作者感谢他们的设计和制造系统，并与准备的稿子援助Valtresa迈尔斯的协助下沙德Schipke和丹尼尔Chesser。

1。组织收获和准备<ol><li>应收集称重后立即死亡的不超过120磅的成年猪主动脉瓣。 </li><li>阀门两次洗净，用无菌磷酸盐缓冲液（PBS）和运输上冰实验室。 </li><li>应在无菌条件下进行的所有后续步骤。 </li><li>确保，单张不显示任何退化的迹象，撕裂或钙化。删除从纤维环削减1 / 3的距离从主动脉根部的宣传单张。 </li><li>放置传单，在六孔板个别井和3毫升贝科改良Eagle 1％anti-biotic/anti-mycotic解决方案，并在37 ° C和5％的CO 2 10％胎牛血清培养基孵育过夜。 </li><li>作为一种替代方法，分离出的细胞可以接种到6孔培养板，压力设备中使用的。如前所述13，14阀内皮细胞和间质细胞的分离，可以进行。 </li></ol> 2。压力研究<ol><li>一个特制的压力系统的设计研究循环压力的主动脉瓣组织15 mechanobiological影响。 </li><li>登录到计算机上，打开LabVIEW程序（图1）。 </li><li>校准： <ol><li>试验之前，该系统应正确校准。 </li><li>连接到电路板的电源。这提供电源的电磁阀控制流入和流出的空气室的流动。 </li><li>确保压缩空气系统连接，并打开送风全速度。 </li><li>开启的信号放大器。确保电压读数是0.00。根据需要调整</li><li> LabVIEW的接口标有“测试/记录”的一个开关。确保将开关设置为“test”。点击标有“空气补给”，打开进水电磁阀“按钮。 </li><li>使用燃气压力调节器，用压缩空气加压室1 PSI。在会议厅内的压力可以读取使用位于室的后端板的数字压力表。一旦压力平衡，记录信号放大器的电压读数。重复2，3，4和5 PSI。 </li><li>构建的压力与电压的校准曲线。压力应该从PSI的转换，以毫米汞柱。从图中的计算公式可以被放置到LabVIEW程序代码</li></ol></li><li>从压力室中删除的铝制前板和喷雾室，用70％乙醇。至少10分钟离开，让残留的乙醇气体消散。 </li><li>将6孔板单张样本含有进入会议厅和更换前端板。确保密封不透气收紧位于四个螺纹杆（位于终板的每个角落）手工坚果。将在37℃孵化器压力室。一个压力室的示意图如图2所示。 </li><li>该界面会提示用户提供的压缩空气输入和输出之间的时间系统周期量。这些应设置为0.6S和0.4秒模仿的舒张压和收缩压的条件下，分别在频率为1Hz，。用户还应该输入数据文件的路径。 </li><li>在LabVIEW中，单击“运行”和“测试/记录”切换开关“录制”。 </li><li>确保在使用LabVIEW的接口图所需的水平压力。压力可以调整使用的燃气调压。 </li><li>运行程序所需的时间长度。 </li><li>一旦实验完成后，单击“基于LabVIEW的停止按钮，关掉供气，打开排气阀上的压力室。 </li><li>从室取出前端板和检索六孔板样品。现在可以分析基因表达，蛋白表达，组织学，力学性能等样品</li></ol> 3。代表性的成果： 压力系统是能够模拟正常血压下观察到的最大跨瓣压力，阶段I和第II阶段的高血压的条件。然而，压力是不能够模仿的收缩压梯度，这基本上是零，在体内。保持以1Hz的频率，0.6S的进气口和排气时间的0.4秒时间。代表从系统中获得的条件正常和压力升高的压力波形图3中可以看出。 <img src="/files/ftp_upload/3316/3316fig1.jpg" alt="图1"/> 图1：LabVIEW的接口的屏幕截图。 <img src="/files/ftp_upload/3316/3316fig2.jpg" alt="图2"/> 图2：A.压力腔压力腔等轴测视图示意图; B.压力室的侧视图;C.压力室的顶视图。 <img src="/files/ftp_upload/3316/3316fig3.jpg" alt="图3"/> 图3：图内的压力腔的压力模拟（一）血压正常，（二）第一阶段高血压，和（C）第二阶段高血压条件。

<ol>
	<li>Freeman, R. V., Otto, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectrum+of+calcific+aortic+valve+disease:+pathogenesis,+disease+progression,+and+treatment+strategies.">Spectrum of calcific aortic valve disease: pathogenesis, disease progression, and treatment strategies.</a> Circulation. 111, 3316-3326 (2005).</li><li>Robicsek, F., Thubrikar, M. J., Fokin, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cause+of+degenerative+disease+of+the+trileaflet+aortic+valve:+review+of+subject+and+presentation+of+a+new+theory.">Cause of degenerative disease of the trileaflet aortic valve: review of subject and presentation of a new theory.</a> Ann Thorac Surg. 73, 1346-1354 (2002).</li><li>Thubrikar, M. J., Aouad, J., Nolan, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+calcific+deposits+in+operatively+excised+stenotic+or+purely+regurgitant+aortic+valves+and+their+relation+to+mechanical+stress.">Patterns of calcific deposits in operatively excised stenotic or purely regurgitant aortic valves and their relation to mechanical stress.</a> Am J Cardiol. 58, 304-308 (1986).</li><li>Agno, F. S., Chinali, M., Bella, J. N., Liu, J. E., Arnett, D. K., Kitzman, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aortic+valve+sclerosis+is+associated+with+preclinical+cardiovascular+disease+in+hypertensive+adults:+the+Hypertension+Genetic+Epidemiology+Network+study.">Aortic valve sclerosis is associated with preclinical cardiovascular disease in hypertensive adults: the Hypertension Genetic Epidemiology Network study.</a> J Hypertens. 23, 867-8673 (2005).</li><li>Cawley, P. J., Otto, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+of+calcific+aortic+valve+stenosis+-+fact+or+fiction.">Prevention of calcific aortic valve stenosis - fact or fiction.</a> Annals of Medicine. 41, 100-108 (2009).</li><li>Durst, C. A., Grande-Allen, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+physical+characterization+of+a+synchronous+multivalve+aortic+valve+culture+system.">Design and physical characterization of a synchronous multivalve aortic valve culture system.</a> Ann Biomed Eng. 38, 319-3125 (2010).</li><li>Engelmayr, G. C., Soletti, L., Vigmostad, S. C., Budilarto, S. G., Federspiel, W. J., Chandran, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+flex-stretch-flow+bioreactor+for+the+study+of+engineered+heart+valve+tissue+mechanobiology.">A novel flex-stretch-flow bioreactor for the study of engineered heart valve tissue mechanobiology.</a> Ann Biomed Eng. 36, 700-712 (2008).</li><li>Sucosky, P., Padala, M., Elhammali, A., Balachandran, K., Jo, H., Yoganathan, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+of+an+ex+vivo+culture+system+to+investigate+the+effects+of+shear+stress+on+cardiovascular+tissue.">Design of an ex vivo culture system to investigate the effects of shear stress on cardiovascular tissue.</a> J Biomech Eng. 130, 035001-03 (2008).</li><li>Syedain, Z. H., Tranquillo, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+cyclic+stretch+bioreactor+for+tissue-engineered+heart+valves.">Controlled cyclic stretch bioreactor for tissue-engineered heart valves.</a> Biomaterials. 30, 4078-4084 (2009).</li><li>Lagana, K., Moretti, M., Dubini, G., Raimondi, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+bioreactor+for+the+controlled+application+of+complex+mechanical+stimuli+for+cartilage+tissue+engineering.">A new bioreactor for the controlled application of complex mechanical stimuli for cartilage tissue engineering.</a> Proc Inst Mech Eng H. 222, 705-715 (2008).</li><li>Wartella, K. A., Wayne, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioreactor+for+biaxial+mechanical+stimulation+to+tissue+engineered+constructs.">Bioreactor for biaxial mechanical stimulation to tissue engineered constructs.</a> J Biomech Eng. 131, 044501-044501 (2009).</li><li>Wallis, M. C., Yeger, H., Cartwright, L., Shou, Z., Radisic, M., Haig, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+study+of+a+novel+urinary+bladder+bioreactor.">Feasibility study of a novel urinary bladder bioreactor.</a> Tissue Eng Part A. 14, 339-348 (2008).</li><li>Butcher, J. T., Nerem, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Valvular+endothelial+cells+regulate+the+phenotype+of+interstitial+cells+in+co-culture:+effects+of+steady+shear+stress.">Valvular endothelial cells regulate the phenotype of interstitial cells in co-culture: effects of steady shear stress.</a> Tissue Eng. 12, 905-915 (2006).</li><li>Metzler, S. A., Pregonero, C. A., Butcher, J. T., Burgess, S. C., Warnock, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+Strain+Regulates+Pro-Inflammatory+Protein+Expression+in+Porcine+Aortic+Valve+Endothelial+Cells.">Cyclic Strain Regulates Pro-Inflammatory Protein Expression in Porcine Aortic Valve Endothelial Cells.</a> J Heart Valve Dis. 17, 571-578 (2008).</li><li>Schipke, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Design of a cyclic pressure bioreactor for the ex vivo study of aortic heart valve mechanobiology. , (2008).</li><li>Smith, K. E., Metzler, S. A., Warnock, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+strain+inhibits+acute+pro-inflammatory+gene+expression+in+aortic+valve+interstitial+cells.">Cyclic strain inhibits acute pro-inflammatory gene expression in aortic valve interstitial cells.</a> Biomechanics and Modeling in Mechanobiology. , (2009).</li><li>Warnock, J. N., Burgess, S. C., Shack, A., Yoganathan, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+immediate-early+gene+responses+to+elevated+pressure+in+porcine+aortic+valve+interstitial+cells.">Differential immediate-early gene responses to elevated pressure in porcine aortic valve interstitial cells.</a> J Heart Valve Dis. 15, 34-41 (2006).</li><li>Brathwaite, D., Weissman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+new+onset+of+atrial+arrhythmias+following+major+noncardiothoracic+surgery+is+associated+with+increased+mortality.">The new onset of atrial arrhythmias following major noncardiothoracic surgery is associated with increased mortality.</a> Chest. 114, 462-468 (1998).</li><li>Walsh, S. R., Oates, J. E., Anderson, J. A., Blair, S. D., Makin, C. A., Walsh, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postoperative+arrhythmias+in+colorectal+surgical+patients:+incidence+and+clinical+correlates.">Postoperative arrhythmias in colorectal surgical patients: incidence and clinical correlates.</a> Colorectal Dis. 8, 212-216 (2006).</li><li>Walsh, S. R., Tang, T., Gaunt, M. E., Schneider, 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=New+arrhythmias+after+non-cardiothoracic+surgery.">New arrhythmias after non-cardiothoracic surgery.</a> BMJ. 7, 333-333 (2006).</li><li>Walsh, S. R., Tang, T., Wijewardena, C., Yarham, S. I., Boyle, J. R., Gaunt, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postoperative+arrhythmias+in+general+surgical+patients.">Postoperative arrhythmias in general surgical patients.</a> Ann R Coll Surg Engl. 89, 91-95 (2007).</li></ol>

没有利益冲突的声明。

压力系统成功地暴露了主动脉瓣舒张期跨瓣压差，分别代表循环压力。然而，这是不能够模仿收缩跨瓣压差，压力仅下降40毫米汞柱。跨瓣压差在升主动脉和左心室之间的压力差。在舒张期，当阀门关闭时，压力差是80mmHg血压正常的条件下和90毫米汞柱，并在第一阶段100mmHg我和第二阶段的高血压，分别为。在收缩过程中，当阀门打开时，左心室和主动脉之间的压力差为零。发生在主动脉瓣病变的?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> DMEM培养液</td><td>西格玛</td><td> D5671 </td><td></td></tr><tr><td>贝科的PBS </td><td>西格玛</td><td> D5652 </td><td></td></tr><tr><td> Anti-mycotic/antibiotic解决方案</td><td>西格玛</td><td> A5955 </td><td></td></tr><tr><td>胎牛血清</td><td> ThermoScientific </td><td> SH30070 </td><td></td></tr><tr><td>氟橡胶膜片电磁阀</td><td>麦克马斯特卡尔</td><td> 4868K11 </td><td></td></tr><tr><td>压力传感器</td><td> Omega工程公司</td><td> PX302 - 200GV </td><td></td></tr><tr><td>称重传感器空调</td><td>喝采电子，公司</td><td> 4025-101 </td><td></td></tr><tr><td>数据采集​​（DAQ）模块</td><td> Measurement Computing公司</td><td> PMD1608 </td><td></td></tr></tbody></table>

design of a cyclic pressure bioreactor for the ex vivo study of aortic heart valves

主动脉瓣，左心室和主动脉之间，允许单向血流量，防止回流入脑室。主动脉瓣叶内细胞外基质（ECM）暂停，内衬内皮细胞单层的间质细胞组成。该阀门可承受苛刻的，动态的环境和不断暴露剪切，前屈，张力和压缩。研究表明病变瓣膜钙化病变发生在机械应力高的地区作为内皮细胞破坏或间质性基质损害 1-3的结果。因此，这并不奇怪，流行病学研究表明高血压，要在一个主要危险因素的主动脉瓣病变 4的发生。 目前瓣膜病唯一的治疗选择是一个bioprosthetic或机械瓣膜病变瓣膜置换手术。物理讲改进阀生物学的了解将有助于阐明阀的发病机制。反过来，这将有助于在非侵入性治疗，如医药干预或预防的发展。几个生物反应器，此前已开发研究本地或工程心脏瓣膜6-9 mechanobiology。脉动生物反应器也已发展到包括软骨，骨 11 10和12膀胱组织的研究范围。这项工作的目的是发展循环压力系统，可以用来澄清压力负荷增加的主动脉瓣叶的生物反应。 该系统由丙烯酸商会在其中放置样品和生产循环的压力，氟橡胶膜片电磁阀控制压力周期的时间，和一台计算机来控制电气设备。压力监测使用的压力传感器，空调使用的信号是一个负载单元空调。一个LabVIEW程序规范使用模拟设备系统，在适当的速度泵入压缩空气的压力。该系统模仿与主动脉瓣跨瓣压差的动态水平;一个锯齿波产生的压力逐渐增加，典型的是目前在舒张期，阀门两端的跨瓣压差梯度，压力急剧描绘阀门的开度在下降收缩。 LabVIEW程序允许用户控制循环压力的大小和频率。该系统能够受到生理和病理的压力条件下的组织样本。此设备可用于增加心脏瓣膜如何应对在当地的机械环境的变化的认识。

Watch this Scientific Journal Video about Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves at JoVE.com

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

循环压力的心脏瓣膜组织遭受的生理和病理的压力条件下的生物反应器已经设计。一个LabVIEW程序允许用户控制压力的大小，幅度和频率。此装置可用于研究心脏瓣膜组织或孤立的细胞mechanobiology。

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 ...

design-cyclic-pressure-bioreactor-for-ex-vivo-study-aortic-heart

Research

Education

JoVE Core

JoVE Journal

Physics

Bioengineering

Unit 1

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

Sources of Magnetic Fields

SCIENTISTS IN ACTION

15.6K 次观看.  Mississippi State University. 循环压力的心脏瓣膜组织遭受的生理和病理的压力条件下的生物反应器已经设计。一个LabVIEW程序允许用户控制压力的大小，幅度和频率。此装置可用于研究心脏瓣膜组织或孤立的细胞mechanobiology。