这项研究是由R-397-000-204-133（新加坡青年科学家奖国立大学）的支持。

<html>注意:所有的软气动夹持器被铸造基于硅氧烷的弹性体的混合物到定制3D印刷模具，其随后的制造过程包括三个步骤制造:成型带嵌入气动通道夹持器臂组件，模制连接到气动通道室元件以及密封填充有空气的腔室部件。 1.弹性体的制备<ol><li>放置在秤为混合器的容器和去皮它。倾部分A和在用1容器中的基于硅氧烷的弹性体的A:1的重量比。 </li><li>覆盖容器和测量的总重量。 </li><li>放置在容器和材料成离心混合器。调整在混频器的重量平衡，以在步骤1.2测量的体重。 </li><li>设置在混合和脱气模式，以2000转2200转分别为30秒。混合弹性体组分充分达到均匀的固化。 </li></ol>乐"&gt; 2，模具设计与制作注意:在模具的几何形状将根据不同的应用的具体要求而变化。下面的步骤说明所必需的制造的模具的腔室和夹持器部件中的CAD软件的一般的关键步骤。 <ol><li>设计采用计算机辅助设计（CAD）软件的模具和封模。参见图1为在本手稿中使用的模具的几何形状和特定的尺寸。 <ol><li>外边界框的设计<ol><li>顶部平面右键单击，然后点击"正常到"按钮正常化顶端平面。 </li><li>点击左上角的"素描"，打开一个"素描"窗口。然后，点击工具栏的左上角的"草图"按钮，绘制室组成的矩形底座。 </li><li>点击"智能尺寸"功能，它位于"草图"按钮旁边，以定义SK蚀刻尺寸。确保草图已经完全定义（ 即所有的画线变成黑色），并在完成后退出草图。 </li><li>点击"功能"窗口。然后，点击"拉伸凸台/基地"功能，在挤出Y方向选择的轮廓。 </li><li>点击模型的顶部表面上预选草图平面。绘制一个矩形，并作为2.1.1.2 2.1.1.3和描述定义尺寸。 </li><li>点击"功能"窗口。然后，点击"拉伸切除"功能来削减挤压铸造弹性体（ 图2A）的腔体。确保壁厚为2.5毫米。 </li></ol></li><li>内腔室的设计<ol><li>在开口面积的Y方向的表面上单击鼠标右键。然后，点击"普通至"正常化到表面。 </li><li>接下来，点击"素描"窗口中绘制室组成一个矩形作为步骤2.1.1.2和描述2.1.1.3。 </li><li>点击"功能"窗口。然后，点击"拉伸凸台/基地"功能，在挤出Y方向（ 图2B）室元件。 注意:在步骤2.1.1.6切口的深度比该挤出基的2.5毫米。 </li></ol></li><li>夹具元件的设计<ol><li>点击X轴负方向的模型表面上预选草图平面的夹持器组件。如步骤2.1.1.2 2.1.1.3和创建描述中的"素描"窗口中的矩形。 </li><li>点击"功能"窗口。然后，点击"拉伸凸台/基地"功能，在挤出负X方向的选择轮廓。 </li><li>点击夹持器组件的顶表面上以预选草图平面。创建在"草图"的窗口（ 图2C）夹持器的形状和退出草图时的尺寸完全定义如步骤2.1.1.2一个ð2.1.1.3。 </li><li>点击"功能"窗口。然后，点击"拉伸切口"切一腔在夹具元件铸造弹性体。确保壁厚为2.5毫米。 </li></ol></li><li>室和夹持器之间的连接的设计<ol><li>如2.1.1.2和2.1.1.3中描述的室件的顶表面上创建的"草图"窗口的矩形。 </li><li>点击"功能"窗口。然后，点击"拉伸切口"创造室和夹持元件（ 图2D）之间的连接。 </li></ol></li><li>气动通道的设计<ol><li>如步骤2.1.1.2和2.1.1.3中所述创建在正X方向上的腔室片表面1.5毫米直径的圆。 </li><li>点击"功能"窗口。然后，单击"剪切挤压"创建线材插入（ 图2E）通道。确保霍尔ES不通过夹持器组件切断。 </li></ol></li></ol></li><li>在一个单独的CAD文件，绘制与长度和宽度的空腔是比夹持器的腔室部件的外尺寸大1mm的密封模具。注意:壁厚为2.5毫米。 <ol><li>点击"素描"窗口创建顶部平面矩形所描述的步骤2.1.1.2 2.1.1.3和。 </li><li>点击"功能"窗口。然后，点击"拉伸凸台/基地"功能，在挤出Y方向选择的轮廓。 </li><li>点击模型的顶面预选草图平面。绘制一个矩形，如步骤2.1.1.2 2.1.1.3和）所述定义尺寸。 </li><li>点击"功能"窗口。然后，点击"拉伸切除"功能来削减挤出一腔浇铸弹性体。确保壁厚为2.5毫米。 </li></ol></li><li>保存每个防霉片为3D打印的.STL文件。 </l我&gt; <li>加载.STL文件与30微米的分辨率的3D打印机和打印模片9。 </li><li>上模片除去任何载体材料，用水洗涤，模具件。 </li></ol> 3.软单/双动臂气动手指<ol><li>成型夹具臂组件嵌入气动渠道<ol><li>为了产生与连接到它的气动通道密封的室插入在左和室元件（ 图3A）的右侧两个3D印刷室块。 </li><li>通过腔室插入两个毫米直径的1.5钛线棒，保持从夹紧提示2毫米的距离以创建气动通道（ 图3A）。注意:使用一个线材为单致动臂夹持器。 </li><li>倾弹性体混合物到模具中以完全填充所述夹持器组件。 </li><li>确保没有明显的气泡存在。 </li><li>的PlacE中的模具放入烘箱中在60℃下固化10分钟。一旦弹性体固化后，取出从烤箱模具。 </li></ol></li><li>连接到气动通道成型室元件<ol><li>拉动线材和两腔室阻挡从模具。 </li><li>放置在夹持器组件的顶部上的三维印刷夹持器块，以创建所述腔室（ 图3B）。插入线材以阻止在模具的壁上的孔。 </li><li>倾弹性体混合物到模具中以填充腔室部件的剩余部分，​​并确保有困在模具无可见的气泡。 </li><li>在60℃下进行10分钟的温度固化的部分。一旦弹性体固化从烤箱中取出模具。 </li><li>取下夹具块，并与脱模腔室结构的完全固化抓手。 </li></ol></li><li>密封充有空气的腔室组件<ol><li>倾弹性体混合物放入密封模具和固化它在60℃下10分钟。 </li><li>在固化2.5毫米密封层刷弹性体材料的层。放置与在涂覆密封层的顶部的腔室结构的固化夹持器和两片结合在一起（ 图3C）。 </li><li>接着，完全固化整个结构在60℃下15分钟。 </li><li>脱模的完全固化的软的机器人夹持装置。 </li></ol></li></ol> 4.插入软机器人气动夹紧装置进入处理工具<ol><li>设计处理工具运用CAD软件的补充文件1描述并将其保存在文件.STL。 见图4和5的工具的尺寸。 </li><li>加载在3D打印机的.STL文件和打印模片9。 注:手动控制处理工具，矩形帽，和可移动的活塞（ 图4）所有的印刷工序可在3小时48分钟内完成。公关inting时间用于制造机械手控制处理工具和矩形帽（ 图5）是1小时56分钟。见三维打印机使用说明书补充文件2。 </li><li>完成打印后脱落的工具，任何支持材料。然后，洗工具水。 </li><li>插入夹持器插入手动控制处理工具（ 图4A）和覆盖的可动矩形帽（ 图4B）的开口面积。 </li><li>插入一个可移动的活塞（ 图4C），以促进腔室压缩。 </li><li>插入夹持器和线性驱动器到机器人控制处理工具（ 图5A）。注意:线性致动器替代在手动控制模式下的可移动的活塞对腔室压缩。 </li><li>覆盖可动矩形帽（ 图5B）的开口面积。 </li></ol> 5.评估和抓地力压缩试验<ol><li>评估软抓手用跳线进行测试扣人心弦的功能。 <ol><li>将餐桌上的跳线。 </li><li>调整夹具，以使金属丝在两夹爪臂之间。 </li><li>移动可移动的活塞，以便致动所述夹具臂以保持导线以压缩室中。 注意:只有手动控制处理工具在握持演示中使用。 </li><li>保持和线移动到位于20厘米的线的原始位置离开的盒子。 </li></ol></li><li>放置夹持器的两个夹爪之间的校准力传感电阻。确保在传感区域中的夹爪抓地力。注意:在传感区域的直径为14.7毫米。 </li><li>压缩所述腔室以促动夹持器手臂上的力感应电阻的抓地力。 </li><li>测量最大的抓地力压缩力，由于在10中描述的软单致动臂和双促动臂气动夹具可以产生。 注意:读出值将显示在笔记本电脑上。最大的抓地力压缩力在那个气动通道可以承受最大压力的点测量。 </li><li>剪出由柔软的双致动臂气动夹具个别弹性夹爪。 </li><li>插入镊子提示入弹性把持爪的气动通道。 </li><li>放置钳子的两个钳口之间的校准力传感电阻。 </li><li>测量由神经外科医生进行的模拟神经外科手术期间通过弹性体涂层的镊子和钳子所产生的压缩力10。 注意:神经外科医生施加力是类似于他的力感测电阻器实际手术过程中通常适用。 </li><li>平均从每个测试五项试验获得的数据。 </li></ol>

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作者什么都没有透露。

我们已经成功地证明，允许的对象符合握持软机器人手爪气动装置，它产生的夹着的物体比弹性体涂层镊子技巧和镊子上低得多的压缩力发挥。镊子是周围神经损伤修复过程中神经操作的必备工具手术11，12，但需要其金属结构，从医生的使用非常谨慎，以防止因过度夹持力和对周围组织的附带损伤神经损伤。取决于损伤的严重程度，各种并发症包括从较不严重的，如疼痛，严重的，如血?...

软机器人已经引起了机器人界极大的研究兴趣，他们在不同的功能任务被使用，如在非结构化环境中1和2握持运动波动。它们主要由软弹性体材料和通过利用不同材料的不同的致动技术，如电活化聚合物（EAP），形状记忆合金（SMA），或压缩流体3进行控制。根据诱导的静电力，以产生活性的菌株，由此产生致动差动电压EAPS功能。的形状记忆合金的固有形状记忆效应被部署到生成期间时的温度变化的相变基于所述力产生所需的致动。最后，压缩的流体致动技术有利于简单的设计策略，以诱导在软的致动器的刚度差异，从而使得多个柔性区域将膨胀在增压。软机器人被设计为扩大传统硬机器人的应用，尤其是在微妙的对象涉及的应用程序。特别是，在本文中，我们提出我们在制定细腻手术操作的软机器人抓手独特的方法。 外科用夹紧是参与许多外科手术如肝，妇科，泌尿科和神经修复手术4,5的一个重要方面，它通常是由刚性的，钢的组织夹紧工具，如钳子和腹腔镜抓紧器用于促进的目的而进行观察，切除，吻合程序等 。然而，由于传统的夹紧工具由金属制成，可能在接触6的点的软组织会导致高应力集中区域的需要格外小心。根据组织损伤的严重程度，各种并发症，如疼痛，病理性瘢痕组织˚Formation，甚至终身残疾，可能会导致。预先研究报告说，在周围神经手术并发症的发生率为3％7。因此，软握持，可以提供安全的抓地力兼容的概念可以是微妙的手术操作有希望的候选人。 这里，我们提出的3D印刷和改性软光刻技术，其通过基于杆的方法中，以制造定制的软机器人气动夹具的组合。基于压缩流体致动的软机器人的传统制造技术需要印有和密封工艺密封通道8个气动频道的模具。然而，这是不适用于需要小气动通道，其中通道的阻塞可在密封过程中容易发生微型软机器人可行的。传统技术需要的气动通道的密封，以通过将其粘结的涂覆密封层来完成。因此，拉弹性体材料的揭掉最初用作接合层洒入的微小通道和封闭这些频道。也不可能在该结构的中间定位的充气通道，并连接到使用传统技术的腔室部件。所提出的方法允许连接到一个空气填充用室杆小型化的气动通道的建立，并且不需要的微小通道的密封。此外，连接到气动通道的腔室作为不需要对压缩流体致动的外部空气源的空气源。它允许手动和通过促进该腔室压缩致动夹紧部件，由此提供用户控制的力，它们是通过夹持器施加的量的选择机械手控制模式。这种方法是高度可定制的，并且可以用于制造各种类型的软夹持器的设计，如单或亩夹具ltiple致动臂。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Weighing Scale</td>
			<td>Severin</td>
			<td>KW3667</td>
			<td>(Step: Preparation of elastomers)</td>
		</tr>
		<tr>
			<td>Ecoflex Supersoft 0030 Elastomer</td>
			<td>Smooth-On</td>
			<td>EF0030</td>
			<td>(Step: Preparation of elastomers)</td>
		</tr>
		<tr>
			<td>Planetary Centrifugal Mixer and Containers</td>
			<td>THINKY USA Inc.</td>
			<td>ARE-310</td>
			<td>(Step: Preparation of elastomers)</td>
		</tr>
		<tr>
			<td>Solidworks CAD</td>
			<td>Dassault Syst&#232;mes&#160;</td>
			<td>Solidworks Research Subscription</td>
			<td>(Step: Soft single/double-actuatable arm pneumatic grippers)</td>
		</tr>
		<tr>
			<td>Objet 3D Printer</td>
			<td>Stratasys</td>
			<td>260 Connex2</td>
			<td>(Step: Soft single/double-actuatable arm pneumatic grippers)</td>
		</tr>
		<tr>
			<td>Titanium Wire Rods</td>
			<td>Titan Engineering</td>
			<td>N/A</td>
			<td>(Step: Soft single/double-actuatable arm pneumatic grippers)</td>
		</tr>
		<tr>
			<td>Natural Convection Oven with Timer</td>
			<td>Thermo Fisher Scientific</td>
			<td>BIN#ED53</td>
			<td>(Step: Soft single/double-actuatable arm pneumatic grippers)</td>
		</tr>
		<tr>
			<td>Linear Actuator</td>
			<td>Firgelli Technologies</td>
			<td>L12</td>
			<td>(Step: Insertion of soft robotic pneumatic gripper device into handling tool)</td>
		</tr>
		<tr>
			<td>Jumper Wire</td>
			<td>sgbotic</td>
			<td>CAB-01146</td>
			<td>(Step: Evaluations and grip compressive test)</td>
		</tr>
		<tr>
			<td>Force Sensing Resistor</td>
			<td>Interlink Electronics</td>
			<td>FSR402</td>
			<td>(Step: Evaluations and grip compressive test)</td>
		</tr>
	</tbody>
</table>

rod-based fabrication of customizable soft robotic pneumatic gripper devices for delicate tissue manipulation

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations at the point of contact. It can be achieved by complementing traditional rigid grippers with soft robotic pneumatic gripper devices. This manuscript describes a rod-based approach that combined both 3D-printing and a modified soft lithography technique to fabricate the soft pneumatic gripper. In brief, the pneumatic featureless mold with chamber component is 3D-printed and the rods were used to create the pneumatic channels that connect to the chamber. This protocol eliminates the risk of channels occluding during the sealing process and the need for external air source or related control circuit. The soft gripper consists of a chamber filled with air, and one or more gripper arms with a pneumatic channel in each arm connected to the chamber. The pneumatic channel is positioned close to the outer wall to create different stiffness in the gripper arm. Upon compression of the chamber which generates pressure on the pneumatic channel, the gripper arm will bend inward to form a close grip posture because the outer wall area is more compliant. The soft gripper can be inserted into a 3D-printed handling tool with two different control modes for chamber compression: manual gripper mode with a movable piston, and robotic gripper mode with a linear actuator. The double-arm gripper with two actuatable arms was able to pick up objects of sizes up to 2 mm and yet generate lower compressive forces as compared to elastomer-coated and non-coated rigid grippers. The feasibility of having other designs, such as single-arm or hook gripper, was also demonstrated, which further highlighted the customizability of the soft gripper device, and it's potential to be used in delicate surgical manipulation to reduce the risk of tissue grip damage.

软机器人气动夹持器装置是能够与在直径达至1.2mm的尺寸（ 图6）拾取对象。由单致动臂产生的最大握压缩力，和双致动臂软夹持器装置分别0.79±0.14 N的0.27±0.07 N和，相对于1.71±0.16 N和2.61±0.22Ñ压缩力在由该弹性体涂覆的镊子和由未涂覆镊子（ 图7）的模拟手术。握持力可根据夹具的几何形状和气动通道的尺寸而变化。弹性体的材料性质将决定的充...

Watch this Scientific Journal Video about Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation at JoVE.com

Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation

This protocol describes a rod-based approach, combining 3D-printing and soft lithography techniques for fabricating the soft gripper devices. This approach eliminates the need for an external air source by incorporating a chamber component and reduces the chance of occlusion during the sealing process, particularly for miniaturized pneumatic channels.

娇嫩的组织操纵定制的软机器人气动设备手爪基于杆制造

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations ...

rod-based-fabrication-customizable-soft-robotic-pneumatic-gripper

Research

JoVE Journal

Bioengineering

8.7K 次观看.  National University of Singapore. This protocol describes a rod-based approach, combining 3D-printing and soft lithography techniques for fabricating the soft gripper devices. This approach eliminates the need for an external air source by incorporating a chamber component and reduces the chance of occlusion during the sealing process, particularly for miniaturized pneumatic channels.

视频: Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications. Briefly, decellularized tissue is lyophilized, milled, enzymatically digested, and then brought to physiological pH. The lyophilization removes all water content from the tissue, resulting in dry ECM that can be ground into a fine powder with a small mill. After milling, the ECM powder is digested with pepsin to form an injectable matrix. After adjustment to pH 7.4, the liquid matrix material can be injected into the myocardium. Results of previous characterization assays have shown that matrix gels produced from decellularized pericardial and myocardial tissue retain native ECM components, including diverse proteins, peptides and glycosaminoglycans. Given the use of this material for tissue engineering, in vivo characterization is especially useful; here, a method for performing an intramural injection into the left ventricular (LV) free wall is presented as a means of analyzing the host response to the matrix gel in a small animal model. Access to the chest cavity is gained through the diaphragm and the injection is made slightly above the apex in the LV free wall. The biologically derived scaffold can be visualized by biotin-labeling before injection and then staining tissue sections with a horse radish peroxidase-conjugated neutravidin and visualizing via diaminobenzidine (DAB) staining. Analysis of the injection region can also be done with histological and immunohistochemical staining. In this way, the previously examined pericardial and myocardial matrix gels were shown to form fibrous, porous networks and promote vessel formation within the injection region.

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

心肌组织工程生物衍生的注射材料的制备

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

科研

生物工程

Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

Engineered muscle tissues can be used for several different purposes, which include the production of tissues for use as a disease model in vitro, e.g. to study pressure ulcers, for regenerative medicine and as a meat alternative 1. The first reported 3D muscle constructs have been made many years ago and pioneers in the field are Vandenburgh and colleagues 2,3. Advances made in muscle tissue engineering are not only the result from the vast gain in knowledge of biochemical factors, stem cells and progenitor cells, but are in particular based on insights gained by researchers that physical factors play essential roles in the control of cell behavior and tissue development. State-of-the-art engineered muscle constructs currently consist of cell-populated hydrogel constructs. In our lab these generally consist of murine myoblast progenitor cells, isolated from murine hind limb muscles or a murine myoblast cell line C2C12, mixed with a mixture of collagen/Matrigel and plated between two anchoring points, mimicking the muscle ligaments. Other cells may be considered as well, e.g. alternative cell lines such as L6 rat myoblasts 4, neonatal muscle derived progenitor cells 5, cells derived from adult muscle tissues from other species such as human 6 or even induced pluripotent stem cells (iPS cells) 7. Cell contractility causes alignment of the cells along the long axis of the construct 8,9 and differentiation of the muscle progenitor cells after approximately one week of culture. Moreover, the application of electrical stimulation can enhance the process of differentiation to some extent 8. Because of its limited size (8 x 2 x 0.5 mm) the complete tissue can be analyzed using confocal microscopy to monitor e.g. viability, differentiation and cell alignment. Depending on the specific application the requirements for the engineered muscle tissue will vary; e.g. use for regenerative medicine requires the up scaling of tissue size and vascularization, while to serve as a meat alternative translation to other species is necessary.

Engineered muscle tissue has great potential in regenerative medicine, as disease model and also as an alternative source for meat. Here we describe the engineering of a muscle construct, in this case from mouse myoblast progenitor cells, and the stimulation by electrical pulses.

工程骨骼肌组织中的小鼠成肌细胞的内皮祖细胞与应用电刺激

Engineered muscle tissues can be used for several different purposes, which include the production of tissues for use as a disease model in vitro, e.g. to ...

Density Gradient Multilayered Polymerization (DGMP): A Novel Technique for Creating Multi-compartment, Customizable Scaffolds for Tissue Engineering

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix structure (e.g. composition, concentration, or stiffness of the matrix) vary over space, would enable a wide range of investigations concerning how these variables affect cell differentiation, migration, and other phenomena. The major challenge in creating layered matrices is maintaining the structural integrity of layer interfaces without diffusion of individual components from each layer1. Current methodologies to achieve this include photopatterning2-3, lithography4, sequential functionalization5, freeze drying6, microfluidics7, or centrifugation8, many of which require sophisticated instrumentation and technical skills. Others rely on sequential attachment of individual layers, which may lead to delamination of layers9. 
DGMP overcomes these issues by using an inert density modifier such as iodixanol to create layers of varying densities10. Since the density modifier can be mixed with any prepolymer or bioactive molecule, DGMP allows each scaffold layer to be customized. Simply varying the concentration of the density modifier prevents mixing of adjacent layers while they remain aqueous. Subsequent single step polymerization gives rise to a structurally continuous multilayered scaffold, in which each layer has distinct chemical and mechanical properties. The density modifier can be easily removed with sufficient rinsing without perturbation of the individual layers or their components. This technique is therefore well suited for creating hydrogels of various sizes, shapes, and materials.
A protocol for fabricating a 2D-polyethylene glycol (PEG) gel, in which alternating layers incorporate RGDS-350, is outlined below. We use PEG because it is biocompatible and inert. RGDS, a cell adhesion peptide11, is used to demonstrate spatial restriction of a biological cue, and the conjugation of a fluorophore (Alexa Fluor 350) enables us to visually distinguish various layers. This procedure can be adapted for other materials (e.g. collagen, hyaluronan, etc.) and can be extended to fabricate 3D gels with some modifications10.

Density Gradient Multilayered Polymerization DGMP: A Novel Technique for Creating Multi-compartment, Customizable Scaffolds for Tissue Engineering

Here we describe a unique strategy for creating biocompatible, layered matrices with continuous interfaces between distinct layers for tissue engineering. Such a scaffold could provide an ideal customizable environment to modulate cell behavior by various biological, chemical or mechanical cues

密度梯度多层聚合（dGMP的）:创建多室的，可定制的一种新技术，组织工程支架

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

人体软骨组织制作采用三维喷墨打印技术

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System

Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized medicine innovations. However, translating these advances into therapeutic applications will require in vitro systems that allow for robust, flexible, and cost effective bioreactor systems. There are several bioreactor systems currently utilized in research and commercial settings; however, many of these systems are not optimal for establishing, expanding, and monitoring the growth of different cell types. The culture parameters most challenging to control in these systems include, minimizing hydrodynamic shear, preventing nutrient gradient formation, establishing uniform culture medium aeration, preventing microbial contamination, and monitoring and adjusting culture conditions in real-time. Using a pneumatic single-use bioreactor system, we demonstrate the assembly and operation of this novel bioreactor for mammalian cells grown on micro-carriers. This bioreactor system eliminates many of the challenges associated with currently available systems by minimizing hydrodynamic shear and nutrient gradient formation, and allowing for uniform culture medium aeration. Moreover, the bioreactor&#8217;s software allows for remote real-time monitoring and adjusting of the bioreactor run parameters. This bioreactor system also has tremendous potential for scale-up of adherent and suspension mammalian cells for production of a variety therapeutic proteins, monoclonal antibodies, stem cells, biosimilars, and vaccines.

Using a pneumatic bioreactor, we demonstrate the assembly, operation, and performance of this single-use bioreactor system for the growth of mammalian cells.

培养哺乳动物细胞使用一次性使用气动生物反应器系统

Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized ...

Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery

In recent years, soft robotics technologies have aroused increasing interest in the medical field due to their intrinsically safe interaction in unstructured environments. At the same time, new procedures and techniques have been developed to reduce the invasiveness of surgical operations. Minimally Invasive Surgery (MIS) has been successfully employed for abdominal interventions, however standard MIS procedures are mainly based on rigid or semi-rigid tools that limit the dexterity of the clinician. This paper presents a soft and high dexterous manipulator for MIS. The manipulator was inspired by the biological capabilities of the octopus arm, and is designed with a modular approach. Each module presents the same functional characteristics, thus achieving high dexterity and versatility when more modules are integrated. The paper details the design, fabrication process and the materials necessary for the development of a single unit, which is fabricated by casting silicone inside specific molds. The result consists in an elastomeric cylinder including three flexible pneumatic actuators that enable elongation and omni-directional bending of the unit. An external braided sheath improves the motion of the module. In the center of each module a granular jamming-based mechanism varies the stiffness of the structure during the tasks. Tests demonstrate that the module is able to bend up to 120&#176; and to elongate up to 66% of the initial length. The module generates a maximum force of 47 N, and its stiffness can increase up to 36%.

This paper describes the design and fabrication of a soft unit for surgical manipulators. The base module includes three flexible fluidic actuators to achieve omnidirectional bending and elongation, and a granular jamming-based mechanism to enable stiffness control. A complete mechanical characterization is also reported.

设计和弹性股软模块化机器人微创手术制造

In recent years, soft robotics technologies have aroused increasing interest in the medical field due to their intrinsically safe interaction in ...

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; represented by &#34;devices&#34; created as combinations of individual genetic components. However, manual assembly of such large numbers of devices is time-intensive, error-prone, and costly. The increasing sophistication and scale of synthetic biology research necessitates an efficient, reproducible way to accommodate large-scale, complex, and high throughput device construction.
Here, a DNA assembly protocol using the Type-IIS restriction endonuclease based Modular Cloning (MoClo) technique is automated on two liquid-handling robotic platforms. Automated liquid-handling robots require careful, often times tedious optimization of pipetting parameters for liquids of different viscosities (e.g. enzymes, DNA, water, buffers), as well as explicit programming to ensure correct aspiration and dispensing of DNA parts and reagents. This makes manual script writing for complex assemblies just as problematic as manual DNA assembly, and necessitates a software tool that can automate script generation. To this end, we have developed a web-based software tool, http://mocloassembly.com, for generating combinatorial DNA device libraries from basic DNA parts uploaded as Genbank files. We provide access to the tool, and an export file from our liquid handler software which includes optimized liquid classes, labware parameters, and deck layout. All DNA parts used are available through Addgene, and their digital maps can be accessed via the Boston University BDC ICE Registry. Together, these elements provide a foundation for other organizations to automate modular cloning experiments and similar protocols.
The automated DNA assembly workflow presented here enables the repeatable, automated, high-throughput production of DNA devices, and reduces the risk of human error arising from repetitive manual pipetting. Sequencing data show the automated DNA assembly reactions generated from this workflow are ~95% correct and require as little as 4% as much hands-on time, compared to manual reaction preparation.

Here, an automated workflow to perform modular DNA &#34;device&#34; assembly using a modular cloning DNA assembly method on liquid-handling robots is presented. The protocol uses an intuitive software tool for generating liquid handler picklists for combinatorial DNA device library generation, which we demonstrate using two liquid handling platforms.

模块化 DNA 设备的自动机器人液体处理组件

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and directed within a layer of paper. Moreover, stacking multiple layers of patterned paper creates sophisticated three-dimensional microfluidic networks that can support the development of analytical and bioanalytical assays. Paper-based microfluidic devices are inexpensive, portable, easy to use, and require no external equipment to operate. As a result, they hold great promise as a platform for point-of-care diagnostics. In order to properly evaluate the utility and analytical performance of paper-based devices, suitable methods must be developed to ensure their manufacture is reproducible and at a scale that is appropriate for laboratory settings. In this manuscript, a method to fabricate a general device architecture that can be used for paper-based immunoassays is described. We use a form of additive manufacturing (multi-layer lamination) to prepare devices that comprise multiple layers of patterned paper and patterned adhesive. In addition to demonstrating the proper use of these three-dimensional paper-based microfluidic devices with an immunoassay for human chorionic gonadotropin (hCG), errors in the manufacturing process that may result in device failures are discussed. We expect this approach to manufacturing paper-based devices will find broad utility in the development of analytical applications designed specifically for limited-resource settings.

We detail a method to fabricate three-dimensional paper-based microfluidic devices for use in the development of immunoassays. Our approach to device assembly is a type of multilayer, additive manufacturing. We demonstrate a sandwich immunoassay to provide representative results for these types of paper-based devices.

三维纸基微流体装置的制造用于免疫分析

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

A Simple and Scalable Fabrication Method for Organic Electronic Devices on Textiles

Today, wearable electronics devices combine a large variety of functional, stretchable, and flexible technologies. However, in many cases, these devices cannot be worn under everyday conditions. Therefore, textiles are commonly considered the best substrate to accommodate electronic devices in wearable use. In this paper, we describe how to selectively pattern organic electroactive materials on textiles from a solution in an easy and scalable manner. This versatile deposition technique enables the fabrication of wearable organic electronic devices on clothes.

In this paper, we present a protocol to selectively deposit organic materials on textiles, which allows for the direct integration of organic electronic devices with wearables. The fabricated devices can be fully integrated in textiles, respecting their mechanical appearance and enabling sensing capabilities.

关于纺织品有机电子设备的简单和可扩展的制作方法

Today, wearable electronics devices combine a large variety of functional, stretchable, and flexible technologies. However, in many cases, these devices ...

Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable microfluidic devices. Here, a protocol for the fabrication of plastic microfluidic devices is demonstrated, where soft lithographic techniques are used to embed transparent Calcium Fluoride (CaF2) view-ports in connection with observation chamber(s). The method is based on a replica casting approach, where a polydimethylsiloxane (PDMS) mold is produced through standard lithographic procedures and then used as the template to produce a plastic device. The plastic device features ultraviolet/visible/infrared (UV/Vis/IR) -transparent windows made of CaF2 to allow for direct observation with visible and IR light. The advantages of the proposed method include: a reduced need for accessing a clean room micro-fabrication facility, multiple view-ports, an easy and versatile connection to an external pumping system through the plastic body, flexibility of the design, e.g., open/closed channels configuration, and the possibility to add sophisticated features such as nanoporous membranes.

A protocol for the fabrication of plastic microfluidic devices with transparent view-ports for visible and infrared light imaging is described.

生产塑料微流控装置与查看端口对可见光和红外透明软光刻过程

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable ...