Este estudio fue apoyado por una subvención (CNUH-BRI-2012-02-005) financiado por el Instituto de Investigación Biomédica del Hospital Universitario Nacional de Chonbuk (CNUH-BRI), República de Corea.

El Comité Institucional de Cuidado y Uso de Animales de la Universidad Nacional de Chonbuk aprobó este estudio. El tratamiento, el uso y la manipulación de los animales siguieron todas las directrices y políticas. Mantener la sala de operaciones a 24 ° C. 1. Anestesia <ol><li> Aclimate a los cerdos miniatura, de 12 meses de edad, en la unidad experimental durante al menos una semana. Realizar un examen clínico que mida la frecuencia respiratoria, el calor del corazón y la temperatura corporal. No alimente a cada cerdo en miniatura durante 12 h antes del procedimiento de anestesia. </li><li> Inyecte atropina (0,05 mg / kg) y ketamina (20 mg / kg) / xilaxina (2 mg / kg) en la región lateral del músculo cervical, detrás de la oreja, para la premedicación. </li><li> Después de la sedación, aplicar fuertemente una banda de goma alrededor de la base de la oreja y limpiar el oído con alcohol tópico. </li><li> Coloque un catéter de plástico sobre la aguja en la vena del oído y retire la goma. Confirme que el catéter está colocado correctamente. GripeSh el catéter con solución salina heparinizada y fijar con cinta adhesiva. </li><li> Para la intubación endotraqueal, coloque el cerdo en miniatura en decúbito esternal. Con la ayuda de un asistente, sostener la mandíbula del cerdo con una honda adecuada y abrir la boca. <ol><li> Pase la punta del laringoscopio hacia la cavidad faríngea para desplazar la epiglotis del paladar blando. Utilice la punta de la hoja del laringoscopio para hacer que las cuerdas vocales sean visibles y avance el tubo endotraqueal hacia la tráquea durante la expiración. </li><li> Sentirse por un paso de aire libre para una intubación adecuada y revisar la auscultación torácica para detectar sonidos respiratorios tanto en el lado izquierdo como en el derecho del cerdo en miniatura. </li></ol></li><li> Llene el manguito con el volumen adecuado de aire del tubo endotraqueal con aire usando una jeringa de 10 ml y fije el tubo al hocico con cinta adhesiva. </li><li> Proporcionar una concentración de 2.0% de isoflurano, un anestésico de inhalación para la anestesia prolongada, a través del endotrTubo de intubación. Pruebe los reflejos corneales y palpebrales para confirmar la anestesia y use un ungüento en los ojos para prevenir la sequedad. </li><li> Monitoree el sistema cardiovascular, el sistema respiratorio y la temperatura corporal durante la anestesia al menos cada 5 min hasta que el cerdo en miniatura se recupere. </li><li> 30 min antes de la cirugía de la columna vertebral, aplique 30 mg / kg de cefazolina IV lentamente, la primera generación de antibióticos cefalosporina. </li><li> Administrar 5-10 mL / kg / h de solución salina caliente (37 ° C) usando una línea intravenosa para mantener la homeostasis y proporcionar 50 μg / kg / min de fentanilo para controlar el dolor. </li><li> Después de la cirugía de la columna vertebral, realizar la extubación cuando un fuerte reflejo de deglución es evidente. </li><li> Tome el cerdo en miniatura en una habitación y el monitor hasta que se recupere de la anestesia. Proporcionar alimentos y agua cuando el cerdo en miniatura está completamente consciente. </li><li> Aplique 3 mg / kg de antibiótico enrofloxacino con 4,4 mg / kg de carprofeno diariamente para controlar el dolor durante los primeros 3 días. </li><li> MoniDiariamente hasta la eliminación de las suturas. </li></ol> 2. Cirugía de la columna vertebral <ol><li> Autoclave los tornillos transpedicular y los sistemas de fijador posterior para la esterilización, siguiendo las directrices del fabricante. </li><li> Añada la parte posterior del cerdo en miniatura, aproximadamente a 10 cm del centro hacia la izquierda o la derecha, usando una afeitadora mientras el cerdo esté en decúbito supino. Limpiar la piel con solución de povidona-yodo y alcohol al 70%. </li><li> Haga una incisión longitudinal longitudinal desde el segundo proceso espinoso de la columna lumbar hasta la primera cresta sacra mediana usando un bisturí. Disecar a través del tejido subcutáneo y la fascia hasta la punta de los procesos espinosos. </li><li> Elevar los músculos paraspinal subperiosteally de las láminas subyacentes usando un elevador de Cobb. Disecar a lo largo del proceso espinoso y de la lámina limitada a las articulaciones facetarias. </li><li> Abrir la corteza superficial del punto de entrada (que es justo inferior al proceso mamilar de L3A L5 en ambos lados) con una rebaba o un rongeur. </li><li> Inserte el pasador de guía en un sitio abierto, paralelo a la placa de extremo superior y en un ángulo de 20 ° con el proceso espinoso. Definir el punto de partida ideal utilizando rayos X portátero-anterior / lateral portadores de brazo C o portátiles. </li><li> Insertar una sonda de pedículo hasta 25 mm según la radiografía. Confirmar la trayectoria intraósea completa por pedículo y palpación corporal utilizando un dispositivo de sondeo pedículo. </li><li> Inserte seis tornillos pediculares en el pedículo preparado de L3 a L5 hasta que la cabeza del tornillo quede bien asentada. Apunte la abertura lateral de la cabeza del implante en la dirección deseada y alinee la posición horizontal con la trayectoria del vástago. </li><li> Inserte dos barras en ambos lados de las cabezas de los tornillos pediculares, respectivamente. Coloque la manga y la tuerca en la cabeza del tornillo pedicular usando un mango universal. </li><li> Apriete la tuerca con una llave de tubo recto y apriete firmemente la tuerca con una llave dinamométrica. </li><li> Confirme las posiciones oF los tornillos pediculares mediante radiografías postero-anteriores / laterales portátiles. NOTA: Espere a que el cerdo en miniatura se despierte y compruebe los patrones de la marcha y la función del motor de la pata trasera para determinar si un tornillo ha sido mal implantado. </li><li> Irrigar el sitio de la cirugía con 3 L de solución salina normal usando una jeringa de irrigación de bulbo con succión. </li><li> Coloque un drenaje de silicona en el sitio de la cirugía y sacar la punta de silicona. Cierre los músculos paraspinal y subcutánea usando suturas absorbibles métricas 1.0. Cierre la piel con suturas de nylon de 2.0 métricas no absorbibles. </li><li> Desinfectar el sitio de sutura con povidona-yodo y aplicar un vendaje con gasa esterilizada y cinta adhesiva. </li></ol> 3. Procedimiento de Cosecha <ol><li> A las 12 semanas de postoperatorio, inyectar xilazina (2 mg / kg) y ketamina (10 mg / kg) en la región lateral del músculo cervical, detrás de la oreja, para la premedicación. </li><li> Después de la sedación, administrar 15 mg / kg de KCl directamente en la vena del oídoCateter para la eutanasia. </li><li> Hacer una incisión longitudinal mediana en la lesión de cicatriz de la cirugía anterior. Disecar los tejidos blandos y los músculos paraspinales. </li><li> Exponer el proceso espinoso de las espinas lumbares, laminas, varillas, tuercas y procesos transversales de L3 a L5. </li><li> Retire las tuercas con una llave dinamométrica y varillas. Corte el espacio del disco L2-3 y el espacio del disco L5-S1 usando una sierra oscilante. </li><li> Disecar ambos lados de la parte media y anterior de la columna L3-5 con un elevador Cobb y una pinza de torre. Después de la cosecha, si la columna vertebral no puede ser probada inmediatamente, envolverla en gasa empapada con solución salina y almacenar 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>

Los autores no tienen nada relacionado con este documento a divulgar.

La evaluación de los tornillos transpediculares en la columna vertebral porcina requiere mucho tiempo y esfuerzo. En primer lugar, el cerdo en miniatura es un animal grande. Para el cuidado de los animales y la anestesia, el investigador necesita un protocolo especializado. En segundo lugar, la cirugía debe mantener un entorno similar al de la cirugía humana. El propósito de evaluar los tornillos pediculares en la columna vertebral porcina es desarrollar un tornillo eficiente que se puede aplicar a los seres humanos...

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 border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<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&#39;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&#160;</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>&#160;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&#160;</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 fijación del tornillo del pedículo es el estándar de oro para el tratamiento de enfermedades espinales. Sin embargo, muchos estudios han informado de la cuestión de aflojar los tornillos pediculares después de la cirugía de la columna vertebral, que es una preocupación seria. Para abordar este problema, se han examinado diversos tipos de tornillos pediculares para identificar aquellos con buena fuerza de fijación y osteointegración en el hueso de la columna vertebral. La columna porcina es una buena alternativa para la columna vertebral humana en la evaluación de los tornillos pediculares debido al tamaño anatómico, características mecánicas y coste. Aunque varios estudios han reportado que los tornillos pediculares son eficientes en el modelo porcino, ningún estudio ha descrito protocolos detallados para la evaluación de un tornillo pedículo utilizando el modelo porcino. En este sentido, se describe un método detallado para la evaluación de tornillos transpedicular utilizando un modelo in vivo de la columna lumbar porcino. Los detalles técnicos para anestesia, cirugía de columna y cosecha proporcionados aquí facilitarán con la evaluación de la tModelo de fijación de tornillo ranspedicular.

Se describe aquí un protocolo detallado para anestesia, cirugía y cosecha para la evaluación de tornillos transpediculares utilizando un modelo de columna lumbar porcina in vivo . Este protocolo es adecuado para una serie de análisis posteriores, incluyendo pruebas mecánicas ( Figura 1 ), cuantitativos de micro-CT evaluación ( Figura 2 ], y la histología ( Figura 3 ]. Pruebas mecánicas representativas ( Figura 1<...

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

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

Aquí, se introduce un método para evaluar los tornillos transpedicular mediante la utilización de un modelo de la columna lumbar porcino in vivo .

Anestesia, Cirugía y Método de Cosecha para la Evaluación de Tornillos Transpediculares Usando un<em&gt; In Vivo</em&gt; Modelo de columna lumbar porcina

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

an-anesthesia-surgery-harvest-method-for-evaluation-transpedicular

Research

Education

JoVE Core

JoVE Journal

Pharmacology

Medicine

Unit 1

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

Local Anesthetics

SCIENTISTS IN ACTION

7.6K vistas.  Chonbuk National University School of Medicine, Chonbuk National University Hospital.  Aquí, se introduce un método para evaluar los tornillos transpedicular mediante la utilización de un modelo de la columna lumbar porcino in vivo . 

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

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Usando cuantitativa en tiempo real PCR para determinar quimerismo de células donantes en un modelo competitivo Trasplante de médula ósea murina

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Investigación

Medicina

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Un método mejorado para la rápida intubación de la tráquea en ratones

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 &#177; 80 &#937; cm2, and paracellular permeability (Papp) for Lucifer Yellow of 0.90 10-6 &#177; 0.13 10-6 cm sec-1 (mean &#177; SEM, n=55). Further evaluation of this pBEC phenotype showed good expression of the tight junctional proteins claudin 5, ZO-1, occludin and adherens junction protein p120 catenin. The model presented can be used for a range of studies of the BBB in health and disease and, with the highly restrictive paracellular permeability, this model is suitable for studies of transport and intracellular trafficking.

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

Mejor método para el establecimiento de un In Vitro Barrera Blood - Brain modelo basado en las células endoteliales de cerebro porcino

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

En Vivo Evaluación del desarrollo del callo de fractura durante la cicatrización ósea en ratones usando un dispositivo de osteosíntesis de MRI-compatible para el fémur de ratón

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

Un modelo preclínico porcino de trasplante ortotópico de corazón

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

Anesthesia is a routine component of cancer care that is used for diagnostic and therapeutic procedures. The anesthetic technique has recently been implicated in impacting long-term cancer outcomes, possibly through modulation of adrenergic-inflammatory responses that impact cancer cell behavior and immune cell function. Emerging evidence suggests that propofol-based total intravenous anesthesia (TIVA) may be beneficial for long-term cancer outcomes when compared to inhaled volatile anesthesia. However, the available clinical findings are inconsistent. Preclinical studies that identify the underlying mechanisms involved are critically needed to guide the design of clinical studies that will expedite insight. Most preclinical models of anesthesia have been extrapolated from the use of anesthesia in in vivo research and are not optimally designed to study the impact of anesthesia itself as the primary endpoint. This paper describes a method for delivering propofol-TIVA anesthesia in a mouse model of breast cancer resection that replicates key aspects of clinical delivery in cancer patients. The model can be used to study mechanisms of action of anesthesia on cancer outcomes in diverse cancer types and can be extrapolated to other non-cancer areas of preclinical anesthesia research.

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

This paper describes a method for modeling total intravenous anesthesia (TIVA) during cancer resection surgery in mice. The goal is to replicate key features of anesthesia delivery to patients with cancer. The method allows investigation of how anesthetic technique affects cancer recurrence after resection surgery.

Un modelo de ratón in vivo de anestesia intravenosa total durante la cirugía de resección del cáncer

Anesthesia is a routine component of cancer care that is used for diagnostic and therapeutic procedures. The anesthetic technique has recently been ...

Establishment and Evaluation of a Porcine Vein Graft Disease Model

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the investigation of disease mechanisms and the development of therapeutic strategies. 
To perform the surgery, we enter the cardiac chamber through the third intercostal space and carefully dissect the internal mammary vein and immerse it in normal saline. The right main coronary artery is then treated for ischemia. The target vessel is incised, a shunt plug is placed, and the distal end of the graft vein is anastomosed. The ascending aorta is partially blocked, and the proximal end of the graft vein is anastomosed after perforation. The graft vein is checked for patency, and the proximal right coronary artery is ligated. 
CABG surgery is performed in minipigs to harvest the left internal mammary vein for its use as a vascular graft. Serum biochemical tests are used to evaluate the physiological status of the animals after surgery. Ultrasound examination shows that the proximal, middle, and distal end of the graft vessel are unobstructed. In the surgical model, turbulent blood flow in the graft is observed upon histological examination after the CABG surgery, and venous graft stenosis associated with intimal hyperplasia is observed in the graft. The study here provides detailed surgical procedures for the establishment of a repeatable CABG-induced VGD model.

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.

Establecimiento y evaluación de un modelo de enfermedad por injerto de vena porcina

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the ...

3D Printing Model of a Patient's Specific Lumbar Vertebra

Selective dorsal rhizotomy (SDR) is a difficult, risky, and sophisticated operation, in which a laminectomy should not only expose an adequate surgical field of view but also protect the patient&#39;s spinal nerves from injury. Digital models play an important role in the pre-and intra-operation of SDR, because they can not only make doctors more familiar with the anatomical structure of the surgical site, but also provide precise surgical navigation coordinates for the manipulator. This study aims to create a 3D digital model of a patient-specific lumbar vertebra that can be used for planning, surgical navigation, and training of the SDR operation. The 3D printing model is also manufactured for more effective work during these processes.
Traditional orthopedic digital models rely almost entirely on computed tomography (CT) data, which is less sensitive to soft tissues. Fusion of the bone structure from CT and the neural structure from magnetic resonance imaging (MRI) is the key element for the model reconstruction in this study. The patient&#39;s specific 3D digital model is reconstructed for the real appearance of the surgical area and shows the accurate measurement of inter-structural distances and regional segmentation, which can effectively help in the preoperative planning and training of SDR. The transparent bone structure material of the 3D-printed model allows surgeons to clearly distinguish the relative relationship between the spinal nerve and the vertebral plate of the operated segment, enhancing their anatomical understanding and spatial sense of the structure. The advantages of the individualized 3D digital model and its accurate relationship between spinal nerve and bone structures make this method a good choice for preoperative planning of SDR surgery.

This study aims to create a 3D-printed model of a patient-specific lumbar vertebra, which contains both the vertebra and spinal nerve models fused from high-resolution computed tomography (HRCT) and MRI-Dixon data.

Modelo de impresión 3D de la vértebra lumbar específica de un paciente

Selective dorsal rhizotomy (SDR) is a difficult, risky, and sophisticated operation, in which a laminectomy should not only expose an adequate surgical ...

Reconstrucción 3D para el diagnóstico y tratamiento de nódulos pulmonares

A 3D Digital Model for the Diagnosis and Treatment of Pulmonary Nodules

The three-dimensional (3D) reconstruction of pulmonary nodules using medical images has introduced new technical approaches for diagnosing and treating pulmonary nodules, and these approaches are progressively being acknowledged and adopted by physicians and patients. Nonetheless, constructing a relatively universal 3D digital model of pulmonary nodules for diagnosis and treatment is challenging due to device differences, shooting times, and nodule types. The objective of this study is to propose a new 3D digital model of pulmonary nodules that serves as a bridge between physicians and patients and is also a cutting-edge tool for pre-diagnosis and prognostic evaluation. Many AI-driven pulmonary nodule detection and recognition methods employ deep learning techniques to capture the radiological features of pulmonary nodules, and these methods can achieve a good area under-the-curve (AUC) performance. However, false positives and false negatives remain a challenge for radiologists and clinicians. The interpretation and expression of features from the perspective of pulmonary nodule classification and examination are still unsatisfactory. In this study, a method of continuous 3D reconstruction of the whole lung in horizontal and coronal positions is proposed by combining existing medical image processing technologies. Compared with other applicable methods, this method allows users to rapidly locate pulmonary nodules and identify their fundamental properties while also observing pulmonary nodules from multiple perspectives, thereby providing a more effective clinical tool for diagnosing and treating pulmonary nodules.

Autor destacado: Un modelo digital 3D para el diagnóstico y tratamiento de nódulos pulmonares  

The objective of this study is to develop a novel 3D digital model of pulmonary nodules that serves as a communication bridge between physicians and patients and is also a cutting-edge tool for pre-diagnosis and prognostic evaluation.

Un modelo digital 3D para el diagnóstico y tratamiento de nódulos pulmonares

The three-dimensional (3D) reconstruction of pulmonary nodules using medical images has introduced new technical approaches for diagnosing and treating ...

Anestesia, Cirugía y Método de Cosecha para la Evaluación de Tornillos Transpediculares Usando un

Resumen

Explorar más videos

Capítulos en este video

Usando cuantitativa en tiempo real PCR para determinar quimerismo de células donantes en un modelo competitivo Trasplante de médula ósea murina

Un método mejorado para la rápida intubación de la tráquea en ratones

Mejor método para el establecimiento de un In Vitro Barrera Blood - Brain modelo basado en las células endoteliales de cerebro porcino

En Vivo Evaluación del desarrollo del callo de fractura durante la cicatrización ósea en ratones usando un dispositivo de osteosíntesis de MRI-compatible para el fémur de ratón

Un modelo preclínico porcino de trasplante ortotópico de corazón

Un modelo de ratón in vivo de anestesia intravenosa total durante la cirugía de resección del cáncer

Establecimiento y evaluación de un modelo de enfermedad por injerto de vena porcina

Modelo de impresión 3D de la vértebra lumbar específica de un paciente

Un modelo digital 3D para el diagnóstico y tratamiento de nódulos pulmonares

Un modelo digital 3D para el diagnóstico y tratamiento de nódulos pulmonares