ASSET parallel imaging with an acceleration factor of 2 and b-value of 800 s/mm&#178;. The scanning time was less than 9 min.
The DTI-data obtained were processed using a FDA approved surgical navigation system program which uses a straightforward fiber-tracking approach known as fiber assignment by continuous tracking (FACT). This is based on the propagation of lines between regions of interest (ROI) which is defined by a physician. A maximum angle of 50, FA start value of 0.10 and ADC stop value of 0.20 mm&#178;/s were the parameters used for tractography.
There are some limitations to this technique. The limited acquisition time frame enforces trade-offs in the image quality. Another important point not to be neglected is the brain shift during surgery. As for the latter intra-operative MRI might be helpful. Furthermore the risk of false positive or false negative tracts needs to be taken into account which might compromise the final results.

Diffusion tensor imaging (DTI) was performed to try to depict the major parts of the visual pathway. The goal was to use an FDA approved standard commercial workstation which could be used for everyday routine in order to try to reduce postoperative damage of the visual pathway in the patients.

DTI der Sehbahn - der weißen Substanz und zerebrale Läsionen

DTI is a technique that identifies white matter tracts (WMT) non-invasively in healthy and non-healthy patients using diffusion measurements. Similar to ...

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.

Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.

Isolierung und Exzision Murine Aorta; Eine vielseitige Technik in das Studium der Herz-Kreislauf-Krankheit

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Eine einfache Kritische große Femoral Defect Modell an der Maus

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier Dysfunction

Ischemia-Reperfusion (IR) injury is known to contribute significantly to the morbidity and mortality associated with ischemic strokes. Ischemic cerebrovascular accidents account for 80% of all strokes. A common cause of IR injury is the rapid inflow of fluids following an acute/chronic occlusion of blood, nutrients, oxygen to the tissue triggering the formation of free radicals.
Ischemic stroke is followed by blood-brain barrier (BBB) dysfunction and vasogenic brain edema. Structurally, tight junctions (TJs) between the endothelial cells play an important role in maintaining the integrity of the blood-brain barrier (BBB). IR injury is an early secondary injury leading to a non-specific, inflammatory response. Oxidative and metabolic stress following inflammation triggers secondary brain damage including BBB permeability and disruption of tight junction (TJ) integrity.
Our protocol presents an in vitro example of oxygen-glucose deprivation and reoxygenation (OGD-R) on rat brain endothelial cell TJ integrity and stress fiber formation. Currently, several experimental in vivo models are used to study the effects of IR injury; however they have several limitations, such as the technical challenges in performing surgeries, gene dependent molecular influences and difficulty in studying mechanistic relationships. However, in vitro models may aid in overcoming many of those limitations. The presented protocol can be used to study the various molecular mechanisms and mechanistic relationships to provide potential therapeutic strategies. However, the results of in vitro studies may differ from standard in vivo studies and should be interpreted with caution.

Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier Dysfunction

Ischemia-Reperfusion (IR) injury is associated with a high rate of morbidity and mortality. The goal of the in vitro model of oxygen-glucose deprivation and reoxygenation (OGD-R) described here is to assess the effects of ischemia reperfusion injury on a variety of cells, particularly in blood-brain barrier (BBB) endothelial cells.

Sauerstoff-Glukose Deprivation und Reoxygenierung als<em&gt; In-vitro-</em&gt; Ischämie-Reperfusion Injury Modell zum Studium der Blut-Hirnschranke Dysfunction

Ischemia-Reperfusion (IR) injury is known to contribute significantly to the morbidity and mortality associated with ischemic strokes. Ischemic ...

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle screws after spinal surgery, which is a serious concern. To address this problem, diverse types of pedicle screws have been examined to identify those with good fixation strength and osseointegration in spine bone. The porcine spine is a good alternative for the human spine in the evaluation of pedicle screws due to the anatomical size, mechanical characteristics, and cost. Although several studies have reported that pedicle screws are efficient in the porcine model, no study has described detailed protocols for the evaluation of a pedicle screw using the porcine model. Here, we describe a detailed method for evaluating transpedicular screws using an in vivo porcine lumbar spine model. The technical details for anesthesia, spine surgery, and harvest provided here will facilitate with the evaluation of the transpedicular screw fixation model.

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Here, we introduce a method to evaluate transpedicular screws by using an in vivo porcine lumbar spine model.

Eine Anästhesie, Chirurgie und Ernteverfahren zur Bewertung von transpedikulären Schrauben mit einem<em&gt; In vivo</em&gt; Schwein Lendenwirbelsäule Modell

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...