Los autores agradecen al Dr. Madsen en la Universidad de Wisconsin-Madison y Cristel Baiu en Gammex Inc. para proporcionar asesoramiento sobre materiales TM. Los autores también agradecen al Dr. Malcolm David Murray para proporcionar los métodos para construir el fantasma.

1. Phantom Diseño Un dibujo de la preclínica multimodal fantasma se muestra en la Figura 1 7, 8. El tamaño de la fantasma es de 38 mm de diámetro y 115 mm de longitud para permitir que el fantasma para ser escaneado en varios escáneres animales. El fantasma contiene cinco tumores que simulan los objetos de ensayo (diámetro: 14, 10, 7, 4 y 2 mm) colocados a una profundidad de 10 mm dentro de la fantasma. 2. Molde del silicón de la construcción Moldes de silicona se preparan para lanzar los objetos de prueba que simulan tumores como se describe en la sección 7. Todas las placas de acrílico y perfiles necesarios para la preparación de los moldes de silicona se cortan con una precisión de 25 micras en el taller de mecánica de la Universidad de Texas Health Science Center en San Antonio (UTHSCSA). <ol><li> Hacer cinco agujeros (diámetro: 14, 10, 7, 4, 2 mm) para los objetos de prueba y otros cinco agujeros (diámetro: 6 mm) para las barras de alineación en dos base acrílicaplacas (tamaño: 4.2 cm x 11.5 cm x 0.9 cm) (Figura 2 A). </li><li> Cortar pares espaciadores con altura de 7, 5, 3,5, 2 y 1 mm (tamaño: 1,0 cm × 5,5 cm) (Figura 2 B). </li><li> Preparar bolas de acero (diámetro: 14, 10, 7, 4 y 2 mm, la precisión: 2,5 micras). </li><li> Coloque dos pares de separadores con altura de 7 mm y una placa base sobre una placa acrílica delgada en secuencia y apretarlos con abrazaderas en C (Figura 2 C). </li><li> Inserte la bola de acero con 14 mm de diámetro en 14 mm agujero de la placa base y la cola con JB KWIK (Figura 2 C). Repetir los procedimientos para el resto de las bolas (Figura 2 D) y para la otra placa de base. Tenga en cuenta que las bolas de acero en dos placas de base se pegan como imágenes de espejo 7. </li><li> Puede conectar cuatro placas de acrílico cm de altura (tamaño: 2,5 cm x 11,5 cm por dos platos y 2,5 cm x 4,2 cm para otras dos placas) 2.5 en cada base con cinta adhesiva como las cercas ( <str<html> ong&gt; Figura 3 A). </li><li> Coloque la placa superior (tamaño: 4,2 cm x 11,5 cm, cinco agujeros con 0,8 cm de diámetro, diez agujeros con 1,2 cm de diámetro) en uno de los conjuntos de placas de base para insertar cinco barras de acrílico (diámetro: 0,8 cm y la longitud: 0,5 cm) con 1 mm de consejos, y para insertar cinco barras de alineación (diámetro: 0,9 cm y longitud: 5,0 cm) y para verter de silicona (Figura 3 A). </li><li> Inserte las barras de acrílico en los orificios de 0,8 cm de la placa superior hasta llegar a la parte superior de las bolas de acero y pegarlas con pegamento de silicona. A continuación, inserte las barras de alineación en los orificios de la placa base a través de los agujeros más grandes en la placa superior (Figura 3 A). </li><li> Mezclar la parte A del compuesto de caucho de silicona con la parte B en la relación de 10 a 1 en peso. </li><li> Verter el compuesto de caucho de silicona en el montaje y secar el conjunto a temperatura ambiente durante aproximadamente 24 h (Figura 3 B). </li></ol> 3. Asamblea Rotator t &quot;&gt; El rotor se prepara a partir de tubos de PVC y un motor del asador. <ol><li> Moler el extremo de un perno para encajar el agujero de un motor del asador. </li><li> Atornillar el perno de tierra hasta el final de la tubería de PVC (longitud: 270 mm y el diámetro interior: 75 mm) utilizando una tuerca y una arandela. </li><li> Doble las placas de metal y pegarlas en una placa de plástico usando JB KWIK para apoyar el tubo de PVC y para ajustar la altura del tubo de PVC 7. </li></ol> 4. TM Preparación de materiales Los protocolos para la preparación de los materiales de TM se modifican a partir de los desarrollados en el laboratorio del Dr. Ernest L. Madsen en la Universidad de Wisconsin, Madison y más detalles están en Lee et al. 8,11. 4.1 Antecedentes preparación del material TM <ol><li> Pasar la leche entera comercial (200 cc) a través de 20 micras y a continuación, filtros de malla 10 micras. </li><li> Disolver timerosal (0,2 g) en la leche filtrada (100 cc). </li><li> Usando vacío de la casa, degas de esta solución de leche durante 30 segundos a temperatura ambiente. </li><li> Disolver agarosa seca (2 g) en agua desionizada (18 mW) (100 cc) a temperatura ambiente. </li><li> A continuación, añadir 1-propanol (7,9 cc) y BaSO4 (1 g) a la solución de agarosa. </li><li> Degas la solución de agarosa y luego calentar en un baño de agua a 95 ° C hasta que la solución de agarosa aclare. </li><li> Si bien la solución de agarosa despeja en el 95 ° C baño de agua, caliente la leche condensada en un baño de agua a 55 ° C. </li><li> Mueva la solución de agarosa fundido al baño de agua 55 ° C que se enfríe. </li><li> Una vez que ambas soluciones están a 55 ° C, la mezcla de solución de agarosa (50 cc) con leche condensada (50 cc) para hacer la relación del 50 al 50 en volumen y, lentamente, se agita la mezcla, seguido de eliminación de burbujas de aire de la superficie. </li><li> A continuación, añadir EDTA (0,103 g) y CuCl 2 · 2H 2 O (0,06 g) a la mezcla de agarosa-leche seguido de suficiente agitación para asegurar la homogeneidad. </li><li> Por último, añadir perlas de vidrio (15- 60 m de diámetro, el diámetro medio: 35 m) (0,1 g) y se agita la mezcla final en varias ocasiones. Antes de su uso, sumerja las perlas de vidrio en ácido nítrico concentrado durante 24 horas para eliminar cualquier impureza y luego enjuagar el ácido. </li></ol> 4.2 Prueba objeto TM preparación de la materia El objeto material de TM de ensayo se prepara de una manera similar como el material de fondo TM excepto por las siguientes diferencias de composición: <ol><li> Pasar la leche entera comercial (20 cc) a través de 20 micras y a continuación, filtros de malla 10 micras. </li><li> Disolver timerosal (0,02 g) en la leche filtrada (10 cc). </li><li> Disolver agarosa seca (0,60 g) en una solución a temperatura ambiente de agua desionizada (10 cc) y 1-propanol (0,79 cc). </li><li> Degas la solución de agarosa y luego calentar en un baño de agua a 95 ° C hasta que la solución de agarosa aclare. </li><li> Si bien la solución de agarosa despeja en el 95 ° C baño de agua, caliente la leche condensada en un baño de agua a 55 ° C. </li><li&gt; Mueva la solución de agarosa fundido al ° C baño de agua 55. </li><li> Una vez que ambas soluciones están a 55 ° C, mezclar la solución de agarosa (5 cc) con leche condensada (5 cc) lentamente y se agita la mezcla, seguido de eliminación de burbujas de aire de la superficie. </li><li> A continuación, añadir EDTA (0,0017 g) y CuCl 2 · 2H 2 O (0,0010 g) a la leche de agarosa seguido de suficiente agitación. </li></ol> 5. Asamblea Phantom multimodalidad Uso de los moldes de silicona, los siguientes pasos se llevan a cabo para construir el fantasma multimodalidad. <ol><li> En el molde de silicona sin agujeros de 1 mm, adjuntar hilo de nylon a lo largo del centro de las esferas y el pegamento en ambos extremos del molde utilizando pegamento de silicona (Figura 4 A). </li><li> El uso de un cepillo suave, aplique grasa de silicona en la superficie de dos moldes (Figura 4 A) y montar dos moldes utilizando varillas de alineación. </li><li> Preparar la prueba objeto material TM como se describe en la sección4,2 y se vierte a través de agujeros de 1 mm del molde de silicona con una aguja de calibre 22 de una jeringa. </li><li> Para permitir que los objetos de prueba para establecer, almacenar los moldes en un refrigerador (5 ° C) durante aproximadamente 30 min. </li><li> En cada lado de un contenedor de medio-cilíndrica (longitud: 115 mm y diámetro: 38 mm), hacer dos agujeros de 1 mm a una profundidad de 10 mm desde la superficie del maniquí con el fin de montar hilo de nylon con objetos de prueba. Haga un agujero adicional de 6 mm para verter material de TM. </li><li> Descarga de los objetos de prueba con hilo de nylon de los moldes (Figura 4 B) y luego montarlos en el recipiente de medio-cilíndrica (Figura 4 C). </li><li> Usando 3M Scotch-Weld DP-100 y la cinta adhesiva de 3M, se adhieren aluminio no conductora delgada (espesor: 0,12 mm) sobre el recipiente de acrílico. Bloquear los orificios de 1 mm en el recipiente acrílico utilizando la misma cola (Figura 4 C). </li><li> Preparar material de TM rápidamente y poco a poco se vierte en el agujero de 6 mm del recipienteel uso de un pequeño embudo de plástico. </li><li> Después de la eliminación de las burbujas de aire, pegamento del agujero 6 mm con 3M Scotch-Weld DP-100. </li><li> Una vez montado, gire el fantasma en 2 rpm en el rotor de 4 a 5 horas a temperatura ambiente. </li><li> Retire el hilo de nylon después de que los materiales de TM en el fantasma se endurece por completo. </li></ol> 6. Imágenes multimodalidad El fantasma se escanea en la ecografía preclínica, CT y MRI e imágenes en tres modalidades se adquieren. Los protocolos de formación de imágenes se describen en detalle en Lee et al. 7, 8.

<ol>
	<li>Prasad, S. R., 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=CT+tumor+measurement+for+therapeutic+response+assessment:+Comparison+of+unidimensional,+bidimensional,+and+volumetric+techniques-Initial+observations.">CT tumor measurement for therapeutic response assessment: Comparison of unidimensional, bidimensional, and volumetric techniques-Initial observations.</a> Radiology. 225 (2), 416-419 (2002).</li><li>Cortes, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+unidimensional+and+bidimensional+measurement+in+metastatic+non-small+cell+lung+cancer.">Comparison of unidimensional and bidimensional measurement in metastatic non-small cell lung cancer.</a> Br. J. Cancer. 87 (2), 158-160 (2002).</li><li>Saini, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiologic+measurement+of+tumor+size+in+clinical+trials:+past,+present,+and+future.">Radiologic measurement of tumor size in clinical trials: past, present, and future.</a> AJR Am. J. Roentgenol. 176 (2), 333-334 (2001).</li><li>Suzuki, C., 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=Radiologic+measurements+of+tumor+response+to+treatment:+practical+approaches+and+limitations.">Radiologic measurements of tumor response to treatment: practical approaches and limitations.</a> Radiographics. 28 (2), 329-344 (2008).</li><li>Therasse, P., 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=RECIST+revisited:+A+review+of+validation+studies+on+tumour+assessment.">RECIST revisited: A review of validation studies on tumour assessment.</a> Eur. J. Cancer. 42 (8), 1031-1039 (2006).</li><li>O'Connor, P. B., 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=Quantitative+imaging+biomarkers+in+the+clinical+development+of+targeted+therapeutics:+current+and+future+perspectives.">Quantitative imaging biomarkers in the clinical development of targeted therapeutics: current and future perspectives.</a> Lancet Oncol. 9 (8), 766-776 (2008).</li><li>Lee, Y. C., 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=Preclinical+multimodality+phantom+design+for+quality+assurance+of+tumor+size+measurement.">Preclinical multimodality phantom design for quality assurance of tumor size measurement.</a> BMC Med. Phys. 11 (1), (2011).</li><li>Lee, Y. C., 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=QA+procedures+for+multimodality+preclinical+tumor+drug+response+testing.">QA procedures for multimodality preclinical tumor drug response testing.</a> Med. Phys. 37 (9), 4806-4816 (2010).</li><li>Park, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+response+in+solid+tumors:+comparison+of+RECIST+and+WHO+response+criteria.">Measuring response in solid tumors: comparison of RECIST and WHO response criteria.</a> Jpn. J. Clin. Oncol. 33 (10), 533-537 (2003).</li><li>Eisenhauer, E. A., 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=New+response+evaluation+criteria+in+solid+tumors:+revised+RECIST+guideline+(version+1.1).">New response evaluation criteria in solid tumors: revised RECIST guideline (version 1.1).</a> Eur. J. Cancer. 45 (2), 228-247 (2009).</li><li>D'Souza, W. D., 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=Tissue+mimicking+materials+for+a+multi-imaging+modality+prostate+phantom.">Tissue mimicking materials for a multi-imaging modality prostate phantom.</a> Med. Phys. 28 (4), 688-700 (2001).</li><li>Mitchell, M. D., 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=Agarose+as+a+tissue+equivalent+phantom+material+for+NMR+images.">Agarose as a tissue equivalent phantom material for NMR images.</a> Magn. Reson. Imaging. 4 (3), 263-266 (1986).</li></ol>

No hay conflictos de interés declarado.

El objetivo de este artículo es proporcionar los métodos para la fabricación de materiales TM para imágenes multimodalidad y la construcción de un fantasma multimodalidad preclínica como una herramienta de control de calidad para medir el tamaño del tumor precisa el uso de diferentes modalidades en múltiples instituciones. Como se mencionó anteriormente, los materiales de TM fueron originalmente desarrollados por el laboratorio del Dr. Ernest L. Madsen en la Universidad de Wisconsin, Madison por una modalidad d...

<html> Evaluación del cambio en el tamaño del tumor es un punto final importante para evaluar la actividad de los fármacos anti-tumorales tanto en la reducción del tumor y la progresión de la enfermedad 9, 10. Organización Mundial de la Salud (OMS) y los Criterios de Evaluación de Respuesta en Tumores Sólidos (RECIST) son los métodos codificados para la evaluación anatómica de las lesiones tumorales en las técnicas de imagen como la ecografía (EE.UU.), la tomografía computarizada (TC) o la resonancia magnética (MRI). Para los criterios de la OMS, el producto del diámetro máximo del tumor y su diámetro mayor perpendicular en el plano transversal de las regiones de destino se calcula 4. En contraste, para los criterios RECIST, la suma de los diámetros más largo en el plano transversal para un número limitado de lesiones diana se calcula 4. A pesar de la continua creciente interés en la evaluación de la respuesta terapéutica tumoral, no ha habido ninguna garantía de calidad preclínico (QA) fantasma / QA procedimientos para el biomarcador de imagen. contenido &quot;&gt; Teniendo en cuenta que la medida del tamaño del tumor en base a criterios de la OMS y / o RECIST es el único biomarcador de imagen aprobado por la Food and Drug Administration (FDA), como punto de partida de la GC para otros biomarcadores de imagen, Lee et al. diseñado y construido UTHSCSA / Gammex Marcos 1 y Mark 2 fantasmas de QA de la medida del tamaño del tumor en colaboración con Gammex Inc 7. The Mark 1 fantasma era una versión revisada de un Gammex comercial EE.UU. fantasma y por lo tanto, el tamaño era demasiado grande para caber en animales escáneres TC y RM. también algunas herramientas en la Marca 1 fantasma eran innecesarios para la medición del tamaño del tumor. La marca 2 fantasma fue diseñado sobre la base de criterios RECIST que es el biomarcador de imágenes aprobado por la FDA más reciente. Sin embargo, el tamaño de la marca 2 fantasma era todavía demasiado grande para escáneres MR, y CT y MR calidad de la imagen de la fantasma no era aceptable para la medición precisa el tamaño del tumor 7. El QA fantasma describe querienda fue re-diseñado para superar las deficiencias de los fantasmas anteriores y construido con materiales modificados tejidos que imitan (TM) y los protocolos desarrollados en nuestro laboratorio. Este documento describe los detalles de los protocolos para la construcción de fantasma: En primer lugar, se introducen los métodos para la preparación de los moldes de silicona necesarios para la fundición de tumor que simulan objetos de prueba y para el montaje de un rotador para la rotación de un fantasma para evitar la sedimentación gravitación. En segundo lugar, los protocolos para la preparación de materiales TM modificados de D&#39;Souza et al. &#39;S para los EE.UU., TC y RM se describen 11. Las propiedades físicas de los materiales de TM fueron probados en cada modalidad para asegurar que los materiales TM representados tejidos blandos humanos como se observa en las imágenes obtenidas con las diferentes modalidades, pero los resultados no se muestran aquí. En tercer lugar, se describe el protocolo para la construcción fantasma. Por último, los Estados Unidos, TC y RM imágenes del fantasma se presentan como resultados.

<table border="1">
	<tbody>
		<tr>
			<td>Reagent/Material</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PVC pipe</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Home Depot</td>
		</tr>
		<tr>
			<td>Bolt, nut, washer and metal plates</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Home Depot</td>
		</tr>
		<tr>
			<td>Acrylic plates and rods</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Plastic supply in San Antonio, TX</td>
		</tr>
		<tr>
			<td>Steel balls</td>
			<td>Nordex, Inc.</td>
			<td>AEC-M2-2, -4, -7, -10 and -14</td>
			<td>2, 4, 7, 10 and 14 mm diameter</td>
		</tr>
		<tr>
			<td>C-clamps</td>
			<td>Adjustable Clamp</td>
			<td>1420-C</td>
			<td>2 inch length</td>
		</tr>
		<tr>
			<td>Masking tape</td>
			<td>3M Industrial Adhesives and Tapes</td>
			<td>2600</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Duct tape</td>
			<td>3M Industrial Adhesives and Tapes</td>
			<td>S-3763SIL</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>J-B KWIK</td>
			<td>J-B WELD Co.</td>
			<td>380238</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>3M Scotch-Weld Epoxy Adhesive</td>
			<td>3M Industrial Adhesives and Tapes</td>
			<td>DP-100</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Silicone grease</td>
			<td>Permatex, Inc.</td>
			<td>22058</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Silicone glue</td>
			<td>DAP, Inc.</td>
			<td>688</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Silicone rubber compound</td>
			<td>Smooth-ON, Inc.</td>
			<td>Smooth-SilTM950 Part A and B</td>
			<td>A:B mix ratio = 10:1 by weight</td>
		</tr>
		<tr>
			<td>Brush</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Hobby Lobby</td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>Becton Dickinson</td>
			<td>309604</td>
			<td>10 ml</td>
		</tr>
		<tr>
			<td>Needle</td>
			<td>Becton Dickinson</td>
			<td>305156</td>
			<td>22-gauge 1.5 inch length</td>
		</tr>
		<tr>
			<td>Funnel</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Mesh filters</td>
			<td>Small parts, Inc.</td>
			<td>CMN-0010-C and CMN-0020-C</td>
			<td>10 and 20 &mu;m</td>
		</tr>
		<tr>
			<td>Whole milk</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>HEB in San Antonio, TX</td>
		</tr>
		<tr>
			<td>Thimerosal</td>
			<td>Sigma-Aldrich Co.</td>
			<td>T5125</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Propanol</td>
			<td>Sigma-Aldrich Co.</td>
			<td>33538</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Sigma-Aldrich Co.</td>
			<td>431788</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>CuCl2</td>
			<td>Sigma-Aldrich Co.</td>
			<td>459097</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Sigma-Aldrich Co.</td>
			<td>A0169</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>BaSO4</td>
			<td>Sigma-Aldrich Co.</td>
			<td>B8675</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Glass beads</td>
			<td>Potters Industries, Inc.</td>
			<td>3000E</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PET/AL/LLDPE*</td>
			<td>Pechiney Plastic Packaging, Inc.</td>
			<td>Pechiney Spec 151</td>
			<td>Phantom cover material</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>*Polyethylene terephthalate/aluminum/linear low density polyethylene</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Rotisserie motor</td>
			<td>Brinkmann</td>
			<td>812-7103-S</td>
			<td>Home Depot</td>
		</tr>
		<tr>
			<td>Water bath 1</td>
			<td>Precision, Inc.</td>
			<td>Model: 282, Serial #: 601091552</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Water bath 2</td>
			<td>VWR, Inc.</td>
			<td>Model: 1212, Serial #: 08119606</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ultrasound</td>
			<td>Visualsonics</td>
			<td>Serial #: 770/120-259</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>CT</td>
			<td>Gamma Medica-Ideas</td>
			<td>Serial #: GR 0050</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>MRI</td>
			<td>Bruker</td>
			<td>Part #: W3301390, Serial #: 0030</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

construction of a preclinical multimodality phantom using tissue-mimicking materials for quality assurance in tumor size measurement

Organización Mundial de la Salud (OMS) y los Criterios de Evaluación de Respuesta en Tumores Sólidos (RECIST) grupos de trabajo abogó por criterios estandarizados para la evaluación radiológica de los tumores sólidos, en respuesta a la terapia con medicamentos anti-tumor en los años 1980 y 1990, respectivamente. Criterios de la OMS miden los tumores sólidos en dos dimensiones, mientras que las mediciones RECIST utilizan sólo una dimensión que se considera que es más reproducible 1, 2, 3,4,5. Estos criterios han sido ampliamente utilizados como el único biomarcador de imagen aprobado por la Food and Drug Administration (FDA) 6. Con el fin de medir la respuesta del tumor a los fármacos anti-tumorales en las imágenes con precisión, por lo tanto, se necesitan unos procedimientos de garantía de calidad (QA sólidos) y de control de calidad correspondiente fantasma. Para hacer frente a esta necesidad, los autores construyeron un multimodalidad preclínica (por ultrasonido (EE.UU.), la tomografía computarizada (TC) y la resonancia magnética (MRI)) fantasma usando tejido que imita (TM)materiales basados ​​en el número limitado de lesiones diana requeridos por RECIST mediante la revisión de una Gammex EE.UU. comercial phantom 7. El Apéndice en Lee et al. Demuestra los procedimientos de fabricación fantasma 7. En este artículo, todos los protocolos se introducen en un paso-a-paso de la manera principio con los procedimientos para la preparación de los moldes de silicona para la fundición de tumor que simulan objetos de prueba en el espectro, seguido por la preparación de materiales de TM para imágenes multimodalidad, y, finalmente, la construcción de la preclínica multimodal QA fantasma. El propósito principal de este trabajo es proporcionar los protocolos para permitir que cualquier persona interesada en la construcción de forma independiente un fantasma para sus propios proyectos. Procedimientos de control de calidad para la medida del tamaño del tumor y el RECIST, la OMS y los resultados de medición de volumen de objetos de prueba realizadas en múltiples instituciones que utilizan este QA fantasma se muestran en detalle en Lee et al. 8.

Figura 3 B y la Figura 5 muestran dos moldes de silicona para convertir los objetos de prueba, y la multimodalidad fantasma, respectivamente. La longitud x anchura x profundidad de cada molde es de 109 mm × 37 mm × 21 mm y dos moldes son idénticas imágenes especulares. Un molde tiene 1 mm Agujeros en el material de TM se puede introducir mediante una aguja fina. Cada molde tiene un cinco agujeros adicionales para las barras de alineación. La longitud x anchura x profundidad del fan...

Watch this Scientific Journal Video about Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement at JoVE.com

Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement

Este documento describe los procedimientos internos de la construcción de una multimodalidad preclínica fantasma hecho de tejido que imita (TM) materiales para la garantía de calidad (QA) de la medida del tamaño del tumor en las modalidades de imágenes de animales como la ecografía (EE.UU.), la tomografía computarizada (CT) y magnético resonancia magnética (MRI).

Construcción de un Phantom multimodalidad preclínicos Utilización de los materiales de tejidos que imitan para la Garantía de la Calidad en la medida del tamaño del tumor

World Health Organization (WHO) and the Response Evaluation Criteria in Solid Tumors (RECIST) working groups advocated standardized criteria for ...

construction-preclinical-multimodality-phantom-using-tissue-mimicking

Research

JoVE Journal

Bioengineering

11.2K vistas.  University of Kansas School of Medicine. Este documento describe los procedimientos internos de la construcción de una multimodalidad preclínica fantasma hecho de tejido que imita (TM) materiales para la garantía de calidad (QA) de la medida del tamaño del tumor en las modalidades de imágenes de animales como la ecografía (EE.UU.), la tomografía computarizada (CT) y magnético resonancia magnética (MRI).

Video: Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement

Viral Nanoparticles for In vivo Tumor Imaging

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being investigated for applications in imaging and therapy. Viral nanoparticles (VNPs) derived from plants can be regarded as self-assembled bionanomaterials with defined sizes and shapes. Plant viruses under investigation in the Steinmetz lab include icosahedral particles formed by Cowpea mosaic virus (CPMV) and Brome mosaic virus (BMV), both of which are 30 nm in diameter. We are also developing rod-shaped and filamentous structures derived from the following plant viruses: Tobacco mosaic virus (TMV), which forms rigid rods with dimensions of 300 nm by 18 nm, and Potato virus X (PVX), which form filamentous particles 515 nm in length and 13 nm in width (the reader is referred to refs. 1 and 2 for further information on VNPs).
From a materials scientist's point of view, VNPs are attractive building blocks for several reasons: the particles are monodisperse, can be produced with ease on large scale in planta, are exceptionally stable, and biocompatible. Also, VNPs are "programmable" units, which can be specifically engineered using genetic modification or chemical bioconjugation methods 3. The structure of VNPs is known to atomic resolution, and modifications can be carried out with spatial precision at the atomic level4, a level of control that cannot be achieved using synthetic nanomaterials with current state-of-the-art technologies.
In this paper, we describe the propagation of CPMV, PVX, TMV, and BMV in Vigna ungiuculata and Nicotiana benthamiana plants. Extraction and purification protocols for each VNP are given. Methods for characterization of purified and chemically-labeled VNPs are described. In this study, we focus on chemical labeling of VNPs with fluorophores (e.g. Alexa Fluor 647) and polyethylene glycol (PEG). The dyes facilitate tracking and detection of the VNPs 5-10, and PEG reduces immunogenicity of the proteinaceous nanoparticles while enhancing their pharmacokinetics 8,11. We demonstrate tumor homing of PEGylated VNPs using a mouse xenograft tumor model. A combination of fluorescence imaging of tissues ex vivo using Maestro Imaging System, fluorescence quantification in homogenized tissues, and confocal microscopy is used to study biodistribution. VNPs are cleared via the reticuloendothelial system (RES); tumor homing is achieved passively via the enhanced permeability and retention (EPR) effect12. The VNP nanotechnology is a powerful plug-and-play technology to image and treat sites of disease in vivo. We are further developing VNPs to carry drug cargos and clinically-relevant imaging moieties, as well as tissue-specific ligands to target molecular receptors overexpressed in cancer and cardiovascular disease.

Viral Nanoparticles for In vivo Tumor Imaging

Plant viral nanoparticles (VNPs) are promising platforms for applications in biomedicine. Here, we describe the procedures for plant VNP propagation, purification, characterization, and bioconjugation. Finally, we show the application of VNPs for tumor homing and imaging using a mouse xenograft model and fluorescence imaging.

Las nanopartículas virales para<em&gt; En vivo</em&gt; Imágenes de tumores

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being ...

Investigación

Bioingeniería

Production of Large Numbers of Size-controlled Tumor Spheroids Using Microwell Plates

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically relevant than conventional adherent-sheet cultures, they more accurately recapitulate the complexity and interactions present in real tumors. In order to harness this model to better assess tumor biology, or the efficacy of novel therapeutic agents, it is necessary to be able to generate spheroids reproducibly, in a controlled manner and in significant numbers.
The AggreWell system consists of a high-density array of pyramid-shaped microwells, into which a suspension of single cells is centrifuged. The numbers of cells clustering at the bottom of each microwell, and the number and ratio of distinct cell types involved depend only on the properties of the suspension introduced by the experimenter. Thus, we are able to generate tumor spheroids of arbitrary size and composition without needing to modify the underlying platform technology. The hundreds of microwells per square centimeter of culture surface area in turn ensure that extremely high production levels may be attained via a straightforward, nonlabor-intensive process. We therefore expect that this protocol will be broadly useful to researchers in the tumor spheroid field.

We introduce a protocol for the generation of large numbers (thousands to hundreds of thousands) of uniform size- and composition-controlled tumor spheroids, using commercially available microwell plates.

La producción de grandes cantidades de esferoides tumorales Tamaño controlados utilizando placas de micropocillos

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically ...

Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and modified as part of a set of complex, two-way mechanosensitive interactions. While the development of biologically-relevant 3D tumor models have facilitated mechanistic studies on the impact of matrix rheology on tumor growth, the inverse problem of mapping changes in the mechanical environment induced by tumors remains challenging. Here, we describe the implementation of particle-tracking microrheology (PTM) in conjunction with 3D models of pancreatic cancer as part of a robust and viable approach for longitudinally monitoring physical changes in the tumor microenvironment, in situ. The methodology described here integrates a system of preparing in vitro 3D models embedded in a model extracellular matrix (ECM) scaffold of Type I collagen with fluorescently labeled probes uniformly distributed for position- and time-dependent microrheology measurements throughout the specimen. In vitro tumors are plated and probed in parallel conditions using multiwell imaging plates. Drawing on established methods, videos of tracer probe movements are transformed via the Generalized Stokes Einstein Relation (GSER) to report the complex frequency-dependent viscoelastic shear modulus, G*(&#969;). Because this approach is imaging-based, mechanical characterization is also mapped onto large transmitted-light spatial fields to simultaneously report qualitative changes in 3D tumor size and phenotype. Representative results showing contrasting mechanical response in sub-regions associated with localized invasion-induced matrix degradation as well as system calibration, validation data are presented. Undesirable outcomes from common experimental errors and troubleshooting of these issues are also presented. The 96-well 3D culture plating format implemented in this protocol is conducive to correlation of microrheology measurements with therapeutic screening assays or molecular imaging to gain new insights into impact of treatments or biochemical stimuli on the mechanical microenvironment.

Particle-tracking microrheology can be used to non-destructively quantify and spatially map changes in extracellular matrix mechanical properties in 3D tumor models.

Medida longitudinal de la matriz extracelular Rigidez en modelos de tumores en 3D utilizando partículas de seguimiento Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and ...

Construction and Characterization of a Novel Vocal Fold Bioreactor

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 &#181;m. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

A novel vocal fold bioreactor capable of delivering physiologically relevant, vibratory stimulation to cultured cells is constructed and characterized. This dynamic culture device, when combined with a fibrous poly(&#949;-caprolactone) scaffold, creates a vocal fold-mimetic environment that modulates the behaviors of mesenchymal stem cells.

Construcción y caracterización de un Fold biorreactor Novela Vocal

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To ...

Design and Construction of Artificial Extracellular Matrix (aECM) Proteins from Escherichia coli for Skin Tissue Engineering

Recombinant technology is a versatile platform to create novel artificial proteins with tunable properties. For the last decade, many artificial proteins that have incorporated functional domains derived from nature (or created de novo) have been reported. In particular, artificial extracellular matrix (aECM) proteins have been developed; these aECM proteins consist of biological domains taken from fibronectin, laminins and collagens and are combined with structural domains including elastin-like repeats, silk and collagen repeats. To date, aECM proteins have been widely investigated for applications in tissue engineering and wound repair. Recently, Tjin and coworkers developed integrin-specific aECM proteins designed for promoting human skin keratinocyte attachment and propagation. In their work, the aECM proteins incorporate cell binding domains taken from fibronectin, laminin-5 and collagen IV, as well as flanking elastin-like repeats. They demonstrated that the aECM proteins developed in their work were promising candidates for use as substrates in artificial skin. Here, we outline the design and construction of such aECM proteins as well as their purification process using the thermo-responsive characteristics of elastin.

Design and Construction of Artificial Extracellular Matrix aECM Proteins from Escherichia coli for Skin Tissue Engineering

Recombinant technologies have enabled material designers to create novel artificial proteins with customized functionalities for tissue engineering applications. For example, artificial extracellular matrix proteins can be designed to incorporate structural and biological domains derived from native ECMs. Here, we describe the construction and purification of aECM proteins containing elastin-like repeats.

Diseño y Construcción de Artificial matriz extracelular (AECM) Las proteínas de<em&gt; Escherichia coli</em&gt; Para la piel Ingeniería de Tejidos

Recombinant technology is a versatile platform to create novel artificial proteins with tunable properties. For the last decade, many artificial proteins ...

Agarose-based Tissue Mimicking Optical Phantoms for Diffuse Reflectance Spectroscopy

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a conventional optical system with an integrating sphere. Measuring systems for the acquisition of the diffuse reflectance and total transmittance spectra are constructed with a broadband white light source, a light guide, an achromatic lens, an integrating sphere, a sample holder, an optical fiber probe, and a multi-channel spectrometer. An acrylic mold consisting of two rectangular acrylic pieces and a U-shaped acrylic piece is constructed to create an epidermal phantom and a dermal phantom with whole blood. The application of a sodium dithionite (Na2S2O4) solution to the dermal phantom enables the researcher to deoxygenate hemoglobin in red blood cells distributed in the dermal phantom. The inverse Monte Carlo simulation with the diffuse reflectance and total transmittance spectra measured by a spectrometer with an integrating sphere is performed to determine the absorption coefficient spectrum &#181;a(&#955;) and the reduced scattering coefficient spectrum &#181;s&#39;(&#955;) of each layer phantom. A two-layered phantom mimicking the diffuse reflectance of human skin tissue is also demonstrated by piling up the epidermal phantom on the dermal phantom.

Here, we demonstrate how agarose-based tissue-mimicking optical phantoms are made and how their optical properties are determined using a conventional optical system with an integrating sphere.

Basado en la agarosa tejido imitando fantasmas ópticos para espectroscopia de reflectancia difusa

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a ...

Manufacturing Abdominal Aorta Hydrogel Tissue-Mimicking Phantoms for Ultrasound Elastography Validation

Ultrasound (US) elastography, or elasticity imaging, is an adjunct imaging technique that utilizes sequential US images of soft tissues to measure the tissue motion and infer or quantify the underlying biomechanical characteristics. For abdominal aortic aneurysms (AAA), biomechanical properties such as changes in the tissue's elastic modulus and estimates of the tissue stress may be essential for assessing the need for the surgical intervention. Abdominal aortic aneurysms US elastography could be a useful tool to monitor AAA progression and identify changes in biomechanical properties characteristic of high-risk patients.
A preliminary goal in the development of an AAA US elastography technique is the validation of the method using a physically relevant model with known material properties. Here we present a process for the production of AAA tissue-mimicking phantoms with physically relevant geometries and spatially modulated material properties. These tissue phantoms aim to mimic the US properties, material modulus, and geometry of the abdominal aortic aneurysms. Tissue phantoms are made using a polyvinyl alcohol cryogel (PVA-c) and molded using 3D printed parts created using computer aided design (CAD) software. The modulus of the phantoms is controlled by altering the concentration of PVA-c and by changing the number of freeze-thaw cycles used to polymerize the cryogel. The AAA phantoms are connected to a hemodynamic pump, designed to deform the phantoms with the physiologic cyclic pressure and flows. Ultra sound image sequences of the deforming phantoms allowed for the spatial calculation of the pressure normalized strain and the identification of mechanical properties of the vessel wall. Representative results of the pressure normalized strain are presented.

Here we describe a method to manufacture aneurysmal, aortic tissue-mimicking phantoms for the use in testing ultrasound elastography. The combined use of computer-aided design (CAD) and 3-dimensional (3D) printing techniques produce aortic phantoms with predictable, complex geometries to validate the elastographic imaging algorithms with controlled experiments.

Fabricación de Aorta Abdominal hidrogel de tejido imitando fantasmas para la validación de la elastografía de ultrasonido

Ultrasound (US) elastography, or elasticity imaging, is an adjunct imaging technique that utilizes sequential US images of soft tissues to measure the ...

Patient-Specific Polyvinyl Alcohol Phantom Fabrication with Ultrasound and X-Ray Contrast for Brain Tumor Surgery Planning

Phantoms are essential tools for clinical training, surgical planning and the development of novel medical devices. However, it is challenging to create anatomically accurate head phantoms with realistic brain imaging properties because standard fabrication methods are not optimized to replicate any patient-specific anatomical detail and 3D printing materials are not optimized for imaging properties. In order to test and validate a novel navigation system for use during brain tumor surgery, an anatomically accurate phantom with realistic imaging and mechanical properties was required. Therefore, a phantom was developed using real patient data as input and 3D printing of molds to fabricate a patient-specific head phantom comprising the skull, brain and tumor with both ultrasound and X-ray contrast. The phantom also had mechanical properties that allowed the phantom tissue to be manipulated in a similar manner to how human brain tissue is handled during surgery. The phantom was successfully tested during a surgical simulation in a virtual operating room.
The phantom fabrication method uses commercially available materials and is easy to reproduce. The 3D printing files can be readily shared, and the technique can be adapted to encompass many different types of tumor.

This protocol describes the fabrication of a patient specific skull, brain and tumor phantom. It uses 3D printing to create molds, and polyvinyl alcohol (PVA-c) is used as the tissue mimicking material.

Fabricación fantasma de alcohol polivinílico específica del paciente con ultrasonido y contraste de rayos X para la planificación de la cirugía de tumor cerebral

Phantoms are essential tools for clinical training, surgical planning and the development of novel medical devices. However, it is challenging to create ...

Generation of Dynamical Environmental Conditions using a High-Throughput Microfluidic Device

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and organs are either highly expensive, like robotics, or lack nanoliter volume and millisecond time accuracy in liquid manipulation. We herein present the design and fabrication of a microfluidic system, which consists of 1,500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. To demonstrate the capacities of the microfluidic device, neural stem cell (NSC) spheres are maintained in the proposed system. We observed that when the NSC sphere is exposed to CXCL in day 1 and EGF in day 2, the round-shaped conformation is well maintained. Variation in the input order of 6 drugs causes morphological changes to the NSC sphere and the expression level representative marker for NSC stemness (i.e., Hes5 and Dcx). These results indicate that dynamic and complex environmental conditions have great effects on NSC differentiation and self-renewal, and the proposed microfluidic device is a suitable platform for high throughput studies on the complex life machinery.

We present a microfluidic system for high throughput studies on complex life machinery, which consists of 1500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. The microfluidic chip allows for the analysis of the highly complex and dynamic micro-environmental conditions in vivo.

Generación de condiciones ambientales dinámicas utilizando un dispositivo microfluídico de alto rendimiento

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and ...

Simple, Affordable, and Modular Patterning of Cells using DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells of the same or different type, micropatterning techniques have proved useful. DNA Programmed Assembly of Cells (DPAC) is a micropatterning technique that targets the adhesion of cells to a substrate or other cells using DNA hybridization. The most basic operations in DPAC begin with decorating cell membranes with lipid-modified oligonucleotides, then flowing them over a substrate that has been patterned with complementary DNA sequences. Cells adhere selectively to the substrate only where they find a complementary DNA sequence. Non-adherent cells are washed away, revealing a pattern of adherent cells. Additional operations include further rounds of cell-substrate or cell-cell adhesion, as well as transferring the patterns formed by DPAC to an embedding hydrogel for long-term culture. Previously, methods for patterning oligonucleotides on surfaces and decorating cells with DNA sequences required specialized equipment and custom DNA synthesis, respectively. We report an updated version of the protocol, utilizing an inexpensive benchtop photolithography setup and commercially available cholesterol modified oligonucleotides (CMOs) deployed using a modular format. CMO-labeled cells adhere with high efficiency to DNA-patterned substrates. This approach can be used to pattern multiple cell types at once with high precision and to create arrays of microtissues embedded within an extracellular matrix. Advantages of this method include its high resolution, ability to embed cells into a three-dimensional microenvironment without disrupting the micropattern, and flexibility in patterning any cell type.

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

Patrones simples, asequibles y modulares de células utilizando ADN

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...