Name: heterotopic mucosal engrafting procedure for direct drug delivery to the brain in mice
Uploaded: 2014-07-16T14:00:00
Description: Watch this Scientific Journal Video about Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice at JoVE.com

这项研究是由该Mcihael J.福克斯基金会帕金森研究2011年快速响应创新奖励计划。资助者在研究设计，数据收集和分析，发布的决定，或准备稿子没有任何作用。

手术前一定要完成所有的程序是由IACUC和任何其他道德或法律部门批准，并使用人性化的动物治疗的做法。这包括使用无菌手术条件下，采用IACUC批准的方法麻醉小鼠，在手术过程中润滑老鼠的眼睛兽医药膏，并提供术后护理。不进行手术，如果有程序方面是否批准的任何问题。在此进行的所有程序批准了美国波士顿大学实验动物管理和使用委员会。 动物1。制备和手术用品<...

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本文所描述的过程中最困难的步骤是一个足够大的粘膜的成功转移到脑表面。这一步是由显著容易，如果收获的鼻中隔足够大，打扫好。如果隔膜的腹侧部分被截断，应获得一个新的移植物。钻孔角度应垂直于鼠标头，以确保粘膜不被钻头损坏。如果宽度大于推荐钻孔通道被接便更难除去，其连接到该隔膜的额外的组织。任何大块的结缔组织应该用锋利的剪刀割伤，而不是试图拉他们松被删除。?...

神经和精神疾病的治疗中是受到严重的血-脑屏障（BBB），防止所有潜在的药物制剂的95％以上到达中枢神经系统1-3的存在阻碍。例如，神经胶质源性神经营养因子（GDNF）已被证明可有效地治疗帕金森氏病时，直接注射到大脑，交付时全身性，因为它不能穿透血脑屏障4-6不过是无效的。 众多走近已经发展到试图绕过这个问题。改善全身递送neurotheraputics的已证实通过使用含有抗体的选择性为位于脑毛细血管内皮转运蛋白药物结合物;然而这种方法并没有被证明是适用于范围广泛的药物7,8。此外，血脑屏障渗透性开放已经临床使用盟友，但这种方法从全身用药剂量患有而不是一个更直接的传递到感兴趣的9大脑区域。大量的努力已投入在优化直接针对大脑10-12的希望鼻交付。虽然有些已经取得成功的，结论性的结果只获得针对药物具有内源性受体，如胰岛素13,14。此外鼻交付机制一直存在争议证据通过嗅觉神经元摄取或通过血液11暗示间接进入大脑。使用植入导管直接的，经颅传递已经实现，但这一程序具有很强的侵袭，并与众多的并发症15,16关联。迄今为止，不存在一般的微创方法，以提供高分子量的化合物进入脑。 本文中所呈现的是一种鼠的外科手术创建与大脑半透界面。这是通过移植粘膜外植体17以上的小鼠外科开颅缺陷来实现的。使用此程序它已经表明，可溶性化合物高达500 kDa的可递送到中枢神经系统（直接进入脑实质以及进入脑脊液）在两个时间和分子量依赖性方式18。绕过血脑屏障这种方法是一款型号为颅底缺损修复的人类，它使用血管黏膜瓣下鼻内镜手术19,20修复颅骨孔。

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			<td height="21" style="height:21px;width:247px;">Name of Material/ Equipment</td>
			<td style="width:220px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:263px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">mice</td>
			<td style="width:220px;">Taconic</td>
			<td>C57BL/6</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Isoflurane</td>
			<td style="width:220px;">Piramal Healthcare</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Student Fine Scissors</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91461-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">pneumatic drill</td>
			<td style="width:220px;">MTI Dental</td>
			<td style="width:119px;">333-CB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">drill bit</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">forcepts</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91106-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">0.9% Sodium Chloride Injection USP</td>
			<td style="width:220px;">Abbott Laboratories</td>
			<td>4925</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Polystyrene Detri Dish</td>
			<td style="width:220px;">Fisher</td>
			<td>08-757-12</td>
			<td>for temporarily storing graft</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bead Sterilizer</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>18000-45</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen/Isoflurane System</td>
			<td style="width:220px;">SurgiVet</td>
			<td>V720100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temperature Control System</td>
			<td style="width:220px;">Physitemp</td>
			<td style="width:119px;">TCAT-2LV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small Animal Stereotaxic Instrument</td>
			<td style="width:220px;">KOPF</td>
			<td style="width:119px;">Model 940</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Eye Ointment</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Electric shaver</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Cotton-Tipped Applicators</td>
			<td style="width:220px;">Fisher</td>
			<td>23-400-106</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7.5% Provadone Iodine</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>Betadine surgical scrub</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% Ethanol</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Surgical Blade Stainless</td>
			<td style="width:220px;">Feather</td>
			<td style="width:119px;">2976#10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Scalpel Handle - #3</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>10003-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3% Hydogen Peroxide</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>for cleaning the skull</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Vetbond Tissue Adhesive</td>
			<td style="width:220px;">3M</td>
			<td>1469SB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">needles</td>
			<td>Becton, Dickinson and Company</td>
			<td>305176</td>
			<td>needle tip cut off and used as well</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">syringes</td>
			<td>Becton, Dickinson and Company</td>
			<td>309597</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">nitrile gloves</td>
			<td style="width:220px;">Denville Scientific Inc</td>
			<td>G4162</td>
			<td>for well closure and protection of graft</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-0 Nylon Suture Thread</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Student Halsey Needle Holder</td>
			<td style="width:220px;">Fine Science Tools</td>
			<td>91201-13</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">cyanoacrylate adhesive</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td>commecially available super glue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dental Cement Kit, 1 lb, Pink Opaque</td>
			<td style="width:220px;">Stoelting</td>
			<td>51458</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">isobutane (2-methylbutane)</td>
			<td style="width:220px;">Aldrich</td>
			<td>M32631</td>
			<td>for dry ice bath</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">dry ice</td>
			<td style="width:220px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

heterotopic mucosal engrafting procedure for direct drug delivery to the brain in mice

递送治疗剂进入脑是通过血 - 脑屏障（BBB）的制约的极性和高分子量化合物从血液进入脑组织的通道的存在阻碍。一些直接交付的成功在人类已通过经颅导管植入已经实现;然而，这种方法具有很强的侵袭，并与众多的并发症。侵入性较低的替代办法是通过外科手术植入，半透膜如用于下列在人类鼻内镜切除肿瘤手术，以修复颅底缺损鼻黏膜剂量的大脑。药物传递虽然这种膜能有效地绕过血脑屏障，并直接扩散到脑和脑脊液。通过这种方法的启发，在小鼠的手术方法的开发，使用捐赠者间隔粘膜栽种了一个多颅外手术血脑屏障的缺陷。该模型已被证明能有效地允许的高分子量化合物的通路进入大脑。由于众多的候选药物都无法穿越血脑屏障，这种模式是对神经系统和精神疾病进行的新疗法临床前试验的价值。

获得足够大的鼻中隔外植体是用于随后的步骤是至关重​​要的。这可以通过在该位置上， 如图1a中所示的供体小鼠的颅骨钻孔来完成。沿着这条路径切割将产生足够大小的外植体， 如图1b所示 。如果钻孔深度不够深，移植将被截断，这将是很难获得足够大的膜覆盖大脑表面。多钻横向比建议的路径是不妥当的，因为更多的组织会留在鼻中​​隔，下一个步骤将是更加?...

Watch this Scientific Journal Video about Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice at JoVE.com

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

异位黏膜移植程序直接给药到小鼠的大脑

已开发人力内镜颅底重建的小鼠模型，会使用鼻腔粘膜移植大脑和鼻子之间的半透界面。这种方法使研究人员能够研究发货到全身给药时，被排除在外，否则由血脑屏障高分子量治疗中枢神经系统。

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科研

医学

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

通过40W的Elvax缓释给药的大鼠视网膜：对慢性疾病的治疗

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

Direct Mouse Trauma/Burn Model of Heterotopic Ossification

异位骨化直接鼠标创伤/烧伤模型

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical ...

Heterotopic Cervical Heart Transplantation in Mice

颈椎异位心脏移植小鼠

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

腔内药物输送到鼠标动静脉瘘内皮

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

Optimized LC-MS/MS Method for the High-throughput Analysis of Clinical Samples of Ivacaftor, Its Major Metabolites, and Lumacaftor in Biological Fluids of Cystic Fibrosis Patients

经优化的 LC/ms 方法对囊性纤维化患者 Ivacaftor 及其主要代谢物和 Lumacaftor 的高通量分析

Defects in the cystic fibrosis trans-membrane conductance regulator (CFTR) are the cause of cystic fibrosis (CF), a disease with life-threatening ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

一种基于猪脑内皮细胞的体外血脑屏障模型的改进方法

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

Trans-Tympanic Drug Delivery for the Treatment of Ototoxicity

经鼓膜药物治疗耳毒性的临床研究

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could ...

In Vivo Photolabeling of Cells in the Colon to Assess Migratory Potential of Hematopoietic Cells in Neonatal Mice

In Vivo Photolabeling of Cells in the Colon to Assess Migratory Potential of Hematopoietic Cells in Neonatal Mice

体内结肠细胞 Photolabeling 对新生小鼠造血细胞迁移潜能的评价

Enteric bacterial communities are established early in life and influence immune cell development and function. The neonatal microbiota is susceptible to ...

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

吸入麻醉下鼻内给药评价鼻腔给脑药物的新方法

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain ...