这项工作是部分由美国社会的试点手奖和美国国立卫生研究院/ NIAMS 1K01AR068386-01（到AEL）和NIAMS / NIH P30AR061307外科支持。

大学委员会的动物研究在罗切斯特大学批准的所有动物实验。十-12周龄雌性C57BL /使用了6J小鼠。 1.屈肌腱手术动物的制备（约15分钟） <ol><li>高压灭菌手术器械消毒，戴整个无菌手套，并保持无菌操作领域。 </li><li>麻醉通过腹膜内注射（IP）用氯胺酮对应于体重的体积（80毫克/千克）和赛拉嗪（10毫克/千克）鼠标。通过没有脚趾捏反射确认镇静深度。 </li><li>通过皮下注射施用先发制人镇痛（丁丙诺啡，0.05-0.1毫克/千克）。应用眼药膏，以防止眼睛干燥。 </li><li>通过裁剪对整个后肢皮毛准备手术部位。通过依次用聚维酮碘，接着用70％乙醇洗涤用聚维酮碘消毒皮肤，并再次。 </li></ol><ol><li>找到趾长屈肌（FDL）肌腱，小腿内侧表面方面可见。使用手术刀，使一个小0.5-1厘米的切口在与微型剪刀将皮肤暴露肌腱。 </li><li>使用镊子的FDL肌腱与周围组织分离并跟踪它的肌腱交界处。切筋在该路口与弹簧剪刀将其释放，注意避免胫后动脉。 </li><li>关闭用5-0尼龙缝合皮肤。 注意事项:应使用立体显微镜来进行FDL横断和修理（步骤2.4）。 </li><li>使在使用弹簧微型剪刀的后爪的后外侧面3毫米切口。轻轻缩回周围的软组织和肌肉用钳子，确保最大限度地减少组织损伤，并确定FDL肌腱。 <ol><li>轻轻抬起FDL肌腱和使用MI完全横切它CRO剪刀。缝合FDL的两端肌腱一起用8-0缝线修改凯斯勒模式，通过用生理盐水润湿定期它避免了肌腱的干燥。 </li><li>更换软组织和肌肉在肌腱，然后关闭用5-0尼龙缝合切口。 </li></ol></li><li>放置在37℃的滑动温暖组小鼠中，以维持体温，直到从麻醉中恢复。 </li><li>监测小鼠手术后的损伤或感染的迹象，包括增加发声和皱皮。注射丁丙诺啡（50微升/鼠标）镇痛每隔12小时，直到小鼠不再显示疼痛或痛苦的迹象。 </li><li>让小鼠愈合继续下一步骤之前期10天，手术后的。 注:细胞可以在任何时间点手术后收获。这里，10天被选择为在此时给定的组织的相对高的细胞构成这些代表性实验。 </li></ol> 3. Labeli骑自行车细胞纳克（约10分钟） <ol><li>制备的EdU（5-乙炔基-2&#39;脱氧尿苷）溶液（10毫克/毫升在无菌磷酸盐缓冲盐水[PBS] + 1％二甲亚砜[DMSO]，以增加溶解度）。 </li><li>通过腹腔注射注射用100μl的EdU（50毫克/公斤）小鼠处死前24小时。 </li></ol> 4.收集细胞细胞学离心（2.5小时，约30分钟的手工操作时间） <ol><li>制备3毫克/毫升胶原酶我于PBS中。 </li><li>安乐死通过二氧化碳窒息小鼠在不超过3升/分钟的流速，以及执行颈椎脱位作为次要措施。 </li><li>使用上述步骤2.4定位修复肌腱，并通过削减约2毫米的受伤部位用微型剪刀的两边隔离肌腱组织。 <ol><li>与在培养皿用1ml的3毫克/毫升胶原酶手术刀切碎组织，然后通过18G的针头几次传混合物，打破了组织。收集混合物到1.5毫升离心管中。 </li&gt; <li>消化腱组织进行1小时，在37℃振荡。 </li><li>自旋组织向下（300×g离心5分钟），吸胶原酶，然后通过上下吹打数次重悬在PBS中500μl的3％牛血清白蛋白（BSA）的细胞/组织。 </li><li>通过一个70微米的细胞滤网过滤细胞悬浮液以除去碎片和大块肌腱，并在37℃培养箱中备用​​。 </li><li>肌腱组织放入另外150毫升离心管中，并用1 ml胶原酶溶液刷新，上下吹打混匀。 </li><li>孵育组织另一小时，在37℃振荡。 </li><li>降速消化肌腱组织（300×g离心5分钟），然后在PBS中500μl的3％BSA中通过上下吹打数次重悬。 </li><li>通过一个70微米的细胞滤网过滤细胞悬液，用悬挂在4.3.3中分离结合起来，再算上用血球细胞。 </li><li>用在PBS中的3％BSA洗涤细胞一次，然后重悬到3-6×10 百分之四微升。 </li></ol></li><li>附加细胞学漏斗至带正电的滑动和锁定到滑动载体。插入整个装置成细胞学离心机和100μl的细胞悬浮液添加到该漏斗中。离心机在300×g离心5分钟以分散到幻灯片的细胞。 </li></ol> 5.免疫细胞化学检测埃杜（2小时，〜45分钟的手工操作时间） 注意:应在黑暗中进行所有的温育步骤，限制荧光团的光漂白。 <ol><li>圈单元与疏水性屏障笔，以减少所需要的以下步骤溶液中，然后立即固定用在PBS中15分钟150μl的3％多聚甲醛在室温（RT）。 <ol><li>为5分钟，在PBS中的150微升3％BSA洗涤两次。 </li><li>通透细胞用0.5％Triton-X的PBS 20分钟在RT 100。 </li><li>重复洗涤步骤为5.1.1。 </li></ol></li><li>准备的EdU反应cocktAIL按试剂盒说明书不超过30分钟后使用。 <ol><li>加入150微升鸡尾酒每一张幻灯片，在室温孵育30分钟。 </li><li>重复洗涤步骤为5.1.1。 </li></ol></li><li> 2000在RT PBS 30分钟:用150微升核染剂Hoechst33342稀释孵育。 <ol><li> 5分钟用150微升1×PBS中洗两次。 </li></ol></li><li>加入1-2滴防褪色的安装介质细胞，再缓慢降低盖玻片防止泡沫。 <ol><li>允许安装介质的RT的黑暗治愈过夜。 </li><li>使用透明指甲油密封盖玻片的幻灯片。 </li></ol></li><li>使用配备有能够检测DAPI（Hoeschst33342）激发/发射滤​​光片的荧光显微镜在40X放大率图像的幻灯片，和德克萨斯红（的Alexa Fluor 594）。 </li></ol>

<ol>
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The authors declare that they have no competing financial interests

为完整横断和屈指长肌腱修复的鼠类模型中的外科手术过程，提出了这项研究。除了聚光用细胞学离心机小细胞群体的一种新颖的应用表明，允许屈肌腱愈合期间蜂窝环境的定量免疫细胞化学分析。屈肌腱修复的此模型表明可再现愈合反应，这可以被用来评估使用基因敲除模型或在治疗过程中的变化与药理干预。手术后，炎性反应开始24小时，在此期间，细胞被招募到损伤部位和纤维组织的愈伤?...

在音乐会的手工作与前臂的屈肌和数字鞘屈肌腱启用的位弯曲和手的抓握功能​​。屈肌腱沿手的掌面运行;这种相对位置表浅创伤手时常常导致伤及肌腱。肌腱愈合通过瘢痕组织反应，而不是正常的肌腱组织1再生。虽然这种疤痕组织提供连续性肌腱，功能显着相对健康的肌腱下降。肌腱瘢痕组织复合材料的特点是受损的机械性能1，使修复后的肌腱更容易破裂。此外，疤痕组织缺乏天然腱胶原纤维结构的组织，从而增加在肌腱尺寸和体积。由于肌腱鞘单元的解剖限制，即使是适度增加肌腱的大小可以大大的红UCE肌腱的滑翔功能，因此，运动和手功能的数字范围。 在此之前的1960年的伤及屈肌腱，特别是那些在手的II区，没有定期修理由于在愈合严重并发症，与这些维修2产生的。手的这个区域被称为"无人区"3。然而，在外科技术，缝合模式和理疗康复协议的改善大大提高了屈肌腱修复2的结果。尽管有这些进展，修理的高达40％造成足够的粘合形成阻碍手功能4。因此，需要一种生物的方法，以改善愈合。遗憾的是，很知之甚少在细胞和分子水平上的腱愈合过程。因此，我们的目标是开发可用于提高基本相互理解鼠模型克屈肌腱愈合和瘢痕形成反应的细胞和分子成分，以识别新的治疗目标，以提高愈合的手段。 更大的动物模型已经在促进屈肌腱愈合过程的认识工具。犬和兔的研究已经证明既屈肌腱5,6的内在和外在的愈合能力，早期控制被动运动的相对粘连形成最小化的固定化7的重要性，以及不同的缝合图形的对愈合过程8的影响，9。此外，犬模型已在测试平移组织工程方法来改善愈合10有用。但是，也有在利用鼠相对于大型动物模型的模型，其中包括的相对成本，鼠特异性试剂的可用性几个重要的优点，以及便于产生全球KNOC的K-切口或组织特异性缺失/过表达构建体。此外，人类和老鼠之间的功能类似相对于肌腱11表明在开发的小鼠模型的潜在效用。 屈肌腱切断术和修复模拟临床愈合的许多方面，包括丰富的疤痕组织和受损的力学性能形成的小鼠模型的建立。这里描述的模型不是临床实践的真实概括由于在肌腱交界处FDL的横断为了保护维修站点。此外，该模型没有考虑滑膜鞘细胞对愈合响应的贡献，因为没有滑膜鞘覆盖，其中修复发生肌腱的中间部分。尽管有这些限制，这种模式有运动限制性粘连发生范围，这还没有的优点在小鼠模型更CLOS待证实伊利近似临床情况。该模型已被用于评估敲除小鼠模型12,13，并以测试不同的药理学方法来改善愈合14-17。这种模式的组织学分析，采用免疫组化和原位杂交，可以愈合期间提供关键基因和蛋白质的本地化的重要见解。然而，组织学仅提供一剖空间分析和不允许在整个组织定量。流式细胞仪代表更加定量的方法，但只是细胞的数量非常有限可在小鼠模型中的愈合腱组织中分离，并且这个数字期间固定，透化和洗涤步骤进一步降低。采取这一到帐户，流式细胞术成为不可行的方法由于是将需要的动物的数量。另一种方法是需要保留的大多数这种小细胞群，以便以进一步表征愈合环境。用于实现此目的的方法，在这里显示的，涉及经由细胞学离心分离的细胞的浓度到载玻片，随后通过免疫细胞化学。在本研究的EdU（5-乙炔基-2&#39;脱氧尿苷，胸苷类似物）掺入和随后的标记被用来确定在愈合部位细胞的相对增殖状态。这种方法可以应用到测试对细胞增殖，基因敲除或过表达的药物治疗的功效，或确定和量化不同的细胞群。

<table><tbody><tr><td>&lt;strong&gt;Surgical preparation&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>C57BL/6J mice&amp;nbsp;</td><td>Jackson Laboratories</td><td>000664</td><td></td></tr><tr><td>Ketamine</td><td>Hospira</td><td>NDC# 0409-2051-05</td><td></td></tr><tr><td>Xylazine</td><td>Lloyd Inc.</td><td>NDC# 61311-482-10</td><td></td></tr><tr><td>Buprenorphine</td><td>Par Pharmaceutical Inc.</td><td>NDC# 42023-179-10</td><td></td></tr><tr><td>0.9% sodium chloride irrigation</td><td>Hospira</td><td>NDC# 0409-6138-03</td><td>For preparation of ketamine/xylazine and buprenorphine solutions</td></tr><tr><td>1 ml syringe</td><td>BD</td><td>309659</td><td></td></tr><tr><td>30 G needle</td><td>BD</td><td>305106</td><td></td></tr><tr><td>Povidone-Iodine solution</td><td>Aplicare</td><td>82-226</td><td></td></tr><tr><td>70% ethanol</td><td></td><td></td><td></td></tr><tr><td>Puralube vet opthalmic ointment</td><td>Dechra Veterinary Products</td><td>NDC# 17033-211-38</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Surgical tools&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Portable balance 200 g</td><td>Ohaus</td><td>SP202</td><td></td></tr><tr><td>Spring scissors</td><td>Fine Science Tools</td><td>15124-12</td><td></td></tr><tr><td>Dumont #5 forceps</td><td>Fine Science Tools</td><td>11251-30</td><td></td></tr><tr><td>Needle holders</td><td>Fine Science Tools</td><td>91201-13</td><td></td></tr><tr><td>Micro spring scissors</td><td>Fine Science Tools</td><td>15003-08</td><td></td></tr><tr><td>Micro needle holders</td><td>Fine Science Tools</td><td>12061-02</td><td></td></tr><tr><td>5-0 nylon sutures</td><td>Ethicon</td><td>668G</td><td></td></tr><tr><td>8-0 microsurgery nylon sutures</td><td>Ethicon</td><td>2808G</td><td></td></tr><tr><td>Lab-Line histology slide warmer</td><td>Barnstead International</td><td>26025</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Cytospin method&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Collagenase Type I, lyophilized</td><td>Life Technologies&amp;nbsp;</td><td>1700-017</td><td></td></tr><tr><td>Bovine Serum Albumin</td><td>Cell Signaling Technologies</td><td>9998S</td><td></td></tr><tr><td>1x PBS</td><td>Thermo Fisher</td><td>10010-023</td><td></td></tr><tr><td>Cytology funnels</td><td>Fisher HealthCare</td><td>10-354</td><td></td></tr><tr><td>HistoBond+ microscope slides</td><td>VWR</td><td>16005-110</td><td></td></tr><tr><td>Cytospin 2 centrifuge</td><td>Shandon</td><td>SH-CYTO2</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Immunocytochemistry&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Slide staining tray with black lid</td><td>IHC World</td><td>M920-2</td><td></td></tr><tr><td>Click-iT Plus EdU Imaging Kit</td><td>Life Technologies&amp;nbsp;</td><td>C10639</td><td>Includes EdU and&amp;nbsp; Hoeschst 33342</td></tr><tr><td>Immedge hydrophobic barrier pen</td><td>Vector Laboratories</td><td>H-4000</td><td></td></tr><tr><td>ProLong Diamond mounting medium</td><td>Thermo Fisher</td><td>P36970</td><td></td></tr><tr><td>Glass coverslips 24 x 50 mm #1.5</td><td></td><td></td><td></td></tr><tr><td>Clear nail polish</td><td></td><td></td><td></td></tr></tbody></table>

murine flexor tendon injury and repair surgery

肌腱连接骨骼肌肉和骨骼，便利几乎整个身体的运动。在手，屈肌腱（FTS）使手指和一般手功能的屈曲。受伤的罚球是常见的，满意的治疗往往是受损由于过度的疤痕组织和肌腱与周围组织粘连之间。然而，知之甚少FT修复的分子和细胞成分。为此目的，该概括在人类愈合的许多方面，包括范围受损的运动和减小的机械性能的FT修复的鼠类模型，已开发和如前所述。这里这种手术过程的深入示范设置，涉及横断和在小鼠的后爪的趾长屈肌（FDL）腱的后续修复。这种技术可以用来进行不同类型的细胞的谱系分析，评估基因增益或功能失的影响，并以测试艾菲在愈合过程中的药理学干预cacy。不过，也有这种模型的两个主要限制:i）如鼠后爪，其中，发生在横断和修理的中间部分的FDL肌腱，不是由滑膜鞘包围。因此，该模型没有考虑护套的给瘢痕形成过程中的潜在贡献。 ⅱ）为了保护修复部位的完整性，对FT被释放在腱交界处，减少腱的机械力，可能有助于瘢痕形成增加。从期间流式细胞分析愈合过程中的FT的肉芽组织足够细胞分离已证明具有挑战性;细胞学离心浓缩这些细胞是用另一种方法，并且允许进行一代细胞制剂的可以在其上进行免疫荧光标记。用这种方法，FT愈合期间的细胞或感兴趣的蛋白质的定量成为可能。

在趾长屈肌（FDL）肌肉，位于小腿，行为经屈肌腱弯曲鼠标后爪的数字（蓝色轮廓图1A和图2A组织学显示），它从肌腱近端运行结并在远端指骨终止。在这个模型中屈肌腱愈合，所述FDL腱中的后爪的数字（红色箭头， 图1A）横切，并在脚中间修复，近端分叉。以防止修复，这将排除实验分析的破裂，肌腱（和相邻肌肉）被切断，或释放...

Watch this Scientific Journal Video about Murine Flexor Tendon Injury and Repair Surgery at JoVE.com

Murine Flexor Tendon Injury and Repair Surgery

在手屈肌腱通常受伤，导致受损的手部功能。然而，疤痕组织愈合反应不能很好地表征。屈肌腱愈合的小鼠模型在这里展示。该模型可以增强愈合过程的全面了解和评估治疗方法，以改善愈合。

鼠屈肌腱损伤修复手术

Tendon connects skeletal muscle and bone, facilitating movement of nearly the entire body. In the hand, flexor tendons (FTs) enable flexion of the fingers ...

murine-flexor-tendon-injury-and-repair-surgery

Research

JoVE Journal

Medicine

13.0K 次观看.  University of Rochester Medical Center. 在手屈肌腱通常受伤，导致受损的手部功能。然而，疤痕组织愈合反应不能很好地表征。屈肌腱愈合的小鼠模型在这里展示。该模型可以增强愈合过程的全面了解和评估治疗方法，以改善愈合。 

视频: Murine Flexor Tendon Injury and Repair Surgery

A Mini-Invasive Internal Fixation Technique for Studying Immobilization-Induced Knee Flexion Contracture in Rats

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to restrict knee joint mobility is a widely accepted model to generate experimental contracture. However, implanting application will inevitably cause surgical trauma to the animals. Aiming to develop a less invasive approach, we combined a muscle-gap separation modus with a previously reported mini-incision skill during the surgical procedure: Two mini skin incisions were made on the lateral thigh and leg, followed by performing muscle-gap separation to expose the bone surface. The rat knee joint was gradually immobilized by a preconstructed internal fixation at approximately 135&#176; knee flexion without interfering essential nerves or blood vessels. As expected, this simple technique permits rapid postoperative rehabilitation in animals. The correct position of the internal fixation was confirmed by an x-ray or micro-CT scanning analysis. The range of motion was significantly restricted in the immobilized knee joint than that observed in the contralateral knee joint demonstrating the effectiveness of this model. Besides, histological analysis revealed the development of fibrous deposition and adhesion in the posterior-superior knee joint capsule over time. Thus, this mini-invasive model may be suitable for mimicking the development of immobilized knee joint contracture.

Here, we present a protocol to describe a minimally invasive technique for knee joint immobilization in a rat model. This reproducible protocol, basing on muscle-gap separation modus and the mini-incision skill, is suitable for studying the underlying molecular mechanism of acquired joint contracture.

研究大鼠固定性诱导膝关节弯曲收缩的微型侵入性内固定技术

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to ...

科研

发育生物学

A Novel Tenorrhaphy Suture Technique with Tissue Engineered Collagen Graft to Repair Large Tendon Defects

Surgical management of large tendon defects with tendon grafts is challenging, as there are a finite number of sites where donors can be readily identified and used. Currently, this gap is filled with tendon auto-, allo-, xeno-, or artificial grafts, but clinical methods to secure them are not necessarily translatable to animals because of the scale. In order to evaluate new biomaterials or study a tendon graft made up of collagen type 1, we have developed a modified suture technique to help maintain the engineered tendon in alignment with the tendon ends. Mechanical properties of these grafts are inferior to the native tendon. To incorporate engineered tendon into clinically relevant models of loaded repair, a strategy was adopted to offload the tissue engineered tendon graft and allow for the maturation and integration of the engineered tendon in vivo until a mechanically sound neo-tendon was formed. We describe this technique using incorporation of the collagen type 1 tissue engineered tendon construct.

In this paper, we present an in vitro and in situ protocol to repair a tendon gap of up to 1.5 cm by filling it with engineered collagen graft. This was performed by developing a modified suture technique to take the mechanical load until the graft matures into the host tissue.

一种新型的腱绦缝合技术，采用组织工程胶原蛋白移植物修复大肌腱缺损

Surgical management of large tendon defects with tendon grafts is challenging, as there are a finite number of sites where donors can be readily ...

生物工程

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 ...

医学

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes. Here, we introduce a clinically relevant mouse model of tibial fracture. The postoperative changes in these mice mimic some of the cognitive impairments commonly observed after routine orthopedic surgery in humans. Briefly, an incision is performed in the right hind limb under strictly aseptic conditions. Muscles are disassociated, and a 0.38-mm stainless steel pin is inserted into the upper crest of the tibia, inside the intramedullary canal. Osteotomy is then performed, and the wound is stapled. We have used this model to investigate the effects of surgical trauma on postoperative neuroinflammation and behavioral changes. By applying this fracture model in combination with parabiosis, a surgical model in which 2 mice are anastomosed, we have studied cells and secreted factors that systemically rejuvenate organ function and tissue regeneration after injury. By following our step-by-step protocol, these models can be reproduced with high fidelity, and can be adapted to interrogate many biologic pathways that are altered by surgical trauma.

This protocol describes a mouse model of orthopedic surgery that has been used to study mechanisms of postoperative neuroinflammation and behavioral changes, and when combined with parabiosis, to study tissue regeneration during aging.

骨科手术的小鼠模型研究术后认知功能障碍与组织再生

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

了解肌腱撞击 – 用于研究机械生物学的小鼠外植体模型

Murine Hind Limb Explant Model for Studying the Mechanobiology of Achilles Tendon Impingement

Tendon impingement upon bone generates a multiaxial mechanical strain environment with markedly elevated transverse compressive strain, which elicits a localized fibrocartilage phenotype characterized by accumulation of glycosaminoglycan (GAG)-rich matrix and remodeling of the collagen network. While fibrocartilage is a normal feature in impinged regions of healthy tendons, excess GAG deposition and disorganization of the collagen network are hallmark features of tendinopathy. Accordingly, impingement is clinically recognized as an important extrinsic factor in the initiation and progression of tendinopathy. Nevertheless, the mechanobiology underlying tendon impingement remains understudied. Prior efforts to elucidate the cellular response to tendon impingement have applied uniaxial compression to cells and excised tendon explants in vitro. However, isolated cells lack a three-dimensional extracellular environment crucial to mechanoresponse, and both in vitro and excised explant studies fail to recapitulate the multiaxial strain environment generated by tendon impingement in vivo, which depends on anatomical features of the impinged region. Moreover, in vivo models of tendon impingement lack control over the mechanical strain environment. To overcome these limitations, we present a novel murine hind limb explant model suitable for studying the mechanobiology of Achilles tendon impingement. This model maintains the Achilles tendon in situ to preserve local anatomy and reproduces the multiaxial strain environment generated by impingement of the Achilles tendon insertion upon the calcaneus during passively applied ankle dorsiflexion while retaining cells within their native environment. We describe a tissue culture protocol integral to this model and present data establishing sustained explant viability over 7 days. The representative results demonstrate enhanced histological GAG staining and decreased collagen fiber alignment secondary to impingement, suggesting elevated fibrocartilage formation. This model can easily be adapted to investigate different mechanical loading regimens and allows for the manipulation of molecular pathways of interest to identify mechanisms mediating phenotypic change in the Achilles tendon in response to impingement.

作者聚焦：揭开肌腱撞击的力学生物学 – 多轴鼠后肢外植体模型 

We present a custom experimental platform and tissue culture protocol that recreates fibrocartilaginous change driven by impingement of the Achilles tendon insertion in murine hind limb explants with sustained cell viability, providing a model suitable for exploring the mechanobiology of tendon impingement.

小鼠后肢外植体研究跟腱撞击力学生物学的模型

Tendon impingement upon bone generates a multiaxial mechanical strain environment with markedly elevated transverse compressive strain, which elicits a ...

Using Q Suture to Enhance Resistance to Gap Formation and Tensile Strength of Repaired Flexor Tendons

Peripheral epitendinous sutures are believed to enhance core suture strength in tendon repair and decrease the risk of gapping between tendon ends. Here Q suture, an alternative to peripheral sutures, is presented for the use in tendon repair. Its effects on gap formation and tensile strength of the repaired tendons were compared with conventional running peripheral sutures. Three 2-strand sutures and three 4-strand sutures were used in repairing porcine tendons. The time required for performing 2Q and running sutures were recorded. The repaired tendons were subjected to a cyclic loading test, and the cycle number, during which a 2-mm gap was formed, was determined. After the cyclic loading, the gap size at the tendon ends and the ultimate strength of the repaired tendons were measured. Augmentation with the Q sutures reduced the number of tendons showing 2-mm gaps at tendon ends during cyclic loading. With addition of Q sutures 2-strand sutures significantly increased the ultimate strength of the repaired tendons and 4-strand sutures decreased the gap distance at the repair site of tendons. The time required for performing 2Q sutures was significantly less than that for running sutures. Therefore, we conclude that the Q suture is efficient in enhancing the tensile resistance and tendon repair strength and can be an alternative to conventional peripheral sutures.

Here, we present a &#8220;Q&#8221; suture technique that can be performed in tendon repair and its effects on the gap formation and tensile strength of the repaired tendons. Q suture is shown to be efficient in enhancing the tensile resistance and tendon repair strength.

使用 Q 缝合增强对修复的屈肌的间隙形成和拉伸强度的抵抗力

Peripheral epitendinous sutures are believed to enhance core suture strength in tendon repair and decrease the risk of gapping between tendon ends. Here Q ...

Polytetrafluoroethylene (PTFE) as a Suture Material in Tendon Surgery

With the evolution of suture materials, there has been a change in paradigms in primary and secondary tendon repair. Improved mechanical properties allow more aggressive rehabilitation and earlier recovery. However, for the repair to hold against higher mechanical demands, more advanced suturing and knotting techniques must be assessed in combination with those materials. In this protocol, the use of polytetrafluoroethylene (PTFE) as a suture material in combination with different repair techniques was investigated. In the first part of the protocol, both linear tension strength and elongation of knotted against not-knotted strands of three different materials used in flexor tendon repair were evaluated. The three different materials are polypropylene (PPL), ultra-high molecular weight polyethylene with a braided jacket of polyester (UHMWPE), and polytetrafluoroethylene (PTFE). In the next part (ex vivo experiments with cadaveric flexor tendons), the behavior of PTFE using different suture techniques was assessed and compared with PPL and UHMWPE.
This experiment is comprised of four steps: harvesting of the flexor tendons from fresh cadaveric hands, transection of the tendons in a standardized manner, tendon repair by four different techniques, mounting, and measurement of the tendon repairs on a standard linear dynamometer. The UHMWPE and PTFE showed comparable mechanical properties and were significantly superior to PPL in terms of linear traction strength. Repairs with four- and six-strand techniques proved stronger than two-strand techniques. Handling and knotting of PTFE are a challenge due to very low surface friction but fastening of the four- or six-strand repair is comparatively easy to achieve. Surgeons routinely use PTFE suture material in cardiovascular surgery and breast surgery. The PTFE strands are suitable for use in tendon surgery, providing a robust tendon repair so that early active motion regimens for rehabilitation can be applied.

Polytetrafluoroethylene PTFE as a Suture Material in Tendon Surgery

The present protocol illustrates a method for assessing the biophysical properties of tendon repairs ex vivo. A polytetrafluoroethylene (PTFE) suture material was evaluated by this method and compared to other materials under different conditions.

聚四氟乙烯（PTFE）作为肌腱手术中的缝合材料

With the evolution of suture materials, there has been a change in paradigms in primary and secondary tendon repair. Improved mechanical properties allow ...

A1 Pulley Reconstruction for Severe Trigger Finger

The aim of this study was to evaluate the effectiveness of A1 pulley reconstruction in treating severe trigger fingers and to compare its outcomes with those of the traditional A1 pulley release technique. A total of 43 patients participated in the study, divided into two groups: 22 patients underwent A1 pulley reconstruction, while the remaining 21 patients received the standard A1 pulley release procedure. The outcomes were assessed using the Michigan Hand Outcomes Questionnaire (MHQ) 1 month post-surgery. The results demonstrated that patients who received the A1 pulley reconstruction reported significantly better outcomes. These included enhanced hand function and quality of life, reduced levels of pain, improved aesthetic appearances, and higher overall satisfaction when compared to the traditional release group. The findings suggest that A1 pulley reconstruction offers superior benefits over the standard release procedure for individuals suffering from severe trigger fingers, highlighting its potential as a more effective surgical intervention.

Here, we present a protocol to compare A1 pulley reconstruction with traditional release surgery in treating a trigger finger, revealing enhanced outcomes and patient satisfaction through the Michigan Hand Outcomes Questionnaire.

重度扳机指的 A1 滑轮重建术

The aim of this study was to evaluate the effectiveness of A1 pulley reconstruction in treating severe trigger fingers and to compare its outcomes with ...

鼠屈肌腱损伤修复手术

摘要

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聚四氟乙烯（PTFE）作为肌腱手术中的缝合材料

重度扳机指的 A1 滑轮重建术