Name: murine flexor tendon injury and repair surgery
Uploaded: 2016-09-19T14:00:00
Description: Watch this Scientific Journal Video about Murine Flexor Tendon Injury and Repair Surgery at JoVE.com

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...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Work+of+flexion+after+flexor+tendon+repair+according+to+the+placement+of+sutures.">Work of flexion after flexor tendon repair according to the placement of sutures.</a> Clin Orthop Relat Res. , 205-210 (1995).</li><li>Zhao, C., 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=Award+for+Outstanding+Orthopaedic+Research:+Engineering+flexor+tendon+repair+with+lubricant,+cells,+and+cytokines+in+a+canine+model.">Award for Outstanding Orthopaedic Research: Engineering flexor tendon repair with lubricant, cells, and cytokines in a canine model.</a> Clin Orthop Relat Res. 472, 2569-2578 (2014).</li><li>Wong, J., Bennett, W., Ferguson, M. W., McGrouther, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+and+histological+examination+of+the+mouse+hindpaw+digit+and+flexor+tendon+arrangement+with+3D+reconstruction.">Microscopic and histological examination of the mouse hindpaw digit and flexor tendon arrangement with 3D reconstruction.</a> J Anat. 209, 533-545 (2006).</li><li>Katzel, E. 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=Impact+of+Smad3+loss+of+function+on+scarring+and+adhesion+formation+during+tendon+healing.">Impact of Smad3 loss of function on scarring and adhesion formation during tendon healing.</a> J. Orthop. Res. 29, 684-693 (2011).</li><li>Loiselle, A. E., 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+marrow-derived+matrix+metalloproteinase-9+is+associated+with+fibrous+adhesion+formation+after+murine+flexor+tendon+injury.">Bone marrow-derived matrix metalloproteinase-9 is associated with fibrous adhesion formation after murine flexor tendon injury.</a> PloS one. 7, e40602 (2012).</li><li>Lee, D. J., 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=Parathyroid+hormone+1-34+enhances+extracellular+matrix+deposition+and+organization+during+flexor+tendon+repair.">Parathyroid hormone 1-34 enhances extracellular matrix deposition and organization during flexor tendon repair.</a> J Orthop Res. 33, 17-24 (2015).</li><li>Geary, M. 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=Systemic+EP4+Inhibition+Increases+Adhesion+Formation+in+a+Murine+Model+of+Flexor+Tendon+Repair.">Systemic EP4 Inhibition Increases Adhesion Formation in a Murine Model of Flexor Tendon Repair.</a> PloS one. 10, e0136351 (2015).</li><li>Loiselle, A. E., 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=Development+of+antisense+oligonucleotide+(ASO)+technology+against+Tgf-beta+signaling+to+prevent+scarring+during+flexor+tendon+repair.">Development of antisense oligonucleotide (ASO) technology against Tgf-beta signaling to prevent scarring during flexor tendon repair.</a> J Orthop Res. 33, 859-866 (2015).</li><li>Orner, C. A., Geary, M. B., Hammert, W. C., O'Keefe, R. J., Loiselle, 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=Low-dose+and+short-duration+Matrix+Metalloproteinase+9+Inhibition+does+not+affect+adhesion+formation+during+murine+flexor+tendon+healing.">Low-dose and short-duration Matrix Metalloproteinase 9 Inhibition does not affect adhesion formation during murine flexor tendon healing.</a> Plast Reconstr Surg. , (2016).</li><li>Loiselle, A. E., 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=Remodeling+of+murine+intrasynovial+tendon+adhesions+following+injury:+MMP+and+neotendon+gene+expression.">Remodeling of murine intrasynovial tendon adhesions following injury: MMP and neotendon gene expression.</a> J Orthop Res. 27, 833-840 (2009).</li><li>Tsubone, T., 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+TGF-beta+inducible+early+gene+deficiency+on+flexor+tendon+healing.">Effect of TGF-beta inducible early gene deficiency on flexor tendon healing.</a> J Orthop Res. 24, 569-575 (2006).</li><li>Beason, D. P., Kuntz, A. F., Hsu, J. E., Miller, K. S., Soslowsky, L. 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K., 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=The+cellular+biology+of+flexor+tendon+adhesion+formation:+an+old+problem+in+a+new+paradigm.">The cellular biology of flexor tendon adhesion formation: an old problem in a new paradigm.</a> Am J Pathol. 175, 1938-1951 (2009).</li></ol>

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 border="0" cellpadding="0" cellspacing="0" width="1047">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical preparation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">C57BL/6J mice&#160;</td>
			<td>Jackson Laboratories</td>
			<td>000664</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine</td>
			<td>Hospira</td>
			<td>NDC# 0409-2051-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine</td>
			<td>Lloyd Inc.</td>
			<td>NDC# 61311-482-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td>Par Pharmaceutical Inc.</td>
			<td>NDC# 42023-179-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">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 height="21">
			<td height="21" style="height:21px;">1ml syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">30G needle</td>
			<td>BD</td>
			<td>305106</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone-Iodine solution</td>
			<td>Aplicare</td>
			<td>82-226</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Puralube vet opthalmic ointment</td>
			<td>Dechra Veterinary Products</td>
			<td>NDC# 17033-211-38</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical tools</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Portable balance 200g</td>
			<td>Ohaus</td>
			<td>SP202</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15124-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #5 forceps</td>
			<td>Fine Science Tools</td>
			<td>11251-30</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle holders</td>
			<td>Fine Science Tools</td>
			<td>91201-13</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15003-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro needle holders</td>
			<td>Fine Science Tools</td>
			<td>12061-02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-0 nylon sutures</td>
			<td>Ethicon</td>
			<td>668G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">8-0 microsurgery nylon sutures</td>
			<td>Ethicon</td>
			<td>2808G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab-Line histology slide warmer</td>
			<td>Barnstead International</td>
			<td>26025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cytospin method</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Collagenase Type I, lyophilized</td>
			<td>Life Technologies&#160;</td>
			<td>1700-017</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bovine Serum Albumin</td>
			<td>Cell Signaling Technologies</td>
			<td>9998S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1X PBS</td>
			<td>Thermo Fisher</td>
			<td>10010-023</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cytology funnels</td>
			<td>Fisher HealthCare</td>
			<td>10-354</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HistoBond+ microscope slides</td>
			<td>VWR</td>
			<td>16005-110</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cytospin 2 centrifuge</td>
			<td>Shandon</td>
			<td>SH-CYTO2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Immunocytochemistry</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
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			<td height="21" style="height:21px;">Click-iT Plus EdU Imaging Kit</td>
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			<td>Vector Laboratories</td>
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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 ...

Research

JoVE Journal

Medicine

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

单侧颈脊髓损伤的挫伤模型，利用无限的地平线冲击

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

一种新的小鼠模型主动脉瘤腔内修复

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Controlled Cortical Impact Model for Traumatic Brain Injury

控制皮层影响模型创伤性脑损伤

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

A Mouse Fetal Skin Model of Scarless Wound Repair

对无瘢痕修复鼠标胎儿皮肤模型

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

动脉再狭窄小鼠模型：股导线损伤的技术方面

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Murine Model of Intestinal Ischemia-reperfusion Injury

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

重复性的动脉剥脱损伤腹主动脉阻断在小鼠模型

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

Evaluation of Stem Cell Therapies in a Bilateral Patellar Tendon Injury Model in Rats

大鼠双侧髌腱损伤模型中干细胞治疗的评价

Regenerative medicine provides novel alternatives to conditions that challenge traditional treatments. The prevalence and morbidity of tendinopathy across ...