Name: a murine tail lymphedema model
Uploaded: 2021-02-10T13:00:00
Description: Watch this Scientific Journal Video about A Murine Tail Lymphedema Model at JoVE.com

这项工作得到了美国整形外科医生协会学术奖学金和国防部W81XWH2110135   向AHH提供的赠款资助的支持。美容外科教育和研究基金会向NIH女士U01DK119099、R01NS042617和R01DK125835授予CKS。

该协议遵循该机构动物研究伦理委员会的指导方针。所有动物实验都得到了印第安纳大学医学院动物护理和使用委员会的批准。动物被安置在12小时的光暗循环下，食物和水的脂肪。
1. 小鼠尾淋巴的手术中断
<ol><li>使用八周大的C57BL/6小鼠，性别分布均等。</li>
	<li>将鼠标置于全身麻醉下，在诱导室中，3-4%的异黄素在100%的氧气中，然后在手术过程中以1-3%的维持饱和度进行。<...

<ol>
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淋巴水肿分为原发性（先天性）或继发性（致畸淋巴）损伤38，39。继发性淋巴水肿占39例病例的99%。继发性淋巴水肿最常由感染（丝虫病）或肿瘤后治疗与淋巴切除术或辐射4，39引起。转化动物模型对继发性淋巴水肿具有挑战性，因为70%接受淋巴切除术和放射治疗的动物不会获得淋巴水肿<su...

淋巴水肿是由淋巴功能障碍引起的四肢肿胀。受影响的肢体由于积液、脂肪和纤维化1而扩大。淋巴水肿影响全球2.5亿人2，3，4。据估计，20-40%接受固体恶性肿瘤治疗的患者，如乳腺癌、黑色素瘤、妇科/泌尿肿瘤或肉瘤，会发展成淋巴水肿2、4、5。淋巴水肿的发病率包括复发性感染、疼痛和畸形6。这种渐进的终身疾病是无法治愈的。目前的疗法是变种有效7，包括压缩，完全排泄治疗由物理治疗师，切除程序，和显微手术，包括血管淋巴结转移和淋巴旁路7，8，9，10，11，12，13，14。淋巴水肿的理想治疗方法尚未被发现。
研究淋巴水肿的机理和治疗是有限的。淋巴损伤后平均延迟一年，15、16和大多数经历辐射和手术的诱导侮辱的人不会发展淋巴水肿4，6，17。虽然大型动物模型，包括犬，羊，和猪已经描述了18，19，20，老鼠尾巴模型已被最广泛的应用，因为方便，成本，和繁殖能力。用于研究淋巴水肿的鼠标模型包括尾部模型、白喉毒素介导淋巴消融，以及轴状或弹出性淋巴结解剖 21、22、23、24、25、26。大多数尾部模型使用焦点，全厚皮肤切除与淋巴通道剪裁，这是在尾部底部附近执行22，导致尾部肿胀和组织特征类似于人类淋巴水肿24，27，28，29。然而，标准的木乃伊尾巴模型通常自发解决在短短20天，并伴有周期性尾坏死30。淋巴水肿鼠尾模型将持续淋巴水肿延长超过15周，演示确认的动脉和静脉轻度，并允许功能淋巴功能障碍评估。
淋巴水肿的阴唇尾部模型允许评估治疗淋巴水肿的新型治疗方法。基因策略已被用于由病毒载体31，32调解的老鼠模型。我们还使用一种新的组织纳米传递技术（TNT）将遗传货物运送到淋巴小鼠尾部。TNT利用纳米通道芯片在快速聚焦电场33、34、35、36中促进直接的、皮下基因的传递。该模型包括使用TNT2.0，使潜在的基因治疗焦点基因传递到小鼠尾部的淋巴损伤部位35。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Surgical Microscope</td>
			<td>Leica, Wetzlar, Germany</td>
			<td>MSV266</td>
			<td></td>
		</tr>
		<tr>
			<td>Adherent Dressing (Tegaderm)</td>
			<td>3M, St. Paul, Minn.</td>
			<td>1626W</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser speckle (Pericam PSI System )</td>
			<td>Perimed AB, Stockholm, Sweden)</td>
			<td>PSIZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Near-infrared laser (LUNA)</td>
			<td>Stryker (Formerly Novadaq Technologies, Toronto, Canada)</td>
			<td>LU3000</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Jackson Laboratories</td>
			<td>000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-Adson Forceps - 1x2 Teeth</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>11019-12</td>
			<td></td>
		</tr>
		<tr>
			<td>V-Hook</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>18052-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel SS NO15</td>
			<td>Fischer Scientific</td>
			<td>29556</td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable Needle 30GX1</td>
			<td>Fischer Scientific</td>
			<td>305128</td>
			<td></td>
		</tr>
		<tr>
			<td>Operating Scissors</td>
			<td>Fischer Scientific</td>
			<td>12-460-796</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgi-Or Jeweler&#39;s Forceps, Sklar 4-1/2 in</td>
			<td>Fischer Scientific</td>
			<td>50-118-4255</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring Scissors - Straight/Sharp-Sharp/8mm Cutting Edge</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>15024-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Cardiogreen</td>
			<td>Sigma</td>
			<td>I2633-25MG</td>
			<td></td>
		</tr>
		<tr>
			<td>IsosulfanBlue (Lymphazurin)&#160; 50 mg/5ml</td>
			<td>Mylan</td>
			<td>67457-220-05</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine tail lymphedema model

淋巴水肿是由淋巴功能障碍引起的四肢肿胀。受影响的肢体由于积液、脂肪和纤维化而扩大。这种疾病没有治愈的方法。老鼠尾巴模型，使用焦点全厚皮肤切除附近的尾巴底部，导致尾部肿胀，已被用于研究淋巴水肿。然而，该模型可能导致血管组成和随后的尾部坏死和早期尾部肿胀的分辨率，限制其临床可翻译性。慢性阴唇尾淋巴水肿模型诱导持续淋巴水肿超过15周和可靠的灌注到尾巴。传统木质尾淋巴水肿模型的增强包括1）使用手术显微镜精确全厚切除和淋巴剪裁，2）使用高分辨率激光斑点确认术后动脉和静脉灌注，3）使用非多氰氨酸绿色近红外激光淋巴扫描进行功能评估。我们还使用组织纳米传递技术 （TNT） 为新的非病毒性、跨皮性、焦点性遗传货物输送到小鼠尾血管。

持续淋巴水肿的鼠标尾部模型技术见 图1。该图展示了鼠标尾部模型的相关解剖。 图2 显示了淋巴水肿诱导后小鼠尾部的渐进肿胀和持续持续淋巴水肿。按截断锥方程计算，鼠标尾部体积在第 4 周达到峰值，在第 6 周达到稳定点，随后逐渐改善，持续到第 15 周。尾部体积可用作结果变量，以评估治疗干预对模型淋巴水肿的影响?...

Watch this Scientific Journal Video about A Murine Tail Lymphedema Model at JoVE.com

A Murine Tail Lymphedema Model

淋巴水肿是由淋巴功能障碍引起的四肢肿胀。我们描述了淋巴水肿的慢性粘液尾部模型，以及组织纳米透射技术 （TNT） 对尾部进行遗传货物输送的新用途。

穆林尾淋巴水肿模型

Lymphedema is extremity swelling caused by lymphatic dysfunction. The affected limb enlarges because of accumulation of fluid, adipose, and fibrosis. ...

淋巴水肿是一种慢性疾病，没有治愈的方法。它影响三分之一的患者谁做了轴状淋巴结手术和辐射治疗乳腺癌。拥有有效的动物模型有助于理解新治疗策略的机制和发展。淋巴水肿的鼠尾模型是可靠和可重复的快速发作。它表现出与人类淋巴水肿一致的组织学变化。演示这个程序的将是加内什·莫汉博士，一个来自我实验室的博士后。首先，将八周大的饱和鼠标放在严厉的位置，并准备70%异丙醇的尾巴。使用卡钳从尾部底部开始，以 5 毫米为增量测量尾径 20 毫米。在距底座20毫米的尾部标记一个3毫米的环切除。在手术显微放大下，进行细致的三毫米全厚皮肤切除，使所有底层血管完好无损。首先通过真皮将优越的环状标记，然后通过环切的全厚切口将三毫米的切口切成第一个切口。做一个垂直全厚度的垂直切口连接两个切口，然后使用齿形精细皮卡抓住领先优势，用微型剪刀解剖血管平面深处的真皮和表面的静脉冒险。将 0.1 毫升 1% 异硫素蓝色皮下近处注射到尾部。识别两个淋巴通道显示蓝色，由于注射毗邻横向尾部静脉。仔细切除淋巴，用直微手术剪刀解剖横静脉和淋巴之间的平面。在淋巴血管和横静脉之间传递一把剪刀刀片的尖端，并关闭刀片以穿过淋巴血管。完成后，用无菌的粘附清晰地址给尾部伤口穿衣。将丁二氨酸绿色 0.1 毫升皮下管理到尖端附近的远小鼠尾。调暗房间灯光，然后将近红外激光血管造影仪置于缓冲环境中，并进行实时成像。将异硫素蓝色插入尾尖后，淋巴片呈现蓝色。淋巴被中断，而相邻的横静脉被保留。淋巴水肿诱导后，小鼠尾部的渐进肿胀和持续持续淋巴水肿如下。按截断的锥体方程计算，鼠标尾部体积在第四周达到峰值，稳定到第六周，随后逐渐改善，一直持续到第15周。高分辨率激光斑点对比成像是为了确认淋巴水肿尾部模型中的鼠尾灌注，该模型显示受伤的横静脉和完好无损的横静脉，以评估尾部血管的粘附度。近红外激光淋巴成像术显示术前完整的淋巴，术后没有ICG转场，从而证实肿胀是由淋巴功能障碍引起的。使用组织纳米转化技术运送遗传货物的效率通过将标有DNA的荧光素输送到木耳尾部来证明。使用适应小鼠尾淋巴水肿模型和ICG近红外激光血管造影术是一个临床翻译和令人兴奋的动物模型与治疗的影响。

Research

JoVE Journal

Medicine

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

一个小鼠模型，通过神经肌肉电刺激肌肉训练

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

科研

医学

Murine Model of Wound Healing

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

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 ...

A Murine Model of Subarachnoid Hemorrhage

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Preclinical Murine Model of Hepatic Metastases

肝转移的临床前小鼠模型

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

的不可逆和可逆单侧输尿管梗阻小鼠模型

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

人前列腺癌的临床前原位小鼠模型

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy

小鼠原位转移性乳腺癌模型的建立及小鼠根治性乳房切除术

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...