Name: cardiac muscle-cell based actuator and self-stabilizing biorobot - part 1
Uploaded: 2017-07-11T13:00:00
Description: Watch this Scientific Journal Video about Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1 at JoVE.com

MT Holley由路易斯安那州理事会研究生研究员计划和C. Danielson由霍华德·休斯医学院教授课程支持。这项研究得到NSF授权号：1530884的支持。作者要感谢高级微结构和器件中心（CAMD）的洁净室的支持。

1.计算PDMS和添加剂的质量<ol><li>使用以下公式找到以下程序中特定高度所需PDMS的质量， M =ρ* V =ρ*高度*面积（1）， “高度”是层的高度，“面积”是PDMS将被固化的容器的面积，“ρ”是混合物的密度，“V”是体积。 注意：高度计算的密度为PDMS = 0.965 g / mL，Ni-PDMS = 1.639 g / mL，MB-PDMS = 0.648 g / mL。 </li><li>使用方程（1）估计给定容器所需的PDMS的质量，以获得生物致动器底座的特定?...

<ol>
	<li>Williams, B., Anand, S., Rajagopalan, J., Saif, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+self-propelled+biohybrid+swimmer+at+low+Reynolds+number.">A self-propelled biohybrid swimmer at low Reynolds number.</a> Nat commun. 5, (2014).</li><li>Nawroth, 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=A+tissue-engineered+jellyfish+with+biomimetic+propulsion.">A tissue-engineered jellyfish with biomimetic propulsion.</a> Nat Biotechnol. 30 (8), 729-797 (2012).</li><li>Huge Herr, ., D, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+swimming+Robot+Actuated+by+Living+Muscle+Tissue.">A swimming Robot Actuated by Living Muscle Tissue.</a> J Neuroeng. Rehabil. 1, (2004).</li><li>Park, S., al, e. l. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phototactic+guidance+of+a+tissue-engineered+soft-robotic+ray.">Phototactic guidance of a tissue-engineered soft-robotic ray.</a> Science. 353 (6295), 158-162 (2016).</li><li>Chan, V., Park, K., Colleens, M., Kong, H., Saif, T., Bashir, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+miniaturized+walking+biological+machines.">Development of miniaturized walking biological machines.</a> Sci. Rep. 2, 857 (2012).</li><li>Cvetkovic, 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=Three-dimensionally+printed+biological+machines+powered+by+skeletal+muscle.">Three-dimensionally printed biological machines powered by skeletal muscle.</a> PNAS. 111, 10125-10130 (2014).</li><li>Xi, J., Scmidt, J., Montemagno, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembled+microdevices+driven+by+muscle.">Self-assembled microdevices driven by muscle.</a> Nat. Mater. 4, 180-184 (2005).</li><li>Kim, 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=Establishment+of+a+fabrication+method+for+a+long-term+actuated+hybrid+cell+robot.">Establishment of a fabrication method for a long-term actuated hybrid cell robot.</a> Lab Chip. 7, 1504-1508 (2007).</li><li>Tanaka, Y., Sato, K., Shimizu, T., Yamato, M., Okano, T., Kitamori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+micro-spherical+heart+pump+powered+by+cultured+cardiomyocytes.">A micro-spherical heart pump powered by cultured cardiomyocytes.</a> Lab Chip. 7, 207-212 (2007).</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=Micro+pumping+with+cardiomyocyte-polymer+hybrid.">Micro pumping with cardiomyocyte-polymer hybrid.</a> Lab Chip. 7, 1367-1370 (2007).</li><li>Akiyama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atmospheric-operable+bioactuator+powered+by+insect+muscle+packaged+with+medium.">Atmospheric-operable bioactuator powered by insect muscle packaged with medium.</a> Lab Chip. 13, 4870-4880 (2013).</li><li>Kabumoto, K., Hoshino, T., Akiyama, Y., Morishima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voluntary+movement+controlled+by+the+surface+EMG+signal+for+tissue-engineered+skeletal+muscle+on+a+gripping+tool.">Voluntary movement controlled by the surface EMG signal for tissue-engineered skeletal muscle on a gripping tool.</a> Tissue Eng. Part A. 19, 1695-1703 (2013).</li><li>Feinberg, A., Feigel, A., Shevkoplyas, S., Sheehy, S., Whitesides, G., Parker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscular+thin+films+for+building+actuators+and+powering+devices.">Muscular thin films for building actuators and powering devices.</a> Science. 317 (5843), 1366-1370 (2007).</li><li>Vannozi, L., 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=Self-assembly+of+polydimethylsiloxane+structures+from+2D+to+3D+for+bio-hybrid+actuation.">Self-assembly of polydimethylsiloxane structures from 2D to 3D for bio-hybrid actuation.</a> Bioinspiration Biomimetics. 10 (5), (2015).</li><li>Holley, M. T., Nagarajan, N., Danielson, C., Zorlutuna, P., Park, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+muscle-based+actuators+for+self-stabilizing+swimming+biorobots.">Development and characterization of muscle-based actuators for self-stabilizing swimming biorobots.</a> Lab Chip. 16, 3473-3484 (2016).</li><li>Sfakiotakis, M., Lane, D., Davies, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+fish+swimming+modes+for+aquatic+locomotion.">Review of fish swimming modes for aquatic locomotion.</a> IEEE J. Oceanic Eng. 24, 237-252 (1999).</li><li>McCain, M., Agarwal, A., Hesmith, H., Nesmith, A., Parker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micromolded+gelatin+hydrogels+for+extended+culture+of+engineered+cardiac+tissues.">Micromolded gelatin hydrogels for extended culture of engineered cardiac tissues.</a> Biomaterials. 35 (21), 5462-5471 (2014).</li></ol>

水中游泳者中可以发现各种运动机制16 。本研究中Biorobot的运动机制采用鳍状运动，特别是鸵鸟运动。鸵鸟式游泳者通过摇摆尾巴（悬臂）并具有刚体（分层底座）推动自己16 。鱼类，如鱼类和鲶鱼使用这种类型的运动。玄武岩型游泳运动员通常很慢，身体尺寸无效。虽然鸵鸟游泳缺乏速度，这种游泳形式允许工程师在基座或身体上实现各种功能（如动态稳定?...

正在积极研究生物执行器和双子座，以便为众多应用提供常规机器人的替代方案。步行5,6,7,8的Biorobots游泳1，2，3，4 ，泵9,10 ，或手柄11，12，13 已经开发。类似地，肌肉细胞可以并入3D卷状PDMS结构14中 。通常，使用具有诸如水凝胶和PDMS（聚二甲基硅氧烷）的材料的软平版印刷技术来制造Biorobot骨架。这些是有吸引力的选择，因为它们的灵活性，biocompatib容易调整刚度。活体肌细胞通常与这些材料结合以通过收缩提供力产生。哺乳动物的心肌细胞（心肌细胞）和骨骼肌细胞主要用于致动。除了这两个之外，昆虫肌肉组织已经用于在室温下操作双子座3 。在这两部分研究中，选择了心肌细胞，因为它们的自发收缩6 。 早期关于biorobots的研究大部分集中在开发生物执行器，而Biorobot架构的优化和Biorobots的基本功能的发展在很大程度上被忽视。最近有一些报告显示了不同游泳模式的实施，这些模式受到自然界中发现的推进模式的启发。这些方法包括PDMS膜和肌肉细胞以模拟各种天然推进方法。例如，已经报道了基于鞭毛的推进1 ，仿生水母推进2 ，生物混合射线4和薄膜PDMS游泳装置13 。 在本文中，我们介绍了可以保持浸入深度以及俯仰和滚动的自稳定游泳芭比娃娃的制作过程。 biorobot具有坚实的基部或身体，其由附着在其表面上的心肌细胞的单个悬臂推动。当心肌细胞收缩时，心肌细胞使得悬臂在纵向方向上弯曲。这种游泳形式被列为鸵鸟游泳。在基地添加额外功能的能力是鸵鸟游泳的独特优势。例如，可以利用该基础来提供过量的浮力以携带用于心肌细胞收缩的附加货物或控制电路。 稳定性在以前的biorobots研究中，biorobot经常被忽视。在本研究中，我们通过使用具有不同质量密度的不同复合PDMS材料设计基体来实现自稳定。因此，biorobot表现出对外部干扰的抵抗力，并保持其浸没深度，俯仰和滚动，无人值守。第一层是微球PDMS（MB-PDMS）， 即与微球混合的PDMS，降低了Biorobot的密度，使其能够漂浮在介质中。第二层是PDMS悬臂，其厚度被定制，使得由心肌细胞产生的力可以使悬臂从45°到90°显着弯曲。底层为镍-PDMS（Ni-PDMS）， 即与镍粉混合的PDMS。该层执行多个功能。它是磁性的，因此在细胞播种期间，使用磁铁，可以将biorobot锚定在培养基的底部。镍混合物的密度比MB-PDMS高中等，并确保漂浮时的biorobot的直立位置。该层的重量在任何间距和滚动条件下都能够在biorobot上产生恢复扭矩。此外，Ni-PDMS和MB-PDMS之间的体积比保持浸没深度。所提出的方案对于对表征肌肉细胞和组织的打击力以及希望建造游泳混合动物的人来说是非常有用的。 功能化生物致动器和双极器件的接种，细胞的机械和生物化学特征以及器件功能的定量分析在这两篇文章的第2部分以及最近的工作中有详细描述。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Polydimethylsiloxane (PDMS)</td>
			<td>Dow Corning</td>
			<td>184 sil elast kit&#160; 0.5kg</td>
			<td>Sylgard 184</td>
		</tr>
		<tr>
			<td>Nickel Powder</td>
			<td>Sigma-Aldrich</td>
			<td>266981-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenolic microballoons</td>
			<td>US Composites</td>
			<td>BJO-0930</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon wafers</td>
			<td></td>
			<td></td>
			<td>4 inch diameter</td>
		</tr>
		<tr>
			<td>PWM101 light-duty spinner</td>
			<td></td>
			<td></td>
			<td>Spin- coater</td>
		</tr>
		<tr>
			<td>Positive photoresist (S1808)</td>
			<td>Dow Corning</td>
			<td>DEM-10018197</td>
			<td></td>
		</tr>
		<tr>
			<td>Hotplate</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum chamber</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>M206 mechanical convection oven</td>
			<td></td>
			<td></td>
			<td>Convection oven</td>
		</tr>
		<tr>
			<td>Laser engraver</td>
			<td>Universal Laser System</td>
			<td>VLS2.30</td>
			<td>Utilizes a 10W, 10.6 &#181;m wavelength, CO2 Laser</td>
		</tr>
		<tr>
			<td>Universal Laser Systems Application</td>
			<td>Universal Laser System</td>
			<td></td>
			<td>Application for running the VLS 2.30</td>
		</tr>
		<tr>
			<td>Matlab</td>
			<td>MathWorks</td>
			<td></td>
			<td>Numerical analysis program</td>
		</tr>
		<tr>
			<td>Scotch Tape</td>
			<td>Scotch Brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Solid-glass beads</td>
			<td>Sigma-Aldrich</td>
			<td>Z265926-1EA</td>
			<td>Soda-lime glass, diameter 3 mm</td>
		</tr>
		<tr>
			<td>Scale</td>
			<td>Mettler Toledo</td>
			<td>EL303</td>
			<td></td>
		</tr>
		<tr>
			<td>BD-20AC Laboratory Corona Treater</td>
			<td>Electrotechnic Products</td>
			<td>12051A</td>
			<td>Corona Discharger</td>
		</tr>
		<tr>
			<td>Ultrasonic Bath 1.9L</td>
			<td>Fisher Scientific</td>
			<td>15-337-402</td>
			<td>40 kHz industrial transducer</td>
		</tr>
		<tr>
			<td>Fibronectin from bovine plasma</td>
			<td>Sigma-Aldrich</td>
			<td>F1141</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Phosphate Buffer (PBS)</td>
			<td>Sigma-Aldrich</td>
			<td>D1408-100ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Modified Eagle Medium (DMEM)</td>
			<td>Hyclone Laboratories</td>
			<td>16750-074</td>
			<td>With 4500 mg/L glucose, 4.0 mM L-glutamine, and 110 mg/L sodium pyruvate.</td>
		</tr>
		<tr>
			<td>Fetalclone III serum</td>
			<td>Hyclone Industries, GE</td>
			<td>16777-240</td>
			<td>Fetal bovine serum</td>
		</tr>
		<tr>
			<td>Penicillin-G sodium salt</td>
			<td>Sigma-Aldrich</td>
			<td>P3032</td>
			<td></td>
		</tr>
	</tbody>
</table>

cardiac muscle-cell based actuator and self-stabilizing biorobot - part 1

生物机器通常被称为双子座，是活体细胞或基于组织的装置，其仅由活体组分的收缩活性来驱动。由于其固有的优势，双子座作为传统全人造机器人的替代品正在获得兴趣。各种研究集中在利用生物执行器的力量，但是最近的研究已经定量地表征了Biorobots的性能，并研究了它们的几何形状以增强功能和效率。在这里，我们展示了一个自我稳定的游泳水族箱的发展，可以在没有外部干预的情况下保持其俯仰，深度和滚动。在第一部分中描述了用于生物致动器和biorobot的PDMS支架的设计和制造，随后用纤连蛋白进行官能化。在这篇两部分文章的第二部分中，我们详细介绍了心肌细胞的结合和表征生物活化ator和biorobot功能。两者都包括产生基于翅片的推进的基部和尾部（悬臂）。尾部由使用PDMS和激光雕刻的软光刻技术构成。将尾部与装置基底结合后，用细胞粘附蛋白功能化，并与心肌细胞融合。生物致动器的基座由具有中心玻璃珠（作为重量）的固体PDMS块组成。 biorobot的基础由两个复合PDMS材料，Ni-PDMS和microballoon-PDMS（MB-PDMS）组成。镍粉（Ni-PDMS）允许细胞播种期间的biorobot的磁控制和运动期间的稳定性。微球（MB-PDMS）降低MB-PDMS的密度，使Biorobot能够稳定地游泳和游泳。使用这两种具有不同质量密度的材料，能够精确控制重量分布，以确保在biorobot的任何角度处的积极恢复力。这个技巧产生一个磁力控制的自稳定游泳混合动力。

生物致动器和双子座具有非常相似的制造工艺，因为biorobot是生物致动器的自然延伸（ 图1 ）。首先开发生物致动器以建立生物机器所需的技术，分析由细胞产生的力并且机械和生物化学地表征细胞成熟，这两者在这两篇文章的第2部分中详细描述就像我们最近出版的作品一样。 评估和调整致动器的弹簧常数，以在...

Watch this Scientific Journal Video about Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1 at JoVE.com

Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1

在这篇两部分的研究中，使用高度柔性的聚二甲基硅氧烷（PDMS）悬臂和活肌细胞（心肌细胞）开发了生物致动器，并进行了表征。将生物致动器与由改性PDMS材料制成的基底结合以构建自稳定的游泳制冷机。

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