作者感谢马德森博士在大学，威斯康星大学麦迪逊分校和CRISTEL梅雨在Gammex Inc TM材料上提供意见。马尔科姆·大卫·穆雷博士也感谢作者提供的方法来构造的幻影。

1。幻影设计的潜伏期的多模态的一个画图虚线示出在图1 7，8。假体的大小是直径为38毫米和115毫米的长度，以允许在各种动物扫描器扫描的幻像。幻象包含五个肿瘤的模拟试验对象放置在虚线内的深度为10毫米（直径：14，10，7，4和2毫米）。 2。模具硅胶建设硅胶模具，准备在本节7投肿瘤模拟试验对象。制备的硅胶模具所需的所有压克力板和棒切割精度为25微米的机加工车间在圣安东尼（UTHSCSA）得克萨斯大学健康科学中心大学。 <ol><li>五孔（直径：14，10，7，4，2毫米）为试验对象，另有五孔（直径：6毫米），对准棒两个丙烯酸基地板（尺寸：4.2厘米×11.5厘米×0.9厘米）（ 图2）。 </li><li>剪切间隔对高度7，5，3.5，2和1毫米（大小：1.0厘米×5.5厘米）（ 图2）。 </li><li>准备钢球（直径：14，10，7，4和2毫米，精度：2.5微米）。 </li><li>将两个间隔对高度7毫米，底板到薄亚克力板顺序，使用C型夹（ 图2 C）并拧紧。 </li><li>插入钢球，直径14毫米到14毫米的孔的基片和胶使用：JB KWIK（ 图2）。重复球的其余部分（ 图2 D）的程序和其他基板。需要注意的是在两块基板粘的钢球作为镜像7。 </li><li>到每个基地，如篱笆用胶带将四个高2.5厘米的亚克力板（尺寸：2.5厘米×11.5厘米的两个板块和2.5厘米×4.2厘米，另外两个板块）（ <str<html> ONG&gt;图3）。 </li><li>将顶板（大小：4.2厘米×11.5厘米，五个孔直径为0.8厘米的10孔直径为1.2厘米的）在一个基板组件插入五丙烯酸棒（直径：0.8厘米，长度：0.5厘米），带1毫米的技巧，并插入五个对准棒（直径：0.9厘米，长5.0厘米）和，倒入硅胶（ 图3 A）。 </li><li>亚克力杆插入到0.8厘米的孔，在顶板一路顶部的钢球和胶水他们使用硅胶胶水。然后插入对应杆插入孔中，在基板通过在顶板的较大的孔（ 图3 A）。 </li><li>混合部分A的硅橡胶化合物与B部分中的比例为10〜1（重量）。 </li><li>倒入到装配体的硅橡胶化合物和装配在室温下干燥约24小时（ 图3 B）。 </li></ol> 3。肩大会T“&gt;从PVC管和烤肉店电机转子准备。 <ol><li>碾末的螺栓，以适应电烤架电动机的孔。 </li><li>接地螺栓拧紧的PVC管（长度：270毫米，内径：75毫米）的端部，用螺母和垫圈。 </li><li>弯曲金属板和胶水他们到一个塑料板，使用JB KWIK支持PVC管和调节高度的PVC管7。 </li></ol> 4。 TM材料制备准备TM材料的协议修改那些在威斯康星州麦迪逊大学的在欧内斯特·L.·马德森博士的实验室和更多的细节开发Lee 等人 8,11。 4.1背景TM材料的制备<ol><li>通过商业全脂牛奶（200毫升），通过20微米和10微米网状过滤器。 </li><li>在过滤牛奶（100毫升），溶解硫柳汞（0.2克）。 </li><li>使用室内真空，气体乳溶液，在室温下持续30秒。 </li><li>将无水琼脂糖（2克）溶解在去离子水（18MΩ）（100毫升），在室温下。 </li><li>添加1 -丙醇（7.9毫升）和硫酸钡（1克）的琼脂糖溶液。 </li><li>脱气琼脂糖溶液，然后在95℃水浴中加热，直到琼脂糖溶液清除。 </li><li>虽然琼脂糖溶液清除，在95℃水浴，炼乳在55℃水浴加热。 </li><li>将熔融的琼脂糖溶液，在55℃的水浴中冷却下来。 </li><li>一旦双方的解决方案是在55℃下，混合的琼脂糖溶液（50毫升），炼乳（50毫升）的比例为50〜50（体积），并慢慢搅拌该混合物中，接着由气泡从表面上移除。 </li><li>添加乙二胺四乙酸（0.103克）和CuCl 2·2H 2 O（0.06克）的混合物中，随后充分搅拌以确保均匀性琼脂糖牛奶。 </li><li>最后，添加玻璃珠（15- 直径为60微米，平均直径35微米）（0.1克）和搅拌最终混合物反复。在使用之前，玻璃珠浸泡在浓硝酸中24小时，以除去任何杂质，然后冲洗掉酸。 </li></ol> 4.2测试对象TM材料的制备测试对象TM材料制备以类似的方式作为背景TM材料，除了下面的组成差异： <ol><li>通过商业全脂牛奶（20毫升），通过20微米和10微米网状过滤器。 </li><li>在过滤牛奶（10毫升），溶解硫柳汞（0.02克）。 </li><li>将干燥的琼脂糖（0.60克）溶解在去离子水（10毫升）和1 - 丙醇（0.79毫升）的溶液中室温下。 </li><li>脱气琼脂糖溶液，然后在95℃水浴中加热，直到琼脂糖溶液清除。 </li><li>虽然琼脂糖溶液清除，在95℃水浴，炼乳在55℃水浴加热。 </li><l&gt;将熔融的琼脂糖溶液，在55℃的水浴中。 </li><li>一旦双方的解决方案是在55℃下，用炼乳（5毫升）的琼脂糖溶液（5毫升）混合，并慢慢搅拌该混合物中，接着由气泡从表面上移除。 </li><li>然后添加EDTA（0.0017克）和CuCl 2·2H 2 O（0.0010克）琼脂糖牛奶充分搅拌。 </li></ol> 5。多模态幻影大会使用硅胶模具，下面的步骤完成的构造的多模态幻像。 <ol><li>无1毫米的孔的模具硅胶，将尼龙线沿球体的中心，然后把它粘在模具使用硅胶胶水（ 甲图4）的两端。 </li><li>使用软毛刷，两个模具的表面上涂抹硅脂（ 图4）和组装两个模具使用对齐棒。 </li><li>准备测试对象TM节中描述的材料4.2倒1毫米的孔的硅胶模具，用22号针头的注射器。 </li><li>以允许测试对象来设置，存储模具在冰箱（5℃）下进行30分钟。 </li><li>在每一侧的半筒状容器（长度：115毫米，直径：38毫米），使假体表面的深度为10毫米的两个孔为1毫米，在为了安装与测试对象的尼龙线。作出额外孔6毫米倒背景TM材料。 </li><li>卸载测试对象用尼龙线从模具中（ 图4），然后将它们安装在半圆柱形的容器（ 图4）。 </li><li>使用3M思高焊接DP-100和3M胶带，坚持薄的非导电铝（厚度0.12毫米）的丙烯酸容器上。座1毫米的孔，在丙烯酸类容器中，使用相同的胶（ 图4）。 </li><li>迅速准备背景TM材料，慢慢倒入容器6毫米的孔用一个小塑料漏斗。 </li><li>去除任何气泡，胶水后6毫米的孔使用3M苏格兰焊DP-100。 </li><li>一旦组装，2 rpm，以4至5小时，在室温下的旋转器的旋转的幻像。 </li><li>取出材料在幻象完全变硬后TM尼龙线。 </li></ol> 6。多模态成像幻象扫描在临床前超声，CT和MRI图像三种方式收购。 Lee 等人7，图 8中详细描述的摄像协议。

<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>

没有利益冲突的声明。

这篇文章的目的是TM材料的多模态成像和构建潜伏期的多模态幻影作为QA工具准确测量肿瘤的大小，用不同的方式在多个机构提供的方法。正如前面所提到的，TM材料最初被开发为多幻影成像模态前列腺在威斯康星州麦迪逊大学的由欧内斯特·L.·马德森博士的实验室。我们修改Madsen的博士的我们自己的目的TM材料协议，以便有足够的测试对象和背景之间的对比度，并代表在美国，CT和MR图像的软组织?...

<html>评估肿瘤大小的变化是一个重要的评价抗肿瘤的药物中的活性肿瘤收缩率和疾病的进展9，10的端点。世界卫生组织（WHO）和固体肿瘤疗效评价标准（RECIST）评估肿瘤病变的解剖成像方式，如超声（US），计算机断层扫描（CT）或磁共振成像（MRI）的编纂方法。 WHO标准，计算肿瘤的最大直径和最大垂直直径为目标区域在横向平面的商品4。 RECIST标准，与此相反，在横向平面的最长直径为有限数目的目标病变的总和来计算。尽管在肿瘤的治疗反应评估的兴趣不断增长，一直没有潜伏期的质量保证（QA）幻影/成像生物标志物的质量保证程序。 内容“考虑到肿瘤的尺寸测量，根据世界卫生组织的标准和/或RECIST成像生物标志物是唯一的，由美国食品和药物管理局（FDA）批准，任何其他成像生物标志物，李某等人的QA作为一个起点。设计和构造UTHSCSA / Gammex马克和马克合作与公司Gammex 7测量肿瘤大小为QA幻影。马克1，一个美国商业Gammex的幻象，幻象的修订版，因此，尺寸过大，以适应动物CT和MR扫描仪。也有一些工具在马克1幻象是不必要的肿瘤大小测量马克2幻像基础上，的RECIST这是最近期FDA批准的成像生物标志物的设计，然而，马克2幻像的大小MR扫描仪，CT和MR图像质量的幻影仍然过大，是不能接受的准确测量肿瘤大小7。 QA幻像形容他收服重新设计以克服以前的幻影和使用修改组织模仿（TM）材料，在我们的实验室开发的协议。本文描述了细节协议虚线建设：首先，制备用于浇铸肿瘤模拟测试对象所需的模具硅胶用于组装旋转体旋转，以防止一个幻象引力沉降方法进行介绍。其次，协议，准备修改D&#39;Souza 等人的“美国，CT和磁共振成像TM材料描述11。的TM材料的物理性能进行了测试，以确保在每一种模式中，TM材料为代表的各种方式获得的图像中所观察到的人体软组织，但结果是这里没有显示。第三，虚线建设的协议进行说明。最后，美国，CT和MR图像的幻影结果。

<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

工作组倡导标准化标准分别为实体瘤的影像学评估，在抗肿瘤的药物治疗，在20世纪80年代和90年代，世界卫生组织（WHO）和实体瘤疗效评价标准（RECIST）。 WHO标准测量实体肿瘤的两维，，而RECIST测量使用只有一个尺寸，被认为是更具有可重复性1，2，3,4,5。这些标准已被广泛地用于成像的生物标志物作为唯一的，经美国食品和药物管理局（FDA）的6。为了测量肿瘤的抗肿瘤药物的反应，准确的图像，因此，需要健全的质量保证（QA）的程序和相应的QA虚线。 为了满足这一需求，构建一个潜伏期的多模态（超声（US），计算机断层扫描（CT）和磁共振成像（MRI））幻象组织模仿（TM）材料的基础上数量有限的靶病变RECIST需要修改美国一个Gammex的商业幻象7。 Lee 等人的附录演示程序的幻像制作7。在本文中，所有的协议都在一步一步地开始准备的硅胶模具铸造肿瘤模拟试验对象在幻象，准备TM材料的多模态成像程序引入，终于建设临床前的多模态QA幻像。本文的主要目的是提供协议允许任何人感兴趣独立构建自己的项目幻象。肿瘤大小测量的质量保证程序，RECIST，世界卫生组织和体积测量结果，测试对象在多个机构，使用这样的QA虚线的详细示出在Lee 等人8。

图3和图5示出两个有机硅模具铸造试验对象，与多模态虚线分别。长×宽×每个模具的深度为109毫米×37毫米×21毫米和两个模具是相同的镜像对称。一模有1毫米的小孔，可以使用细针插入TM材料。每个模具有一个额外的五孔对齐棒。长×宽×深度的幻影是115毫米×38毫米×24毫米，其初始质量101.02克。假体的大小是足够的，以适应临床前的扫描仪。 收购。U...

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

本文介绍了内部程序构建一个潜伏期多模态影像（TM）质量保证（QA）肿瘤大小测量动物的成像方式，如超声（US），计算机断层扫描（CT）和磁材料制成的组织模仿磁共振成像（MRI）。

幻影使用肿瘤尺寸测量的质量保证组织模仿材料的潜伏期多模工程

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 次观看.  University of Kansas School of Medicine. 本文介绍了内部程序构建一个潜伏期多模态影像（TM）质量保证（QA）肿瘤大小测量动物的成像方式，如超声（US），计算机断层扫描（CT）和磁材料制成的组织模仿磁共振成像（MRI）。

视频: 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.

病毒粒子为<em在体内</em&gt;肿瘤显像

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

科研

生物工程

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.

生产尺寸控制肿瘤细胞球体采用微孔板大数

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.

细胞外基质刚性三维肿瘤模型的纵向测量用粒子跟踪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.

一种新型声带生物反应器的构建与鉴定

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.

设计和人工细胞外基质的构建（AECM）蛋白质<em&gt;大肠杆菌</em&gt;对皮肤组织工程

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.

基于琼脂糖组织模拟光学幽灵的漫反射光谱研究

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.

超声弹性成像验证腹主动脉水凝胶组织模拟幻影的研制

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.

患者特异性聚乙烯酒精幻影制造与超声波和X射线对比脑肿瘤手术规划

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.

使用高通量微流体设备生成动态环境条件

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.

使用DNA对细胞进行简单、经济、模块化的图案化

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