Support for this work was obtained by the VA Rehabilitation Research &amp; Development National Center of Excellence for the Medical Consequences of Spinal Cord Injury (VA RR&amp;D #B9212C). Two of the four powered exoskeleton devices were used on a loaner basis at no cost from ReWalk Robotics, Inc. (Marlborough, Massachusetts). Additionally a portion of participants obtained Orthopedic shoes which were donated by Aetrex Worldwide Inc. (Teaneck, New Jersey).
Assistance from Denis Doyle-Green was invaluable during the training program and we thank him for this. We would also like to thank the physical therapists from the Rehabilitation and Spinal Cord Injury Services at the James J. Peters VA Medical Center for their advisement and consultations.

注意:这个手稿中描述的培训协议题为试点项目中的开发:在ClinicalTrials.gov标识符NCT01454570注册"为截瘫者的ReWalk外骨骼步行系统"。制定了培训方案，但是没有这一试点项目的目标;进行这项研究的过程中，培训计划的发展变化。该研究方案和知情同意书进行了审查，并由JJPVAMC的机构审查委员会（IRB）的批准。整个研究和程序，每个研究参与者进行了解释。潜在的参加者有机会提出问题，并鼓励采取之前同意根据需要尽可能多的时间。 1.受试者招募<ol><li>与潜在的参与者进行预筛选评估。 <ol><li>简要说明训练过程的细节（研究的长度，每周次，每天小时）。说明参与的已知风险（皮肤擦伤，如果一个人落在潜在的伤害和不可预见的事件的可能性）。 注:在本协议中的训练课发生每周3次，60间一直持续到90分钟。培训课程的具体频率和/或持续时间，以不需要为参会人员学习使用动力服。 <ol><li>描述医疗限制的潜在参与者，如:在髋关节或膝关节骨密度低，近期骨折史，无法忍受站立，弱的上肢力量和薄弱躯干控制。审查，以便继续筛选过程中入选和排除标准。鼓励潜在的参与者发问。提供了充足的时间来解决任何问题并回答问题。 注意:此处所用的具体动力服的人体测量限制被施加到inclusi对标准:身高&lt;160&gt;190厘米和重量&lt;100公斤。 </li></ol></li></ol></li><li>如果该预筛选评估是成功的，然后提供该研究的详细说明，并启动筛选过程。 <ol><li>执行双能X线骨密度仪（DXA）扫描到的髋关节和膝关节双边评估骨密度。 注意:该设施具有有限的使用外骨骼的人员谁具有在全髋关节和股骨颈的T分数为大于-3.5和骨密度在近侧胫骨和股骨远端的是超过0.60克/厘米2。这些值不消除骨折的危险，但在努力减少的风险被选择。鼓励临床医生检讨相关文献，并根据其信息31-33解释调整值。 </li><li>为执行SCI（ISNSCI）的神经学分类国际标准37考试EVAL审视你们的伤害程度，运动功能和感觉。 注意:在此试点研究性学习的参与者损伤的不同级别被列入并表1给出截瘫的人是主要用户。然而与谁具有用于各个肌肉群的4或更好的上肢运动评分，并能保持平衡与拐杖可以是候选使用该外骨骼，以及其他动力服颈椎损伤的人。 </li><li>获得下肢领域一般病史和身体评估，其中也包括肩部，臀部和膝盖的运动范围，和皮肤检查和下背部接触的动力服。 注:有限的髋关节和20度屈曲或以上在任一关节活动范围膝人被排除在外。此外，肩膀必须有足够的运动范围，以达到合适的拐杖的位置进行静坐于─站，站到坐演习。参与者还必须是自由的下肢的任何压力性溃疡，尤其是任何与外骨骼直接接触面积。这些标准可以针对每个设备而异，该临床医师应参考的制造商为特定于该动力服的要求。 </li></ol></li></ol> 2.配件注意:拟合程序是由装置的制造开发的。一个人嵌合到该装置的方法也将不同的外骨骼而异。临床医生应该是指每个具体制造商的程序。 <ol><li>放置参与者在仰卧位置。使用灵活的卷尺，确定骨盆宽，大腿长度和小腿的长度和纪录厘米。 <ol><li>从臀部到膝关节线的大转子的最突出的点测量的上腿的长度。测量行吟诗人ř肢中相同的方式。记录所有的肢体长度差异。根据在每个参与者的上腿部的长度测量的距离调节从臀部轴的中心的供电外骨骼到膝盖轴的中心。 </li><li>测量从膝关节线脚的底部小腿的长度。重复测量的其它小腿的长度。调整从脚板的底部的长度到膝盖轴的上，用于根据从参与者测得的距离的每个下肢的动力服的中心。 </li><li>调整使用不同大小的骨盆带已启动外骨骼的宽度。通过将参与者坐姿椅子或板凳上，一开回选择骨盆带。在大小最接近的骨盆带的地方，后面的人参与者的骨盆宽度慢慢移动转发到测试它的一个合适的。允许可达1厘米的间隙对骨盆带的两侧。 注意:其他动力服都以不同的方式调整，并适当调整应根据制造商的规范来获得。 </li><li>选择合适的骨盆带的大小后，贴上骨盆带在空档或中央位置上的胸部立柱。后参与是在如果需要的话，使转子与所述髋关节的旋转排队位置，适当地调节前/后位置。 注意:骨盆带可以被设定为使得它可以以前方或后方推臀部。空档或中央位置的骨盆带至它允许前方或后方等量的调整的设置。 </li></ol></li><li>适合与通过去除参与者的鞋，从鞋除去鞋垫调整脚踏板，然后将最大脚板可能进入鞋。放置在底板的顶部的鞋垫。调节脚板的背屈援助通过微调在脚踝处的弹簧机构的张力。 </li><li>在完成所有的测量后，系统现在准备好由参与者穿上。 </li></ol> 3.穿戴注意:佩戴程序是由装置的制造开发的。穿上一个人的动力服的方法，可以在不同的设备之间的变化和临床医生应该是指制造商的程序。 <ol><li>将外骨骼与魁梧开放的椅子上坐着的位置。 注:理想的座椅具有很宽的软垫座椅，并且必须没有扶手或车轮。 <ol><li>指导参与者发生在一个很小的角度旁边坐着的外骨骼的轮椅。 </li><li>向参与者通过对他们的轮椅放置一只手在设备上，另一传送到设备中。确保参与者执行在一个连续的运动的传递到设备中。如果日Ë参与者无法完成传输于一体的运动，鼓励他们的外骨骼上"大腿"部分暂时休息，并恢复与第二运动转移。 注意:如果需要，可设置有转印援助。 </li></ol></li><li>参与者在装置正确就位后，指示参与者第一放置在鞋的脚，然后继续紧固带子开始在最远侧点和近端移动身体，与胸带精加工。 <ol><li>如果需要，使用手动控制功能，可以稍微弯曲髋关节和伸膝关节，以使脚更容易放置到鞋子。 </li><li>小心地将脚伸进鞋，采取特殊照顾，以保证脚趾不卷曲。一旦脚是正确的鞋子，使用手动控制移动腿和脚放回地面，然后收紧鞋。按照同样的步骤，以确保妥善安置<html>第二脚伸进鞋。 </li><li>在鞋固定脚后，固定带正下方的膝盖，其次是固定膝盖和那些大腿上部上面带。注意避免衣物褶皱肩带下方，以避免不必要的摩擦和/或接触的压力点。固定在下和上胸部背带最后。 </li></ol></li><li>一旦参与者绑到设备中，检查是否有任何不当接触，皱巴巴的衣服或压力点。 <ol><li>以下的压力点的识别，通过调整配合和添加或移除填充适当释放压力。 注意:检查合身而坐，站立和行走了几步之后。站立和行走这可能导致可能通过重新检查被识别的接触的附加分之后当参加者可能会稍微移位。 </li></ol></li></ol> 4.常委ENT"&gt;注:要站立起来的过程是由设备的制造和开发不同的外骨骼之间可能会有所不同临床医生应该是指制造商的程序。 <ol><li>嵌合后，引入与会者到外骨骼的整体功能。解释特定于设备的控制器。通知S中的参与者，他/她将有望与外骨骼变得尽可能的独立。解释说，独立将通过学习来控制设备的功能和有短期和长期的目标，成为独立完成。 注意:最初，训练者操作控制以启动动力服的所需运动，然而重要的是，该用户被引入到设备并意识到它如何尽早在训练过程中发挥作用。一旦在设备舒适，用户接管控制，并开始自己的动作。 </li><li>装备ŧ他参加者与一组前臂拐杖以协助装置的平衡和可操作性。而坐在外骨骼，指导参与者放置拐杖的提示后方允许他们在自己的脚推自己的体重的能力的方式。这确保了外骨骼可以延长髋关节和膝关节立起机动期间执行的大部分工作。 需要在包括站立，行走，转弯，坐在该设备的所有演习拐杖:注意。参赛者不得使用外骨骼没有拐杖。有些动力服可以允许使用学步车或拐杖以保持平衡。 </li><li>解释静坐到站姿过程的参与者。有一个教练从用户背后协助，并从前面的另一个后卫。指示参与者站在自己只有用教练的援助需要。 <ol><li>指导参与者向后放置拐杖和身体前倾，同时推动了拐杖协助设备按"定海神针"命令后，站在他们。 注意:首先，鼓励用户专心适当拐杖放置而训练者使用所述控制器启动动力服到站立。 </li></ol></li></ol> 5.站立平衡注:站立平衡过程是由研究人员在开发JJPVAMC。可能有一些程序，针对于当前使用，但大多数的程序应翻译成其他动力服的设备。 <ol><li>有一个教练从后面站着守卫后，有第二个教练站在用户面前，并展示了站立平衡的目标。 注意:在训练会话期间站在周期性识别如果体位性低血压或自主神经异常反射情节被使用者经历后测量血压。 </li><li>前ttempting走路，确保参与者演示了以下功能: <ol><li>有与会者演示使用双拐来保持平衡（ 图1），在"家"的位置站立的能力。 注:放置在参与者面前的一面镜子提供视觉反馈，并帮助改正不正确的倾斜，以及保持"家"定位自己的直立平衡。 </li><li>有自己的体重的参与者实践轻微横向转移和向后理解的位置，感受家的位置。 </li><li>指示参与者保持只用一个拐杖（ 图2）的平衡。指导参与者分别抬起拐杖掉在地上，并保持这一立场长达1分钟的时间练习这个动作。指导参与者实行额外的单手平衡运动。 注意:此动作类似于上一个，但与添加的复杂<html>有一个手臂平衡而对侧手臂达到了触摸平衡臂的手腕，模拟的控制器上的操作选择性。 <ol><li>重复这些练习，以保证参与者能够进行使用或者手臂保持平衡这些演习。 </li></ol></li><li>练习站立平衡的拐杖技能后，教参与者重心偏移横向，让一只脚卸载，其目标是完全抬起脚离地面5秒。指导用户重复这个练习，试图用另一条腿卸载。 </li><li>请参与者重量前后方向转变，同时妥善安置拐杖前后保持平衡。 </li><li>重复练习5.2.2 - 第一届会议期间5.2.5五至十倍。继续，直到用户感到舒适与他们在接下来的会话练习这些练习。 </li></ol></li></ol>"&gt; 6。散步注意:行走过程是与由工作人员在JJPVAMC开发过程的混合物中的设备的制造。的行走内置入动力服，并在装置中使用的双拐杖图案的机理是由制造开发的;然而，教参与者如何正确执行走路的办法，提供援助和用于记录的援助水平的测量结果的机制是研究人员在JPVAMC的努力。虽然，一些程序是特定于所使用的动力服，大部分的程序翻译到使用拐杖保持平衡其他动力服。 <ol><li>指示参与者与动力服行走的机制。使用的特定动力服，需要参与者自己的体重转嫁到左脚，同时失重右脚。使用控制器，训练者选择"步行"模式，并要求参与者稍微向前移动（到预定的目标）;这将引发右腿向前摆动。 <ol><li>指示，一旦右腿已完成摆动，向前推进他们的拐杖，同时为了保持平衡，而踏上了右脚，和失重左脚转移自己的体重向前，向右用户。解释该装置，检测参加者的运动，将启动向前左腿的摆动。 </li><li>通过重复着crutching运动和重量转移序列，每条腿连续进行连续行走。 </li></ol></li><li>鼓励教员根据需要提供援助，但这样做最低限度。 注:救助水平，由功能独立措施（FIM）38确定的，由培训师进行评估和记录。 <ol><li>通过抓住在p发现用户owered外骨骼或参与者在需要时提供支持。更正用户为他/她进行适当的重量边走边转移。 </li><li>如果需要的话，有一个第二训练，其中用户具有完整的感觉（如肩膀）的主体的区域提供援助和战术反馈。 注意:因为用户通常不能够感觉到的协助下，这可能导致难以在学习调整自己的上身在设备正常走动教练劝阻通过动力服或低于损伤的水平提供帮助。 </li></ol></li><li>向用户讲解了动力服停止行走的机制。所用的具体动力服被触发时停止它不会感觉到任何更向前移动到对侧肢体，或者如果用户不提供一个适当的重量偏移允许摆动腿以与地面接触。 注:随意或持平停止满足特殊位置和实行的是包括培训方案的技能之一。 </li></ol> 7.行动训练的渐进目标注意:所述移动性训练的目标均在JJPVAMC开发的标准内掺入用于评估能力由制造使用动力服在家庭环境中。 <ol><li>解释和描述的移动性技能的列表来实施作为训练（ 图3）的一部分。 <ol><li>指示参与者使用动力服的控制器和使用该供电外骨骼变得尽可能独立。 注意:在此试验中使用的动力服具有集成在戴在手腕上的控制器控制。 </li><li>教参与者，使90和180度的转弯行走在系统一段时间。 </li><li>指导参与者通过停止墙边导航休息在墙上和转弯所以他们回来倚靠它。 注意:这允许人休息，而不需要依靠拐杖平衡。 </li><li>在培训课程，使参与者的做法额外的表面行走，如地毯（ 图4），混凝土，沥青和草（ 图5）把不同的行走表面。 </li><li>对表面参与者走了不同的斜坡，例如一个斜坡，下斜坡，路缘切口和不平的表面（ 图6）。然后，在一个嘈杂的环境中参加步行路程，如与其他行人走廊。 注意:走在嘈杂的环境中可能对某些人来说是一个挑战，因为他们无法听到它提供了一个音频阙一个合适的时机，以重心移动马达的声音。 </li><li>有与会者停止命令或随意。 </li><li>门口的阈值的做法导航，打开和摆动做闭幕ORS，开从不同的侧面关门，并穿行自动和/或旋转门（ 图7和图 8）。 注意:在执行这些额外的行动技能的能力被评定为"能"或"不能"来执行动作。 </li><li>包含更多的活动，如达到头顶到机柜（ 图9）或外部坐在和公园的长椅上站起来（ 图10）。 </li></ol></li></ol> 8.散步的评估注意:使用的步行评估是已经由他人建立标准的临床试验。 <ol><li>执行6分钟步行试验（6MWT）。 <ol><li>有参与者发起的步行和指导参与者继续走。 </li><li> 6分钟后请参与者停止。 注:6MWT 39,40是参与者能够感到的距离在未来6分钟周期时间的动力服布拉特。如果参与者不慎触发装置6MWT期间停止行走，时钟继续记录时间和参与者被鼓励恢复他/她的平衡，镇定，并重新启动设备继续尽可能快地行走。 </li></ol></li><li>一个专门为斑点教练和使用测量轮确定的距离的额外培训师和秒表测量经过的时间进行测试。 </li><li>表达6MWT在米中6分钟行走并计算平均步行速度（总米6分钟/ 360秒走），并将它表示为米/秒。 注:6MWT是一个在6分钟定时期间走过的总距离，并通过培训项目的过程中获得。该6MWT是用于确定在所述外骨骼步行领域的进展的主要评估。只要参与者理解机甲执行6MWT测试与动力服行走和NISM能够采取多个步骤。 </li><li>覆盖10米的距离，记录10米时间后使用秒表的单圈功能6MWT中。识别并记录6MWT期间所取得的十佳米计时。 注:10米步行试验（10MWT）40是尽力而为时间（秒），它需要参与者多走10米的距离，虽然人进行6MWT被记录下来。 </li><li>使用定时向上-走（TUG）40,41试验为多少地位的一个指标，走车，坐在功能个人拥有。 <ol><li>通过测量它需要参与者从坐姿站起来，走10英尺转身，往回走，再次坐下来的时候进行测试TUG。开始的时候人一旦启动设备站起来的时候停止，一旦人被安全地坐在位子。 注意:这种测量的结果并不代表（繁体）的TIONAL TUG时间，因为它包含分配给合适的拐杖放置的时间后，将模式选择指示的地位是需要的。拖船测量代表了人的，因为它结合了移动性的多个方面的设备使用外骨骼系统的能力。 </li></ol></li></ol> 9.坐注意:程序坐下由装置的制造进行了开发和不同外骨骼之间可能有所不同。临床医生应该是指制造商的程序。 <ol><li>将用户后面的椅子时，他或她准备坐。使用外骨骼的控制器，放置在仰卧模式外骨骼。 注意:最初，训练者操作动力服的坐运动期间，控制器，然而，与站立，重要的是用户被引入到控制器和训练亲期间意识到其功能尽可能早公克。一旦在设备舒适，要求用户操作控制器，并启动动作。 </li><li>激活后/按静坐命令有一个5秒的延迟。在此期间，请参与者向后将他们的拐杖，以保持其资产负债搭在椅子的中心。有与会者练习拐杖安置任务，如果这是执行坐函数的第几次。 5秒延迟已经过去之后，所述外骨骼降低用户向下直到坐在椅子上。 </li><li>在坐在进程的用户将开始向前弯曲在臀部保持在脚的平衡。有需要的教练协助参与者。 注:最初，练习坐了两个教练，从背后的一个斑点，另一个在前面。当用户成为熟练的动作，并能够与自信和独立完成动作，只需要一个教练。 </li></ol> 10. DoffiNG 注意:落纱程序是由装置的制造开发的。落纱的动力服的方法可以在不同的设备而异。临床医生应该是指制造商的程序。 <ol><li>休息之后，脱下该装置类似的，但在三节的穿上了设备前面所讨论的相反的方式。 <ol><li>发行版开始与胸部和臀部和进步对脚的带子。从设备中删除参与者的脚。鼓励参与者试图自行转移到他们的轮椅，但需要提供援助。 </li></ol></li><li>一旦回到自己的轮椅，检查参与者的脚，下肢和腰部任何瘀伤或擦伤。 </li><li>教参与者定期检查他们的下肢的压力点的迹象，他们已经完成了他们行走会议之后。 </li></ol>

The authors have nothing to disclose.

在过去的五年中，我们的小组已开发成功的筛选和参与者使用需要拐杖动力服的类型的培训计划。我们已经训练了与电机完全瘫痪，以及那些有不全瘫的个体。这个训练程序必须进行修改和在用需要使用拐杖，或在现有设备的较​​新版本的附加设备内置的潜力。 一个培训项目的标准化是非常重要的，以保证参与者的安全，成功地利用该装置，发现人力资源，并获得一致的结果。在一个良好的培训计划，重点包括适当人选的选择，设备的适当配合，适当的技能的进展，并在肩上，或与完整感的地区提供援助，以使用户识别所需要的力和运动，促进适应在他们的动向随后加紧行动。它练习教练和用户之间的这一战略的舞蹈，以尽量减少教练的支持，从而有助于在设备的用户获取专业知识和独立性是很重要的。培训师应该避免，因为在困难这一行动的结果成为独立于外骨骼帮助之下轰动的参与者的水平。另一个关键点，以改善行走技能是走在各种表面，并在不同的环境挑战的参与者。与会者认为在室内行走和在医疗中心平坦/光滑的表面会比走动在地毯更容易。走在铺着地毯的地板，反过来，据报道，比在不平整的表面，如混凝土或沥青户外行走更容易。走来走去不同坡度迫使参与者，因为重量转移的方法变得更具挑战性调整自己的战略，行走由于平衡prese的改变中心由斜坡nted。所有这些挑战性的环境中，通常在社区内遇到的，因此，是实践在受控环境好好准备参加非常重要的。 目前已在脊髓损伤谁已经学会了使用动力服安全地走动地上16-19,21,36者的几份报告。许多这些报告中的参与者几乎没有在他们的下肢无残留功能或感觉。无严重不良事件从这些研究报告和设备被认为安全的适当的培训来使用。报告的不良反应包括皮肤擦伤，青紫或皮肤发红，上肢乏力，尤其是在最初的培训课程16,19,36。有人指出，随着持续的培训，学员注意到减少与设备的更好拟合很快就解决了上肢疲劳和皮肤擦伤。福TURE青紫，红肿避免了与肩带调整和周围的患处额外填充的战略布局。 熟练使用的装置是由以实现更快下地的速度，减少的援助水平，并在不同的环境中安全行走的能力来确定。的行走能力此前的报告显示，那些谁更独立会走动谁比那些需要援助的速度更快。报告由Van Hedel 等分为步行者为"辅助步行者"如果他们能与0.44±0.14米/秒的最小速度走动。与那些谁选择了与援助在使用他们的轮椅42户外散步有关的速度。这个步行 ​​速度类似于中风的人报告的有限社区ambulators的0.40米/秒的速度。43虽然只有少数研究使用机器人exoskelet报道行走速度和辅助级别插件，这些研究表明，许多参与者都能够实现这些以前的报告中提到的0.40米/秒的行走速度。使用动力服的一份报告显示，7月12日的参与者们能够更快地走动0.40米/秒18。使用不同的动力服另一项调查能够说明16 6的参与者成功走动大于0.40米/秒36。虽然使用第三个动力服的报告没有证实为0.40米/秒22,44，未来的报告可能显示增加行走速度与该装置进一步培训和/或自适应速度行走。到目前为止，使用动力服所有的研究都报道那些需要援助的更高水平走在较慢的速度。在这些报告中讨论的一个想法是，尽管一些与会者没有走动0.40米/秒的速度之上，他们能够在"监管"的水平走动作为FI定义中号的规模。这些报道表明，用另外的训练或修改的装置，行走在这些更快的速度，可以实现。 通过氧消耗测量的能量支出已被证明与外骨骼辅助行走增加，但不高于即过分疲劳的阈值。八参与者谁在动力服在0.22±0.11微米的平均步伐ambulated /秒证明行走的氧消耗率11.2±1.7毫升/千克/分钟，118±21的bmp（48％±16％心脏速率储备的心脏速率），而这两者都是从坐在和站立17，但显著低于最大预测值的显著增加。使用不同的动力服另一报告中，评价氧消耗在5与会者在2较量行走并报告9.5±0.8毫升/千克/分钟，在0.19行走时±0.01米/秒和11.5±1.4毫升/公斤/时在步行分钟0.277; 0.05米/秒21。这两项研究表明，在参加中等强度走动均高于通过运动医学的美国学院确定的最低训练强度阈值心肺受益45是有效的。这表明，这些设备具有被用于更长的时间的可能性，提供活动，如果定期执行预期可能导致在用户的健身，身体组成和脂质分布的改进的形式。 该动力服提供站立和行走地上与上肢功能者修改独立性（六级由FIM定义）的一种形式。未来的设备可以被设计以更快的速度走动或提供更大的改变所需的行走速度的能力。未来外骨骼也可以被设计为那些具有有限的手和臂的功能（例如与四肢瘫痪）由maintainin克用户与附加躯干支撑和比保持一个拐杖用于保持平衡提供了另一种机制平衡。大脑控制的进步也许有一天可以被纳入到控制步行运动20。在这个新兴的领域，提出了基本训练概念可以适用于当前和未来的动力服，而应针对用户和正在使用的外骨骼。 标准化的培训战略，目前用于成功的参与者外骨骼辅助行走;这些设备的将来的修改可能需要adaptions到训练模式。教学中的素质SCI保健专业人员适当训练脊髓损伤的人来执行需要对这些设备的持续使用和处方外骨骼辅助行走。前途是光明的这些设备;用SCI的人使用动力服会成为与T更为广泛他建立的培训计划在全球医疗和康复中心。此外，未来的研究可以表明，经常外骨骼辅助步行改善许多与脊髓损伤静止和麻痹二级医疗并发症。

脊髓损伤（SCI）的许多人都无法站立，并使用或不使用辅助设备和物质帮助的走动。几个世纪以来，对于那些严重SCI唯一的移动选项已经轮椅1。在过去的几十年里，随着SCI人不得不通过补充采用被动矫形器其流动性的选择，如各种往复式步行矫形器（RGO）2-7。这些装置，但是，并没有由于由用户使用这些设备走动所需的体力变得更广泛地使用。该RGOS也有爬楼梯，站起来，坐 ​​下3,7的能力限制。已经作出努力通过将功能性电刺激（FES）供电的移动，并有助于促进肢体的向前摆动以提高这些装置的效率;然而，这些努力没有进展超出了概念或原型8-12。在20世纪70年代，马达用矫形器整合到髋关节和膝关节的运动力量和成功地允许与SCI的人采取措施13。然而，时间电池和计算机技术不足限制了设备的范围，并进一步发展被放弃10,13。 随着近来的技术进步，若干动力服已开发以实现与各种病症的人到地上走动。这些动力服的设备已被研究人员与卒中14,15，与导致减少了他们的下肢25-27控制残疾完全和不完全SCI 16-24，和其他人的人。虽然设备不同，每一个需要用户安全性能的训练和练习。所引用的设备的三个要求使用拐杖的走动和保持平衡。第四个保持balance和稳定性，因为它的大踏板和大规模扩大其支持基础并降低重心20的中心。那些需要crutching三个设备利用相同的原理，即使有一些变型与力学和控制所需的操作的方法，由于在装置的设计上的差异。 的培训计划是由一组由生物医学工程师，生理学家，理疗师，运动生理学家，神经科医师和物理治疗师的研究人员在詹姆斯·J·彼得斯VA医学中心（JJPVAMC），布朗克斯，纽约发展。该培训项目与一个特定的动力服研制前面所述17,18，但它采用了两套技能，适用于需要一套拐杖来保持平衡其他动力服。所有潜在的参与者参加了循序渐进的训练计划前筛查。的重要性筛选在脊髓损伤的人是要确保没有可能抑制安全使用这些设备的禁忌医疗并发症。关注的一个领域是低骨矿物质密度（BMD）。脊髓损伤者受损28,29后立即剧烈的骨质流失可能继续在他们的生活30。骨密度这种损失导致长骨骨折的危险性高。目前，有减轻为那些与完整的电机的SCI的骨损失没有有效的治疗方法。此外，对于人SCI患者建立的骨折阈不存在，但已经进行了努力，以确定其可与临床判断和骨折史一起使用作为指导31-33标准。其他常见的禁忌症可被处理和解决的，如运动（ROM）中的范围的限制34和压疮35。每个不同的动力服可能需要资格不同的条件，诸如ROM的标准，是一个甘地日期使用该设备，其中大部分已被描述17-19,21,22,36。 一个人一旦成功通过所有的筛选标准，装修设备到用户和培训可以继续。该装置的正确配合是很重要的，以避免与外骨骼下肢不适当的接触，因为装修不好可能会导致瘀伤和/或皮肤擦伤16。用户可能具有限制或无下肢感觉和本体;这种缺乏从脚感觉和触觉反馈可以促进整体缺乏资产负债的中心意识，减缓用户的掌握设备的能力。这种缺乏平衡的中心的意识也可能导致与适当重量困难衡量向前的程度移位这种挑战和步态周期和不适当定时重量中横向偏移必要换档，导致过量使用的负重上该武器和平衡维护拐杖。一旦站立平衡和重量转移被收购的基本机制，用户传授给走在设备中。需要多个会话，以改善步行和其它移动技能。起初，表面是平坦和医疗中心内平滑用于训练。然而，具有改进的技术水平，用户通过引入不同行走表面如地毯，沥青，混凝土，草，有不同程度的斜坡的不平整表面的挑战与增量更加困难的任务。 这份手稿的目的是报告的筛选标准，正确的安装和培训程序使用了地上行走一个动力服。这项计划是专门为一台设备，由他人16-18描述的发展，但它解决了常见的员工培训师和人与SCI谁参与方面的挑战外骨骼辅助ED走这可以用另一个动力服的方案。这个协议的某些方面是特定的JJPVAMC使用的设备。此外，一些培训计划的组​​成部分是由它包括设备元件的方向，对于一个适当的配合和基础地位的基本准则和坐在技能说明制造开发。在JJPVAMC的研究人员开发，一旦用户执行的所有培训活动中站立起来。这些措施包括增强的地位和坐姿训练指示，站立平衡技巧，室内步行发展技能，户外散步进展技能，并为达到，停止，转弯等机动任务，及各类门/门限导航。

<table border="0" cellpadding="0" cellspacing="0" width="464">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:464px;">Powered Exoskeleton such as ReWalk&#8482;, Ekso&#8482;, REX&#174;, and Indego&#174;, etc.&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Loft strand Crutches</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Comfortable sneakers</td>
		</tr>
	</tbody>
</table>

training persons with spinal cord injury to ambulate using a powered exoskeleton

Powered exoskeletons have become available for overground ambulation in persons with paralyses due to spinal cord injury (SCI) who have intact upper extremity function and are able to maintain upright balance using forearm crutches. To ambulate in an exoskeleton, the user must acquire the ability to maintain balance while standing, sitting and appropriate weight shifting with each step. This can be a challenging task for those with deficits in sensation and proprioception in their lower extremities. This manuscript describes screening criteria and a training program developed at the James J. Peters VA Medical Center, Bronx, NY to teach users the skills needed to utilize these devices in institutional, home or community environments. Before training can begin, potential users are screened for appropriate range of motion of the hip, knee and ankle joints. Persons with SCI are at an increased risk of sustaining lower extremity fractures, even with minimal strain or trauma, therefore a bone mineral density assessment is performed to reduce the risk of fracture. Also, as part of screening, a physical examination is performed in order to identify additional health-related contraindications. 
Once the person has successfully passed all screening requirements, they are cleared to begin the training program. The device is properly adjusted to fit the user. A series of static and dynamic balance tasks are taught and performed by the user before learning to walk. The person is taught to ambulate in various environments ranging from indoor level surfaces to outdoors over uneven or changing surfaces. Once skilled enough to be a candidate for home use with the exoskeleton, the user is then required to designate a companion-walker who will train alongside them. Together, the pair must demonstrate the ability to perform various advanced tasks in order to be permitted to use the exoskeleton in their home/community environment.

以下测量在整个训练获得。双手和单手拐杖平衡能力各自评估1分钟为"能"或"不能"，以保持平衡（ 图2）。时间和距离的散步评估在整个使用6MWT，10MWT和TUG培训课程获得。外骨骼辅助行走通常遇到的表面被在室内进行测试（ 图3和4）和室外（ 图5-6）。其他的移动技能，如导航门（ 图7和图 8），达到了头到机柜（ 图9），并坐在外面公园的长椅上（ 图10）被评定为"能"履行或者"不能"来执行。 在10 SE的10MWT期间的平均行走速度裂变间隔为第60届被描绘（ 图11）。本图显示的参与者必须使用用户中改进的动力服和变化的速率变化的初步能力。最佳拟合线的斜率的平均值±标准偏差是0.0048±0.004米/秒和值从0.00026介于0.015米/秒。这表明，尽管每个参与者浮动利率提高他们走0.0048米/秒的更快的每个会话的平均值。最佳拟合截距的平均值±标准差为0.16±1.8米/秒，并且值从-0.026范围至0.50米/秒。这表明，对平均参与者0.16米/秒的平均初始速度;具有几乎没有能力走动一些与会者和其他人在培训初期非常好的能力。 培训师的援助会影响性能;那些谁需要assistan更高水平CE走路比那些谁在使用该系统18个熟练和独立的慢。这三个走的测试测量，虽然类似，提供不同水平的信息。的10MWT为速度（m /秒）的尽力而为的指示，用户能够在设备中走动。该6MWT距离，换算为米/秒的速度时，提供的平均行走速度是走在了外骨骼的一致性的指示。由于定时器仍然当用户无意中停止行走，从6MWT速度更接近尽力而为10MWT表示对方有一致的行走和更少的停止。拖船需要在连续的组合进行很多技巧。拖船是人的合并站立，行走，转弯，停止，并在动力服坐下整体能力的度量。的6MWT，10MWT和TUG测量的概述已经由杨1先前描述<html> 8和在表1中的参与者的病人的人口统计信息一起呈现。 <img alt="图1" src="/files/ftp_upload/54071/54071fig1.jpg" /> 图1. 双手拄着拐杖的平衡。这个数字说明了一个人呆呆地站在那里，用双拐平衡。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig1large.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/54071/54071fig2.jpg" /> 图2.单手拄着拐杖的平衡。这个数字说明了一个人呆呆地站在那里，只有1拐杖平衡。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> ove_content"FO:保together.within页="1"&gt; <img alt="图3" src="/files/ftp_upload/54071/54071fig3.jpg" /> 图3. 在室内行走在光滑的表面。这个数字说明了一个人在一个平面上行走在室内， <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig3large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54071/54071fig4.jpg" /> 图4. 走在地毯上。这个数字说明了一个人在地毯表面行走在室内， <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig4large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54071/54071fig5.jpg" /> 图5。 </stro<html> NG&gt; 户外行 ​​走在草地上。这一数字说明了一个人在草地户外散步。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/54071/54071fig6.jpg" /> 图6. 在斜坡上行走。这个数字说明了一个人在户外走路边缺口。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig6large.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/54071/54071fig7.jpg" /> 图7. 导航电梯。这个数字说明了一个人走了定时门的设置，如电梯门。jove.com/files/ftp_upload/54071/54071fig7large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图8" src="/files/ftp_upload/54071/54071fig8.jpg" /> 图8. 走出旋转门的。这个数字说明了一个人走了旋转门。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图9" src="/files/ftp_upload/54071/54071fig9.jpg" /> 图9. 架空柜和台面深远。这个数字说明了一个人，取放物品出来的开销柜。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig9large.jpg" target="_blank">请点击此处查看该figu的放大版本</a><html>回覆。 <img alt="图10" src="/files/ftp_upload/54071/54071fig10.jpg" /> 图10.公园的长椅上坐外面这个数字说明了一个人在公园的长椅上坐外面。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig10large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图11" src="/files/ftp_upload/54071/54071fig11.jpg" /> 图11. 场均十个交易日10MWT速度。数据表明由十届平均间隔训练前60届的10MWT速度。 x轴描述了会话和y轴描述了从参与者训练过程中获得的10MWT结果计算出的平均速度（m /秒）。线性最佳拟合线覆盖在每个参与者的结果。="htt​​ps://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54071/54071fig11large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td colspan="8"> 人口统计学特征 </td><td colspan="7"> 步行试验（WT）和援助水平（LOA） </td></tr><tr><td rowspan="2"> SID </td><td rowspan="2"> 年龄 （Y） </td><td rowspan="2"> HT （厘米） </td><td rowspan="2"> 重量 （公斤） </td><td rowspan="2"> 性别 </td><td rowspan="2"> DOI （Y） </td><td rowspan="2"> LOI </td><td rowspan="2"> AIS </td><td colspan="2"> 10米WT </td><td colspan="2"> 6分钟WT </td><td> 拖船 </td><td rowspan="2"&gt;（LOA） </td><td rowspan="2"> 评估- 换货会议 </td></tr><tr><td> （秒） </td><td> （米/秒） </td><td> （米） </td><td> （米/秒） </td><td> （秒） </td></tr><tr><td> 1 </td><td> 34 </td><td> 173 </td><td> 66.7 </td><td>男</td><td> 9 </td><td> T4 </td><td>乙</td><td> 39 </td><td> 0.26 </td><td> 90 </td><td> 0.25 </td><td> 83 </td><td>敏</td><td> 89 </td></tr><tr><td> 2 </td><td> 48 </td><td> 168 </td><td> 68 </td><td>男</td><td> 4 </td><td> T10 </td><td>一个</td><td> 62 </td><td> 0.16 </td><td> 51 </td><td> 0.14 </td><td> NP </td><td>敏</td><td> 18 </td></tr><tr><td> 3 </td><td> 44 </td><td> 183 </td><td> 77.1 </td><td>男</td><td> 4.5 </td><td> T4 </td><td>一个</td><td> 20 </td> &lt;TD&gt; 0.50 </td><td> 209 </td><td> 0.58 </td><td> 56 </td><td> MI </td><td> 63 </td></tr><tr><td> 4 </td><td> 58 </td><td> 160 </td><td> 64.4 </td><td>女</td><td> 1.5 </td><td> C8 / T8 </td><td> A（NT） </td><td> 24 </td><td> 0.42 </td><td> 139 </td><td> 0.39 </td><td> 59 </td><td> MI </td><td> 43 </td></tr><tr><td> 五 </td><td> 61 </td><td> 175 </td><td> 72.6 </td><td>男</td><td> 14 </td><td> T11 </td><td>一个</td><td> 23 </td><td> 0.44 </td><td> 137 </td><td> 0.38 </td><td> 66 </td><td> MI </td><td> 37 </td></tr><tr><td> 6 </td><td> 24 </td><td> 185 </td><td> 74.8 </td><td>男</td><td>五</td><td> T5 </td><td>一个</td><td> 56 </td><td> 0.18 </td><td> 60 </td><td> 0.17 </td><td> NP </td><td>敏</td><td> 12 </td></tr><tr><td> 7 </td><td> 40 </td><td> 183 </td><td> 88.5 </tD&gt; <td>男</td><td> 1.5 </td><td> T1 </td><td>乙</td><td> 61 </td><td> 0.16 </td><td> 51 </td><td> 0.14 </td><td> 70 </td><td>小号</td><td> 102 </td></tr><tr><td> 8 </td><td> 56 </td><td> 175 </td><td> 83.9 </td><td>男</td><td> 3 </td><td> T9 </td><td>一个</td><td> 22 </td><td> 0.46 </td><td> 151 </td><td> 0.42 </td><td> 116 </td><td>小号</td><td> 51 </td></tr><tr><td> 9 </td><td> 50 </td><td> 183 </td><td> 99.8 </td><td>男</td><td> 11 </td><td> T7 </td><td>一个</td><td> 17 </td><td> 0.59 </td><td> 208 </td><td> 0.58 </td><td> 56 </td><td> MI </td><td> 56 </td></tr><tr><td> 10 </td><td> 37 </td><td> 170 </td><td> 65.8 </td><td>男</td><td> 6 </td><td> T2 </td><td>一个</td><td> 22 </td><td> 0.46 </td><td> 150 </td><td> 0.42 </td><td> 63 </td><td>敏</td><td> 59 </td></tr><tr><td> 11 </td><td> 64 </td><td> 173 </td><td> 72.8 </td><td>男</td><td> 3 </td><td> T2 </td><td>一个</td><td> 78 </td><td> 0.13 </td><td> 46 </td><td> 0.13 </td><td> NP </td><td> MOD </td><td> 28 </td></tr><tr><td> 12 </td><td> 37 </td><td> 152 </td><td> 65.8 </td><td>女</td><td> 19 </td><td> C8 </td><td> C（NT） </td><td> 14 </td><td> 0.71 </td><td> 256 </td><td> 0.71 </td><td> 42 </td><td> MI </td><td> 39 </td></tr></tbody></table> 表1.参与者和步行测试结果的特性 SID =对象的识别号码。 Y =年; CM =厘米;公斤=公斤; DOI =伤害的持续时间; LOI =损伤水平; AIS =美国脊髓损伤协会障碍量表; LOA =援助的水平; s =秒; M =米; NP =未-演出和NT =非外伤性脊髓损伤。 LOA从FIM改编为下列之一:温和辅助器（Mod） - 参与者执行任务的50％至74％;最低限度的援助（最小值） - 用户执行任务的75％或更多;监督（S） - 教练员未接触到的参与者，但足够接近达成以提供平衡或指导需要的支持;和改良的独立性（MI） - 培训师不提供任何帮助和参与者是完全独立行走的设备，同时。重新打印在脊髓损伤的人一个动力服许可，杨A，阿斯兰P，克内热维奇S，Kornfeld S，院内行走速度和援助水平庞根A.评估。前脊髓已经来到Rehabil。 2015年; 21（2）:100-109。版权所有（C）2015年托马斯土地出版公司

Watch this Scientific Journal Video about Training Persons with Spinal Cord Injury to Ambulate Using a Powered Exoskeleton at JoVE.com

Training Persons with Spinal Cord Injury to Ambulate Using a Powered Exoskeleton

Training a person with paralysis to ambulate using a powered exoskeleton may present challenges. The goals are to present the candidate selection criteria and the training procedures for exoskeletal-assisted walking and other mobility skills that can be progressed as the participant's skill level improves.

训练脊髓损伤者走动使用动力服

Powered exoskeletons have become available for overground ambulation in persons with paralyses due to spinal cord injury (SCI) who have intact upper ...

training-persons-with-spinal-cord-injury-to-ambulate-using-powered

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

20.5K 次观看.  James J. Peters VA Medical Center. Training a person with paralysis to ambulate using a powered exoskeleton may present challenges. The goals are to present the candidate selection criteria and the training procedures for exoskeletal-assisted walking and other mobility skills that can be progressed as the participant's skill level improves. 

视频: Training Persons with Spinal Cord Injury to Ambulate Using a Powered Exoskeleton

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic valve leaflets are composed of interstitial cells suspended within an extracellular matrix (ECM) and are lined with an endothelial cell monolayer. The valve withstands a harsh, dynamic environment and is constantly exposed to shear, flexion, tension, and compression. Research has shown calcific lesions in diseased valves occur in areas of high mechanical stress as a result of endothelial disruption or interstitial matrix damage1-3. Hence, it is not surprising that epidemiological studies have shown high blood pressure to be a leading risk factor in the onset of aortic valve disease4. 
The only treatment option currently available for valve disease is surgical replacement of the diseased valve with a bioprosthetic or mechanical valve5. Improved understanding of valve biology in response to physical stresses would help elucidate the mechanisms of valve pathogenesis. In turn, this could help in the development of non-invasive therapies such as pharmaceutical intervention or prevention. Several bioreactors have been previously developed to study the mechanobiology of native or engineered heart valves6-9. Pulsatile bioreactors have also been developed to study a range of tissues including cartilage10, bone11 and bladder12. The aim of this work was to develop a cyclic pressure system that could be used to elucidate the biological response of aortic valve leaflets to increased pressure loads. 
The system consisted of an acrylic chamber in which to place samples and produce cyclic pressure, viton diaphragm solenoid valves to control the timing of the pressure cycle, and a computer to control electrical devices. The pressure was monitored using a pressure transducer, and the signal was conditioned using a load cell conditioner. A LabVIEW program regulated the pressure using an analog device to pump compressed air into the system at the appropriate rate. The system mimicked the dynamic transvalvular pressure levels associated with the aortic valve; a saw tooth wave produced a gradual increase in pressure, typical of the transvalvular pressure gradient that is present across the valve during diastole, followed by a sharp pressure drop depicting valve opening in systole. The LabVIEW program allowed users to control the magnitude and frequency of cyclic pressure. The system was able to subject tissue samples to physiological and pathological pressure conditions. This device can be used to increase our understanding of how heart valves respond to changes in the local mechanical environment.

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

A cyclic pressure bioreactor capable of subjecting heart valve tissue to physiological and pathological pressure conditions has been designed. A LabVIEW program allows users to control pressure magnitude, amplitude and frequency. This device can be used to study the mechanobiology of heart valve tissue or isolated cells.

循环压力为生物反应器的设计体外主动脉心脏瓣膜的研究

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

科研

生物工程

Microfluidic Fabrication of Polymeric and Biohybrid Fibers with Predesigned Size and Shape

A &#8220;sheath&#8221; fluid passing through a microfluidic channel at low Reynolds number can be directed around another &#8220;core&#8221; stream and used to dictate the shape as well as the diameter of a core stream.&#160;Grooves in the top and bottom of a microfluidic channel were designed to direct the sheath fluid and shape the core fluid.&#160;By matching the viscosity and hydrophilicity of the sheath and core fluids, the interfacial effects are minimized and complex fluid shapes can be formed.&#160;Controlling the relative flow rates of the sheath and core fluids determines the cross-sectional area of the core fluid.&#160;Fibers have been produced with sizes ranging from 300 nm to ~1 mm, and fiber cross-sections can be round, flat, square, or complex as in the case with double anchor fibers. Polymerization of the core fluid downstream from the shaping region solidifies the fibers.&#160;Photoinitiated click chemistries are well suited for rapid polymerization of the core fluid by irradiation with ultraviolet light.&#160;Fibers with a wide variety of shapes have been produced from a list of polymers including liquid crystals, poly(methylmethacrylate), thiol-ene and thiol-yne resins, polyethylene glycol, and hydrogel derivatives. Minimal shear during the shaping process and mild polymerization conditions also makes the fabrication process well suited for encapsulation of cells and other biological components.

Two adjacent fluids passing through a grooved microfluidic channel can be directed to form a sheath around a prepolymer core; thereby&#160;determining both shape and cross-section. Photoinitiated polymerization, such as thiol click chemistry, is well suited for rapidly solidifying the core fluid into a microfiber with predetermined size and shape.

预设计尺寸和形状的聚合物纤维和生物贿赂纤维的微流体制造

A &#8220;sheath&#8221; fluid passing through a microfluidic channel at low Reynolds number can be directed around another &#8220;core&#8221; stream and ...

Using Microfluidics Chips for Live Imaging and Study of Injury Responses in Drosophila Larvae

Live imaging is an important technique for studying cell biological processes, however this can be challenging in live animals. The translucent cuticle of the Drosophila larva makes it an attractive model organism for live imaging studies. However, an important challenge for live imaging techniques is to noninvasively immobilize and position an animal on the microscope. This protocol presents a simple and easy to use method for immobilizing and imaging Drosophila larvae on a polydimethylsiloxane (PDMS) microfluidic device, which we call the 'larva chip'. The larva chip is comprised of a snug-fitting PDMS microchamber that is attached to a thin glass coverslip, which, upon application of a vacuum via a syringe, immobilizes the animal and brings ventral structures such as the nerve cord, segmental nerves, and body wall muscles, within close proximity to the coverslip. This allows for high-resolution imaging, and importantly, avoids the use of anesthetics and chemicals, which facilitates the study of a broad range of physiological processes. Since larvae recover easily from the immobilization, they can be readily subjected to multiple imaging sessions. This allows for longitudinal studies over time courses ranging from hours to days. This protocol describes step-by-step how to prepare the chip and how to utilize the chip for live imaging of neuronal events in 3rd instar larvae. These events include the rapid transport of organelles in axons, calcium responses to injury, and time-lapse studies of the trafficking of photo-convertible proteins over long distances and time scales. Another application of the chip is to study regenerative and degenerative responses to axonal injury, so the second part of this protocol describes a new and simple procedure for injuring axons within peripheral nerves by a segmental nerve crush.

Using Microfluidics Chips for Live Imaging and Study of Injury Responses in Drosophila Larvae

Drosophila larvae are an attractive model system for live imaging due to their translucent cuticle and powerful genetics. This protocol describes how to utilize a single-layer PDMS device, called the &#39;larva chip&#39; for live imaging of cellular processes within neurons of 3rd instar Drosophila larvae.

使用微流体芯片实时成像和损伤研究的响应<em&gt;果蝇</em&gt;幼虫

Live imaging is an important technique for studying cell biological processes, however this can be challenging in live animals. The translucent cuticle of ...

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 ...

Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion

3D organotypic cultures of epithelial cells on a matrix embedded with mesenchymal cells are widely used to study epithelial cell differentiation and invasion. Rat tail type I collagen and/or matrix derived from Engelbreth-Holm-Swarm mouse sarcoma cells have been traditionally employed as the substrates to model the matrix or stromal microenvironment into which mesenchymal cells (usually fibroblasts) are populated. Although experiments using such matrices are very informative, it can be argued that due to an overriding presence of a single protein (such as in type I Collagen) or a high content of basement membrane components and growth factors (such as in matrix derived from mouse sarcoma cells), these substrates do not best reflect the contribution to matrix composition made by the stromal cells themselves. To study native matrices produced by primary dermal fibroblasts isolated from patients with a tumor prone, genetic blistering disorder (recessive dystrophic epidermolysis bullosa), we have adapted an existing native matrix protocol to study tumor cell invasion. Fibroblasts are induced to produce their own matrix over a prolonged period in culture. This native matrix is then detached from the culture dish and epithelial cells are seeded onto it before the entire coculture is raised to the air-liquid interface. Cellular differentiation and/or invasion can then be assessed over time. This technique provides the ability to assess epithelial-mesenchymal cell interactions in a 3D setting without the need for a synthetic or foreign matrix with the only disadvantage being the prolonged period of time required to produce the native matrix. Here we describe the application of this technique to assess the ability of a single molecule expressed by fibroblasts, type VII collagen, to inhibit tumor cell invasion.

Tissue engineered fibroblast-derived native matrix is an emerging tool to generate a stromal substrate which supports epithelial cell proliferation and differentiation. Here a protocol applying this methodology to assess the impact of different stromal cell types on tumor cell biology is presented.

肿瘤微环境基质与应用癌细胞侵袭组织工程

3D organotypic cultures of epithelial cells on a matrix embedded with mesenchymal cells are widely used to study epithelial cell differentiation and ...

Fabrication of a Functionalized Magnetic Bacterial Nanocellulose with Iron Oxide Nanoparticles

In this study, bacterial nanocellulose (BNC) produced by the bacteria Gluconacetobacter xylinus is synthesized and impregnated in situ with iron oxide nanoparticles (IONP) (Fe3O4) to yield a magnetic bacterial nanocellulose (MBNC). The synthesis of MBNC is a precise and specifically designed multi-step process. Briefly, bacterial nanocellulose (BNC) pellicles are formed from preserved G. xylinus strain according to our experimental requirements of size and morphology. A solution of iron(III) chloride hexahydrate (FeCl3&#183;6H2O) and iron(II) chloride tetrahydrate (FeCl2&#183;4H2O) with a 2:1 molar ratio is prepared and diluted in deoxygenated high purity water. A BNC pellicle is then introduced in the vessel with the reactants. This mixture is stirred and heated at 80 &#176;C in a silicon oil bath and ammonium hydroxide (14%) is then added by dropping to precipitate the ferrous ions into the BNC mesh. This last step allows forming in situ magnetite nanoparticles (Fe3O4) inside the bacterial nanocellulose mesh to confer magnetic properties to BNC pellicle. A toxicological assay was used to evaluate the biocompatibility of the BNC-IONP pellicle. Polyethylene glycol (PEG) was used to cover the IONPs in order to improve their biocompatibility. Scanning electron microscopy (SEM) images showed that the IONP were located preferentially in the fibril interlacing spaces of the BNC matrix, but some of them were also found along the BNC ribbons. Magnetic force microscope measurements performed on the MBNC detected the presence magnetic domains with high and weak intensity magnetic field, confirming the magnetic nature of the MBNC pellicle. Young&#39;s modulus values obtained in this work are also in a reasonable agreement with those reported for several blood vessels in previous studies.

Here, we present a protocol to make a bacterial nanocellulose (BNC) magnetic for applications in damaged blood vessel reconstruction. The BNC was synthesized by G. xylinus strain. On the other hand, magnetization of the BNC was realized through in situ precipitation of Fe2+ and Fe3+ ferrous ions inside the BNC mesh.

一机能磁性纳米纤维素的细菌与氧化铁纳米粒子的制备

In this study, bacterial nanocellulose (BNC) produced by the bacteria Gluconacetobacter xylinus is synthesized and impregnated in situ with iron oxide ...

Using Synthetic Biology to Engineer Living Cells That Interface with Programmable Materials

We have developed an abiotic-biotic interface that allows engineered cells to control the material properties of a functionalized surface. This system is made by creating two modules: a synthetically engineered strain of E. coli cells and a functionalized material interface. Within this paper, we detail a protocol for genetically engineering selected behaviors within a strain of E. coli using molecular cloning strategies. Once developed, this strain produces elevated levels of biotin when exposed to a chemical inducer. Additionally, we detail protocols for creating two different functionalized surfaces, each of which is able to respond to cell-synthesized biotin. Taken together, we present a methodology for creating a linked, abiotic-biotic system that allows engineered cells to control material composition and assembly on nonliving substrates.

This paper presents a series of protocols for developing engineered cells and functionalized surfaces that enable synthetically engineered E. coli to control and manipulate programmable material surfaces.

利用合成生物学工程师活细胞接口，具有可编程材料

We have developed an abiotic-biotic interface that allows engineered cells to control the material properties of a functionalized surface. This system is ...

Three-dimensional Tissue Engineered Aligned Astrocyte Networks to Recapitulate Developmental Mechanisms and Facilitate Nervous System Regeneration

Neurotrauma and neurodegenerative disease often result in lasting neurological deficits due to the limited capacity of the central nervous system (CNS) to replace lost neurons and regenerate axonal pathways. However, during nervous system development, neuronal migration and axonal extension often occur along pathways formed by other cells, referred to as &#34;living scaffolds&#34;. Seeking to emulate these mechanisms and to design a strategy that circumvents the inhibitory environment of the CNS, this manuscript presents a protocol to fabricate tissue engineered astrocyte-based &#34;living scaffolds&#34;. To create these constructs, we employed a novel biomaterial encasement scheme to induce astrocytes to self-assemble into dense three-dimensional bundles of bipolar longitudinally-aligned somata and processes. First, hollow hydrogel micro-columns were assembled, and the inner lumen was coated with collagen extracellular-matrix. Dissociated cerebral cortical astrocytes were then delivered into the lumen of the cylindrical micro-column and, at a critical inner diameter of &#60;350 &#181;m, spontaneously self-aligned and contracted to produce long fiber-like cables consisting of dense bundles of astrocyte processes and collagen fibrils measuring &#60;150 &#181;m in diameter yet extending several cm in length. These engineered living scaffolds exhibited &#62;97% cell viability and were virtually exclusively comprised of astrocytes expressing a combination of the intermediate filament proteins glial-fibrillary acidic protein (GFAP), vimentin, and nestin. These aligned astrocyte networks were found to provide a permissive substrate for neuronal attachment and aligned neurite extension. Moreover, these constructs maintain integrity and alignment when extracted from the hydrogel encasement, making them suitable for CNS implantation. These preformed constructs structurally emulate key cytoarchitectural elements of naturally occurring glial-based &#34;living scaffolds&#34; in vivo. As such, these engineered living scaffolds may serve as test-beds to study neurodevelopmental mechanisms in vitro or facilitate neuroregeneration by directing neuronal migration and/or axonal pathfinding following CNS degeneration in vivo.

We showcase the development of self-assembled, three-dimensional bundles of longitudinally aligned astrocytic somata and processes within a novel biomaterial encasement. These engineered &#34;living scaffolds&#34;, exhibiting micron-scale diameter yet extending centimeters in length, may serve as test-beds to study neurodevelopmental mechanisms or facilitate neuroregeneration by directing neuronal migration and/or axonal pathfinding.

三维组织工程化排列的星形胶质细胞网络, 以概括发展机制和促进神经系统再生

Neurotrauma and neurodegenerative disease often result in lasting neurological deficits due to the limited capacity of the central nervous system (CNS) to ...

A Test Bed to Examine Helmet Fit and Retention and Biomechanical Measures of Head and Neck Injury in Simulated Impact

Conventional wisdom and the language in international helmet testing and certification standards suggest that appropriate helmet fit and retention during an impact are important factors in protecting the helmet wearer from impact-induced injury. This manuscript aims to investigate impact-induced injury mechanisms in different helmet fit scenarios through analysis of simulated helmeted impacts with an anthropometric test device (ATD), an array of headform acceleration transducers and neck force/moment transducers, a dual high speed camera system, and helmet-fit force sensors developed in our research group based on Bragg gratings in optical fiber. To simulate impacts, an instrumented headform and flexible neck fall along a linear guide rail onto an anvil. The test bed allows simulation of head impact at speeds up to 8.3 m/s, onto impact surfaces that are both flat and angled. The headform is fit with a crash helmet and several fit scenarios can be simulated by making context specific adjustments to the helmet position index and/or helmet size. To quantify helmet retention, the movement of the helmet on the head is quantified using post-hoc image analysis. To quantify head and neck injury potential, biomechanical measures based on headform acceleration and neck force/moment are measured. These biomechanical measures, through comparison with established human tolerance curves, can estimate the risk of severe life threatening and/or mild diffuse brain injury and osteoligamentous neck injury. To our knowledge, the presented test-bed is the first developed specifically to assess biomechanical effects on head and neck injury relative to helmet fit and retention.

Using an anthropometric head and neck, optical fiber-based fit force transducers, an array of head acceleration and neck force/moment transducers, and a dual high speed camera system, we present a test bed to study helmet retention and effects on biomechanical measures of head and neck injury secondary to head impact.

测试床, 检查头盔的拟合和保留, 以及在模拟撞击时头颈部损伤的生物力学措施

Conventional wisdom and the language in international helmet testing and certification standards suggest that appropriate helmet fit and retention during ...

Mouse Model of Pressure Ulcers After Spinal Cord Injury

Pressure ulcers (PUs) are common debilitating complications of traumatic spinal cord injury (SCI) and tend to occur in soft tissues around bony prominences. There is, however, little known about the impact of SCI on skin wound healing in the context of animal models in controlled experimental settings. In this study, a simple, non-invasive, reproducible and clinically relevant mouse model of PUs in the context of complete SCI is presented. Adult male mice (Balb/c, 10 weeks old) were shaved and depilated. Post-depilation (24 h), mice were subjected to laminectomy followed by complete spinal cord transection (T9-T10 vertebrae). Immediately after, a skin fold on the back of the mice was lifted and sandwiched between two magnetic discs held in place for next 12 h, thus creating an ischemic area that developed into a PU over the following days. The wounded areas demonstrated tissue edema and epidermal disappearance by day 3 post-magnet application. PUs spontaneously developed and healed. Healing was, however, slower in the SCI mice compared to control non-SCI mice when the wound was created below the level of SCI. Conversely, no difference in healing was seen between SCI and control non-SCI mice when the wound was created above the level of SCI. This model is a potentially useful tool to study the dynamics of skin PU development and healing after SCI, as well as to test therapeutic approaches that may help heal such wounds.

Here, we describe a simple method to induce clinically relevant skin pressure ulcers (PUs) in a mouse model of spinal cord injury (SCI). This model can be used in pre-clinical studies to screen for different therapeutics for healing PUs in SCI patients.

训练脊髓损伤者走动使用动力服

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