作者想感谢的资金来源:陆军UNL的创伤力学中心（美国国防部/ ARO，W911NF-11-1-0033，PI:适合本国的缓解钱德拉博士），UNL的门外汉奖（26-1110-0033 -001 PI:LIM）。

1。神经元细胞培养<ol><li>可以使用包括神经细胞，星形胶质细胞，和它们的共培养的脑细胞作为细胞模型。作为一个可行性论证，细胞系的神经细胞的脉冲增压。 </li><li>直径为18毫米盖玻片上培养SH-SY5Y人神经母细胞瘤细胞（ATCC CRL-2266）。细胞接种密度为3×10 3细胞/ cm 2使用组成的DMEM生长培养基补充有10％胎牛血清和1％青霉素-链霉素。细胞培养玻片保持在5％CO 2培养箱中，在37℃下的</li><li>要获得的SH-SY5Y细胞的神经元细胞的分化，细胞进一步补充用10μM - 视黄酸（RA）为7天，与媒体的媒体处理改变每两天。在第7天，细胞是准备被加压。 </li></ol> 2。加压设备:Kolsky酒吧<ol><li> Kolsky酒吧，于1949年1 Kolsky开发的，已用于测量材料的力学性能在一个非常高的负荷率。该装置由两个线条之间接触放置在与样品的酒吧。事件栏上创建，传播的应力波，波分裂成一个反射和透射波的样品。样品中的应力是正比于发射波。在这项研究中，我们利用一个单元格被放置的加压室之间的两个杆的Kolsky设置。 </li><li>通过对齐的黄铜轴承的两个铝合金棒（每6米长）悬浮，和一个被夹持在体外细胞加压室之间的横杠。上游酒吧是酒吧，一端夹到了130磅的质量事件。一种摩擦夹紧装置被放置的位置处，将得到所需的脉冲持续时间。从事钳，拧紧剪式千斤顶的质量和一个加载的支持，钳质量部分的事件栏前Compressed到所需的水平。强制锁定夹具带有液压系统突然断裂的切口螺栓迅速释放预先存储的压缩，从而产生一个压力脉冲。这通过传播事件栏，驱动测试室的活塞，加压液和细胞内室的冲动。反过来加压室启动压力脉冲在传输的酒吧下游传播。该菌株的压力脉冲在酒吧与复合高电阻应变计被激发到45伏。计信号与数字示波器在1 MHz的频率作为一个加载历史记录。 </li><li>在体外细胞加压室的活塞缸组成。缸具有2.6厘米内径和3.8厘​​米外径，并有一个小孔，在腔体的基础上抽头。孔作为在细胞样品安装过程中的空气和多余的液体通气孔。活塞是7.5厘米长的并且是由相同的棒材。活塞是包裹着两到三层的水喉匠（聚四氟乙烯）磁带作为低摩擦密封。端盖是32毫米长，有两个小不锈钢螺丝固定在玻片上（细胞培养），在加载。 </li></ol> 3。神经细胞的脉冲增压<ol><li>全部加压室部使用高压釜进行灭菌，并保持在UV光下。的组件的腔室的内部进行的细胞培养罩。腔室的排气螺钉是松散从事的通气孔中在基地的空腔。预先温热的（37℃）新鲜的生长培养基被吸移到未做气泡室。然后，将细胞培养载玻片被拾起，被放置在所述活塞帽（面朝外的细胞）。小的固定螺丝拧紧在幻灯片上举行的地方。用培养细胞的载玻片上的活塞被插入了一个小i置身于腔的腔室，并且装配倾斜时，在最高点与通气。除去排气螺钉和活塞推入空腔首先迫使气泡，然后过量的流体。一个引用标记或夹具是用来为指导，以确保相同的培养基体积为每一个测试。腔室中的介质的轴向尺寸是约6毫米。消毒通风的螺丝和更换，使室内的水紧。腔室不应该泄漏此静载荷下，如果这样做，聚四氟乙烯包裹需要更换或加强。 </li><li>在这一点上，的Kolsky杆系统复位。沉重的质量被移回原来的位置，并且一个新的锁紧螺栓替换所使用的（破碎）1。约200磅的液压搞新的锁定螺栓。用剪刀插孔压缩预装载部分的入射杆的压力略高于所需的值，然后回到它关闭，以充分的摩擦接合杰克的螺丝。现在手持数据采集。 </li><li>组装的细胞加压室被安装到系统中。它被放置在一个V形块由小型实验室剪刀插孔支持，并与两个杆对齐。润滑脂每个接口的光油脂层和摩擦的对接面，以消除空气之间的差距。 </li><li>现在已经准备好进行测试。夹锁紧螺栓强制打破，迅速泵送液压钳驱动程序。钳将分离和应显示的数据采集的结果。从发送巴（表压）的测量值确定的细胞加压历史。如果传输的酒吧是足够长的时间，这应该只从第一干扰的测量结果表明负压。所发送的脉冲的持续时间可能会比入射脉冲短，如果气泡被困住。的幅度可能不到达该入射脉冲，但它应该有足够离子克起卸前的高原。否则，无论是一个大的气泡，有可能发生在密封腔或错位之间的装配和酒吧。 </li><li>然后除去细胞加压室内部的细胞培养罩和拆卸。除去排气螺钉和活塞被拉到免费腔室。取自所述活塞的端部的细胞培养的载玻片。 </li></ol> 4。评估增压细胞生物学特性<ol><li>在加压后，细胞可以立即检查，或进一步培养，以便以后检查。通过适当的无菌操作过程中，更长期的培养是可能的。 </li><li>加压的细胞可以检查所有的分子和细胞生物学技术。特别是神经细胞增压，调查的细胞和分子生理学在TBI条件，在突起生长的压力引起的变化，微管细胞骨架的变化，神经基因expre的分析评估裂变，细胞凋亡，等等，都可以被执行。作为对照细胞样品，培养的细胞相同，保持在相同的时间期间内的加压室，但不被加压的使用（所谓的，腔室控制）。 </li><li>为了评估在突起生长的变化中，加压室的控制细胞后立即加压和1和24小时的孵育后用光学显微镜检查。神经元细胞的图像的一个例子示出在&#39;代表结果"部分（图2）。可以量化的肌动蛋白免疫荧光染色和图像分析突起长度的变化。固定细胞，用4％w / v的多聚甲醛溶液，用0.05％体积/体积的Tween-20洗涤缓冲液漂洗，并用0.1％体积/体积的Triton X-100溶液，透。封闭后用1％w / v的牛血清白蛋白溶液，四甲基罗丹明异硫氰酸酯（TRITC）共轭的鬼笔环肽的细胞孵育。采取的细胞的免疫荧光图像使用荧光显微镜和加压和控制细胞的突起的长度使用ImageJ软件（NIH）的量化。 </li><li>可以被认定为轴突或树突的突起。为了评估轴突或树突中的形态变化，上述实验可以重复通过检测每个结构的抗体，即，神经丝（neurofilament，NF）抗体轴突和微管相关蛋白（MAP2）抗体树突。 </li><li>微管是细胞骨架的主要成分的神经元之一，和与微管 ​​的破坏已被用作一个标记的神经元损伤的2微管免疫荧光可以可视化使用的β-微管蛋白抗体。加压后0和24小时后，固定细胞与β-微管蛋白抗体染色，并通过荧光显微镜观察到的。 </li><li>为了评估脉冲增压对神经元和神经细胞凋亡基因表达的影响，压力和控制细胞的总RNA提取。根E的表达进行定量RT-PCR和实时RT-PCR，我们公布的协议，可以检查。 </li></ol> 5。代表性的成果: <img src="/files/ftp_upload/2723/2723fig1.jpg" alt="图1"/> 图1，一种压力更新的例子适用于细胞内的加压室。 Kolsky酒吧设备成功地产生2 MPa级，持续时间约为0.7毫秒，单脉冲型脉冲增压。 <img src="/files/ftp_upload/2723/2723fig2.jpg" alt="图2"/> 图2。神经元细胞的一个例子，响应于脉冲加压。在2兆帕显示出明显的神经轴突/轴突击穿相对的腔体控制细胞SH-SY5Y细胞暴露于冲动增压。 6。难点与对策<ol><li>腔室的密封是要克服的主要障碍之一。这是fo&gt;公司&gt;，与O形圈的宽磁盘有一个刨等增加的摩擦问题。为了消除摩擦的影响，以及防止欺诈特富龙水喉匠的活塞录音磁带，一个简单的方法是使用。这解决了问题，并产生所需的压力水平和持续时间。 </li><li>加压神经元细胞的行为，尤其是在评估二次的TBI机制或长期的影响，应进行检查，经过了漫长的孵化时间。含细胞的腔室暴露于潜在的未消毒的条件，移动的同时的Kolsky栏，加压，和移动回到细胞培养罩。随着预灭菌的腔室部件和正确的操作，在细胞培养的滑动和腔室内部的细胞培养罩，组装/拆卸较长期的孵育后是可能长达数周。 </li><li>的数据的可再现性是一个重要的参数，在蜂窝的机械性刺激实验。我们的设备，reproducibility在神经细胞的反应确定所需的脉冲增压个人资料如何，在这两个压力的程度及持续时间，重复获得。由于我们记录每个升压得到的压力分布，细胞的反应不必要的压力分布数据，可以手动排除之后。 </li><li>整个加压步骤，包括室组件，转让和安装在Kolsky酒吧，增压，调回，和室拆卸，用了不到10分钟。加压下一个细胞培养幻灯片可以紧接前一后的加压。因此，足够数量的加压试验，可以有效地完成。我们的设置采用了直径为18毫米盖玻片。一个加压实验并不能提供足够的蛋白质，Western免疫印迹由于基板尺寸（也有一定的压力，因为有些细胞都死了）。关于三个重复的的加压实验​​提供immunobl所需的蛋白量otting。同样地，再现性检查每次用的压力轮廓。 </li></ol>

<ol>
	<li>Kolsky, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+investigation+of+mechanical+properties+of+materials+at+very+high+rates+of+loading.">An investigation of mechanical properties of materials at very high rates of loading.</a> Prec. Phys. Soc. London. 62, 676-700 (1949).</li><li>Tang-Schomer, M. D., Patel, A. R., Baas, P. W., Smith, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+breaking+of+microtubules+in+axons+during+dynamic+stretch+injury+underlies+delayed+elasticity,+microtubule+disassembly,+and+axon+degeneration.">Mechanical breaking of microtubules in axons during dynamic stretch injury underlies delayed elasticity, microtubule disassembly, and axon degeneration.</a> FASEB. J. 24, 1401-1410 (2010).</li><li>Lim, J. Y., Taylor, A. F., Li, Z., Vogler, E. A., Donahue, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrin+expression+and+osteopontin+regulation+in+human+fetal+osteoblastic+cells+mediated+by+substratum+surface+characteristics.">Integrin expression and osteopontin regulation in human fetal osteoblastic cells mediated by substratum surface characteristics.</a> Tissue. Eng. 11, 19-29 (2005).</li><li>Chen, Y. C., Smith, D. H., Meaney, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+approaches+for+studying+blast-induced+traumatic+brain+injury.">In vitro approaches for studying blast-induced traumatic brain injury.</a> J. Neurotrauma. 26, 861-876 (2009).</li><li>Ling, G., Bandak, F., Armonda, R., Grant, G., Ecklund, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Explosive+blast+neurotrauma.">Explosive blast neurotrauma.</a> J. Neurotrauma. 26, 815-825 (2009).</li><li>Shepard, S. R., Ghajar, J. B. G., Giannuzzi, R., Kupferman, S., Hariri, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluid+percussion+barotrauma+chamber:+a+new+in+vitro+model+for+traumatic+brain+injury.">Fluid percussion barotrauma chamber: a new in vitro model for traumatic brain injury.</a> J. Surg. Res. 51, 417-424 (1991).</li><li>VandeVord, P. J., Leung, L. Y., Hardy, W., Mason, M., Yang, K. H., King, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Up-regulation+of+reactivity+and+survival+genes+in+astrocytes+after+exposure+to+short+duration+overpressure.">Up-regulation of reactivity and survival genes in astrocytes after exposure to short duration overpressure.</a> Neurosci. Lett. 434, 247-252 (2008).</li><li>Huang, H., Feng, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+study+of+the+dynamic+tribological+response+of+closed+fracture+surface+pairs+by+Kolsky-bar+compression-shear+experiment.">A study of the dynamic tribological response of closed fracture surface pairs by Kolsky-bar compression-shear experiment.</a> Int. J. Solids. Struct. 41, 2821-2835 (2004).</li><li>Song, B., Chen, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Split+Hopkinson+pressure+bar+techniques+for+characterizing+soft+materials.">Split Hopkinson pressure bar techniques for characterizing soft materials.</a> Lat. Am. J. Solids. Struct. 2, 113-152 (2005).</li></ol>

没有利益冲突的声明。

已经尝试了许多在体外实验技术研究脑细胞损伤TBI条件。这些措施包括神经元/胶质细胞拉伸，流引起的剪切应力，体重下降，手写笔裂伤，激光切断术，碎石等4,5它已被认为持续时间较短的超压主要物理因素TBI尤其是6,7 TBI的爆炸冲击波有一个脉冲持续时间为毫秒级。基于这种假设和观察，气压计室的瞬时增压已经制定和评估脑细胞行为在TBI的条件下使用。例如，在流体中的打击乐器气压伤室20-30毫秒的持续时间和长的峰值压力高达0.5MPa的压力脉冲被用来研究人类的神经胶质细胞的行为下TBI 6最近，VandeVord等7中使用的金属击球的晴雨表室检查的星形胶质细胞根据TBI的行为，但金属球惊人的多重压力脉冲。在这项研究中，我们已经开发了一种新的细胞产生单脉冲型脉冲加压的加压装置。装置的基础上的Kolsky杆设置，它已被用于作为材料试验装置，在我们的8和其它的研究开发。1,9的Kolsky杆材料测试能力已被修改，以便能够在非常高的负荷率应用单脉冲型超压的培养细胞内的加压室（图1）。我们可以成功地控制的幅度和持续时间的脉冲的超压。作为一个代表性的细胞反应，我们表明，在2 MPa压力下SH-SY5Y神经细胞显示显着的神经轴突和轴突损失相比，在毫无压力的腔体控制细胞。 总之，我们制定了一个新的冲动细胞加压装置，采用特殊设计的Kolsky在TBI条件研究神经细胞的反应，酒吧设备，并展示了其潜在的使用。我们还注意到，这种技术是非常灵活，可以接触到任何类型的细胞，在不同的程度和持续时间的冲动的压力。因此，该设备可以被用来研究的脉冲压力诱导的细胞的机械力行为，不仅对受损的细胞，但也为积极刺激细胞在它们的功能和命运决定。

<ul><li> SH-SY5Y（ATCC CRL-2266）人类神经母细胞瘤</li><li>直径为18毫米的玻璃盖玻片</li><li>细胞培养基:DMEM，10％胎牛血清，1％青霉素 - 链霉素</li><li>视黄酸（10μM）诱导神经发生</li><li> Kolsky酒吧脉冲增压装置</li><li>活塞缸细胞加压室</li><li>不锈钢螺钉固定在活塞上的细胞培养盖玻片</li><li>润滑脂之间的细胞室和Kolsky的酒吧接口的</li><li>一般分子生物学用品为评估细胞反应增压（固色剂，抗体，荧光染料，PCR耗材等） </li></ul>

impulsive pressurization of neuronal cells for traumatic brain injury study

一种新型的冲动的细胞加压实验已开发使用的Kolsky酒吧的移动设备以调查爆炸引起的创伤性脑损伤（TBI）。我们展现在爆炸TBI相关的脉冲增压视频文章如何适用于神经元细胞在体外。这是通过使用良好控制的压力脉冲创建由一个专门Kolsky的杆装置，与完整的压力历史记录细胞内的加压室。加压神经细胞加压后立即检查，或继续培养，研究长期影响的冲动加压在神经突起/轴突生长，神经元的基因表达，细胞凋亡，等我们观察到，冲动增压约2兆帕引起不同的神经轴突损失相对在毫无压力的细胞。我们的技术提供了一种新的方法来研究分子/细胞机制的爆炸TBI，，通过脉冲增压脑细胞控制的公关essure的幅度和持续时间。

Watch this Scientific Journal Video about Impulsive Pressurization of Neuronal Cells for Traumatic Brain Injury Study at JoVE.com

Impulsive Pressurization of Neuronal Cells for Traumatic Brain Injury Study

一种新型的冲动的细胞加压实验已开发使用Kolsky酒吧的移动设备以调查爆炸引起的外伤性脑损伤的分子/细胞的机制。

创伤性脑损伤研究神经细胞的脉冲增压

A novel impulsive cell pressurization experiment has been developed using a Kolsky bar device to investigate blast-induced traumatic brain injury (TBI). ...

impulsive-pressurization-neuronal-cells-for-traumatic-brain-injury

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10.2K 次观看.  University of Nebraska-Lincoln. 一种新型的冲动的细胞加压实验已开发使用Kolsky酒吧的移动设备以调查爆炸引起的外伤性脑损伤的分子/细胞的机制。

视频: Impulsive Pressurization of Neuronal Cells for Traumatic Brain Injury Study

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

侧向液压冲击:创伤性脑损伤小鼠模型

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

科研

神经科学

Stretch in Brain Microvascular Endothelial Cells (cEND) as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying mechanisms involved in it are useful for treatment. There are both in vivo and in vitro methods available for this purpose. In vivo models can mimic actual head injury as it occurs during TBI. However, in vivo techniques may not be exploited for studies at the cell physiology level. Hence, in vitro methods are more advantageous for this purpose since they provide easier access to the cells and the extracellular environment for manipulation.
Our protocol presents an in vitro model of TBI using stretch injury in brain microvascular endothelial cells. It utilizes pressure applied to the cells cultured in flexible-bottomed wells. The pressure applied may easily be controlled and can produce injury that ranges from low to severe. The murine brain microvascular endothelial cells (cEND) generated in our laboratory is a well-suited model for the blood brain barrier (BBB) thus providing an advantage to other systems that employ a similar technique. In addition, due to the simplicity of the method, experimental set-ups are easily duplicated. Thus, this model can be used in studying the cellular and molecular mechanisms involved in TBI at the BBB.

Stretch in Brain Microvascular Endothelial Cells cEND as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

In vitro traumatic brain injury models are being developed to reproduce in vivo brain deformation. Stretch-induced injury has been employed for astrocytes, neurons, glial cells, aortic, and brain endothelial cells. However, our system uses a blood brain barrier (BBB) model that possesses properties constituting a legitimate model of the BBB to establish an in vitro&#160;TBI model.

作为脑微血管内皮细胞（CEND）的伸展<em&gt;体外</em创伤性脑损伤模型血脑屏障

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying ...

医学

Controlled Cortical Impact Model of Mouse Brain Injury with Therapeutic Transplantation of Human Induced Pluripotent Stem Cell-Derived Neural Cells

Traumatic brain injury (TBI) is a leading cause of morbidity and mortality worldwide. Disease pathology due to TBI progresses from the primary mechanical insult to secondary injury processes, including apoptosis and inflammation. Animal modeling has been valuable in the search to unravel injury mechanisms and evaluate potential neuroprotective therapies. This protocol describes the controlled cortical impact (CCI) model of focal, open-head TBI. Specifically, parameters for producing a mild unilateral cortical injury are described. Behavioral consequences of CCI are analyzed using the adhesive tape removal test of bilateral sensorimotor integration. Regarding experimental therapy for TBI pathology, this protocol also illustrates a process for transplanting cultured cells into the brain. Neural cell cultures derived from human induced pluripotent stem cells (hiPSCs) were chosen for their potential to show superior functional restoration in human TBI patients. Chronic survival of hiPSCs in the host mouse brain tissue is detected using a modified DAB immunohistochemical process.

This protocol demonstrates methodologies for a mouse model of open-skull traumatic brain injury and transplantation of cultured human induced pluripotent stem cell-derived cells into the injury site. Behavioral and histologic tests of outcomes from these procedures are also described in brief.

人类诱导多能干细胞衍生神经细胞治疗移植小鼠脑损伤控制皮质影响模型

Traumatic brain injury (TBI) is a leading cause of morbidity and mortality worldwide. Disease pathology due to TBI progresses from the primary mechanical ...

发育生物学

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

控制皮层影响模型创伤性脑损伤

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the detonation of explosive devices, however, the mechanisms that underlie the brain damage resulting from blast overpressure exposure are not entirely understood and are believed to be unique to this type of brain injury. Preclinical models are crucial tools that contribute to better understand blast-induced brain injury. A novel in vitro blast TBI model was developed using an open-ended shock tube to simulate real-life open-field blast waves modelled by the Friedlander waveform. C57BL/6N mouse organotypic hippocampal slice cultures were exposed to single shock waves and the development of injury was characterized up to 72 h using propidium iodide, a well-established fluorescent marker of cell damage that only penetrates cells with compromised cellular membranes. Propidium iodide fluorescence was significantly higher in the slices exposed to a blast wave when compared with sham slices throughout the duration of the protocol. The brain tissue injury is very reproducible and proportional to the peak overpressure of the shock wave applied.

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

一种新的脑外伤体损伤模型

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral vascular effects. Impact (i.e., non-blast) traumatic brain injury (TBI) is known to decrease pressure autoregulation in the cerebral vasculature in both humans and experimental animals. The hypothesis that blast-induced traumatic brain injury (bTBI), like impact TBI, results in impaired cerebral vascular reactivity was tested by measuring myogenic dilatory responses to reduced intravascular pressure in rodent middle cerebral arterial (MCA) segments from rats subjected to mild bTBI using an Advanced Blast Simulator (ABS) shock tube. Adult, male Sprague-Dawley rats were anesthetized, intubated, ventilated and prepared for Sham bTBI (identical manipulation and anesthesia except for blast injury) or mild bTBI. Rats were randomly assigned to receive Sham bTBI or mild bTBI followed by sacrifice 30 or 60 min post-injury. Immediately after bTBI, righting reflex (RR) suppression times were assessed, euthanasia at the time points post-injury was completed, the brain was harvested and the individual MCA segments were collected, mounted and pressurized. As the intraluminal pressure perfused through the arterial segments was reduced in 20 mmHg increments from 100 to 20 mmHg, MCA diameters were measured and recorded. With decreasing intraluminal pressure, MCA diameters steadily increased significantly above baseline in the Sham bTBI groups while MCA dilator responses were significantly reduced (p &#60; 0.05) in both bTBI groups as evidenced by the impaired, smaller MCA diameters recorded for the bTBI groups. In addition, RR suppression in the bTBI groups was significantly (p &#60; 0.05) higher than in the Sham bTBI groups. MCA&#39;s collected from the Sham bTBI groups exhibited typical vasodilatory properties to decreases in intraluminal pressure while MCA&#39;s collected following bTBI exhibited significantly impaired myogenic vasodilatory responses to reduced pressure that persisted for at least 60 min after bTBI.

Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

爆炸性神经损伤对压生啮鼠中脑动脉的影响

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

创伤性脑损伤诱导受控皮质冲击的母模型

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

Low-intensity Blast Wave Model for Preclinical Assessment of Closed-head Mild Traumatic Brain Injury in Rodents

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of medical visits in the United States. There are currently no FDA-approved treatments available for TBI. The increased incidence of military-related, blast-induced TBI further accentuates the urgent need for effective TBI treatments. Therefore, new preclinical TBI animal models that recapitulate aspects of human blast-related TBI will greatly advance the research efforts into the neurobiological and pathophysiological processes underlying mild to moderate TBI as well as the development of novel therapeutic strategies for TBI.
Here we present a reliable, reproducible model for the investigation of the molecular, cellular, and behavioral effects of mild to moderate blast-induced TBI. We describe a step-by-step protocol for closed-head, blast-induced mild TBI in rodents using a bench-top setup consisting of a gas-driven shock tube equipped with piezoelectric pressure sensors to ensure consistent test conditions. The benefits of the setup that we have established are its relative low-cost, ease of installation, ease of use and high-throughput capacity. Further advantages of this non-invasive TBI model include the scalability of the blast peak overpressure and the generation of controlled reproducible outcomes. The reproducibility and relevance of this TBI model has been evaluated in a number of downstream applications, including neurobiological, neuropathological, neurophysiological and behavioral analyses, supporting the use of this model for the characterization of processes underlying the etiology of mild to moderate TBI.

We present here a protocol of a blast wave model for rodents to investigate neurobiological and pathophysiological effects of mild to moderate traumatic brain injury. We established a gas-driven, bench-top setup equipped with pressure sensors allowing for reliable and reproducible generation of blast-induced mild to moderate traumatic brain injury.

低强度冲击波模型用于啮齿动物闭头轻度创伤性脑损伤的临床前评估

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of ...

A Preclinical Controlled Cortical Impact Model for Traumatic Hemorrhage Contusion and Neuroinflammation

Cerebral contusion is a severe medical problem affecting millions of people worldwide each year. There is an urgent need to understand the pathophysiological mechanism and to develop effective therapeutic strategy for this devastating neurological disorder. Intraparenchymal hemorrhage and post-traumatic inflammatory response induced by initial physical impact can aggravate microglia/macrophage activation and neuroinflammation which subsequently worsen brain pathology. We provide here a controlled cortical impact (CCI) protocol that can reproduce experimental cortical contusion in mice by using a pneumatic impactor system to deliver mechanical force with controllable magnitude and velocity onto the dural surface. This preclinical model allows researchers to induce moderately severe focal cerebral contusion in mice and to investigate a wide range of post-traumatic pathological progressions including hemorrhage contusion, microglia/macrophage activation, iron toxicity, axonal injury, as well as short-term and long-term neurobehavioral deficits. The present protocol can be useful for exploring the long-term effects of and potential interventions for cerebral contusion.

<meta charset="utf8"></head><body>创伤性出血挫伤和神经炎症的临床前受控皮质影响模型

Intraparenchymal hemorrhage and neuroinflammation accompanied by cerebral contusion can trigger severe secondary brain injury. This protocol details a mouse controlled cortical impact (CCI) model allowing researchers to study hemorrhage contusion and post-traumatic immune responses and explore potential therapeutics.