Os autores agradecem ao Dr. Madsen na Universidade de Wisconsin-Madison e Cristel baiu em Gammex Inc. para fornecer conselhos sobre materiais de TM. Os autores também são gratos ao Dr. Malcolm David Murray para fornecer os métodos para construir o fantasma.

1. Fantasma projeto Um desenho do pré multimodalidade fantasma é mostrado na Figura 1 7, 8. O tamanho do simulador é de 38 mm de diâmetro e 115 mm de comprimento para permitir que o fantasma para ser digitalizado em vários scanners animais. O fantasma contém cinco tumor simulando objetos de teste (diâmetro: 14, 10, 7, 4 e 2 mm) colocados a uma profundidade de 10 mm para dentro da fantasma. 2. Silicone Mold Construção Moldes de silicone estão preparados para lançar os objetos de teste, simulando tumor, conforme descrito nesta secção 7. Todas as placas de acrílico e barras necessárias para a preparação dos moldes de silicone são cortadas com uma precisão de 25 m de a oficina mecânica na University of Texas Health Science Center em San Antonio (UTHSCSA). <ol><li> Adicione cinco buracos (diâmetro: 14, 10, 7, 4, 2 mm) para os objectos de teste e mais cinco orifícios (diâmetro de 6 mm) para as hastes de alinhamento em dois base acrílicaplacas (tamanho: 4,2 centímetros x 11,5 centímetros x 0,9 centímetros) (Figura 2 A). </li><li> Cortar pares espaçador com a altura de 7, 5, 3,5, 2 e 1 mm (tamanho: 1,0 centímetros x 5,5 centímetros) (Figura 2 B). </li><li> Prepara esferas de aço (diâmetro: 14, 10, 7, 4 e 2 mm, a precisão: 2,5 mM). </li><li> Coloque dois pares de espaçadores com altura de 7 mm e uma placa de base para uma placa de acrílico fino em seqüência e apertá-los usando C-braçadeiras (Figura 2 C). </li><li> Coloque a bola de aço com 14 mm de diâmetro por 14 mm de orifício da placa de base e colá-la utilizando JB KWIK (Figura 2 C). Repita os procedimentos para o resto das bolas (figura 2, D) e para a outra placa de base. Note-se que as esferas de aço de duas placas de base são coladas como imagens no espelho 7. </li><li> Ligue quatro placas de 2,5 cm de altura em acrílico (tamanho: 2,5 centímetros x 11,5 centímetros de duas placas e 2,5 cm × 4,2 centímetros por mais duas placas) em cada base com fita adesiva como cercas ( <str<html> ong&gt; Figura 3 A). </li><li> Fixe a placa de topo (Tamanho: 4.2 cm × 11,5 centímetros, cinco furos, com 0,8 cm de diâmetro, dez buracos com 1,2 cm de diâmetro) em um dos conjuntos da placa de base para inserir cinco hastes de acrílico (diâmetro: 0,8 cm e comprimento: 0,5 cm) com 1 mm de pontas, e para inserir cinco barras de alinhamento (diâmetro: 0,9 cm e comprimento: 5.0 cm) e silicone para derramar (Figura 3 A). </li><li> Insira as hastes de acrílico nos orifícios 0,8 centímetros na placa superior por todo o caminho até o topo de bolas de aço e cola-los com cola de silicone. Em seguida, inserir barras de alinhamento nos orifícios na placa de base através dos furos maiores da placa de topo (Figura 3 A). </li><li> Misture a parte A do composto de borracha de silicone com a parte B na proporção de 10 para 1, em peso. </li><li> Deite o composto de borracha de silicone para a montagem e secar o conjunto à temperatura ambiente durante cerca de 24 horas (Figura 3 B). </li></ol> 3. Assembléia Rotator t &quot;&gt; O rotador é preparada a partir de tubo de PVC e um motor de rotisserie. <ol><li> Moer a extremidade de um parafuso de ajuste para o furo de um motor rotativo. </li><li> Parafuso do parafuso do solo para a extremidade do tubo de PVC (comprimento: 270 mm e diâmetro interno: 75 mm), utilizando uma porca e uma anilha. </li><li> Dobrar chapas de metal e cola-los em um prato de plástico com JB Kwik para apoiar o tubo de PVC e para ajustar a altura do tubo de PVC 7. </li></ol> 4. TM preparação do material Os protocolos para a preparação dos materiais de TM são modificados daqueles desenvolvidos no laboratório do Dr. Ernest L. Madsen, da Universidade de Wisconsin, Madison e mais detalhes são em Lee et al. 8,11. 4.1 Introdução TM preparação do material <ol><li> Passe leite integral comercial (200 cc) através de 20 mM e, em seguida, 10 filtros de malha mM. </li><li> Dissolve-se Thimerosal (0,2 g) no leite filtrado (100 cc). </li><li> Usando o vácuo casa, degás esta solução de leite durante 30 segundos à temperatura ambiente. </li><li> Dissolve-se a seco de agarose (2 g) em água desionizada (18 mohms) (100 cc) à temperatura ambiente. </li><li> Em seguida, adicionar 1-propanol (7,9 cc) e BaSO 4 (1 g) à solução de agarose. </li><li> Desgaseificar a solução de agarose e depois aquecê-lo num banho de água a 95 ° C até que a solução de agarose limpa. </li><li> Embora a solução de agarose limpa no 95 ° C, banho de água, aquecer o leite condensado num banho de água a 55 ° C. </li><li> Mova a solução de agarose fundida ao banho de água 55 ° C para esfriar. </li><li> Uma vez que ambas as soluções são a 55 ° C, a solução de mistura de agarose (50 cc) com leite condensado (50 cc) para tornar a proporção de 50 para 50 até volume e agita-se a mistura lentamente, seguido de remoção de ar a partir da superfície da bolha. </li><li> Em seguida, adicionar EDTA (0,103 g) e CuCl 2 · 2H 2 O (0,06 g) à mistura de agarose de leite seguida de agitação suficiente para assegurar a homogeneidade. </li><li> Por último, adicione pérolas de vidro (15- 60 um de diâmetro, a média do diâmetro: 35 mm) (0,1 g) e agita-se a mistura final repetidamente. Antes de usar, embeber os grânulos de vidro, em ácido nítrico concentrado durante 24 horas para remover as impurezas e depois enxaguar o ácido. </li></ol> 4.2 Teste objeto TM preparação do material O objecto de teste de material TM é preparada de uma forma semelhante como o material de fundo TM, excepto para as seguintes diferenças de composição: <ol><li> Passe leite integral comercial (20 cc) através de 20 mM e, em seguida, 10 filtros de malha mM. </li><li> Dissolve-se Thimerosal (0,02 g) no leite filtrado (10 cc). </li><li> Dissolve-se a seco de agarose (0,60 g) numa solução de água desionizada (10 cc) e 1-propanol (0,79 cc) à temperatura ambiente. </li><li> Desgaseificar a solução de agarose e depois aquecê-lo num banho de água a 95 ° C até que a solução de agarose limpa. </li><li> Embora a solução de agarose limpa no 95 ° C, banho de água, aquecer o leite condensado num banho de água a 55 ° C. </li><li&gt; Mover para a solução de agarose fundida ao ° C banho de água 55. </li><li> Uma vez que ambas as soluções são a 55 ° C, misturar a solução de agarose (5 cc) com leite condensado (5 ml) e agita-se a mistura lentamente, seguido de remoção de ar a partir da superfície da bolha. </li><li> Em seguida, adicionar EDTA (0,0017 g) e CuCl 2 · 2H 2 O (0,0010 g), para o leite de agarose seguido de agitação suficiente. </li></ol> 5. Multimodalidade Fantasma Assembleia Utilizando os moldes de silicone, as seguintes etapas são efectuadas para construir o fantasma multimodalidade. <ol><li> No molde de silicone sem furos 1 mM, prender fio de nylon ao longo do centro das esferas de cola e que em ambas as extremidades do molde, utilizando cola de silicone (Figura 4 A). </li><li> Utilizando uma escova macia, aplicar lubrificante de silicone na superfície de dois moldes (Figura 4 A) e montar dois moldes usando varetas de alinhamento. </li><li> Prepare teste objeto material TM como descrito na seção4,2 e despeje através de 1 mm de orifícios do molde de silicone, utilizando uma agulha de calibre 22 numa seringa. </li><li> Para permitir que os objetos de teste para definir, armazenar os moldes em um refrigerador (5 ° C) por cerca de 30 min. </li><li> Em cada lado de um recipiente semi-cilíndrica (comprimento: 115 mm, diâmetro: 38 mm), fazer dois furos de 1 mm a uma profundidade de 10 mm a partir da superfície do simulador, a fim de montar de fio de nylon com objectos de teste. Faça um buraco adicional de 6 mm a derramar material de fundo TM. </li><li> Descarregar os objectos de teste com fio de nylon a partir dos moldes (Figura 4 B), e em seguida montá-los no recipiente semi-cilíndrica (Fig. 4 C). </li><li> Usando 3M Scotch-Weld DP-100 e 3M fita adesiva, aderir fina de alumínio não-condutor (espessura de 0,12 mm) no recipiente de acrílico. Bloquear os orifícios de 1 mm no recipiente acrílico usando a mesma cola (Figura 4 C). </li><li> Preparar material de fundo TM rapidamente e despeje lentamente-lo no buraco 6 milímetros do recipienteusando um pequeno funil de plástico. </li><li> Após a remoção de todas as bolhas de ar, cola no buraco 6 milímetros usando 3M Scotch-Weld DP-100. </li><li> Uma vez montada, a rodar a 2 rpm fantasma no rotador durante 4 a 5 horas à temperatura ambiente. </li><li> Retire o fio de nylon após os materiais TM no fantasma endurece completamente. </li></ol> 6. Multimodalidade de imagem O fantasma é digitalizada no ultra-som pré-clínica, tomografia computadorizada e ressonância magnética e imagens em três modalidades são adquiridos. Os protocolos de imagem são descritos em detalhe em 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>

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O objetivo deste artigo foi o de fornecer os métodos para a fabricação de materiais de TM para imagens multimodalidade e construção de um fantasma multimodalidade pré-clínica como uma ferramenta de controle de qualidade para a medição do tamanho do tumor preciso utilizar diferentes modalidades em várias instituições. Como mencionado anteriormente, os materiais de TM foram originalmente desenvolvido no laboratório do Dr. Ernest L. Madsen, da Universidade de Wisconsin, Madison para uma modalidade de imagem mu...

<html> Avaliação da alteração do tamanho do tumor é um parâmetro importante para avaliar a actividade de fármacos anti-tumorais, tanto o encolhimento do tumor e a progressão da doença 9, 10. Organização Mundial de Saúde (OMS) e Critérios de Avaliação de resposta em tumores sólidos (RECIST) são os métodos codificados para a avaliação anatômica das lesões tumorais nas modalidades de imagem, como ultra-som (EUA), tomografia computadorizada (TC) ou ressonância magnética (MRI). Para os critérios da OMS, o produto do diâmetro máximo do tumor e o seu maior diâmetro perpendicular no plano transversal para as regiões alvo é calculado 4. Em contraste, por RECIST, a soma dos diâmetros maior no plano transversal para um número limitado de lesões alvo é calculado 4. Apesar continuamente crescente interesse na avaliação da resposta terapêutica do tumor, não houve nenhuma garantia de qualidade pré-clínica (QA) fantasma / QA procedimentos para o biomarcador imagiologia. conteúdo &quot;&gt; Considerando-se que a medição do tamanho do tumor com base em critérios da OMS e / ou RECIST é o único biomarcador de imagem aprovados pela United States Food and Drug Administration (FDA), como ponto de partida de QA para quaisquer outros biomarcadores de imagem, Lee et al. projetado e construído UTHSCSA / Gammex Mark 1 e Mark 2 fantasmas para QA de medição do tamanho do tumor, em colaboração com Gammex Inc 7. a marca de 1 fantasma era uma versão revisada de um Gammex comercial EUA fantasma e, portanto, o tamanho era muito grande para caber em animais CT e MR scanners. Além disso, algumas ferramentas na marca um fantasma eram desnecessários para a medição do tamanho do tumor. The Mark 2 phantom foi projetado com base no RECIST que é o mais recente biomarcador de imagem FDA-approved. No entanto, o tamanho do Mark 2 phantom ainda era muito grande para MR scanners e TC e RM qualidade de imagem do fantasma não era aceitável para a medição do tamanho do tumor precisa 7. A QA fantasma descrito elerédea foi re-projetado para superar deficiências dos fantasmas anteriores e construído usando tecidos que imitam (TM) de materiais e protocolos modificados desenvolvidos em nosso laboratório. Este artigo descreve os detalhes dos protocolos para a construção phantom: Primeiro, os métodos são introduzidas para a preparação de moldes de silicone necessários para a fundição de tumor que simulam a objectos de teste e para a montagem de um rotor para rodar um fantasma para evitar a sedimentação gravitacional. Em segundo lugar, os protocolos para a preparação de materiais de TM modificados de D&#39;Souza et al. &#39;S para os EUA, tomografia computadorizada e ressonância magnética são descritos 11. As propriedades físicas dos materiais de TM foram testados em cada modalidade para garantir que os materiais TM representada tecidos moles humanos como observado nas imagens obtidas com as várias modalidades, mas os resultados não são mostradas aqui. Em terceiro lugar, o protocolo de construção é descrito fantasma. Por último, US, TC e RM imagens do phantom são apresentados 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

Organização Mundial de Saúde (OMS) e os critérios de avaliação de resposta em tumores sólidos (RECIST) grupos de trabalho defendido critérios padronizados para a avaliação radiológica de tumores sólidos em resposta à terapia medicamentosa anti-tumor na década de 1980 e 1990, respectivamente. Critérios da OMS medir tumores sólidos em duas dimensões, enquanto que as medições RECIST utilizar apenas uma dimensão que é considerado ser mais reprodutível 1, 2, 3,4,5. Estes critérios têm sido amplamente utilizados como o único biomarcador imagiologia aprovado pela United States Food and Drug Administration (FDA), 6. A fim de medir a resposta do tumor às drogas anti-tumorais em imagens com precisão, por isso, são necessárias uma garantia robustos (QA) procedimentos de qualidade e correspondente QA fantasmas. Para atender a essa necessidade, os autores construíram uma multimodalidade pré-clínico (por ultra-som (EUA), tomografia computadorizada (TC) e ressonância magnética (MRI)) fantasma usando tecido imitando (TM)materiais em função do número limitado de lesões-alvo exigidos pela RECIST revisando um Gammex EUA comercial fantasma 7. O apêndice em Lee et al. Demonstra os procedimentos do fantasma fabricação 7. Neste artigo, os protocolos são introduzidas de uma forma início passo-a-passo com os procedimentos para preparar os moldes para o vazamento de silicone tumor simulando objectos de ensaio no simulador, seguido de preparação de materiais de TM para imagiologia multimodalidade, e, finalmente, a construção do pré multimodalidade QA fantasma. O objetivo principal deste artigo é fornecer os protocolos para permitir que qualquer pessoa interessada em construir de forma independente um fantasma para seus próprios projetos. Procedimentos de controle de qualidade para a medição do tamanho do tumor, e RECIST, a OMS e os resultados dos objetos de teste feitas em várias instituições que utilizam este QA fantasma de medição de volume são mostrados em detalhes no Lee et al. 8.

Figura 3 B e Figura 5 mostram dois moldes de silicone para fundição de objetos de teste, ea multimodalidade fantasma, respectivamente. O comprimento x largura x profundidade de cada molde é de 109 mm x 37 milímetros x 21 mm e dois moldes são idênticos imagens especulares. Um molde tem um milímetro buracos onde o material TM podem ser inseridos usando uma agulha fina. Cada molde possui um adicional de cinco furos para as hastes de alinhamento. O comprimento x largura x profundidad...

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 artigo descreve os procedimentos internos de construção de uma multimodalidade pré fantasma feito de tecido imitando (TM) materiais para garantia de qualidade (QA) de medição de tamanho de tumor em modalidades de imagem animal, tais como ultra-som (EUA), tomografia computadorizada (TC) e magnético resonance imaging (MRI).

Construção de um fantasma Multimodalidade pré-clínicos com materiais que imitam tecido para a Garantia da Qualidade no Size Measurement 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 visualizações.  University of Kansas School of Medicine. Este artigo descreve os procedimentos internos de construção de uma multimodalidade pré fantasma feito de tecido imitando (TM) materiais para garantia de qualidade (QA) de medição de tamanho de tumor em modalidades de imagem animal, tais como ultra-som (EUA), tomografia computadorizada (TC) e magnético resonance imaging (MRI).

Vídeo: 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.

Nanopartículas virais para<em&gt; Em vivo</em&gt; Imaging Tumor

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

Bioengenharia

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.

Produção de grande número de controladas pelo tamanho Spheroids tumorais usando placas de micropoços

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.

Medição Longitudinal de Matriz Extracelular Rigidez em 3D tumorais modelos com o uso de rastreamento de partículas 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.

Construção e Caracterização de um biorreator Fold Novel 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.

Design e Construção de Matriz Extracelular Artificial (MECa) Proteínas de<em&gt; Escherichia coli</em&gt; Para engenharia de tecidos da pele

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.

Baseado em agarose tecido imitando Phantoms ópticos por espectroscopia de reflectância 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.

Fabricação de Aorta Abdominal hidrogel tecido-imitando Phantoms por ultra-som Elastography validação

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.

Fabricação fantasma do álcool polivinyl específico do paciente com ultrassom e contraste de raios-X para planejamento de cirurgia 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.

Geração de Condições Ambientais Dinâmicas usando um dispositivo microfluido de alto rendimento

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.

Padronização simples, acessível e modular de células usando DNA

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