This work is supported by Multi-University Research Initiative through the Air Force Office of Scientific Research of the United States (AFOSR-FA9550-15-1-0009) (M. B. F., S. E. N., J.-Y. J., J. M). Collection of pink sea urchins was supported by the University of California Ship Funds and the US National Marine Fisheries Service (K.N.S., J.R.A.T). The authors acknowledge the following people: Prof. Jerry Tustaniwskyj for helpful suggestions during development of the bioinspired Aristotle's lantern sampler, Prof. Marc A. Meyers (UCSD, Dept. of Mechanical and Aerospace Engineering, Materials Science and Engineering Program), Prof. Robert L. Sah and Esther Cory (UCSD, Dept. of Bioengineering), and Dr. Maya deVries (Marine Biology Research Division, Scripps Institution of Oceanography). We also thank undergraduate students Sze Hei Siu, Jerry Ng and Ivan Torres for polishing urchin teeth cross-sections.

1.生物材料科学<ol><li>穿戴个人防护装备（ 如手套，防护眼镜和白大褂），并遵守所有适用的安全程序使用解剖工具。 </li><li>冲洗掉镊子并用蒸馏水手术刀用于清扫。 </li><li>解冻在RT冻结粉海胆1小时。放置在具有足够空间的玻璃盘一解冻样品，以便能够操纵海胆和切削工具。打开顽童倒挂使牙齿尖端朝上。 <ol><li>切掉周围的亚里士多德的灯笼用手术刀的周边结缔组织并小心了灯笼。冲洗灯笼了与运行的蒸馏水。在一个适当的废物处理容器丢弃不用的顽童部分。 </li><li>打开亚里士多德的灯笼一遍所以齿的齿尖朝下。找到每个齿（相对于尖）的莲子结束朝上，然后使用镊子小心完全从灯笼滑出个别牙齿。 </li></ol></li><li>准备环氧锅牙齿。称取g树脂，并添加1.15克固化剂（ 如 100份树脂至23重量份的固化剂）在浅一次性塑料托盘。慢慢地混合在一起的内容不会形成泡沫。 注意:不要把吃剩的混合环氧树脂与曝光不足的气氛容器中。固化过程是放热的，可以点燃附近易燃物。保留所有剩余的环氧混合在一个通风良好的通风橱来自易燃物品离开。 <ol><li>润滑2.5 DRAM塑料管（内径22mm，38 mm长）使用，用手指加凡士林擦去多余用纸巾。填补中途管混合环氧树脂。 </li><li>使用镊子拿起一颗牙齿，仔细与弧形凹面朝上环氧淹没它。让在RT环氧树脂固化24小时。 注:从漂流到触摸防止齿尖塑料管壁，因为这环氧树脂固化会使抛光尖更加困难。 </li></ol></li><li>将塑料管固定在台钳固化的环氧树脂。慢慢地拧紧台钳，直到裂缝在塑料管​​的。剥离远离环氧树脂表面残留的塑料。 <ol><li>用切片锯向下切环氧牙齿周围到一个较小的块（1 毫升 ）。 </li></ol></li><li>准备一个干净的区域进行抛光，并成立了平工作站用硬塑料板。补喷瓶蒸馏水。 <ol><li>开始的最低砂纸（ 例如，120）和从洗涤瓶到砂纸挤的少量水。利用光压，擦5分钟一回来回方向（ 例如，左，右）的样品。 </li><li>样品的表面洗掉在一个水槽和一个自由颗粒组织擦去。除去压缩空气任何剩余的砂纸砂粒，持续15秒。 </li><li>使用逐渐提高砂纸（如 600 2400）重复步骤协议1.6.1和1.6.2。利用光的压力，擦垂直于前面的抛光步骤在后样品和前后方向（ 例如，上下，左右）。 注:在放大20倍使用光学显微镜看到垂直划痕每个砂砾水平相交（ 例如，120，600 2400）。移动到下一个更高的砂纸时从先前砂砾水平划痕消失。 </li><li>准备一个喷瓶，在一个1 3微米的钻石抛光悬挂系统:1蒸馏水的解决方案。使用金​​刚石悬浮液重复步骤协议1.6.1和1.6.2抛光布。 </li><li>准备一个喷瓶，在1 0.5微米的氧化铝抛光悬挂系统:1蒸馏水的解决方案。使用microcloth抛光表面重复步骤协议1.6.1和1.6.2。 注:细划痕从协议的步骤和1.6.4 1.6.5不会visib乐在20X的放大倍率。对于这些协议的步骤，抛光5分钟一个来回运动删除所有以前的划痕。 </li><li>清洁用压缩空气仔细干燥蒸馏水和使用无颗粒组织的抛光表面。与无颗粒组织包裹，以保持镜面抛光光洁度。 注:干所有抛光表面面朝下放在大无颗粒的组织。在一个塑料套筒储存，以避免沉降抛光时间之间的表面上的灰尘颗粒。 </li></ol></li><li>表征利用海胆牙齿显微扫描电子显微镜（SEM）。使用溅射涂布机以溅射用85毫安，持续10秒的沉积电流铱到抛光的牙齿表面为20nm的〜涂层厚度。 <ol><li>在获取250X显微图像 - 用SEM 4,000×放大倍率。 注意:使用5千伏的扫描电子（SE）模式，并在背散射电子（BSE）的模式15千伏。使用BSE模式，以确定方解石FIBERS穿插镁丰富的多晶基质。 </li></ol></li><li>执行微型计算机断层扫描（μ-CT）整体粉红色海胆的扫描和新鲜解剖亚里士多德的灯笼。密闭腔容器内的各解冻样品与湿润的组织定位，以提供在扫描时的潮湿环境。 <ol><li>通过扫描μ-CT整个海胆和亚里士多德的灯笼与36.00微米和9.06微米的各向同性体素的大小，分别为。应用100 kVp的的电势和70 kVp的与百毫安和141毫安的电流，对整个海胆和亚里士多德的灯笼，分别使用这两个1.0mm的铝过滤器。 </li><li>图像重建期间应用束硬化校正算法以考虑从μ-CT x射线源发射使用制造商的协议的多个能量的X射线导致射束硬化伪影。 </li></ol></li><li>使用图像处理软件来完善IMAGE分割，并获得了亚里士多德的灯笼结构的三角形网格模型。 <ol><li>加载和预览从μ-CT扫描亚里士多德的灯笼图像数据。匹配体素尺寸（9.06微米），以从微CT扫描的值。 </li><li>使用体绘制功能形象化亚里士多德在三维空间灯笼。调整的边界框模块二维正交切片，调整与体绘制模块阈值/颜色。 </li><li>使面具片段使用分割编辑的感兴趣区域（ 例如，海胆齿）。选择XY，YZ，和XZ平面和三维立体图。使用魔棒（黑色箭头）在亚里士多德的灯笼结构简单（牙与金字塔）区别开来。 </li><li>从提取的面具片段重建模型表面。选择曲面生成模块和应用。取消批量渲染设置有明显的顶面消失。添加表面观模块显示表面的结果。 </li><li>通过减少面的数目为&lt;18000简化模型表面。 </li><li>根据需要修改模型表面上单个三角形网格。保存模型作为立体光刻（STL）文件导出带计算机辅助设计（CAD）建模软件的使用。 </li></ol></li></ol> 2.仿生设计<ol><li>从微观CT扫描使用亚里士多德的灯笼作为参考，可与CAD建模软件设计的仿生。 注:仿生设计有五个弧形齿高6厘米，直径为8厘米为封闭的灯笼。它是从天然亚里士多德的灯笼规模扩大了5倍〜。 </li><li>保存STL文件的部分到闪存驱动器和文件上传到熔融沉积成型（FDM）的3D打印机。 <ol><li>装载丙烯腈 - 丁二烯 - 苯乙烯（ABS）塑料和支持塑料盒放入3D p的相应插槽rinter。 </li><li>将z平台上的造型基础，并与金属托盘插槽对齐的标签。 </li><li>打开每个STL文件部分，然后按照屏幕上显示的步骤，同时打印所有灯笼部件。 注意:灯零件必须符合建筑物外壳（25×25×30 毫升 ）对所述三维打印机内。所有五个齿布置在建模底座和与齿尖朝上同时打印。构建速率是每小时16厘米3，总建造时间是约8小时。 </li><li>从释放的选项卡建模的基础时​​，打印的所有文件部分和底座出沿托盘导向的三维打印机中滑出。 </li><li>用金属铲撬下基底和金属文件中的所有部件拖垮任何额外的塑料连接到的部分。 </li><li>印刷部件放入温水洗澡基地直到支撑塑料材质溶解。 </li></ol></li><li>拧紧每颗牙齿的联合的手臂里NK杆和两侧两架E-扣环。 注:参见图6为仿生亚里士多德的灯笼组装。 </li></ol> 3. Bioexploration <ol><li>使用CAD文件的仿生牙做了有限元模型（FEM）应力分析测试。 <ol><li>打开文件（xx.sldprt）做进一步的工程分析。上面的"产品展示厅"选项卡，选中"的SolidWorks Simulation"按钮。 </li><li>上面的"模拟"选项卡中，选择"学习顾问"按钮，然后在下拉选项"新学"。 </li><li>选择模拟试验的类型通过选择"静态"运行。 </li><li>在静态测试列表，关于"灯具"单击鼠标右键，选择"固定几何"。 </li><li>点击内面为灯具添加到安装孔，其中销会。 </li><li>在静态测试列表，关于"外部载荷"和SELEC右击T"强制"。 </li><li>点击牙齿磨尖面向应用45牛顿的力向四周扩散。 </li><li>在静态测试列表，关于"外部载荷"单击鼠标右键，选择"万有引力"。 </li><li>指示"顶面"为施加垂直于该平面的重力。 </li><li>在静态测试列表，关于"网格"单击鼠标右键，选择"创建网格"。 </li><li>对于移动"网格密度"一路为"精细"右侧的比例尺。 </li><li>在静态测试列表，关于"静"，单击鼠标右键，选择"运行"来运行测试。 注:最高压力和"屈服强度"的领域彩色比例尺。 </li></ol></li><li>比较应力分析测试结果为具有和不具有龙骨的仿生齿。 </li></ol>

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Zool., London. 220, 345-366 (1990).</li><li>Ellers, O., Telford, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forces+generated+by+the+jaws+of+Clypeasteroids+(Echinodermata:+Echionoidea).">Forces generated by the jaws of Clypeasteroids (Echinodermata: Echionoidea).</a> J. Exp. Biol. 155, 585-603 (1991).</li><li>Candia Carnevali, M. D., Wilkie, I. C., Lucca, E., Andrietti, F., Melone, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Aristotle's+lantern+of+the+sea-urchin+Stylocidaris+affinis+(Echinoida,+Cidaridae):+functional+morphology+of+the+musculo-skeletal+system.">The Aristotle's lantern of the sea-urchin Stylocidaris affinis (Echinoida, Cidaridae): functional morphology of the musculo-skeletal system.</a> Zoomorphology. 113 (3), 173-189 (1993).</li><li>Wilkie, I. C., Candia Carnevali, M. D., Andrietti, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+of+sea-urchin+lantern+muscles:+a+comparative+investigation+of+intact+muscle+groups+in+Paracentrotus+lividus+(Lam)+and+Stylocidaris+affinis+(Phil)+(Echinodermata,+Echinoidea).">Mechanical properties of sea-urchin lantern muscles: a comparative investigation of intact muscle groups in Paracentrotus lividus (Lam) and Stylocidaris affinis (Phil) (Echinodermata, Echinoidea).</a> J. Comp. Physiol. B. 168 (3), 204-212 (1998).</li><li>Witze, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NASA+plans+Mars+sample-return+rover.">NASA plans Mars sample-return rover.</a> Nature. 509 (7500), 272 (2014).</li></ol>

We have nothing to disclose.

海胆用亚里士多德的灯笼（ 图1A）的各种功能（进料，镗孔，旋转等 ）。化石记录表明，灯笼形状和功能已经发展从最原始的cidaroid类型到camarodont型现代海胆14。 Cidaroid灯笼已经沿纵向开槽的齿（ 图1B，上图 ）和非分离肌肉附着到其金字塔结构。这限制了它们的上下运动和剥夺由横向运动，这在更现代camarodont灯笼（ 图1B，下图 ）观察到所产生的较大的刮功率它们。生物学家们推测，在camarodonts进化的龙骨牙（ 图1C，D），以加强在刮硬质承印18,20,23所产生的强烈的张力牙齿。 该仿生设计方案在这个工作相结合生物学，生物材料科学，仿生设计和bioexploration（ 图3），以制定一个仿生设备与采样沉积物的特定功能。亚里士多德的灯笼（ 图4）的μ-CT扫描导入为仅供参考，因为最终的采样设计没有模仿的自然结构复杂的肌肉附着STL文件。代替仿生设计采用与可以很容易地由一个三维打印机装配到亚里士多德的灯笼采样来制造零件较简单的开闭机构。总体而言，我们用于仿生设计的圆形的方法，因为允许新结论bioexploration步骤必须从天然生物学绘制。该仿生设计的潜在修改可以解决除了沉积物取样不同的应用程序。此协议的一个限制是，它被聚焦在仿生方法的一个具体应用为基础的装置在亚里士多德的灯笼。然而，这里概述的协议可以被应用到分析，开发和基于生物样品其他仿生设计最终制造。 造成这种情况的主要应用组装仿生亚里士多德的灯笼采样器（ 图6）是用于收集松散和压实砂（ 图7）。展望未来，美国航空航天局有一个计划带回火星样品地球中使用任务长年累月29继承后样品送回月球车。用仿生亚里士多德的灯笼采样器配备一个样品送回月球车可能对今后的任务是有益的。类似于自然亚里士多德的灯笼规模较小的采样也可用于其他用途。在天然海胆齿硬度的各向异性，而在它自己右边吸引，并没有在这个仿生设计并入。 k的BioexplorationEELED与非翻转齿确认在自然海胆龙骨重要结构目的（ 图8）。该bioexploration结果提供的数据，有助于解释为什么现代海胆演变龙骨结构。我们承认，波特17,18是第一个提出在这项工作中，这是利用工程分析方法来量化海胆牙齿龙骨结构的机械优势必不可少的应用bioexploration一步。未来的仿生设计，连接自然的观察，生物材料科学，仿生设计和bioexploration可以是结合自然的设计原则更深的植根熟悉有利。

生物学领域，生物材料科学，生物材料，生物工程和生化聘请首映的科学技术和思想，试图提供令人难以置信的自然世界有更深的了解。这项研究解释了许多最惊人的生物结构和生物;从人骨1,2的内在韧性大嘴巨嘴鸟3。然而，大部分的这方面的知识，很难以一种方式，可以提供一个好处社会采用。其结果是，bioinspiration的切线野采用从自然学会了现代材料的经验教训，以解决共同问题。例子包括莲花启发超疏水表面留下4-6，由壁虎的脚鼓舞粘接面和昆虫7,8，由9-11鲍鱼和海胆的喉舌启发活检收割机的珍珠层的启发强硬陶瓷，也知道n作为亚里士多德的灯笼12,13。 海胆是长满刺的栖息地最常见的包括在海底的岩石床的无脊椎动物。人体中最大的海胆物种（称为测试）可以超过18厘米，直径;在这项研究考察了粉红色的海胆测试尺寸（ 海胆脆弱 ），可以长到直径10厘米。亚里士多德的灯笼是由由封闭所有，但牙齿（ 图1A）的远端粉碎提示矿化组织的组成和排列为半球状形成金字塔结构，支助了五个主要为碳酸钙的牙齿。 颌骨肌肉结构能够有效的咀嚼和拼抢甚至对硬盘的海洋岩石和珊瑚。当夹爪打开，牙齿向外突出并在爪闭合，牙齿在一个平滑的运动向内缩回。 primitiv的比较E（上）和现代（下）海胆牙齿横截面（ 图1B）表明一个龙骨状齿演变对硬磨底物时，强化牙齿。每个单独的齿具有由于纵向附龙骨（ 图1C，D） ​​的稍微凸起的曲率和在横向平面（垂直于生长方向）的T形的形态。 Bioinspiration始于有趣的自然现象，如亚里士多德在海胆灯笼高效率咀嚼运动的观察。这自然结构最初迷住亚里士多德，因为这让他想起喇叭灯笼与喇叭的窗格冷落。两千多年后，斯卡帕被亚里士多德的灯笼的复杂性，他后来Trogu模仿只使用纸和橡胶带（ 图2A）15,16天然咀嚼运动迷住了。同样，耶利内克是由C仿生砍伐了亚里士多德的灯笼的运动，并制定了更好的活检收割机，可以安全地隔离肿瘤组织，不扩散的癌细胞（ 图2B，C）12,13。在这种情况下，被利用仿生设计，使适合用于期望的应用的特定需要的生物医学装置。 这里描述的设计协议适用于海胆仿生沉积物取样。通过生物材料科学，亚里士多德的灯笼的自然结构的特点。仿生设计识别潜在的应用那里的自然机制，可以通过使用现代材料和制造技术的提高。通过bioexploration的棱镜的最终设计重新检查，以了解自然的牙齿结构如何演变 （ 图3）。最后一步bioexploration，波特提出的17,18，采用工程分析方法到e智能与解释生物现象。所有bioinspiration过程中的重要步骤，表示为用于利用技术，预先核准的本质，可用于解决现代问题的一个例子。我们的协议，所提交由Arzt 7特定应用程序的早期bioinspiration程序的动机，是针对生物学家，工程师和其他人谁是灵感来自于大自然。

<table>
	<tbody>
		<tr>
			<td>BUEHLERMET II 8 PLN 600/P1200</td>
			<td>Buehler</td>
			<td>305308600102</td>
			<td>Abrasive paper for polishing</td>
		</tr>
		<tr>
			<td>TRIDENT POLISH CLOTH 8&#34; PSA</td>
			<td>Buehler</td>
			<td>407518</td>
			<td>Polish cloth for 3 um suspension</td>
		</tr>
		<tr>
			<td>METADI SUPREME POLY SUSP,3MIC&#160;</td>
			<td>Buehler</td>
			<td>406631</td>
			<td>Polish suspension (3 um)</td>
		</tr>
		<tr>
			<td>MICROCLOTH FOR 8 IN WHEEL PSA</td>
			<td>Buehler</td>
			<td>407218</td>
			<td>Polish cloth for 50 nm suspension</td>
		</tr>
		<tr>
			<td>MASTERPREP SUSPENSION, 6 OZ</td>
			<td>Buehler</td>
			<td>636377006</td>
			<td>Polish suspension (50 nm)</td>
		</tr>
		<tr>
			<td>Skyscan 1076 micro-CT Scanner</td>
			<td>Bruker</td>
			<td></td>
			<td>Micro-CT scanner equipment</td>
		</tr>
		<tr>
			<td>Amira software</td>
			<td>FEI Visualization Sciences Group</td>
			<td></td>
			<td>Software for 3D manipulation of Micro-CT scans</td>
		</tr>
		<tr>
			<td>FEI Philips XL30</td>
			<td>FEI Philips</td>
			<td></td>
			<td>ESEM equipment for characterization of polished tooth cross-sections</td>
		</tr>
		<tr>
			<td>SolidWorks Design software</td>
			<td>Dassault Systems</td>
			<td></td>
			<td>Design software for CAD drawing bioinspired device</td>
		</tr>
		<tr>
			<td>SolidWorks Simulation software</td>
			<td>Dassault Systems</td>
			<td></td>
			<td>Simulation software for stress test of CAD drawing bioinspired device</td>
		</tr>
		<tr>
			<td>Dimension 1200es</td>
			<td>Stratasys</td>
			<td></td>
			<td>3D printer for fabrication of bioinspired device from CAD drawing</td>
		</tr>
		<tr>
			<td>ABSplus</td>
			<td>Stratasys</td>
			<td></td>
			<td>3D printer plastic</td>
		</tr>
	</tbody>
</table>

a protocol for bioinspired design: a ground sampler based on sea urchin jaws

Bioinspired design is an emerging field that takes inspiration from nature to develop high-performance materials and devices. The sea urchin mouthpiece, known as the Aristotle's lantern, is a compelling source of bioinspiration with an intricate network of musculature and calcareous teeth that can scrape, cut, chew food and bore holes into rocky substrates. We describe the bioinspiration process as including animal observation, specimen characterization, device fabrication and mechanism bioexploration. The last step of bioexploration allows for a deeper understanding of the initial biology. The design architecture of the Aristotle's lantern is analyzed with micro-computed tomography and individual teeth are examined with scanning electron microscopy to identify the microstructure. Bioinspired designs are fabricated with a 3D printer, assembled and tested to determine the most efficient lantern opening and closing mechanism. Teeth from the bioinspired lantern design are bioexplored via finite element analysis to explain from a mechanical perspective why keeled tooth structures evolved in the modern sea urchins we observed. This circular approach allows for new conclusions to be drawn from biology and nature.

亚里士多德的灯笼采样装置的设计仿生很大程度上取决于所采用的表征方法的质量。像μ-CT非侵入性技术是用于分析整个灯和单独的齿申请应用特定的增强的仿生设计（ 图4）有帮助。同时，齿微结构可以通过二次电子和一个个人齿（ 图5）的抛光横截面的背散射电子显微照片加以探讨。较暗的灰色区域是磨齿尖的硬石部件和具有多达该取代钙原子为40摩尔％的镁原子。 齿显微疯牛病-SEM（ 图5）的分析确认了Mg的富集石部件的在牙齿磨尖的结构的重要性。板和纤维表面RY元件（方解石单晶，在图5C浅灰色）由次级元件（方解石和碳酸镁多晶体，在图5C深灰色），构成齿磨尖的最硬石区域的基质连接在一起的。 该仿生灯笼的设计采用CAD软件，3D印刷和装配（ 图6）为砂收集在海滩（ 图7）。应力分析试验用于计算两齿设计中，一个没有龙骨（ 图8A）和另一个与龙骨（ 图8B）的冯米塞斯应力。四面体组成的实体网格被雇用了牙齿的几何形状。选择（45 N）的力值在沙滩匹配从试验测量穿透深1厘米到硬砂与灯笼齿垂直于表面。 图7A，B）。在强调龙骨提供比下降的质量增加很小。在应力的降低说明了这一点仿生设计为龙骨状区域内的应力集中的效果。 <img alt="图1" src="/files/ftp_upload/53554/53554fig1.jpg" /> 图1.海胆亚里士多德的灯笼，牙齿形态。（A）海胆（左）和亚里士多德的灯笼（右）13的腹面的特写。 （B）的横截面一个原始cidaroid海胆（上）和一个现代camarodont海胆（底部）14的龙骨齿的槽的齿。 （C）从它与前端（底部）侧看的分离的齿和指示龙骨（左侧）20。一个抛光的牙截面与所指示的龙骨（底部）20（D）的SEM图像。改编自用于表示引用（A），（B），（C）和（D）的图像。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53554/53554fig2.jpg" /> 图2.仿生设计的基础上，亚里士多德的灯笼。（A）对亚里士多德的灯笼的仿生模型，具有3D打印塑料P A绘图等轴测图由橡胶带（未示出），用于附加的肌肉组织16连接第（B，C）的亚里士多德的灯笼担任活检收割机13生物灵感。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53554/53554fig3.jpg" /> 图3.四个步骤bioinspiration的过程。该过程bioinspiration（左起顺时针方向）开始通过粉红色的海胆和亚里士多德的灯笼观察大自然学习。海胆（顶部）分析和μ-CT扫描（左）亚里士多德的灯笼结构。 （右）累计结果被用来生成一个仿生设计原型。 （底）工程分析方法应用于探索生物啉omena和仿生设计17,18。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/53554/53554fig4.jpg" /> 图4.微计算的亚里士多德的灯笼结构的断层分析金字塔结构，这有助于支持牙齿的（A）侧视图。 （B）的海胆齿堆叠在彼此的顶部，并表现出五倍对称性。被删除（C）远端末梢部分显示所有五个齿纵向附加龙骨结构。 （D）的一个单独的齿和龙骨（蓝色）与对应金字塔（黄色）示出以及在（C）的指示。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig4large.jpg" target="_blank">请</a>此处查看该图的放大版本。 <img alt="图5" src="/files/ftp_upload/53554/53554fig5.jpg" /> 海胆的牙齿微观结构图5.扫描电子显微镜（SEM）分析。（A）抛光牙齿横截面着淡淡的条纹石地区和龙骨（下）的SEM照片显示。 （B，C）的距离（A）显示弯板和位于上方的密集镁丰富的多晶基质（深灰色）圆形纤维方解石主要元素的紫色和橙色盒子背散射电子SEM照片。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig5large.jpg" target="_blank">请点击此处查看大图这一数字。</a> <img alt="图6" src="/files/ftp_upload/53554/53554fig6.jpg" /> 数字6.组装3D打印机仿生亚里士多德的灯笼部分。（A）E-扣环和链路棒被用来在三个关节的位置紧固3D打印的齿部。 （B）组装仿生亚里士多德的灯笼与一个齿去除。 （ 三 ）龙骨个别牙齿的视图，并在灯笼是部分（左）和全开（右）不断变化的共同立场。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图7" src="/files/ftp_upload/53554/53554fig7.jpg" /> 图7.仿生亚里士多德的彩灯设计和使用在海滩上。（A，B）的仿生亚里士多德的灯笼，而分别关闭和完全打开，计算机辅助设计的图像。 （C）的3D打印机仿生亚里士多德的灯笼收集不同类型的沙滩上沙子。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图8" src="/files/ftp_upload/53554/53554fig8.jpg" /> 图8.仿生海胆牙齿应力分析检验。（A，B）有限元分析表明非龙骨（A）与力时在牙齿边缘应用龙骨（B）的牙齿。在龙骨状齿设计经验少〜16％的压力，由于增加了龙骨。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53554/53554fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about A Protocol for Bioinspired Design: A Ground Sampler Based on Sea Urchin Jaws at JoVE.com

A Protocol for Bioinspired Design: A Ground Sampler Based on Sea Urchin Jaws

A protocol for bioinspired design is described for a sampling device based on the jaws of a sea urchin. The bioinspiration process includes observing the sea urchins, characterizing the mouthpiece, 3D printing of the teeth and their assembly, and bioexploring the tooth structure.

基于海胆大白鲨A接地采样:仿生为设计一个协议

Bioinspired design is an emerging field that takes inspiration from nature to develop high-performance materials and devices. The sea urchin mouthpiece, ...

a-protocol-for-bioinspired-design-ground-sampler-based-on-sea-urchin

Nanyang Technological University

Research

JoVE Journal

Bioengineering

11.0K 次观看.  University of California, San Diego. A protocol for bioinspired design is described for a sampling device based on the jaws of a sea urchin. The bioinspiration process includes observing the sea urchins, characterizing the mouthpiece, 3D printing of the teeth and their assembly, and bioexploring the tooth structure. 

视频: A Protocol for Bioinspired Design: A Ground Sampler Based on Sea Urchin Jaws

Mass Cytometry: Protocol for Daily Tuning and Running Cell Samples on a CyTOF Mass Cytometer

In recent years, the rapid analysis of single cells has commonly been performed using flow cytometry and fluorescently-labeled antibodies. However, the issue of spectral overlap of fluorophore emissions has limited the number of simultaneous probes. In contrast, the new CyTOF mass cytometer by DVS Sciences couples a liquid single-cell introduction system to an ICP-MS.1 Rather than fluorophores, chelating polymers containing highly-enriched metal isotopes are coupled to antibodies or other specific probes.2-5 Because of the metal purity and mass resolution of the mass cytometer, there is no "spectral overlap" from neighboring isotopes, and therefore no need for compensation matrices. Additionally, due to the use of lanthanide metals, there is no biological background and therefore no equivalent of autofluorescence. With a mass window spanning atomic mass 103-203, theoretically up to 100 labels could be distinguished simultaneously. Currently, more than 35 channels are available using the chelating reagents available from DVS Sciences, allowing unprecedented dissection of the immunological profile of samples.6-7
Disadvantages to mass cytometry include the strict requirement for a separate metal isotope per probe (no equivalent of forward or side scatter), and the fact that it is a destructive technique (no possibility of sorting recovery). The current configuration of the mass cytometer also has a cell transmission rate of only ~25%, thus requiring a higher input number of cells.
Optimal daily performance of the mass cytometer requires several steps. The basic goal of the optimization is to maximize the measured signal intensity of the desired metal isotopes (M) while minimizing the formation of oxides (M+16) that will decrease the M signal intensity and interfere with any desired signal at M+16. The first step is to warm up the machine so a hot, stable ICP plasma has been established. Second, the settings for current and make-up gas flow rate must be optimized on a daily basis. During sample collection, the maximum cell event rate is limited by detector efficiency and processing speed to 1000 cells/sec. However, depending on the sample quality, a slower cell event rate (300-500 cells/sec) is usually desirable to allow better resolution between cells events and thus maximize intact singlets over doublets and debris. Finally, adequate cleaning of the machine at the end of the day helps minimize background signal due to free metal.

The steps necessary for daily tuning and optimization of the performance of a CyTOF mass cytometer are described. Comments on optimal sample preparation and flow rate are discussed

质量流式细胞术:协议，每日调整和执行细胞的样品上一个CyTOF的质量流式细胞仪

In recent years, the rapid analysis of single cells has commonly been performed using flow cytometry and fluorescently-labeled antibodies. However, the ...

科研

生物工程

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

We designed a loading device that is capable of applying uniaxial or biaxial mechanical strain to a tissue engineered biocomposites fabricated for transplantation. While the device primarily functions as a bioreactor that mimics the native mechanical strains, it is also outfitted with a load cell for providing force feedback or mechanical testing of the constructs. The device subjects engineered cartilage constructs to biaxial mechanical loading with great precision of loading dose (amplitude and frequency) and is compact enough to fit inside a standard tissue culture incubator. It loads samples directly in a tissue culture plate, and multiple plate sizes are compatible with the system. The device has been designed using components manufactured for precision-guided laser applications. Bi-axial loading is accomplished by two orthogonal stages. The stages have a 50 mm travel range and are driven independently by stepper motor actuators, controlled by a closed-loop stepper motor driver that features micro-stepping capabilities, enabling step sizes of less than 50 nm. A polysulfone loading platen is coupled to the bi-axial moving platform. Movements of the stages are controlled by Thor-labs Advanced Positioning Technology (APT) software. The stepper motor driver is used with the software to adjust load parameters of frequency and amplitude of both shear and compression independently and simultaneously. Positional feedback is provided by linear optical encoders that have a bidirectional repeatability of 0.1 &mu;m and a resolution of 20 nm, translating to a positional accuracy of less than 3 &mu;m over the full 50 mm of travel. These encoders provide the necessary position feedback to the drive electronics to ensure true nanopositioning capabilities. In order to provide the force feedback to detect contact and evaluate loading responses, a precision miniature load cell is positioned between the loading platen and the moving platform. The load cell has high accuracies of 0.15% to 0.25% full scale.

We designed a novel mechanical loading bioreactor that can apply uniaxial or biaxial mechanical strain to a cartilage biocomposite prior to transplantation into an articular cartilage defect.

组织工程生物反应器的双轴机械装载设计

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy

Cellulosomes are discrete multienzyme complexes used by a subset of anaerobic bacteria and fungi to digest lignocellulosic substrates. Assembly of the enzymes onto the noncatalytic scaffold protein is directed by interactions among a family of related receptor-ligand pairs comprising interacting cohesin and dockerin modules. The extremely strong binding between cohesin and dockerin modules results in dissociation constants in the low picomolar to nanomolar range, which may hamper accurate off-rate measurements with conventional bulk methods. Single-molecule force spectroscopy (SMFS) with the atomic force microscope measures the response of individual biomolecules to force, and in contrast to other single-molecule manipulation methods (i.e.&#160;optical tweezers), is optimal for studying high-affinity receptor-ligand interactions because of its ability to probe the high-force regime (&#62;120 pN). Here we present our complete protocol for studying cellulosomal protein assemblies at the single-molecule level. Using a protein topology derived from the native cellulosome, we worked with enzyme-dockerin and carbohydrate binding module-cohesin (CBM-cohesin) fusion proteins, each with an accessible free thiol group at an engineered cysteine residue. We present our site-specific surface immobilization protocol, along with our measurement and data analysis procedure for obtaining detailed binding parameters for the high-affinity complex. We demonstrate how to quantify single subdomain unfolding forces, complex rupture forces, kinetic off-rates, and potential widths of the binding well. The successful application of these methods in characterizing the cohesin-dockerin interaction responsible for assembly of multidomain cellulolytic complexes is further described.

Cellulosomes are multienzyme complexes designed for digesting cellulose. AFM-based SMFS was used to study the mechanical properties and folding configuration of cellulosome-associated protein assemblies. We present a complete workflow for protein immobilization, data acquisition, and data analysis to study the interactions of individual receptor-ligand complexes involved in cellulosome assembly.

调查纤维素酶与基于AFM的单分子力谱受体 - 配体系统

Cellulosomes are discrete multienzyme complexes used by a subset of anaerobic bacteria and fungi to digest lignocellulosic substrates. Assembly of the ...

Engineering Platform and Experimental Protocol for Design and Evaluation of a Neurally-controlled Powered Transfemoral Prosthesis

To enable intuitive operation of powered artificial legs, an interface between user and prosthesis that can recognize the user's movement intent is desired. A novel neural-machine interface (NMI) based on neuromuscular-mechanical fusion developed in our previous study has demonstrated a great potential to accurately identify the intended movement of transfemoral amputees. However, this interface has not yet been integrated with a powered prosthetic leg for true neural control. This study aimed to report (1) a flexible platform to implement and optimize neural control of powered lower limb prosthesis and (2) an experimental setup and protocol to evaluate neural prosthesis control on patients with lower limb amputations. First a platform based on a PC and a visual programming environment were developed to implement the prosthesis control algorithms, including NMI training algorithm, NMI online testing algorithm, and intrinsic control algorithm. To demonstrate the function of this platform, in this study the NMI based on neuromuscular-mechanical fusion was hierarchically integrated with intrinsic control of a prototypical transfemoral prosthesis. One patient with a unilateral transfemoral amputation was recruited to evaluate our implemented neural controller when performing activities, such as standing, level-ground walking, ramp ascent, and ramp descent continuously in the laboratory. A novel experimental setup and protocol were developed in order to test the new prosthesis control safely and efficiently. The presented proof-of-concept platform and experimental setup and protocol could aid the future development and application of neurally-controlled powered artificial legs.

Neural-machine interfaces (NMI) have been developed to identify the user's locomotion mode. These NMIs are potentially useful for neural control of powered artificial legs, but have not been fully demonstrated. This paper presented (1) our designed engineering platform for easy implementation and development of neural control for powered lower limb prostheses and (2) an experimental setup and protocol in a laboratory environment to evaluate neurally-controlled artificial legs on patients with lower limb amputations safely and efficiently.

工程平台和设计一个Neurally控制供电采用经假体与评价实验方案

To enable intuitive operation of powered artificial legs, an interface between user and prosthesis that can recognize the user's movement intent is ...

Synthesis of Keratin-based Nanofiber for Biomedical Engineering

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of these porous nanofibers is of great interest due to their unique physiochemical properties. Here we elaborate on the fabrication of keratin containing poly (&#949;-caprolactone) (PCL) nanofibers (i.e., PCL/keratin composite fiber). Water soluble keratin was first extracted from human hair and mixed with PCL in different ratios. The blended solution of PCL/keratin was transformed into nanofibrous membranes using a laboratory designed electrospinning set up. Fiber morphology and mechanical properties of the obtained nanofiber were observed and measured using scanning electron microscopy and tensile tester. Furthermore, degradability and chemical properties of the nanofiber were studied by FTIR. SEM images showed uniform surface morphology for PCL/keratin fibers of different compositions. These PCL/keratin fibers also showed excellent mechanical properties such as Young&#39;s modulus and failure point. Fibroblast cells were able to attach and proliferate thus proving good cell viability. Based on the characteristics discussed above, we can strongly argue that the blended nanofibers of natural and synthetic polymers can represent an excellent development of composite materials that can be used for different biomedical applications.

Electrospun nanofibers have a high surface area to weight ratio, excellent mechanical integrity, and support cell growth and proliferation. These nanofibers have a wide range of biomedical applications. Here we fabricate keratin/ PCL nanofibers, using the electrospinning technique, and characterize the fibers for possible applications in tissue engineering.

基于角蛋白纳米纤维生物医学工程合成

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of ...

Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest regarding the use of bioinspired nanoscale hydroxyapatite (nHA). However, biological apatite is known to be calcium-deficient and carbonate-substituted with a nanoscale platelet-like morphology. Bioinspired nHA has the potential to stimulate optimal bone tissue regeneration due to its similarity to bone and tooth enamel mineral. Many of the methods currently used to fabricate nHA both in the laboratory and commercially, involve lengthy processes and complex equipment. Therefore, the aim of this study was to develop a rapid and reliable method to prepare high quality bioinspired nHA. The rapid mixing method developed was based upon an acid-base reaction involving calcium hydroxide and phosphoric acid. Briefly, a phosphoric acid solution was poured into a calcium hydroxide solution followed by stirring, washing and drying stages. Part of the batch was sintered at 1,000 &#176;C for 2 h in order to investigate the products&#39; high temperature stability. X-ray diffraction analysis showed the successful formation of HA, which showed thermal decomposition to &#946;-tricalcium phosphate after high temperature processing, which is typical for calcium-deficient HA. Fourier transform infrared spectroscopy showed the presence of carbonate groups in the precipitated product. The nHA particles had a low aspect ratio with approximate dimensions of 50 x 30 nm, close to the dimensions of biological apatite. The material was also calcium deficient with a Ca:P molar ratio of 1.63, which like biological apatite is lower than the stoichiometric HA ratio of 1.67. This new method is therefore a reliable and far more convenient process for the manufacture of bioinspired nHA, overcoming the need for lengthy titrations and complex equipment. The resulting bioinspired HA product is suitable for use in a wide variety of medical and consumer health applications.

This paper describes a novel method for the rapid manufacture of high quality bioinspired nanoscale hydroxyapatite. This biomaterial is of great significance in the manufacture of a wide range of innovative medical devices for clinical applications in orthopedics, craniofacial surgery and dentistry.

仿生纳米羟基磷灰石的快速混合制备生物医学应用

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest ...

A Protocol for Decellularizing Mouse Cochleae for Inner Ear Tissue Engineering

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current regenerative medicine strategies have focused on transplanting stem cells or genetic manipulation of surrounding support cells in the inner ear to encourage replacement of damaged stem cells to correct hearing loss. Yet, the extracellular matrix (ECM) may play a vital role in inducing and maintaining function of hair cells, and has not been well investigated. Using the cochlear ECM as a scaffold to grow adult stem cells may provide unique insights into how the composition and architecture of the extracellular environment aids cells in sustaining hearing function. Here we present a method for isolating and decellularizing cochleae from mice to use as scaffolds accepting perfused adult stem cells. In the current protocol, cochleae are isolated from euthanized mice, decellularized, and decalcified. Afterward, human Wharton&#39;s jelly cells (hWJCs) that were isolated from the umbilical cord were carefully perfused into each cochlea. The cochleae were used as bioreactors, and cells were cultured for 30 days before undergoing processing for analysis. Decellularized cochleae retained identifiable extracellular structures, but did not reveal the presence of cells or noticeable fragments of DNA. Cells perfused into the cochlea invaded most of the interior and exterior of the cochlea and grew without incident over a duration of 30 days. Thus, the current method can be used to study how cochlear ECM affects cell development and behavior.

The goal of this protocol is to demonstrate an effective method to decellularize and decalcify mouse cochleae for utilization as scaffolds for tissue engineering applications.

用于内耳组织工程的 Decellularizing 小鼠耳蜗协议

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current ...

A Protocol for Real-time 3D Single Particle Tracking

Real-time three-dimensional single particle tracking (RT-3D-SPT) has the potential to shed light on fast, 3D processes in cellular systems. Although various RT-3D-SPT methods have been put forward in recent years, tracking high speed 3D diffusing particles at low photon count rates remains a challenge. Moreover, RT-3D-SPT setups are generally complex and difficult to implement, limiting their widespread application to biological problems. This protocol presents a RT-3D-SPT system named 3D Dynamic Photon Localization Tracking (3D-DyPLoT), which can track particles with high diffusive speed (up to 20 &#181;m2/s) at low photon count rates (down to 10 kHz). 3D-DyPLoT employs a 2D electro-optic deflector (2D-EOD) and a tunable acoustic gradient (TAG) lens to drive a single focused laser spot dynamically in 3D. Combined with an optimized position estimation algorithm, 3D-DyPLoT can lock onto single particles with high tracking speed and high localization precision. Owing to the single excitation and single detection path layout, 3D-DyPLoT is robust and easy to set up. This protocol discusses how to build 3D-DyPLoT step by step. First, the optical layout is described. Next, the system is calibrated and optimized by raster scanning a 190 nm fluorescent bead with the piezoelectric nanopositioner. Finally, to demonstrate real-time 3D tracking ability, 110 nm fluorescent beads are tracked in water.

This protocol details the construction and operation of a real-time 3D single particle tracking microscope capable of tracking nanoscale fluorescent probes at high diffusive speeds and low photon count rates.

一种实时3D 单粒子跟踪协议

Real-time three-dimensional single particle tracking (RT-3D-SPT) has the potential to shed light on fast, 3D processes in cellular systems. Although ...

A Macrophage Reporter Cell Assay to Examine Toll-Like Receptor-Mediated NF-kB/AP-1 Signaling on Adsorbed Protein Layers on Polymeric Surfaces

The persistent inflammatory host response to an implanted biomaterial, known as the foreign body reaction, is a significant challenge in the development and implementation of biomedical devices and tissue engineering constructs. Macrophages, an innate immune cell, are key players in the foreign body reaction because they remain at the implant site for the lifetime of the device, and are commonly studied to gain an understanding of this detrimental host response. Many biomaterials researchers have shown that adsorbed protein layers on implanted materials influence macrophage behavior, and subsequently impact the host response. The methods in this paper describe an in vitro model using adsorbed protein layers containing cellular damage molecules on polymer biomaterial surfaces to assess macrophage responses. An NF-&#1082;B/AP-1 reporter macrophage cell line and the associated colorimetric alkaline phosphatase assay were used as a rapid method to indirectly examine NF-&#1082;B/AP-1 transcription factor activity in response to complex adsorbed protein layers containing blood proteins and damage-associated molecular patterns, as a model of the complex adsorbed protein layers formed on biomaterial surfaces in vivo.

This protocol provides researchers with a rapid, indirect method of measuring TLR-dependent NF-&#1082;B/AP-1 transcription factor activity in a murine macrophage cell line in response to a variety of polymeric surfaces and adsorbed protein layers that model the biomaterial implant microenvironment.

巨噬细胞报告器细胞分析，以检查聚合物表面的吸附蛋白层上的收费类似受体介导NF-kB/AP-1信号

The persistent inflammatory host response to an implanted biomaterial, known as the foreign body reaction, is a significant challenge in the development ...

Bioinspired Soft Robot with Incorporated Microelectrodes

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics paradigm, especially in biomedical research. Recreating artificial life-like actuation dynamics is crucial for a soft-robotic system. However, the precise control and tuning of actuation behavior still represents one of the main challenges of modern soft robotic systems. This method describes a low-cost, highly scalable, and easy-to-use procedure to fabricate an electrically controllable soft robot with life-like movements that is activated and controlled by the contraction of cardiac muscle tissue on a micropatterned sting ray-like hydrogel scaffold. The use of soft photolithography methods makes it possible to successfully integrate multiple components in the soft robotic system, including micropatterned hydrogel-based scaffolds with carbon nanotubes (CNTs) embedded gelatin methacryloyl (CNT-GelMA), poly(ethylene glycol) diacrylate (PEGDA), flexible gold (Au) microelectrodes, and cardiac muscle tissue. In particular, the hydrogels alignment and micropattern are designed to mimic the muscle and cartilage structure of the sting ray. The electrically conductive CNT-GelMA hydrogel acts as a cell scaffold that improves the maturation and contraction behavior of cardiomyocytes, while the mechanically robust PEGDA hydrogel provides structural cartilage-like support to the whole soft robot. To overcome the hard and brittle nature of metal-based microelectrodes, we designed a serpentine pattern that has high flexibility and can avoid hampering the beating dynamics of cardiomyocytes. The incorporated flexible Au microelectrodes provide electrical stimulation across the soft robot, making it easier to control the contraction behavior of cardiac tissue.

A bioinspired scaffold is fabricated by a soft photolithography technique using mechanically robust and electrically conductive hydrogels. The micropatterned hydrogels provide directional cardiomyocyte cell alignment, resulting in a tailored direction of actuation. Flexible microelectrodes are also integrated into the scaffold to bring electrical controllability for a self-actuating cardiac tissue.

生物灵感软机器人与合并微电极

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics ...