Name: an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model
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Description: Watch this Scientific Journal Video about An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model at JoVE.com

Questo studio è stato sostenuto da una sovvenzione (CNUH-BRI-2012-02-005) finanziata dall&#39;Istituto di Ricerca Biomedica dell&#39;ospedale universitario di Chonbuk (CNUH-BRI), Repubblica di Corea.

Il comitato istituzionale per la cura e l&#39;uso degli animali della Chonbuk National University ha approvato questo studio. Il trattamento, l&#39;uso e la manipolazione degli animali hanno seguito tutte le linee guida e le politiche. Mantenere la sala operatoria a 24 ° C. 1. Anestesia <ol><li> Acclimatizzare i maiali in miniatura, di età compresa tra 12 mesi, nell&#39;unità sperimentale per almeno una settimana. Esegui un esame clinico che misura la frequenza respiratoria, il cuore e la temperatura corporea. Non alimentare ogni maiale in miniatura per 12 ore prima della procedura di anestesia. </li><li> Iniettare l&#39;atropina (0,05 mg / kg) e il ketamina (20 mg / kg) / xilaxina (2 mg / kg) nella regione muscolare laterale del muscolo cervicale, dietro l&#39;orecchio, per la premedicazione. </li><li> Dopo la sedazione, applicare strettamente una fascia di gomma intorno alla base dell&#39;orecchio e pulire l&#39;orecchio con l&#39;alcool attivo. </li><li> Posizionare un catetere plastico sopra l&#39;ago nella vena dell&#39;orecchio e rimuovere la fascia di gomma. Verificare che il catetere sia posizionato correttamente. InfluenzaIl catetere con salina eparinizzata e fissare con nastro. </li><li> Per l&#39;intubazione endotracheale, posizionare il maiale in miniatura nella ricostruzione sternale. Con l&#39;aiuto di un assistente, tenere la mascella del maiale con una fionda adatta e aprire la bocca. <ol><li> Passare la punta del laringoscopio nella cavità faringea per spostare l&#39;epiglotta dal palato morbido. Utilizzare la punta della lama del laringoscopio per rendere visibili i cavi vocali e far avanzare il tubo endotracheale nella trachea durante la scadenza. </li><li> Sentitevi per un libero passaggio di aria per una corretta intubazione e controllate l&#39;auscultazione del petto per i suoni della respirazione nei lati sinistro e destro del maiale in miniatura. </li></ol></li><li> Riempire il bracciale con un adeguato volume d&#39;aria del tubo endotracheale in aria utilizzando una siringa da 10 ml e fissare il tubo al muso utilizzando nastro adesivo. </li><li> Fornire una concentrazione del 2,0% di isoflurano, un anestetico per inalazione per anestesia prolungata, attraverso l&#39;endotrTubo di intubazione acheal. Testare i riflessi corneali e palpebrali per confermare l&#39;anestetizzazione e utilizzare unguento sugli occhi per prevenire la secchezza. </li><li> Monitorare il sistema cardiovascolare, il sistema respiratorio e la temperatura corporea durante l&#39;anestesia almeno ogni 5 minuti fino a quando il maiale in miniatura recupera. </li><li> 30 minuti prima della chirurgia della colonna vertebrale, applicare lentamente 30 mg / kg di cefazolina IV, la prima generazione di antibiotici di cefalosporina. </li><li> Amministrare 5-10 ml / kg / h di salina calda (37 ° C) usando una linea IV per mantenere l&#39;omeostasi e fornire 50 μg / kg / min di fentanile per controllare il dolore. </li><li> Dopo la chirurgia della spina, eseguire l&#39;estubazione quando è evidente un forte riflesso di deglutizione. </li><li> Prendi il maiale in miniatura in una stanza e in un monitor fino a ricuperare l&#39;anestesia. Fornire cibo e acqua quando il maiale in miniatura è pienamente consapevole. </li><li> Applicare 3 mg / kg di antibiotico enrofloxacin con 4,4 mg / kg carprofen al giorno per il controllo del dolore durante i primi 3 giorni. </li><li> MoniIl maiale al giorno fino alla rimozione delle suture. </li></ol> 2. Chirurgia della colonna vertebrale <ol><li> Autoclave le viti transpedicolari e i sistemi di fissaggio posteriori per la sterilizzazione, seguendo le linee guida del produttore. </li><li> Rasare la parte posteriore del maiale in miniatura, circa 10 cm dal centro verso sinistra o verso destra, utilizzando un rasoio mentre il maiale è in posizione supina. Pulire la pelle con soluzione povidone-iodio e alcool al 70%. </li><li> Fai un&#39;incisione mediana longitudinale dal secondo processo spinoso della colonna lombare alla prima cresta sacrale mediana usando un bisturi. Dissect attraverso il tessuto sottocutaneo e fascia alla punta dei processi spinosi. </li><li> Elevate i muscoli paraspinalini sottoperialmente dalla lamina sottostante usando un ascensore Cobb. Dissect lungo il processo spinoso e lamina limitata ai giunti del facet. </li><li> Aprire la corteccia superficiale del punto di entrata (che è appena inferiore al processo mammillario da L3A L5 su entrambi i lati) con una burr o un rongeur. </li><li> Inserire il perno di guida in un luogo aperto, parallelo alla lastra superiore e ad un angolo di 20 ° rispetto al processo spinoso. Definire il punto di partenza ideale utilizzando i bracci a C oi raggi X postero-anteriori / laterali portatili. </li><li> Inserire una sonda a pedale fino a 25 mm secondo il raggio X. Confermare la traiettoria intraossea completa dalla peduncolo e dalla palpazione del corpo usando un dispositivo di suonatura pedicle. </li><li> Inserire sei viti di peduncolo nel pedone preparato da L3 a L5 finché la testa della vite non è ben posizionata. Puntare l&#39;apertura laterale della testa dell&#39;impianto nella direzione desiderata e allineare la posizione orizzontale con la traiettoria dell&#39;asta. </li><li> Inserire rispettivamente due bacchette su entrambi i lati delle teste a vite. Inserire il manicotto e il dado alla testa della vite di pedice utilizzando una maniglia universale. </li><li> Serrare a fondo il dado con una chiave a filo diritto e stringere saldamente il dado utilizzando una chiave a contatore torcente. </li><li> Confermare le posizioni oLe viti a pedale che utilizzano raggi X posteriori postero-anteriori / laterali. NOTA: attendere che il maiale in miniatura si svegli e controllare i pattern di andatura e la funzione motore dell&#39;arto posteriore per determinare se una vite è stata impigliata male. </li><li> Irrigare il sito di chirurgia con 3 L di salina normale utilizzando una siringa per irrigazione a bulbo con aspirazione. </li><li> Mettere un drenante in silicone nel sito di chirurgia e togliere la punta del silicone. Chiudere i muscoli paraspinal e sottocutanei utilizzando 1,0 suture assorbibili metriche. Chiudere la pelle con suture di nylon non assorbibili di 2,0 metri. </li><li> Disinfettare il sito di sutura con povidone-iodio e applicare una medicazione usando la garza sterilizzata e il nastro. </li></ol> 3. Procedura di raccolta <ol><li> A 12 settimane dopo l&#39;intervento, iniettare xilazina (2 mg / kg) e ketamina (10 mg / kg) nella regione muscolare laterale del muscolo cervicale, dietro l&#39;orecchio, per la premedicazione. </li><li> Dopo la sedazione, somministrare 15 mg / kg di KCl direttamente nella vena dell&#39;orecchioCatetere per l&#39;eutanasia. </li><li> Effettuare un&#39;incisione midline longitudinale alla precedente lesione della cicatrice. Disseccare i tessuti molli e i muscoli paraspinal. </li><li> Esponete il processo spinoso delle spine lombari, delle lamelle, delle aste, dei dadi e dei processi trasversali da L3 a L5. </li><li> Rimuovere i dadi usando una chiave e tiranti di coppia. Tagliare lo spazio del disco L2-3 e lo spazio del disco L5-S1 utilizzando una sega oscillante. </li><li> Disseccare entrambi i lati della parte centrale e anteriore della colonna vertebrale L3-5 con un&#39;aspirazione Cobb e una pinza a torre. Dopo la raccolta, se la colonna vertebrale non può essere testata immediatamente, avvolgerla in garza imbevuta di soluzione salina e conservare a -20 ° C. </li></ol>

<ol>
	<li>Greenfield, R. T., Grant, R. E., Bryant, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pedicle+screw+fixation+in+the+management+of+unstable+thoracolumbar+spine+injuries.">Pedicle screw fixation in the management of unstable thoracolumbar spine injuries.</a> Orthop Rev. 21 (6), 701-706 (1992).</li><li>Upasani, V. 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=Pedicle+screw+surface+coatings+improve+fixation+in+nonfusion+spinal+constructs.">Pedicle screw surface coatings improve fixation in nonfusion spinal constructs.</a> Spine. 34 (4), 335-343 (2009).</li><li>Halvorson, T. L., Kelley, L. A., Thomas, K. A., Whitecloud, T. S., Cook, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+bone+mineral+density+on+pedicle+screw+fixation.">Effects of bone mineral density on pedicle screw fixation.</a> Spine. 19 (21), 2415-2420 (1994).</li><li>Weinstein, J. N., Spratt, K. F., Spengler, D., Brick, C., Reid, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+pedicle+fixation:+reliability+and+validity+of+roentgenogram-based+assessment+and+surgical+factors+on+successful+screw+placement.">Spinal pedicle fixation: reliability and validity of roentgenogram-based assessment and surgical factors on successful screw placement.</a> Spine. 13 (9), 1012-1018 (1988).</li><li>Fini, 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=Biological+assessment+of+the+bone-screw+interface+after+insertion+of+uncoated+and+hydroxyapatite-coated+pedicular+screws+in+the+osteopenic+sheep.">Biological assessment of the bone-screw interface after insertion of uncoated and hydroxyapatite-coated pedicular screws in the osteopenic sheep.</a> J Biomed Mater Res A. 66 (1), 176-183 (2003).</li><li>Kim, D. 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=Evaluation+of+Titanium-Coated+Pedicle+Screws:+In+Vivo+Porcine+Lumbar+Spine+Model.">Evaluation of Titanium-Coated Pedicle Screws: In Vivo Porcine Lumbar Spine Model.</a> World Neurosurg. 91, 163-171 (2016).</li><li>Upasani, V. 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=Pedicle+screw+surface+coatings+improve+fixation+in+nonfusion+spinal+constructs.">Pedicle screw surface coatings improve fixation in nonfusion spinal constructs.</a> Spine. 34 (4), 335-343 (2009).</li><li>Smit, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+a+quadrupted+as+an+in+vivo+model+for+the+study+of+the+spine-biomechanical+considrations.">The use of a quadrupted as an in vivo model for the study of the spine-biomechanical considrations.</a> Eur Spine J. 11 (2), 137-144 (2002).</li><li>Aldini, N. 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=Pedicular+fixation+in+the+osteoporotic+spine:+a+pilot+in+vivo+study+on+long-term+ovariectomized+sheep.">Pedicular fixation in the osteoporotic spine: a pilot in vivo study on long-term ovariectomized sheep.</a> J Orthop Res. 20 (6), 1217-1224 (2002).</li><li>Fini, 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=Biological+assessment+of+the+bone-screw+interface+after+insertion+of+uncoated+and+hydroxyapatite-coated+pedicular+screws+in+the+osteopenic+sheep.">Biological assessment of the bone-screw interface after insertion of uncoated and hydroxyapatite-coated pedicular screws in the osteopenic sheep.</a> J Biomed Mater Res A. 66 (1), 176-183 (2003).</li><li>Branemark, R., Ohrnell, L. O., Skalak, R., Carlsson, L., Branemark, P. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanical+characterization+of+osseointegration:+an+experimental+in+vivo+investigation+in+the+beagle+dog.">Biomechanical characterization of osseointegration: an experimental in vivo investigation in the beagle dog.</a> J Orthop Res. 16 (1), 61-69 (1998).</li><li>McLain, R. F., Yerby, S. A., Moseley, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+morphometry+of+L4+vertebrae:+comparison+of+large+animal+models+for+the+human+lumbar+spine.">Comparative morphometry of L4 vertebrae: comparison of large animal models for the human lumbar spine.</a> Spine. 27 (8), E200-E206 (2002).</li><li>Giavaresi, G., 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=In+vivo+preclinical+evaluation+of+the+influence+of+osteoporosis+on+the+anchorage+of+different+pedicle+screw+designs.">In vivo preclinical evaluation of the influence of osteoporosis on the anchorage of different pedicle screw designs.</a> Eur Spine J. 20 (8), 1289-1296 (2011).</li><li>Hasegawa, T., 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=Hydroxyapatite-coating+of+pedicle+screws+improves+resistance+against+pull-out+force+in+the+osteoporotic+canine+lumbar+spine+model:+a+pilot+study.">Hydroxyapatite-coating of pedicle screws improves resistance against pull-out force in the osteoporotic canine lumbar spine model: a pilot study.</a> Spine J. 5 (3), 239-243 (2005).</li><li>Smorgick, 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=Single-+versus+multilevel+fusion+for+single-level+degenerative+spondylolisthesis+and+multilevel+lumbar+stenosis:+four-year+results+of+the+spine+patient+outcomes+research+trial.">Single- versus multilevel fusion for single-level degenerative spondylolisthesis and multilevel lumbar stenosis: four-year results of the spine patient outcomes research trial.</a> Spine. 38 (10), 797-805 (2013).</li><li>Busscher, I., Ploegmakers, J. J., Verkerke, G. J., Veldhuizen, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+anatomical+dimensions+of+the+complete+human+and+porcine+spine.">Comparative anatomical dimensions of the complete human and porcine spine.</a> Eur Spine J. 19 (7), 1104-1114 (2010).</li></ol>

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La valutazione delle viti transpedicolari nella colonna vertebrale richiede molto tempo e fatica. In primo luogo, il maiale in miniatura è un grande animale. Per la cura degli animali e l&#39;anestesia, il ricercatore necessita di un protocollo specializzato. In secondo luogo, la chirurgia dovrebbe mantenere un ambiente simile a quello della chirurgia umana. Lo scopo di valutare le viti di peduncolo nella colonna vertebrale è quello di sviluppare una vite efficiente applicabile agli esseri umani. In terzo luogo, la va...

Transpedicular screw fixation is a gold-standard treatment for degenerative lumbar spine and bursting fracture because it involves three columns of the spine and achieves stabilization1,2. However, most patients who undergo such surgery also have osteoporosis3,4. Many studies have evaluated the fixation strength and the osseointegration status of transpedicular screws, because the loosening of pedicle screws currently in use has been reported in patients with osteoporosis5,6.
The porcine spine is similar to the human spine in terms of size. It is less expensive compared to a primate model7. Furthermore, an in vivo mechanical study has demonstrated that the quadruped porcine spine is essentially loaded in the same way as that of the human spine8, which is why many researchers use porcine spines for studies on the prevention of pedicle screw loosening. However, it takes several months to study pedicle screws in the porcine spine because identifying the long-term stability of pedicle screws takes times. In order to compare different types of screws in the vertebral body, it is necessary to insert the screws in similar positions. Therefore, researchers should be well-acquainted with proper anesthesia techniques, standardized surgical protocols, and harvest procedures before performing any experiments. Here, we describe a detailed method for anesthesia, surgery, and harvest for the evaluation of pedicle screw fixation using a porcine spine model, including ex vivo imaging, histology, and strength testing.

<table><tbody><tr><td>Miniature pig</td><td>OrientBio</td><td></td><td></td></tr><tr><td>Atropine</td><td>Jeil pharmaceutical</td><td>A04900241</td><td>Anesthesia</td></tr><tr><td>Over-the needle plastic catheter</td><td>BD</td><td>REF382412</td><td>Maintenance of IV line</td></tr><tr><td>Ketamine</td><td>Yuhan</td><td>A04502441</td><td>Anesthesia</td></tr><tr><td>Xylazine</td><td>Bayer Korea</td><td>A00800071</td><td>Anesthesia</td></tr><tr><td>Laryngoscope</td><td>Karl storz</td><td></td><td>Intubation</td></tr><tr><td>Endotracheal tube</td><td>Covidien</td><td></td><td>Intubation</td></tr><tr><td>Isoflurane</td><td>JW pharmaceutical Co</td><td>A02104781</td><td>Anesthesia</td></tr><tr><td>Eye ointment</td><td>Hanlim pharma</td><td>A37851721</td><td>Protection of pig&#039;s eye</td></tr><tr><td>Cefazolin</td><td>Donga pharma</td><td>A01503951</td><td>Antibiotics</td></tr><tr><td>Saline</td><td>JW pharmaceutical Co</td><td>A02151392</td><td>Maintenance of fluid homeostasis</td></tr><tr><td>Fentanyl</td><td>Hana pharm</td><td>C03200032</td><td>Pain control</td></tr><tr><td>Enrofloxacin</td><td>Bayer</td><td>93106-60-6&amp;nbsp;</td><td>Antibiotics</td></tr><tr><td>Morphine</td><td>Myungmoon pharma</td><td>C03700091</td><td>Pain control</td></tr><tr><td>Meloxicam</td><td>Boehringer Ingelheim</td><td>A07600711</td><td>Antibiotics</td></tr><tr><td>Povidone-iodine</td><td>Hyundai pharma</td><td></td><td>Wound dressing</td></tr><tr><td>Scalpel blade size 15</td><td>Braun</td><td>&amp;nbsp;I1 BB515</td><td>Skin incision</td></tr><tr><td>Cobb elevator</td><td>Codman</td><td>65-2546</td><td>Dissection of muscle</td></tr><tr><td>Burr</td><td>Medtronic</td><td></td><td>Making of starting point of screw</td></tr><tr><td>Rongeur</td><td>Aesculap</td><td>FO515R</td><td>Making of starting point of screw</td></tr><tr><td>Guide pin (K-wire)</td><td>CE</td><td>01067803</td><td>Guidance of screw trajectory</td></tr><tr><td>C-arm</td><td>GE</td><td>OEC 9800 plus</td><td>Guidance of screw trajectory</td></tr><tr><td>Portable X-ray</td><td>Siemens</td><td>Mobile XP hybrid</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle probe</td><td>OtisBiotech</td><td>SPI-02-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle sounding device</td><td>OtisBiotech</td><td>SPI-03-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle screw</td><td>OtisBiotech</td><td>MS-40025</td><td></td></tr><tr><td>Posterior fixator systems</td><td>OtisBiotech</td><td></td><td></td></tr><tr><td>Rod&amp;nbsp;</td><td>OtisBiotech</td><td>ROD-60140</td><td>Rigid fixation between screws</td></tr><tr><td>Universal handle</td><td>OtisBiotech</td><td>SPI-08-01</td><td>To fix the screws to the rod</td></tr><tr><td>Straight socket wrench</td><td>OtisBiotech</td><td>SPI-06-01</td><td>To fix the screws to the rod</td></tr><tr><td>counter torque wrench</td><td>OtisBiotech</td><td>SPI-07-01</td><td>To fix the screws to the rod</td></tr><tr><td>Bulb irrigation syringe</td><td>Hyupsug medical</td><td>HS-IR-140</td><td>Irrigation</td></tr><tr><td>Silicone drain</td><td>Sewon medical</td><td>2205-006</td><td>To drain the fluid at the surgical site</td></tr><tr><td>3.0 metric absorbable suture</td><td>Ethicon</td><td>BA1673H</td><td>Muscle suture</td></tr><tr><td>2.0 metric nonabsorbable nylon suture</td><td>Ethicon</td><td>W1626T</td><td>Skin suture</td></tr><tr><td>Gauze</td><td>Kingphar Korea</td><td>KP120-06</td><td></td></tr><tr><td>Pentobarbital</td><td>Hanlim pharma</td><td>645301221</td><td>Euthanasia</td></tr><tr><td>Oscillating saw</td><td>Zimmer</td><td></td><td>Harvest spine</td></tr><tr><td>Tower forceps</td><td>Aesculap</td><td>BF461R</td><td>Harvest spine</td></tr></tbody></table>

an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model

La fissazione a vite a pedicella è il gold standard per il trattamento delle malattie spinali. Tuttavia, molti studi hanno riportato la questione dell&#39;allentamento delle viti di peduncolo dopo la chirurgia spinale, che è una preoccupazione grave. Per risolvere questo problema, sono stati esaminati diversi tipi di viti di peduncolo per identificare quelli con buona forza di fissazione e osseointegrazione nell&#39;osso della colonna vertebrale. La colonna vertebrale è una buona alternativa per la colonna vertebrale umana nella valutazione delle viti peduncolari a causa della dimensione anatomica, delle caratteristiche meccaniche e dei costi. Sebbene diversi studi abbiano riportato che le viti di peduncolo sono efficaci nel modello della suina, nessuno studio ha descritto protocolli dettagliati per la valutazione di una vite a peduncolo usando il modello porcino. Qui descriviamo un metodo dettagliato per valutare le viti transpedicolari usando un modello di colonna vertebrale lombare in vivo . I dettagli tecnici per l&#39;anestesia, la chirurgia della colonna vertebrale e il raccolto forniti qui faciliteranno con la valutazione del tModello di fissaggio a vite ranspedrici.

Qui viene descritto un protocollo dettagliato per l&#39;anestesia, la chirurgia e il raccolto per la valutazione delle viti transpedicolari utilizzando un modello di colonna vertebrale lombare in vivo . Questo protocollo è adatto a numerose analisi a valle, inclusi i test meccanici ( Figura 1 ), la valutazione micro-CT quantitativa ( Figura 2 ) e l&#39;istologia ( Figura 3 ). La prova meccanica rappresentativa ( figura ...

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

Qui introduciamo un metodo per valutare le viti transpediciche utilizzando un modello di colonna vertebrale lombare in vivo .

Anestesia, chirurgia e metodo di raccolta per la valutazione delle viti transpedicolari utilizzando un<em&gt; In Vivo</em&gt; Modello della colonna vertebrale della deriva

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

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

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7.6K Views.  Chonbuk National University School of Medicine, Chonbuk National University Hospital.  Qui introduciamo un metodo per valutare le viti transpediciche utilizzando un modello di colonna vertebrale lombare in vivo . 

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

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has significantly broadened our knowledge of physiological and pathological processes in numerous tissues in vivo 4-7. In studies of the central nervous system (CNS), there has been a broad application of in vivo imaging in the brain, which has produced a plethora of novel and often unexpected findings about the behavior of cells such as neurons, astrocytes, microglia, under physiological or pathological conditions 8-17. However, mostly technical complications have limited the implementation of in vivo imaging in studies of the living mouse spinal cord. In particular, the anatomical proximity of the spinal cord to the lungs and heart generates significant movement artifact that makes imaging the living spinal cord a challenging task.
We developed a novel method that overcomes the inherent limitations of spinal cord imaging by stabilizing the spinal column, reducing respiratory-induced movements and thereby facilitating the use of two-photon microscopy to image the mouse spinal cord in vivo. This is achieved by combining a customized spinal stabilization device with a method of deep anesthesia, resulting in a significant reduction of respiratory-induced movements. This video protocol shows how to expose a small area of the living spinal cord that can be maintained under stable physiological conditions over extended periods of time by keeping tissue injury and bleeding to a minimum. Representative raw images acquired in vivo detail in high resolution the close relationship between microglia and the vasculature. A timelapse sequence shows the dynamic behavior of microglial processes in the living mouse spinal cord. Moreover, a continuous scan of the same z-frame demonstrates the outstanding stability that this method can achieve to generate stacks of images and/or timelapse movies that do not require image alignment post-acquisition. Finally, we show how this method can be used to revisit and reimage the same area of the spinal cord at later timepoints, allowing for longitudinal studies of ongoing physiological or pathological processes in vivo.

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

A minimally invasive protocol to stabilize the mouse spinal column and perform repetitive in vivo spinal cord imaging using two-photon microscopy is described. This method combines a spinal stabilization device and an anesthetic regimen to minimize respiratory-induced movements and produce raw imaging data that require no alignment or other post-processing.

Imaging in vivo del midollo spinale di topo Utilizzo di due fotoni Microscopia

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

Ricerca

Neuroscienze

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

Una procedura di impianto del Camera spinale per longitudinale<em&gt; In Vivo</em&gt; Imaging del cavo del mouse spinale

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive targets for injection of novel therapeutics aimed at treating chronic pain. In small animal models, laminectomy has been used to facilitate DRG injection because it involves surgical removal of the vertebral bone surrounding each DRG. We demonstrate a technique for intraganglionic injection of lumbar DRG in a large animal species, namely, swine. Laminotomy is performed to allow direct access to DRG using standard neurosurgical techniques, instruments, and materials. Compared with more extensive bone removal via laminectomy, we implement laminotomy to conserve spinal anatomy while achieving sufficient DRG access. Intraoperative progress of DRG injection is monitored using a non-toxic dye. Following euthanasia on post-operative day 21, the success of injection is determined by histology for intraganglionic distribution of 4&#39;,6-diamidino-2-phenylindole (DAPI). We inject a biologically inactive solution to demonstrate the protocol. This method could be applied in future preclinical studies to target therapeutic solutions to DRG. Our methodology should facilitate testing the translatability of intraganglionic small animal paradigms in a large animal species. Additionally, this protocol may serve as a key resource for those planning preclinical studies of DRG injection in swine.

We describe a method for laminotomy in swine that provides access to lumbar dorsal root ganglia (DRG) for intraganglionic injection. Injection progress is monitored intraoperatively and histologically confirmed up to 21 days after surgery. This protocol could be used for future preclinical studies involving DRG injection.

Laminotomia per accesso del ganglio di radice dorsale lombare e l'iniezione nei suini

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

Laminectomia e impianto della finestra del cavo spinale nel mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

In Vivo Mouse Model of Spinal Implant Infection

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The purpose of this method is to describe a novel mouse model of spinal implant infection (SII) that was created to provide an inexpensive, rapid, and accurate in vivo tool to test potential therapeutics and treatment strategies for spinal implant infections.
In this method, we present a model of posterior-approach spinal surgery in which a stainless-steel k-wire is transfixed into the L4 spinous process of 12-week old C57BL/6J wild-type mice and inoculated with 1 x 103 CFU of a bioluminescent strain of Staphylococcus aureus Xen36 bacteria. Mice are then longitudinally imaged for bioluminescence in vivo on post-operative days 0, 1, 3, 5, 7, 10, 14, 18, 21, 25, 28, and 35. Bioluminescence imaging (BLI) signals from a standardized field of view are quantified to measure in vivo bacterial burden.
To quantify bacteria adhering to implants and peri-implant tissue, mice are euthanized and the implant and surrounding soft tissue are harvested. Bacteria are detached from the implant by sonication, cultured overnight and then colony forming units (CFUs) are counted. The results acquired from this method include longitudinal bacterial counts as measured by in vivo S. aureus bioluminescence (mean maximum flux) and CFU counts following euthanasia.
While prior animal models of instrumented spine infection have involved invasive, ex vivo tissue analysis, the mouse model of SII presented in this paper leverages noninvasive, real time in vivo optical imaging of bioluminescent bacteria to replace static tissue study. Applications of the model are broad and may include utilizing alternative bioluminescent bacterial strains, incorporating other types of genetically engineered mice to contemporaneously study host immune response, and evaluating current or investigating new diagnostic and therapeutic modalities such as antibiotics or implant coatings.

The protocol describes a novel in vivo mouse model of spinal implant infection where a stainless-steel k-wire implant is infected with bioluminescent Staphylococcus aureus Xen36. Bacterial burden is monitored longitudinally with bioluminescent imaging and confirmed with colony forming unit counts after euthanasia.

Modello murino in vivo di infezione da impianto spinale

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The ...

Medicina

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

Valutazione in tempo reale della microperfusione del midollo spinale in un modello suino di ischemia/riperfusione

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

Surgical Technique for Lumbar Spinal Catheter Insertion in Pigs Enabling Continuous Access to the Thecal Sac in a Terminal Setup

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine and human central nervous systems (CNS). However, accessing the cerebrospinal fluid (CSF) in larger pig breeds by conventional lumbar puncture techniques can be challenging due to an oblique orientation of the spinal spinous processes and a limited interlaminar space. Accordingly, an open surgical procedure for inserting a lumbar spinal catheter for continuous CSF sampling at the L4/L5 level in pigs is thoroughly described in this work. After positioning the pig and identifying the anatomical landmarks, a dorsal midline surgical incision is made to expose the spinous processes. By advancing the introducer needle, the spinal catheter is inserted inside the thecal sac of the spinal canal while leaving the bone structures of the spine intact. This method allows continuous infusion into or sampling from the porcine thecal sac with minimal bleeding or CSF leakage. The procedure is simple, time-efficient, and reproducible across different experimental setups, offering significant potential for various pre-clinical studies, including pharmacokinetic research, surgical training, and spinal cord injury models.

We present a technique for inserting a lumbar spinal catheter at the L4-L5 level in a 3-month-old Danish Landrace pig as part of a terminal research protocol, enabling continuous infusion or CSF sampling from the thecal sac.

Tecnica chirurgica per l'inserimento di catetere spinale lombare nei suini che consente l'accesso continuo al sacco tecale in una configurazione terminale

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine ...

A Mouse Model of Lumbar Spine Instability

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding the pathological processes and evaluating new drugs. Here, we introduced a surgically induced lumbar spine instability (LSI) mouse model that develops IDD starting from 1 week post operation. In detail, the mouse under anesthesia was operated by low back skin incision, L3&#8211;L5 spinous processes exposure, detachment of paraspinous muscles, resection of processes and ligaments, and skin closure. L4&#8211;L5 IVDs were chosen for the observation. The LSI model develops lumbar IDD by porosity and hypertrophy in endplates at an early stage, decrease in intervertebral disc volume, shrinkage in nucleus pulposus at an intermediate stage, and bone loss in lumbar vertebrae (L5) at a later stage. The LSI mouse model has the advantages of strong operability, no requirement of special equipment, reproducibility, inexpensive, and relatively short period of IDD development. However, LSI operation is still a trauma that causes inflammation within the first week post operation. Thus, this animal model is suitable for study of lumbar IDD.

We developed a lumbar intervertebral disc degeneration mouse model by resection of L3&#8211;L5 spinous processes along with supra- and inter-spinous ligaments and detachment of paraspinous muscles.

Un modello di topo di instabilità della colonna lombare

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding ...

Biologia

Pedicle Screw Placement Using an Augmented Reality Head-Mounted Display in a Porcine Model

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) for minimally invasive pedicle screw placement. The cadaveric porcine specimens were placed on a surgical table and draped with sterile covers. The levels of interest were identified using fluoroscopy, and a dynamic reference frame was attached to the spinous process of a vertebra in the region of interest. Cone beam computerized tomography (CBCT) was performed, and a 3D rendering was automatically generated, which was used for the subsequent planning of the pedicle screw placements. Each surgeon was fitted with an HMD that was individually eye-calibrated and connected to the spinal navigation system.
Navigated instruments, tracked by the navigation system and displayed in 2D and 3D in the HMD, were used for 33 pedicle cannulations, each with a diameter of 4.5 mm. Postprocedural CBCT scans were assessed by an independent reviewer to measure the technical (deviation from the planned path) and clinical (Gertzbein grade) accuracy of each cannulation. The navigation time for each cannulation was measured. The technical accuracy was 1.0 mm &#177; 0.5 mm at the entry point and 0.8 mm &#177; 0.1 mm at the target. The angular deviation was 1.5&#176; &#177; 0.6&#176;, and the mean insertion time per cannulation was 141 s &#177; 71 s. The clinical accuracy was 100% according to the Gertzbein grading scale (32 grade 0; 1 grade 1). When used for minimally invasive pedicle cannulations in a porcine model, submillimeter technical accuracy and 100% clinical accuracy could be achieved with this protocol.

The augmented reality head-mounted display, Magic Leap, was used in combination with a conventional navigation system to place pedicle screws in a porcine model by adhering to a novel workflow. With a median insertion time of &lt;2.5 min, submillimeter technical accuracy and 100% clinical accuracy were achieved according to Gertzbein.

Posizionamento della vite peduncolare utilizzando un display montato sulla testa a realtà aumentata in un modello suino

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) ...

<meta charset="utf8"></head><body>Tecnica del sistema a vite di regolazione sagittale per fratture da scoppio toracolombare

Minimally Invasive Treatment for Thoracolumbar Burst Fracture Using Sagittal Alignment Screws and A Trauma Reduction Device

Thoracolumbar (TL) burst fracture is one of the most common indications for minimally invasive percutaneous pedicle screw fixation. Although the indication for surgical treatment of neurologically intact TL fractures remains under debate, studies have demonstrated that posttraumatic malalignment may lead to a deterioration in the patient&#39;s quality of life. For burst fractures with malalignment or fragments in the spinal canal, a reduction technique using ligamentotaxis is commonly used to improve long-term outcomes.
The sagittal adjusting screw (SAS) system is a monoaxial screw system with a fixed head and concave sliding saddle that allows lordotic sliding of the rod in the sagittal plane after screw insertion. SAS also has a percutaneous option and has been used for TL spine fractures. Notably, the SAS only allows motion on the sagittal plane, allowing both secure fixation and angular reduction. The SAS has certain advantages over the conventional Schanz screw system or normal mono-/multiaxial pedicle screws for TL spine fracture treatment. In addition, specialized trauma reduction devices are available for the SAS system. In this video protocol, we discuss the indication for the SAS system in TL burst fracture and describe a technique of TL burst fracture reduction and fixation using the SAS system. Additionally, we describe our recent case series with radiological evaluation, including regional kyphotic angle and percent loss of anterior vertebral body height, to evaluate the newly introduced trauma reduction device.

<meta charset="utf8"></head><body>Trattamento minimamente invasivo per la frattura da scoppio toracolombare utilizzando viti di allineamento sagittale e un dispositivo di riduzione del trauma

The sagittal adjusting screw (SAS) system has been widely used for thoracolumbar (TL) spinal trauma. Percutaneous options and specialized trauma reduction devices are also available for the SAS system. We describe a technique for reducing TL burst fractures using an SAS system and a newly introduced trauma reduction device.

Trattamento minimamente invasivo per la frattura da scoppio toracolombare utilizzando viti di allineamento sagittale e un dispositivo di riduzione del trauma

Anestesia, chirurgia e metodo di raccolta per la valutazione delle viti transpedicolari utilizzando un

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