这项工作是由AO基金会（S-08-42G）和RISystem公司的支持。 我们想扩展一个非常大的"谢谢！"给Stephan Zeiter队在敖研究所达沃斯，瑞士正在让我们用自己的或设施为这个手术的拍摄如此宽松。

动物保健和实验方案如下，根据美国国立卫生研究院的指导方针，并经贝斯以色列女执事医疗中心实验动物管理和使用委员会，马萨诸塞州波士顿。 （协议编号:098-2009） 1，准备手术材料和仪器<ol><li>消毒用于执行使用前手术所有手术材料和仪器。包所需材料，使用或不使用工具托盘，折叠的布或纸包裹内有密封胶带釜蒸汽灭菌。将高压釜的温度应在125-135°C下的灭菌时间20-25分钟，干燥时间，然后10-15分钟。 </li><li>确保手术的时间只为200-250克。这是非常重要的，因为如果只在尺寸更重，那么不同尺寸的固定器应该被使用。对于只应使用重量超过250 GA较大的外固定器系统的版本。 </li> &lt;/ OL&gt; 2，手术过程和外固定器的应用<ol><li>任何动物的认证供应商购买只SD（或任何其他株）大鼠（男或女，200〜250克）。按照相应的动物护理和实验方案，根据已批准调查的机构动物照顾及使用委员会国家准则。允许在手术前至少48小时的适应期。 </li><li>对于手术，运送鼠到专用的手术房。 </li><li>麻醉与第一通过感应腔室的异氟醚的大鼠，然后继续在1-1.5升O 2 /分钟连接到麻醉机以1.5-2％的速度面罩。在手术开始时确保动物是在深度麻醉。要做到这一点，使用由延长肢体捏脚趾之间的网络用手指踏板反射技术（ 而不是脚趾本身！）。动物是不是苏如果肢体被撤回，肌肉抽搐发生，或者如果该动物使得噪声fficiently麻醉。 </li><li>大鼠是在深全身麻醉后进行手术，注射抗生素（头孢唑啉，20毫克/千克）和止痛的丁丙诺啡（剂量0.08毫克/千克）肌内右腿。避免了在手术过程中大量的流体损失，在之前的手术的身体重量3-5％的施用温暖的无菌盐水皮下，并且如果有必要在最后。应用无菌眼药膏眼睛，让他们水合，防止角膜损伤。 </li><li>下列药物注射剂，剃须和清洁用chlorohezadine或其他消毒溶液的大鼠的整个右后腿和动物转移到手术台。 （在腿部动作必须是相同的，将其用注射的人。） </li><li>将动物在俯卧位（ 图1A）加热表面。确保口罩留在鼻子和嘴向后ER转移到手术台，并保持在2.3中提到的麻醉方案）。用无菌窗孔被单悬垂的外科手术的区域，以便只腿用于手术被暴露。 </li><li>通过皮肤运行craniolateral从大转子到用手术刀（ 图1C）的膝关节的髁区域的右股骨的表面上作出的3-4厘米（ 图1B）的近似切口。通过轻柔解剖分离阔筋膜，并确保该肌肉组织不被切割露出股骨的轴。在这之后，分离分开的分枝股外侧肌和M.股二头肌和解除M.阔筋膜张肌，显露股骨的全长（在确认坐骨神经被保留; 图1D）。 </li><li>在截骨术的计划区域中，通过从FE释放周边的肌肉组织制备股骨沿骨干的中途区域MUR。首先，通过将Henahan电梯垂直于股骨的暴露表面，然后用手术刀时，释放在相邻区域中的肌肉启动。 <ol><li>通过向前推进继续到处去股骨，保持接近骨表面，直到所有的周边的肌肉组织从骨（如该缺陷将被创建）的整个中间部分被释放时，以及肌肉组织被完全清洗从骨。而这样做，这是非常重要的，以贴近骨表面，以避免切割任何主要血管。 </li></ol></li><li>对于一个5毫米大的骨缺损，环2个吉利线锯（0.22 MM）周围骨的内-外方向（ 图1E，F）。后循环的线锯，位置一块在股骨的远端侧，靠近膝关节，以及第二片上的基端侧靠近髋关节。夹紧吉利线锯，每边条采用S形弯曲的解剖和超级联赛茜镊子，使得它停留在所需的位置。如果一个切口截骨计划，那么只使用一块线锯。 </li><li>使用外固定器板作为模板，以确定所述植入物的精确位置。的外固定器的位置必须尽可能地靠近股骨的中心。 <ol><li>定位的外固定器板对骨的外侧表面。这是通过从外部旋转股骨实现。在该位置上的软组织层是在其最薄，从而防止过度的软组织张力固定器板下后的伤口闭合。 </li><li>然后稍稍抬起的外固定器板脱骨表面，以确保该板的孔中心向骨表面。持的外固定器具有小钳位为它留下平行于骨的纵向轴线，然后再使用电动工具或手钻预钻的第一孔上股骨与0.79的基端侧毫米的钻头。在向前发展，确保钻头的尖端仍集中于骨面。 </li><li>如果钻头的前端一直滑动，使用1.00毫米计数器沉降片（ 图8F），使第一个孔的位置。计数器沉降应用于定位所有剩余的安装销。这将确保在钻孔的相对于骨表面的精确对准和固定器板。 </li></ol></li></ol> 3，外固定支架植入方法采用SAW指南<ol><li>确保外固定器的板上锯导裁剪之前没有安装颠倒。通过比较在板中的孔的大小来确定。正确的一面是大孔的直径朝上。如果固定器内的孔的大小之间的差异并不明显，使用计数器沉降。 <ol><li>将反伸卡球的尖端插入孔上的一个固定器板，如果计数器沉降片容易装配到孔则是这种固定器的上侧，但是，如果计数器沉降片的前端不适合，那么这是固定器的底侧，并具有可翻转对于植入相应。 （ 重要提示:请确认钻垂直于骨的纵轴，因为这将确保固定器的完美定位于骨面的第一个钻孔的方向决定于骨骼的固定器的最终取向记住安装管脚具有相同的长度，并且如果固定器是不平行于骨的纵向轴线固定器与骨之间的距离会发生变化太多，可能会阻止所有四个安装管脚穿透两个皮层的能力。） </li><li>后的取向被证实，夹在板上的锯导（ 图2A，B），然后夹在装置上的骨，使得第一预钻孔对准在板（ 图2C）的第一孔。使用插入手电钻0.70平方毫米的套筒扳手带动第​​一安装销插入孔。这样做将允许可重复定位其余的安装引脚。 </li><li>后的第一个安装销是否到位，然后钻从在远端侧的第一安装销最远孔，并驱动所述第二安装销插入孔。的两个中间安装管脚注入顺序并不重要。 </li></ol></li></ol> 4，外固定支架植入方法没有锯指南: 也可以进行外固定器的应用程序，而无需使用该锯导。的外固定器植入的起始步骤是相同的​​，直到该单元与锯导被夹在骨头上（步骤3.1）。如果不使用该锯导，这是非常重要的，以保持该固定器板在正确的方向中整个应用程序。股骨需要在外侧方向外旋。 <ol><li>持的外固定器板用小夹子或S形弯曲的解剖和结扎镊子，使得它平行于骨（ 图3A）的纵向轴线。第一安装销的应用将确定固定器的对准，因此，骨的旋转必须被保留，直到第一销被插入时（ 图3B）。后的第一脚到位，小心使用镊子保持固定器板，以作为演练指导。 </li><li>插入钻头进入第二孔-这是最远侧孔的计划截骨间隙（ 图3C）。钻之前，检查并确保第二孔具有相同的方向作为第一个孔;还要确保后钻完成后，双方皮质参透。 </li><li>将0.70毫米见方的微博点¯x扳手的手钻，然后插入安装销插入小费。小心地将其插入到外固定器的板，而不会失去所述第一预钻孔的对齐。 </li><li>一旦该尖端与骨接触，开始转向下施加于手电钻的近端连续的轴向载荷的扳手。经过约5个圈，确保在固定销的近端线捕捉外固定架板的尸体。这个线程锁定系统。停止转动时，骨螺纹的端部是靠近骨（ 图3D）的顶表面上。 </li><li>后在最远端和近端侧上的插针的地方，预钻剩余的两个中间的孔。的中间的两个销植入顺序并不重要（ 图3C）。 </li><li>之后的外固定支架到位后，使用0.22毫米吉利线锯导演锯导，使节段性缺损（ 造型玩具ê4A）。如果选择后一种方法，该锯导板制造缺陷之前裁剪。 <ol><li>对于这一点，通过0.22mm的吉利线锯穿过股骨（ 图5A）下方的2槽使用充分灌溉（用5毫升注射器，免除盐水创造了往复运动一个5毫米的节段性缺损来回（ 图5B）缺陷创建的时间）。为了避免损坏软组织，切锯丝靠近骨的一侧完成截骨后。移除锯导板（ 图4B）。 </li></ol></li><li>缺陷或截骨被创建后，取出锯导和闭合伤口中的层时，肌肉第一（ 图4C），然后在皮肤（ 图4D）。在伤口闭合，治疗的缺陷按计划在研究方案。关闭肌层和使用Ethibond薇乔缝线4-0的阔筋膜，用爱惜康Monocryl 3-0 S中的皮肤uture。避免拖动缝合材料在非无菌表面，同时缝合伤口。注意:为了避免咬的伤口，缝合必须没有结束远端下植入。同样，可用于代替缝合皮肤胶。 </li><li>在前三术后几天，给大鼠镇痛每12小时以及抗生素每24个小时。当然，手术后的治疗方案的药物将取决于所使用的每个研究者的药物的制造商和品牌而有所不同（参照药物说明书中说明）。 </li><li>手术后经常监测动物，以确保他们从麻醉中恢复，然后才返回他们的住房设施。提供单独的住房，第几天手术后，确保没有任何并发​​症。 </li><li>手术后监测水，食物摄取和体重，以确保动物是不疼痛和痛苦。如果动物显示出活动水平降低，走动困难（可能植入失败），ataxiA，蓬头垢面苔白腻，眼睛周围和鼻孔，弯腰驼背的姿势，呼吸窘迫，食物和水的摄入减少等卟啉染色请教兽医。 </li></ol> 5，变更外固定支架刚度体内 <ol><li>若研究协议要求固定架刚度期间体内这个愈合过程的改变是通过改变固定使用附于手电钻0.5mm见方扳手盒的特殊互锁螺钉的连接元件来实现的。在此过程中，麻醉大鼠（参考在协议2.3），并给予镇痛（均指在协议2.4）只有一次的过程（ 图6A）的时间。 <ol><li>镇静的大鼠，然后插入0.50平方毫米的套筒扳手的前端进入安装于组装固定器侧的连动螺钉，并开始小心逆时针旋转，直到销是半出路（ 图6B </stron克&gt;）。之后这部分是完整的，则重复该过程为第二销即上的外固定器板（ 图6C）的同一侧。 </li><li>当两个引脚上的同一侧的一半出路，使用镊子或夹子来用温和的运动（ 图6D）除去在相对 ​​侧上的连接元件。该连接元件应该脱落容易，如果没有，则执行额外的夫妇匝上都互锁螺钉，以确保所述互锁螺钉的前端不会在连接元件埋入。 </li><li>该连接元件被移除之后，在地方的除去1（ 图6E）滑动所需的刚性连接元件，并从相反侧用方形框扳手开始旋转，直到互锁螺钉是从相反侧的一半（ 图6F）。重复同样的步骤进行第二联锁螺丝（ 图6G）。重要提示:次是将需要切换到该板的相对侧，以确保两个互锁螺钉是半出路在该处的连接元件被替换（图6H，I）的一侧。 </li><li>后此部分成功完成后，除去所述第二连接元件（ 图6J），它具有相同的刚度连接元件作为一个替代的相反侧（ 图6K）取代。后的第二连接元件是否到位，开始驱动互锁螺钉，直到联锁螺杆端离开该板的相对侧，并且所述互锁螺钉尖已经退出同量在每一侧（ 图6L）。重复同样的步骤进行第二联锁螺丝（ 图6M，N）。此过程需要约15分钟才能完成。 </li></ol></li></ol>

<ol>
	<li>Einhorn, T. A., Lane, J. M., Burstein, A. H., Kopman, C. R., Vigorita, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+healing+of+segmental+bone+defects+induced+by+demineralized+bone+matrix.+A+radiographic+and+biomechanical+study.">The healing of segmental bone defects induced by demineralized bone matrix. A radiographic and biomechanical study.</a> J Bone Joint Surg Am. 66, 274-279 (1984).</li><li>Feighan, J. E., Davy, D., Prewett, A. B., Stevenson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+bone+by+a+demineralized+bone+matrix+gel:+a+study+in+a+rat+femoral+defect+model.">Induction of bone by a demineralized bone matrix gel: a study in a rat femoral defect model.</a> J Orthop Res. 13, 881-891 (1995).</li><li>Hunt, T. R., Schwappach, J. R., Anderson, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Healing+of+a+segmental+defect+in+the+rat+femur+with+use+of+an+extract+from+a+cultured+human+osteosarcoma+cell-line+(Saos-2).+A+preliminary+report.">Healing of a segmental defect in the rat femur with use of an extract from a cultured human osteosarcoma cell-line (Saos-2). A preliminary report.</a> J Bone Joint Surg Am. 78, 41-48 (1996).</li><li>Jazrawi, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+and+cartilage+formation+in+an+experimental+model+of+distraction+osteogenesis.">Bone and cartilage formation in an experimental model of distraction osteogenesis.</a> J Orthop Trauma. 12, 111-116 (1998).</li><li>Probst, A., Jansen, H., Ladas, A., Spiegel, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Callus+formation+and+fixation+rigidity:+a+fracture+model+in+rats.">Callus formation and fixation rigidity: a fracture model in rats.</a> J Orthop Res. 17, 256-260 (1999).</li><li>Richards, M., Huibregtse, B. A., Caplan, A. I., Goulet, J. A., Goldstein, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marrow-derived+progenitor+cell+injections+enhance+new+bone+formation+during+distraction.">Marrow-derived progenitor cell injections enhance new bone formation during distraction.</a> J Orthop Res. 17, 900-908 (1999).</li><li>Aro, H. T., Chao, E. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone-healing+patterns+affected+by+loading,+fracture+fragment+stability,+fracture+type,+and+fracture+site+compression.">Bone-healing patterns affected by loading, fracture fragment stability, fracture type, and fracture site compression.</a> Clin Orthop Relat Res. , 8-17 (1993).</li><li>Augat, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shear+movement+at+the+fracture+site+delays+healing+in+a+diaphyseal+fracture+model.">Shear movement at the fracture site delays healing in a diaphyseal fracture model.</a> J Orthop Res. 21, 1011-1017 (2003).</li><li>Augat, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+tissue+properties+in+bone+healing:+influence+of+size+and+stability+of+the+osteotomy+gap.">Local tissue properties in bone healing: influence of size and stability of the osteotomy gap.</a> J Orthop Res. 16, 475-481 (1998).</li><li>Claes, L., Augat, P., Suger, G., Wilke, 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=Influence+of+size+and+stability+of+the+osteotomy+gap+on+the+success+of+fracture+healing.">Influence of size and stability of the osteotomy gap on the success of fracture healing.</a> J Orthop Res. 15, 577-584 (1997).</li><li>Claes, L., Eckert-Hubner, K., Augat, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fracture+gap+size+influences+the+local+vascularization+and+tissue+differentiation+in+callus+healing.">The fracture gap size influences the local vascularization and tissue differentiation in callus healing.</a> Langenbecks Arch Surg. 388, 316-322 (2003).</li><li>Duda, G. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interfragmentary+motion+in+tibial+osteotomies+stabilized+with+ring+fixators.">Interfragmentary motion in tibial osteotomies stabilized with ring fixators.</a> Clin Orthop Relat Res. , 163-172 (2002).</li><li>Goodship, A. E., Watkins, P. E., Rigby, H. S., Kenwright, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+fixator+frame+stiffness+in+the+control+of+fracture+healing.+An+experimental+study.">The role of fixator frame stiffness in the control of fracture healing. An experimental study.</a> J Biomech. 26, 1027-1035 (1993).</li><li>Williams, E. A., Rand, J. A., An, K. N., Chao, E. Y., Kelly, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+early+healing+of+tibial+osteotomies+stabilized+by+one-plane+or+two-plane+external+fixation.">The early healing of tibial osteotomies stabilized by one-plane or two-plane external fixation.</a> J Bone Joint Surg Am. 69, 355-365 (1987).</li><li>Wu, J. J., Shyr, H. S., Chao, E. Y., Kelly, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+osteotomy+healing+under+external+fixation+devices+with+different+stiffness+characteristics.">Comparison of osteotomy healing under external fixation devices with different stiffness characteristics.</a> J Bone Joint Surg Am. 66, 1258-1264 (1984).</li><li>Harrison, L. J., Cunningham, J. L., Stromberg, L., Goodship, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+induction+of+a+pseudarthrosis:+a+study+using+a+rodent+model.">Controlled induction of a pseudarthrosis: a study using a rodent model.</a> J Orthop Trauma. 17, 11-21 (2003).</li><li>Kaspar, K., Schell, H., Toben, D., Matziolis, G., Bail, 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=An+easily+reproducible+and+biomechanically+standardized+model+to+investigate+bone+healing+in+rats,+using+external+fixation.">An easily reproducible and biomechanically standardized model to investigate bone healing in rats, using external fixation.</a> Biomed Tech (Berl). 52, 383-390 (2007).</li><li>Mark, H., Bergholm, J., Nilsson, A., Rydevik, B., Stromberg, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+external+fixation+method+and+device+to+study+fracture+healing+in+rats.">An external fixation method and device to study fracture healing in rats.</a> Acta Orthop Scand. 74, 476-482 (2003).</li><li>Mark, H., Nilsson, A., Nannmark, U., Rydevik, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+fracture+fixation+stability+on+ossification+in+healing+fractures.">Effects of fracture fixation stability on ossification in healing fractures.</a> Clin Orthop Relat. Res. , 245-250 (2004).</li><li>Mark, H., Rydevik, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Torsional+stiffness+in+healing+fractures:+influence+of+ossification:+an+experimental+study+in+rats.">Torsional stiffness in healing fractures: influence of ossification: an experimental study in rats.</a> Acta Orthop. 76, 428-433 (2005).</li><li>McCann, R. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+osteoporosis+on+bone+mineral+density+and+fracture+repair+in+a+rat+femoral+fracture+model.">Effect of osteoporosis on bone mineral density and fracture repair in a rat femoral fracture model.</a> J Orthop Res. 26, 384-393 (2008).</li><li>Betz, O. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+percutaneous+gene+delivery+to+enhance+healing+of+segmental+bone+defects.">Direct percutaneous gene delivery to enhance healing of segmental bone defects.</a> J Bone Joint Surg Am. 88, 355-365 (2006).</li><li>Cullinane, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+a+neoarthrosis+by+precisely+controlled+motion+in+an+experimental+mid-femoral+defect.">Induction of a neoarthrosis by precisely controlled motion in an experimental mid-femoral defect.</a> J Orthop Res. 20, 579-586 (2002).</li><li>Dickson, G. R., Geddis, C., Fazzalari, N., Marsh, D., Parkinson, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcomputed+tomography+imaging+in+a+rat+model+of+delayed+union/non-union+fracture.">Microcomputed tomography imaging in a rat model of delayed union/non-union fracture.</a> J Orthop Res. 26, 729-736 (2008).</li><li>Jager, M., Sager, M., Lensing-Hohn, S., Krauspe, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+critical+size+bony+defect+in+a+small+animal+for+bone+healing+studies+(II):+implant+evolution+and+surgical+technique+on+a+rat's+femur.">The critical size bony defect in a small animal for bone healing studies (II): implant evolution and surgical technique on a rat's femur.</a> Biomed Tech (Berl). 50, 137-142 (2005).</li><li>Betz, V. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Healing+of+segmental+bone+defects+by+direct+percutaneous+gene+delivery:+effect+of+vector+dose.">Healing of segmental bone defects by direct percutaneous gene delivery: effect of vector dose.</a> Hum Gene Ther. 18, 907-915 (2007).</li><li>Glatt, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ability+of+recombinant+human+bone+morphogenetic+protein+2+to+enhance+bone+healing+in+the+presence+of+tobramycin:+evaluation+in+a+rat+segmental+defect+model.">Ability of recombinant human bone morphogenetic protein 2 to enhance bone healing in the presence of tobramycin: evaluation in a rat segmental defect model.</a> J Orthop Trauma. 23, 693-701 (2009).</li><li>Willie, B., Adkins, K., Zheng, X., Simon, U., Claes, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+characterization+of+external+fixator+stiffness+for+a+rat+femoral+fracture+model.">Mechanical characterization of external fixator stiffness for a rat femoral fracture model.</a> J Orthop Res. 27, 687-693 (2009).</li><li>Hess, T., Hopf, T., Fritsch, E., Mittelmeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+biomechanical+studies+of+conventional+and+self-tapping+cortical+bone+screws.">Comparative biomechanical studies of conventional and self-tapping cortical bone screws.</a> Z Orthop Ihre Grenzgeb. 129, 278-282 (1991).</li><li>Glatt, V., Evans, C. H., Matthys, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design,+characterisation+and+in+vivo+testing+of+a+new,+adjustable+stiffness,+external+fixator+for+the+rat+femur.">Design, characterisation and in vivo testing of a new, adjustable stiffness, external fixator for the rat femur.</a> Eur Cell Mater. 23, 289-298 (2012).</li><li>Glatt, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+healing+of+large+segmental+defects+in+the+rat+femur+by+reverse+dynamization+in+the+presence+of+bone+morphogenetic+protein-2.">Improved healing of large segmental defects in the rat femur by reverse dynamization in the presence of bone morphogenetic protein-2.</a> J Bone Joint Surg Am. 94, 2063-2073 (2012).</li></ol>

笔者罗马诺马蒂斯是RISystem公司瑞士达沃斯产生的植入物，本文中使用的植入特定的仪器和耗材的雇员。笔者瓦伊达格拉特没有竞争的经济利益。

手术过程中最关键的步骤来创建一个大的骨缺损是:1）选择适当的体重的大鼠相匹配的外固定器的大小; 2）维持在手术过程中的无菌环境;和3）以下的外科手术过程的协议。 本研究的主要目标是设计，制造和对大鼠股骨大缺陷模型表征一个新的，可变刚性的外固定器，并使用此固定器在确定在愈合过程中的生物和机械因素之间的相互作用。新的定位器的机械性能进行了检查，在三级和固定器的特征是发表在一个不同的原稿30。的固定器也被施加到大鼠股骨和其在体内的性能监测X线8周使用和不使用的治疗30,31。 主I该固定架nnovation是其交换的稳定性酒吧连接元件选择不同的标准化刚度的能力。因为稳定性栏的连接元件可以在设备安装于动物进行交换，刚度，可以在不同的阶段在愈合过程中进行调整。连接元件在一个时间交换1，以防止缺陷的边缘和新形成的组织的破坏不对准在协议中说明。目前，四种不同的刚度是可用的，但附加的刚度可以通过简单地通过将植入物系统的生产者订货不同厚度的不同的连接元件来实现的。 安装引脚和主框架是从谭和PEEK，分别作了，因为已经用于骨科植入物这些材料在人类和他们的生物相容性，是公认的。这些材料还允许在体内成像骨折修复以最小的失真，而且感染的发生率降低早期阶段。 体内实验证实，固定器让成像清晰，并保持一个5毫米段差距至少8周，无感染或销松动。 作为一个额外的设计特征，所述固定器具有预先设定为6mm从骨表面到稳定条不论偏移的刚度连接元件使用。这一特点使得固定器植入很重复的。以上文献中的1,18,26,27描述的替代设计的另一个重要优点是，在新的外固定器被设计成具有最小的质量（0.32 g）至避免不受控制的加载由于惯性。此外，皮肤的注入和缝合之后，植入物横杆与皮肤之间的间隙只有约2mm。这样靠近皮肤表面最小化的瞬间力，从而防止的可能性的缺陷比一个目标从外固定器等之内的额外负载。此外，为了保持手术创伤低，常规和旋转锯不被认为是作为工具，用于创建或大或小的截骨。这种锯或者切成相邻组织或剥离骨膜时的组织缩回。在过去，我们已经使用4.5 mm的牙科毛刺看到创建5毫米缺陷和发现，这是不可能创造出平行两端22,26,27精确和可重复大小的缺陷。为了避免这些问题，我们采取了0.22毫米吉利线锯的优点。锯指南可重复创建平行两端精确的缺陷被开发。 有使用这种技术时，有一些限制。一种使用该外固定器时，主要关心的问题是在结果部分所提到的，该大鼠可能咀嚼通过外固定器板，它是由聚醚醚酮的可能性。然而，一个特定人铁盖最近开发的固定器，以防止这种情况发生的生产商。同样，伊丽莎白项圈可用于最初几周的手术后防止动物咀嚼。一个附加的问题是，如果一个空的骨缺损是用于研究，存在安装销可以在手术后的骨几周拔出的机会。另外，至关重要的是，固定器植入在被在该协议中概述的精确定位。如果该指令不小心接着，有很大的风险，即由特定刚性固定器提供的机械环境不会像意，并且将引入的误差，提供虚假的结果。 本文中所描述的固定器让研究者进行所必需的确定各种力学环境和/或机械（刚度）调制上的b经验的影响，实验1治疗大缺陷或截骨30,31。另外，外固定器技术可以在不同的药物和生物材料进行了测试，发现在以加速愈合过程的新疗法不仅复杂骨折，但也适用于标准治疗骨折的各种研究中使用。

大量的研究已经提高了参与骨组织的修复1-6的生物学机制的理解。对骨修复的机械条件，如轴向，剪切和interfragmentary运动（的IFM）的影响已经被广泛地7-15研究。在过去的几年中，越来越多的研究开始出现用骨折，截骨术和大节段性骨缺损的体内模型，描述在骨愈合的力学环境的影响。因此，需要可靠的固定方法来获得可重复和可靠的研究成果。 周围的骨折愈合的力学环境是非常重要的，因为它决定的方式，将骨折愈合。因此，固定装置的选择是非常重要的，应根据不同的研究设计进行仔细选择，以及其它因素，如间隙大小和骨折的类型。该固定装置的机械性能再更重要的，当学习的大块骨缺损的骨性愈合，以建立一个固定的提供不仅遍及全重量的试验期带有一定的间隙大小，但也是一个理想的力学环境对骨愈合。外固定器在骨折和大骨缺损愈合的实验模型常用的，因为他们有比其他固定设备的优势。外固定器的主要优点是，它们允许在缺损部位在生物体内的机械环境的变化而二次干预，这可以通过在实验的过程中改变或调整该装置的稳定性杆来实现骨愈合的进展。此外，它允许对特定局部机械刺激的应用，以提高骨的修复，并且还提供了测量愈伤组织在体内的刚性的可能性。然而，这些设备也有一些缺点包括:软组织，感染和销断裂刺激。 不幸的是，这样的植入物是不提供"现成的"在植入物开发的时间，并调查被迫定制设计自己的固定器用于预期用途。因此，研究这方面的一个约束是缺乏实验控制中的大段缺损的局部力学环境，以及截骨术，因为它愈合。的外固定器的机械特性是由所定义，并且可以通过进行调制，大量的变量，包括:销，销的直径，销材料之间的距离，销，固定器杆的长度，固定器栏数的数目，固定架棒材，固定器杆的厚度和从骨表面到固定器杆（偏移）的距离。令人惊奇的是，研究仅缺乏可以发现，已经研究了各组分的机械贡献在啮齿类动物的研究16,18,28使用固定器或整体框架结构。例如，一项研究的结果表明，主要的因素，在确定的固定结构的总刚度之一是相对于它们的偏移量，直径和材料特性28把持销的灵活性。从上述研究结果清楚地表明，知道由固定装置提供的机械环境中是非常重要的，然而，在许多情况下，不详细调查。本文报道的设计，规格和植入体内的外固定器，解决这个问题的。此固定器还允许机械环境愈合的进展，一个属性，可以在体内治疗过程中的不同阶段的机械性刺激的敏感性的研究的调制。此外，还有强加控制的和可重复的当地技工人的环境中，其辅助功能也允许这样的环境中的骨愈合的不同阶段的调制。 我们设计的固定器是基于外固定，因而被广泛用于骨折固定16-21和大型缺损模型的实验动物22-27。我们的外固定架和文献报道的其他现有设计之间的不同之处在于它们的稳定性条用螺丝紧固有得紧用克氏针（克氏针）。这种类型的设计，需要螺钉被每两周重新拧紧（有时甚至每周一次），以确保该偏移的距离保持为加载是通过负重，以防止稳定性杆的松动施加。如果发生这样的松动，它允许不必要的额外负荷条件，如角，横向和扭转剪切动作，愈合骨（根据个人经验，与researche通信RS），认识到这一点，外部固定器被设计为使得需要改变固定器的刚度时，它会通过除去附着在那里的安装销被嵌入在主模块的连接元件来实现的。与新的外固定器样机进行体内试验试验，以确保其符合之前的大批量制造的所有提出的要求。 其主要目的为本文是提出一种新的手术方法，用于大的骨缺损和骨切开术的大鼠与在愈合过程中改变刚度体内的能力的外固定器。这种固定方法是在体内应用对大鼠股骨。

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			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.390.130</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Drill Bit 0.79 mm</td>
			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.593.203</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Gigly wire saw 0.22 mm</td>
			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.590.100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Square box wrench 0.70 mm</td>
			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.590.112</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Square box wrench 0.50 mm</td>
			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.590.111</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Centering bit 1.00 mm</td>
			<td>RISystem AG Davos, Switzerland</td>
			<td style="width:117px;">RIS.592.205</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Scalpel Blade handle</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Scalpel Blade (Size 15)</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Tissue Forceps</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Scissors</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Retractor</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Needle Holder</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Henahan Elevator</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">S-shape curved dissecting and ligature forceps&#160;</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td>2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Dressing Forceps</td>
			<td style="width:283px;">Fine Science tools</td>
			<td style="width:117px;"></td>
			<td>2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Sterile Fenestrated drape</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>for surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile gauze</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>for surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5 ml syringe&#160;</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>&#160;for irrigation of defect</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">24-27G needle&#160;</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>&#160;for irrigation of defect</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1cc Insulin syringes&#160;</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>for drug injections</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sterile saline&#160;</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>for bone defect irrigation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sterile gloves</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>to perform surgeries</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">chlorohezadine</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>disinfecting solution for surgical site</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vicryl suture 4-0 with SH-1</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>to suture muscle&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethibond suture 3-0&#160;</td>
			<td style="width:283px;">Fisher Scientific</td>
			<td style="width:117px;"></td>
			<td>to suture skin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isofluorine</td>
			<td style="width:283px;">Sigma-Aldrich</td>
			<td style="width:117px;"></td>
			<td>for anesthesia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td style="width:283px;">Sigma-Aldrich</td>
			<td style="width:117px;"></td>
			<td>analgesia during and after the surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cefazolin</td>
			<td style="width:283px;">Sigma-Aldrich</td>
			<td style="width:117px;"></td>
			<td>antibiotic during and after the surgery&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sprague-Dawley Rats or any other strain</td>
			<td>Charles River Laboratories International, Inc. (Wilmington, MA USA)&#160;</td>
			<td style="width:117px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

adjustable stiffness, external fixator for the rat femur osteotomy and segmental bone defect models

围绕断骨的愈合的力学环境是非常重要的，因为它决定的方式，将骨折愈合。在过去的十年里一直在通过病灶周围的固定稳定性改变力学环境改善骨愈合重要的临床利益。临床前动物研究在这方面的一个制约因素是缺乏实验控制了当地的力学环境中的大段缺损，以及他们截骨愈合的。在本文中，我们的设计和使用的外固定器的汇报，研究了大段骨缺损或截骨愈合。此设备不仅允许对骨病变可控制的轴向刚度，因为它可以治愈，但是它也使刚度在体内治疗过程中的变化的进行的实验表明，该固定器能够保持一个5毫米的股骨缺损间隙在不受限制的笼中老鼠体内活性为至少8周。同样，我们没有观察到变形或感染，包括在整个治疗期间销感染。这些结果表明，我们的新研制的外固定器能够实现可再现的和标准化的稳定化，并在大鼠体内大的骨缺损和各种尺寸截骨术的机械环境的改变。这证实了外固定器非常适合于使用在骨再生和修复的领域的大鼠模型的临床前研究调查。

设计规格大鼠股骨与外部固定系统的稳定化使得从0.5至5mm的创建截骨的。外固定支架系统是由聚醚醚酮（PEEK - [主体]）锁定外固定器和钛 - 铝 - 铌合金（TAN - [固定栓]），它提供了一个简单，重复性好，可调节的设计，并在四个不同的刚度可供选择:10，40，70和100％（100％为标准，最严格的固定器（ 图7）根据各研究者的研究需求，无论他们将不得不做植入刚度调节体内的。骨愈合的进展，外固定器板自带无论是作为一种固体片（ 图8）或具有两个连接元件（ 图9A）和两个主要模块（ 图9B）固定有两个互锁螺钉 （ 网络gure 9C）而必须在手术之前（ 图10A-F）的组装。该连接元件是不同厚度的，并因此刚度，和开发，以实现固定刚度相当于10％（0.75毫米厚），40％（1.70毫米厚），70％（2.10毫米厚）和100％（2.50毫米厚; 图7）。的100％的外固定器的刚度基于200克近似体重，一个成熟的大鼠进行了计算，然后通过一个因子4相乘，以质量等于为800g。这样做是为了确保在创建一个5毫米的缺陷之后，固定器是能够承受负重的动物，从而保持对准并且防止缺陷片段的错位。剩下的三个固定支架的刚度分别降低从最高（100％）30％有多种刚度用于各种目的的研究。 各主要模块有两个孔，其中安装引脚是樱雪rted。固定器刚度，同时它仍然附着在活的动物，通过改变使用连接在手电钻（ 图9K）0.5平方毫米扳手框（ 图9H），固定的特殊互锁螺钉（ 图9C）的连接元件来改变。 TAN（钛合金）被用来制作用于安装销（ 图9D），以确保稳定性栏到股骨（ 图7）。固定器有四种片，需要在使用前进行组装，如果刚度变化旨在用于研究（ 图10A-F），如果没有，应该使用一个单一的固体拼凑固定架。外螺钉之间的距离为16毫米，而中间螺钉之间的距离为11毫米。所有的孔都用0.79毫米钻头（ 图9E）预钻。螺钉被锁定在对应于主固定支架框架，其平行于骨表面的孔和设置在6mm的距离来回米的骨（ 图7）。 一种锯导向的开发是为了支持创建一个精确的，可重复的，五毫米段的股骨（ 图9I）缺陷;它也作为定位导向装置的外固定器的安装。的外固定器的主框架被夹在所述锯导板，然后将整个体系被夹在骨， 如图2B所示，C。 5毫米左右间隙用0.22mm的吉利线锯（ 图9J）产生。无论是锯导板和吉利线锯可在134ºC高压灭菌。如果不同大小的截骨的目的是为研究对象，设计了一个自定义的锯导是可用的。由于外固定器的体积小，一组特殊的植入器械的设计和收购;定制0.79毫米钻头（ 图9E），对于孔（ 图9F）的预钻1.00毫米反伸卡球，0.7毫米SQUA再对应用的安装于手电钻（ 图9G），用于应用程序的互锁螺钉（ 图9H），手电钻（ 图9K）0.5平方毫米扳手盒安装销扳手方块。 一个累加器笔钻（ 图9L）也被开发。各安装销的芯直径比钻头0.02毫米更大，以保证安装销正确地安装到骨上。当使用自切螺钉头一起使用，这已被证明，以防止在所述骨螺钉接口29松开，由于骨表面吸收。钻头（ 图9E）是由产生2500转500毫瓦（ 图9L）的功率的微型电机的Accu钻笔操作。 体内实验影像学检查证实，所有刚度的固定器保持1毫米（未显示）或一个5毫米的铁在整个8周的实验（ 图11）道德缺陷。这是为5mm的临界尺寸的缺陷，其中自发愈合，不会发生特别重要。没有失真或感染，包括销感染，观察和销松动缺席，如果应用程序的说明进行操作30。采用外固定器的并发症，如果看到老鼠在手术时的体重已经突破2.5克，和一个更小的尺寸板使用。在其中的一些实例，在安装管脚装载增大到临界电平，以使销pullouts分别在股骨的远端侧的任意位置从一周的手术（ 图12）之后所发生的两周。除此之外，如果一个较大尺寸的动物的情况下，周围的股骨肌肉组织是相对厚的，从而产生皮肤张力，在皮肤结束后，植入物的附近。由于膨胀的张力，当皮肤S挞治愈它创建了一个瘙痒的感觉，以使一些老鼠咬了固定器的动物。由于固定器采用PEEK材料，基本上是高密度塑料，在少数情况下创建的，一些老鼠被称为嚼过它。再次，为了避免这种情况，这是非常重要的，选择推荐的体重为动物研究或切换到外固定器的放大版本。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig1highres.jpg" width="500" /> 图1的手术大鼠股骨制备。 （A）大鼠置于俯卧位（B）显示在股骨上的切口的方向上（c）示出切口，在皮肤，暴露肌肉（d）示出切口穿过肌肉以暴露股骨（ E）是一个小夹子放置骨下通过千兆李丝（F）显示通过骨下方吉利线。 <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig2highres.jpg" width="500" /> 图2（A）锯导（B）外固定器夹在锯导（C）锯导板与外固定器夹在股骨。 <img alt="图3" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig3highres.jpg" width="500" /> 图外固定器3的应用。 （a）示出了第一安装销与所述板的正确应用斜倚前-侧面和平行于骨-绿色的手，和不正确的应用-红手（b）示出插入所述第一安装销的外远端位置（c）示出的插入的<html> 。其余安装管脚先从最基端的位置，接着是两个中间安装管脚（d）表示插入安装销的-在协议4.4节更详细描述的。 <img alt="图4" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig4highres.jpg" width="500" /> 图对大鼠股骨外固定架4手术植入。 （一）演示手术过程，在地方与吉利线外固定器完成。（二）示范创建5毫米节段性缺损（C）演示缝合肌肉层，暴露外固定架的稳定性吧。（四）示范缝合皮肤露外固定架的稳定性吧。 558fig5highres.jpg"WIDTH ="500"/&gt; 吉利丝产生缺陷（二）图5（A）的初始位置。图像显示出吉利丝往复运动。 <img alt="图6" fo:content-width="3.8in" src="/files/ftp_upload/51558/51558fig6highres.jpg" width="500" /> 图6变化外固定支架的刚度体内。（一）外部固定器植入股骨（B）小心地逆时针旋转，直到脚是半出路显示除去第一互锁螺钉。（c）所示拆除（D）的所述第二互锁螺钉通过仔细逆时针旋转，直到销是出了一半。演示除去在相对 ​​侧上的连接元件（E）展示来代替移除的一个替换所需的刚性连接元件。 （F），演示如何确保第一从相反侧替换连接元件转动的方盒子扳手直至联锁螺钉是半出路的相反侧的，（G），演示如何从相反侧固定在第二替换连接元件通过将正方形框扳手直至联锁螺钉是半出路的相反侧的，（H，I）显示出在切换到板的另一侧，以确保两个互锁螺钉是半出路上侧，其中连接元件代替。（J）展示去除所述第二连接元件（K）阐明置换代替移除的一个第二刚性连接元件（L，M）阐明驱动两个互锁螺钉，直到联锁螺纹端退出该板的相对侧（N）表明已完成的程序。 </P&gt; <img alt="图7" fo:content-width="6.5in" src="/files/ftp_upload/51558/51558fig7highres-replacement.jpg" width="650" /> 图中的外固定器的7组件左:刚度是由具有不同厚度的连接元件来确定。固定器连接到骨与钛合金的安装销。右:组装固定器代替对大鼠股骨5毫米节段性缺损。 <img alt="图8" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig8highres-replacement.jpg" width="500" /> 图8。外固定器作为一个单位。 <img alt="图9" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig9highres.jpg" width="500" /> 图9部件和仪器设计用于与外部固定器的使用。 （一）两个连接元件。（二）两个主要模块（C）两个联锁螺丝（D）四个固定栓（E）0.79毫米钻头。（F）的孔的预钻à1.00毫米计数器坠子。（七）0.7平方毫米套筒扳手，用于安装销的应用。（H）使用0.5平方毫米套筒扳手互锁螺钉的应用（一）一个5毫米锯导。（J）0.22mm的吉利线锯的创作缺陷。 （K）手电钻的钻头，0.70和0.50平方毫米的套筒扳手的连接（L）AccuPen 6V +（微型电子笔钻头）来驱动钻头。 <img alt="图10" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig10highres-replacement.jpg" width="500" /> 外固定器的图10大会。 （一）70％STiffness连接元件（B）的连接元件与主模块中的一个（C）展示了如何的连接元件的内部的主要模块的幻灯片（D）展示了如何两个主模块的滑动连接元件的内（E）演示了这两种主要的模块，并在这两个地方连接元件（F）的演示完全组装的稳定性吧-主要功能模块，并用环环相扣的螺丝连接元件。 <img alt="图11" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig11highres.jpg" width="500" /> 图11 中的缺陷，在大鼠体内后的X射线图像立即手术后和8周的所有3刚度外固定器通过外科手术植入在创建大鼠股骨和5mm节段性缺损。造成的缺陷为手术后立即的X光检查（t = 0时），并在weeklÝ间隔直到8周（T = 8周）时终止实验。转载自ECM杂志（一种许可<a href="http://www.ecmjournal.org">http://www.ecmjournal.org</a> ）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/51558/51558fig11highres.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图12" fo:content-width="5in" src="/files/ftp_upload/51558/51558fig12highres.jpg" width="500" /> 图12 在大鼠缺陷9天手术后与远端针拔出（在手术后体重大鼠为340克的时间）的体内 X射线图像。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/51558/51558fig12highres.jpg" target="_blank">请点击这里查看大图这个版本的人物。</a>

Watch this Scientific Journal Video about Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models at JoVE.com

Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

One constraint of preclinical research in the field of bone repair is the lack of experimental control over the local mechanical environment within a healing bone lesion. We report the design and use of an external fixator for bone repair with the ability to change fixator stiffness in vivo.

可调节刚度，外固定支架的大鼠股骨截骨和节段性骨缺损模型

The mechanical environment around the healing of broken bone is very important as it determines the way the fracture will heal. Over the past decade there ...

adjustable-stiffness-external-fixator-for-rat-femur-osteotomy

Nanyang Technological University

Research

JoVE Journal

Medicine

22.4K 次观看.  Queensland University of Technology. One constraint of preclinical research in the field of bone repair is the lack of experimental control over the local mechanical environment within a healing bone lesion. We report the design and use of an external fixator for bone repair with the ability to change fixator stiffness in vivo.

视频: Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

Models of Bone Metastasis

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible for significant morbidity and mortality1. Thus, there is a significant need to understand the molecular mechanisms controlling the establishment, growth and activity of tumors in bone. Several in vivo models have been established to study these events and each has specific benefits and limitations. The most commonly used model utilizes intracardiac inoculation of tumor cells directly into the arterial blood supply of athymic (nude) BalbC mice. This procedure can be applied to many different tumor types (including PC-3 prostate cancer, lung carcinoma, and mouse mammary fat pad tumors); however, in this manuscript we will focus on the breast cancer model, MDA-MB-231. In this model we utilize a highly bone-selective clone, originally derived in Dr. Mundy's group in San Antonio2, that has since been transfected for GFP expression and re-cloned by our group3. This clone is a bone metastatic variant with a high rate of osteotropism and very little metastasis to lung, liver, or adrenal glands. While intracardiac injections are most commonly used for studies of bone metastasis2, in certain instances intratibial4 or mammary fat pad injections are more appropriate. Intracardiac injections are typically performed when using human tumor cells with the goal of monitoring later stages of metastasis, specifically the ability of cancer cells to arrest in bone, survive, proliferate, and establish tumors that develop into cancer-induced bone disease. Intratibial injections are performed if focusing on the relationship of cancer cells and bone after a tumor has metastasized to bone, which correlates roughly to established metastatic bone disease. Neither of these models recapitulates early steps in the metastatic process prior to embolism and entry of tumor cells into the circulation. If monitoring primary tumor growth or metastasis from the primary site to bone, then mammary fat pad inoculations are usually preferred; however, very few tumor cell lines will consistently metastasize to bone from the primary site, with 4T1 bone-preferential clones, a mouse mammary carcinoma, being the exception 5,6.
This manuscript details inoculation procedures and highlights key steps in post inoculation analyses. Specifically, it includes cell culture, tumor cell inoculation procedures for intracardiac and intratibial inoculations, as well as brief information regarding weekly monitoring by x-ray, fluorescence and histomorphometric analyses.

Animal models are frequently utilized to study cancer metastasis to bone. In this protocol we will describe two common methods of tumor inoculation for bone metastasis studies and briefly describe some of the analyses utilized to monitor and quantify these models.

骨转移模型

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible ...

科研

医学

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal hyperplasia (IH) that eventually leads to vessel occlusion and graft failure. Mechanical forces, particularly low shear stress and high wall tension, are thought to initiate and to sustain these cellular and molecular changes, but their exact contribution remains to be unraveled. To selectively evaluate the role of pressure and shear stress on the biology of IH, an ex&#160;vivo perfusion system (EVPS) was created to perfuse segments of human saphenous veins under arterial regimen (high shear stress and high pressure). Further technical innovations allowed the simultaneous perfusion of two segments from the same vein, one reinforced with an external mesh. Veins were harvested using a no-touch technique and immediately transferred to the laboratory for assembly in the EVPS. One segment of the freshly isolated vein was not perfused (control, day 0). The two others segments were perfused for up to 7 days, one being completely sheltered with a 4 mm (diameter) external mesh. The pressure, flow velocity, and pulse rate were continuously monitored and adjusted to mimic the hemodynamic conditions prevailing in the femoral artery. Upon completion of the perfusion, veins were dismounted and used for histological and molecular analysis. Under ex&#160;vivo conditions, high pressure perfusion (arterial, mean = 100 mm Hg) is sufficient to generate IH and remodeling of human veins. These alterations are reduced in the presence of an external polyester mesh.

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mechanisms leading to the development of intimal hyperplasia (IH) and vein graft failure are still poorly understood. This study describes an ex&#160;vivo system to perfuse human veins under controlled flow and pressure. Furthermore the efficiency of external mesh reinforcement to limit the development of IH was evaluated.

步骤人类隐静脉<em&gt;前体内</em&gt;灌注和外部强化

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

鼠肾移植:移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Establishment of a Segmental Femoral Critical-size Defect Model in Mice Stabilized by Plate Osteosynthesis

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a model organism, the mouse has already been widely used in bone-related research. Large diaphyseal bone defect models in mice, however, are sparse and often use bone fixation which fills the bone marrow cavity and does not provide optimal mechanical stability. The objectives of the current study were to develop a critical-size, segmental, femoral defect in nude mice. A 3.5-mm mid-diaphyseal femoral ostectomy (approximately 25% of the femur length) was performed using a dedicated jig, and was stabilized with an anterior located locking plate and 4 locking screws. The bone defect was subsequently either left empty or filled with a bone substitute (syngenic bone graft or coralline scaffold). Bone healing was monitored noninvasively using radiography and in vivo micro-computed-tomography and was subsequently assessed by ex vivo micro-computed-tomography and undecalcified histology after animal sacrifice, 10 weeks postoperatively. The recovery of all mice was excellent, a full-weight-bearing was observed within one day following the surgical procedure. Furthermore, stable bone fixation and consistent fixation of the implanted materials were achieved in all animals tested throughout the study. When the bone defects were left empty, non-union was consistently obtained. In contrast, when the bone defects were filled with syngenic bone grafts, bone union was always observed. When the bone defects were filled with coralline scaffolds, newly-formed bone was observed in the interface between bone resection edges and the scaffold, as well as within a short distance within the scaffold. 
The present model describes a reproducible critical-size femoral defect stabilized by plate osteosynthesis with low morbidity in mice. The new load-bearing segmental bone defect model could be useful for studying the underlying mechanisms in bone regeneration pertinent to orthopaedic applications.

Although mouse models are invaluable tools for bone tissue engineering, models of long bone defects are sparse. This need motivated development of the present protocol which uses a locking plate with four screws and a dedicated jig to perform and stabilize a reproducible, femoral, critical-size defect with low morbidity.

在小鼠节段性股临界尺寸缺陷模型的建立接骨板稳定

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a ...

An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming more commonly used in trauma research. They offer a large number of mutant strains and antibodies for the analysis of the molecular mechanisms behind the highly differentiated process of bone healing. To control the biomechanical environment, standardized and well-characterized osteosynthesis techniques are mandatory in mice. Here, we report on the design and use of an intramedullary nail to stabilize open femur osteotomies in mice. The nail, made of medical-grade stainless steel, provides high axial and rotational stiffness. The implant further allows the creation of defined, constant osteotomy gap sizes from 0.00 mm to 2.00 mm. Intramedullary locking nail stabilization of femur osteotomies with gap sizes of 0.00 mm and 0.25 mm result in adequate bone healing through endochondral and intramembranous ossification. Stabilization of femur osteotomies with a gap size of 2.00 mm results in atrophic non-union. Thus, the intramedullary locking nail can be used in healing and non-healing models. A further advantage of the use of the nail compared to other open bone healing models is the possibility to adequately fix bone substitutes and scaffolds in order to study the process of osseous integration. A disadvantage of the use of the intramedullary nail is the more invasive surgical procedure, inherent to all open procedures compared to closed models. A further disadvantage may be the induction of some damage to the intramedullary cavity, inherent to all intramedullary stabilization techniques compared to extramedullary stabilization procedures.

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

髓内锁钉股骨截骨标准化固定用于分析正常和有缺陷的骨愈合小鼠

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

直接节段性肝内交付使用大鼠肝门部夹具模型方法

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

在体内MRI 对小鼠股骨骨愈合过程中骨折愈伤组织发育的评价

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models

In translational medical research, porcine models have steadily become more popular. Considering the high value of individual animals, particularly of genetically modified pig models, and the often-limited number of available animals of these models, establishment of (biobank) collections of adequately processed tissue samples suited for a broad spectrum of subsequent analyses methods, including analyses not specified at the time point of sampling, represent meaningful approaches to take full advantage of the translational value of the model. With respect to the peculiarities of porcine anatomy, comprehensive guidelines have recently been established for standardized generation of representative, high-quality samples from different porcine organs and tissues. These guidelines are essential prerequisites for the reproducibility of results and their comparability between different studies and investigators. The recording of basic data, such as organ weights and volumes, the determination of the sampling locations and of the numbers of tissue samples to be generated, as well as their orientation, size, processing and trimming directions, are relevant factors determining the generalizability and usability of the specimen for molecular, qualitative, and quantitative morphological analyses. Here, an illustrative, practical, step-by-step demonstration of the most important techniques for generation of representative, multi-purpose biobank specimen from porcine tissues is presented. The methods described here include determination of organ/tissue volumes and densities, the application of a volume-weighted systematic random sampling procedure for parenchymal organs by point-counting, determination of the extent of tissue shrinkage related to histological embedding of samples, and generation of randomly oriented samples for quantitative stereological analyses, such as isotropic uniform random (IUR) sections generated by the &#34;Orientator&#34; and &#34;Isector&#34; methods, and vertical uniform random (VUR) sections.

The practical application and performance of methods for generation of representative tissue samples of porcine animal models for a broad spectrum of downstream analyses in biobank projects are demonstrated, including volumetry, systematic random sampling, and differential processing of tissue samples for qualitative and quantitative morphologic and molecular analyses types.

猪生物医学模型生物库组织样品的取样策略与处理

In translational medical research, porcine models have steadily become more popular. Considering the high value of individual animals, particularly of ...

Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ROM is called a joint contracture and results in increased morbidity. Due to the difficulty of reversing established knee contractures, early detection is important, and hence, knowing risk factors for their development is essential. The rat represents a good model with which the effect of an intervention can be studied due to the similarity of rat knee anatomy to that of humans, the rat's ability to tolerate long durations of knee immobilization in flexion, and because mechanical data can be correlated with histologic and biochemical analysis of knee tissue.
Using an automated arthrometer, we demonstrate a validated, precise, reproducible, user-independent method of measuring the extension ROM of the rat knee joint at specific torques. This arthrometer can be used to determine the effects of interventions on knee joint ROM in the rat.

The goal of the protocol is to measure the extension range of motion of the rat knee. The effects of various diseases that increase the stiffness of the knee joint and the effectiveness of treatments can be quantified.

用膝关节关节仪评价大鼠膝关节屈曲收缩的特定组织贡献

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

大鼠胡萝卜动脉压力控制段气球损伤与腹腔ventit 治疗应用

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...