Name: intratracheal administration of dry powder formulation in mice
Uploaded: 2020-07-25T15:00:00
Description: Watch this Scientific Journal Video about Intratracheal Administration of Dry Powder Formulation in Mice at JoVE.com

作者要感谢李光耀先生、梁振英先生和苏华莱士先生，感谢他们在制造光源和粉末充气器方面给予的善意协助：和动物成像援助的学院核心设施。这项工作得到了香港研究资助委员会（17300319）的支持。

本研究的实验已获香港大学活体动物教学及研究委员会（CULATR）批准。本研究使用喷雾冷冻干燥（SFD）制备的干粉配方，其中含有0.5%的红素酶信使RNA（mRNA）、5%的合成肽PEG12KL4和94.5%的曼尼托（w/w），用于演示肺16中的mRNA表达。SFD 粉末的质中空气动力学直径 （MMAD） 为 2.4μm。喷雾干燥（SD）曼尼托粉用于研究粉末分散16中使用的空气量的影响。SD 粉末的...

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本文介绍了用于干粉窒息和体内插管的定制设备。在粉末装载步骤中，干粉末被加载到 200 μL 凝胶加载移液器尖端。重要的是轻轻敲击尖端，使粉末在尖端的狭窄端松散包装。但是，如果粉末包装太紧，它们会卡在尖端，无法正确分散。建议中和粉末和移液器尖端的静电荷，以方便粉末装载，特别是低密度和低相对湿度的粉末。引导管是设备的关键部件。它用于促进将装有粉末的移液器尖端插入?...

肺病的施政途径为局部和系统行动提供治疗提供各种好处。对于肺病的治疗，可以通过肺分娩实现高局部药物浓度，从而减少所需剂量，降低全身副作用的发生率。此外，肺部相对较低的酶活性可以减少过早的药物代谢。肺部也有效地吸收药物的全身作用，由于大和良好的灌注表面积，极薄的上皮细胞层和肺毛细血管1的高血量。
吸入干粉配方已被广泛研究，用于预防和治疗各种疾病，如哮喘，慢性阻塞性肺病，糖尿病和肺疫苗接种2，3，4。固体状态下的药物通常比液体形式更稳定，干粉吸入器比雾化器5、6更便携、更方便用户。在开发吸入干粉配方时，需要在肺管理7后，在药物前动物模型中评估其安全性、药效和疗效。与能主动吸入干粉的人类不同，肺向小动物输送干粉具有挑战性。有必要建立一个有效的协议，将干粉送到动物的肺部。
老鼠被广泛用作研究动物模型，因为它们经济，繁殖良好。它们也易于处理，许多疾病模型都已建立起来。给小鼠肺部施用干粉有两种主要方法：吸入和体内管理。为了吸入，小鼠被放置在全身或鼻子专用室，其中干粉被气溶胶化，动物呼吸气溶胶没有安定8，9。设备昂贵，药品输送效率低。虽然全身腔在技术上可能不那么具有挑战性，但仅鼻腔暴露室可以最大限度地减少药物暴露在身体表面。无论如何，仍然很难精确控制和确定交付到肺部的剂量。干粉主要沉积在鼻咽部位，其中粘膜清除突出10。此外，在管理过程中，室内老鼠承受着巨大的压力，因为它们受到限制，得不到食物和水的供应。对于体内管理，它通常是指将物质直接引入气管。有两种不同的技术来实现这一点：气管切除术和骨质管内分。前者需要外科手术，使气管切口，这是侵入性的，很少用于粉末管理。这里只描述了第二种技术。与吸入法相比，体内输病是小鼠肺输配的常用方法，因为其输送效率高，药物损失最小。这是一种简单快捷的方法，可在几毫克内精确地将少量粉末输送到鼠标。虽然小鼠在解剖学和生理上与人类不同，在插管过程中需要麻醉，但内科管理绕过上呼吸道，为评估干粉配方的生物活动（如肺吸收、生物利用和治疗效果）提供了更有效的方法。
要在内经管理干粉，必须插管小鼠，这可能具有挑战性。本文介绍了定制干粉灌装器和管内装置的制造情况。演示了在小鼠肺部进行干燥粉末的灌接和窒息过程。

<table>
	<tbody>
		<tr>
			<td>BALB/c mouse</td>
			<td></td>
			<td></td>
			<td>Female; 7-9 weeks old; Body weight 20-25 g</td>
		</tr>
		<tr>
			<td>CleanCap Firefly Luciferase mRNA</td>
			<td>TriLink Biotechnology</td>
			<td>L-7602</td>
			<td></td>
		</tr>
		<tr>
			<td>Dry Powder Insufflator</td>
			<td>PennCentury</td>
			<td>Model DP-4M</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine 10%</td>
			<td>Alfasan International B.V.</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Light emitting diode (LED) torch</td>
			<td>Unilite Internation</td>
			<td>PS-K1</td>
			<td></td>
		</tr>
		<tr>
			<td>Mannitol (Pearlitol 160C)</td>
			<td>Roquette</td>
			<td>450001</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-filter round gel loading pipette tip (200 &#181;L)</td>
			<td>Labcon</td>
			<td>1034-800-000</td>
			<td></td>
		</tr>
		<tr>
			<td>Nylon floss</td>
			<td>Reach</td>
			<td>30017050</td>
			<td></td>
		</tr>
		<tr>
			<td>One milliliter syringe without needle</td>
			<td>Terumo</td>
			<td>SS-01T</td>
			<td></td>
		</tr>
		<tr>
			<td>Optical fibre</td>
			<td>Fibre Data</td>
			<td>OMPF1000</td>
			<td></td>
		</tr>
		<tr>
			<td>PEG12KL4 peptide</td>
			<td>EZ Biolab</td>
			<td></td>
			<td>(PEG12)-KLLLLKLLLLKLLLLKLLLLK-NH2</td>
		</tr>
		<tr>
			<td>Plastic Pasteur fine tip pipette</td>
			<td>Alpha Labotatories</td>
			<td>LW4061</td>
			<td></td>
		</tr>
		<tr>
			<td>Three-way stopcock</td>
			<td>Braun</td>
			<td>D201</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine 2%</td>
			<td>Alfasan International B.V.</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Zerostat 3 anti-static gun</td>
			<td>MILTY</td>
			<td>5036694022153</td>
			<td></td>
		</tr>
	</tbody>
</table>

intratracheal administration of dry powder formulation in mice

在开发可吸入干粉配方时，必须评估其在前层动物模型中的生物活动。本文介绍了一种在小鼠体内输送干粉制剂的非侵入性方法。提供由 200 μL 凝胶装载移液器尖端组成的干粉装装置，通过三向塞孔连接到 1 mL 注射器。少量干粉（1-2毫克）被装入移液器尖端，并在注射器中通过0.6毫升的空气分散。由于移液器尖端是一次性的，价格低廉，因此可以提前将不同的干粉配方加载到不同的提示中。在同一动物实验中，无需设备清洁和剂量补充，即可评估各种配方，从而节省时间并消除残留粉末交叉污染的风险。粉末分散的程度可以通过移液器尖端残留的粉末量来检查。包括一个带有定制光源和导引管的鼠标内导管协议。适当的插管是影响干粉配方在健康内输送到小鼠深肺区域的关键因素之一。

当使用干粉充气器将粉末气溶胶输送到动物的肺部时，所使用的空气量至关重要，因为它会影响粉末的安全性以及粉末分散效率。为了优化该方法，使用不同体积的空气（0.3 mL、0.6 mL 和 1.0 mL）来分散干粉（1 毫克喷雾干曼尼托），并在给药后监测小鼠的重量 48 小时（图 6）。使用0.3 mL和0.6 mL的空气不会导致小鼠在施用后体重减轻至48小时。将粉末与 1 mL 空气分散，在 24 小时...

Watch this Scientific Journal Video about Intratracheal Administration of Dry Powder Formulation in Mice at JoVE.com

Intratracheal Administration of Dry Powder Formulation in Mice

吸入干粉配方在治疗呼吸系统疾病方面有很大的潜力。在进入人类研究之前，有必要在前科研究中评估干粉配方的功效。提出了一种简单而非侵入性的方法，通过体外路线对小鼠的干粉进行管理。

小鼠干粉配方的内经管理

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper ...

Research

JoVE Journal

Medicine

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

一种新的手术方法气管内注射生物活性剂在胎鼠模型

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

科研

医学

Intranasal Administration of CNS Therapeutics to Awake Mice

清醒小鼠滴鼻的CNS药物治疗管理

Intranasal administration is a method of delivering  therapeutic agents to the central nervous system (CNS). It is  non-invasive and allows large ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Dry Powder and Nebulized Aerosol Inhalation of Pharmaceuticals Delivered to Mice Using a Nose-only Exposure System

干粉和药品的雾化吸入气雾剂递送小鼠使用鼻只曝光系统

Obstructive respiratory diseases like asthma and chronic obstructive pulmonary disease (COPD) are currently treated by inhaled anti-inflammatory and ...

A Pleural Effusion Model in Rats by Intratracheal Instillation of Polyacrylate/Nanosilica

聚丙烯酸/纳米水的气管内灌注积液模型

Pleural effusion is a prevalent clinical finding of many pulmonary diseases. Having a useful animal pleural effusion model is very important to study ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

通过直接内切内脂多糖灌输诱导小鼠急性肺损伤

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...

Establishment of a Severe Dry Eye Model Using Complete Dacryoadenectomy in Rabbits

在兔子中使用完全的白眼切除术建立严重干眼模型

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic ...

Intratracheal Instillation of Stem Cells in Term Neonatal Rats

期新生儿大鼠干细胞的内切内培养

Prolonged exposure to high concentrations of oxygen leads to inflammation and acute lung injury, which is similar to human bronchopulmonary dysplasia ...

A Minimally Invasive Method for Intratracheal Instillation of Drugs in Neonatal Rodents to Treat Lung Disease

一种新生儿啮齿动物气管内滴注药物治疗肺部疾病的微创方法

Treatment of neonatal rodent with drugs instilled directly into the trachea could serve as a valuable tool to study the impact of a locally administered ...