Name: manufacture of concentrated, lipid-based oxygen microbubble emulsions by high shear homogenization and serial concentration
Uploaded: 2014-05-26T14:00:00
Description: Watch this Scientific Journal Video about Manufacture of Concentrated, Lipid-based Oxygen Microbubble Emulsions by High Shear Homogenization and Serial Concentration at JoVE.com

资金：美国陆军医学研究和装备司令部（USAMRMC）及远程医疗与先进技术研究中心管理。顺溪籍提供如这里所描述的注射器的修改。

1，系统设置了该系统由配有单级混合器，在线高剪切均化器，辊子泵来移动HCT和均化器之间的流体，和一个热交换器（图1）的保持和浓缩罐（HCT）的。 <ol><li>放置消毒，广口4升的玻璃集装上2底座端口和3个侧端口的单级混频器下方容器。降低混频器头在容器的口部，并确保气密接头用橡胶密封件或带（为了防止环境空气污染的头部空间）。 </li><li>适合的基地?...

<ol>
	<li>Lentacker, I., De Smedt, S. C., Sanders, N. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+loaded+microbubble+design+for+ultrasound+triggered+delivery.">Drug loaded microbubble design for ultrasound triggered delivery.</a> Soft Matter. 5 (11), 2161-2170 (2009).</li><li>Ren, J. L., 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=A+Novel+Ultrasound+Microbubble+Carrying+Gene+and+Tat+Peptide:+Preparation+and+Characterization.">A Novel Ultrasound Microbubble Carrying Gene and Tat Peptide: Preparation and Characterization.</a> Academic Radiology. 16 (12), 1457-1465 (2009).</li><li>Tinkov, S., 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=Microbubbles+as+Ultrasound+Triggered+Drug+Carriers.">Microbubbles as Ultrasound Triggered Drug Carriers.</a> Journal of Pharmaceutical Sciences. 98 (6), 1935-1961 (2009).</li><li>Hernot, S., Klibanov, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbubbles+in+ultrasound-triggered+drug+and+gene+delivery.">Microbubbles in ultrasound-triggered drug and gene delivery.</a> Advanced Drug Delivery Reviews. 60 (10), 1153-1166 (2008).</li><li>Lanza, G. M., 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=A+novel+site-targeted+ultrasonic+contrast+agent+with+broad+biomedical+application.">A novel site-targeted ultrasonic contrast agent with broad biomedical application.</a> Circulation. 94 (12), 3334-3340 (1996).</li><li>Cavalli, R., 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=Preparation+and+characterization+of+dextran+nanobubbles+for+oxygen+delivery.">Preparation and characterization of dextran nanobubbles for oxygen delivery.</a> International Journal of Pharmaceutics. 381 (2), 160-165 (2009).</li><li>Kheir, J. N., 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=Oxygen+Gas-Filled+Microparticles+Provide+Intravenous+Oxygen+Delivery.">Oxygen Gas-Filled Microparticles Provide Intravenous Oxygen Delivery.</a> Science Translational Medicine. 4 (140), (2012).</li><li>Lindner, J. R., 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=Microvascular+rheology+of+definity+microbubbles+after+intra-arterial+and+intravenous+administration.">Microvascular rheology of definity microbubbles after intra-arterial and intravenous administration.</a> Journal of the American Society of Echocardiography. 15 (5), 396-403 (2002).</li><li>Zhao, Y. Z., 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=Preparation,+characterization+and+in+vivo+observation+of+phospholipid-based+gas-filled+microbubbles+containing+hirudin.">Preparation, characterization and in vivo observation of phospholipid-based gas-filled microbubbles containing hirudin.</a> Ultrasound in Medicine and Biology. 31 (9), 1237-1243 (2005).</li><li>Seo, M., 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=Microfluidic+Assembly+of+Monodisperse,+Nanoparticle-Incorporated+Perfluorocarbon+Microbubbles+for+Medical+Imaging+and+Therapy.">Microfluidic Assembly of Monodisperse, Nanoparticle-Incorporated Perfluorocarbon Microbubbles for Medical Imaging and Therapy.</a> Langmuir. 26 (17), 13855-13860 (2010).</li><li>Wan, J. D., Stone, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coated+Gas+Bubbles+for+the+Continuous+Synthesis+of+Hollow+Inorganic+Particles.">Coated Gas Bubbles for the Continuous Synthesis of Hollow Inorganic Particles.</a> Langmuir. 28 (1), 37-41 (2012).</li><li>Duncanson, W. 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=Monodisperse+Gas-Filled+Microparticles+from+Reactions+in+Double+Emulsions.">Monodisperse Gas-Filled Microparticles from Reactions in Double Emulsions.</a> Langmuir. 28 (17), 6742-6745 (2012).</li><li>Kendall, M. R., 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=Scaled-Up+Production+of+Monodisperse,+Dual+Layer+Microbubbles+Using+Multi-Array+Microfluidic+Module+for+Medical+Imaging+and+Drug+Delivery.">Scaled-Up Production of Monodisperse, Dual Layer Microbubbles Using Multi-Array Microfluidic Module for Medical Imaging and Drug Delivery.</a> Bubble Science Engineering and Technology. 4 (1), 12-20 (2012).</li><li>Szijjarto, 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=Effects+of+Perfluorocarbon+Gases+on+the+Size+and+Stability+Characteristics+of+Phospholipid-Coated+Microbubbles:+Osmotic+Effect+versus+Interfacial+Film+Stabilization.">Effects of Perfluorocarbon Gases on the Size and Stability Characteristics of Phospholipid-Coated Microbubbles: Osmotic Effect versus Interfacial Film Stabilization.</a> Langmuir. 28 (2), 1182-1189 (2012).</li><li>Rossi, S., Waton, G., Krafft, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phospholipid-Coated+Gas+Bubble+Engineering:+Key+Parameters+for+Size+and+Stability+Control,+as+Determined+by+an+Acoustical+Method.">Phospholipid-Coated Gas Bubble Engineering: Key Parameters for Size and Stability Control, as Determined by an Acoustical Method.</a> Langmuir. 26 (3), 1649-1655 (2010).</li><li>Swanson, E. 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=Phospholipid-Stabilized+Microbubble+Foam+for+Injectable+Oxygen+Delivery.">Phospholipid-Stabilized Microbubble Foam for Injectable Oxygen Delivery.</a> Langmuir. 26 (20), 15726-15729 (2010).</li><li>Kv&#229;le, S., 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=Size+fractionation+of+gas-filled+microspheres+by+flotation.">Size fractionation of gas-filled microspheres by flotation.</a> Separations Technology. 6 (4), 219-226 (1996).</li><li>Dressaire, 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=Interfacial+polygonal+nanopatterning+of+stable+microbubbles.">Interfacial polygonal nanopatterning of stable microbubbles.</a> Science. 320 (5880), 1198-1201 (2008).</li><li>Kheir, J. N., 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=Bulk+Manufacture+of+Concentrated+Oxygen+Gas-Filled+Microparticles+for+Intravenous+Oxygen+Delivery.">Bulk Manufacture of Concentrated Oxygen Gas-Filled Microparticles for Intravenous Oxygen Delivery.</a> Advanced Healthcare Materials. , (2013).</li></ol>

最重要的步骤，以建立集中的，高度氧化的LOM包括：1）确保HCT内的顶部空间保持完全氧化; 2）确保所述脂质赋形剂的纯度为最佳（包括储存条件和使用GMP的产品）; 3）确保粉状脂质一起吸系统之前水相混匀;和4）密切关注的HCT内增加的气体馏分，以确保气体的体积分数不超过70％。 我们在这里描述的方法，利用高剪切均质创建空心微。两亲性脂质悬浮的水相，它是用于承载脂...

微泡的蛋白质，聚合物和脂质壳的组成已经被开发作为药物递送载体，基因治疗，和超声造影剂1-5。因为这些治疗用途需要血管内微泡的持久性，这样的微泡通常填充有惰性的，高分子量的气体如全氟化碳6，它具有低的溶解度在血液和稳定的气泡3,4。 最近，基于脂质的氧气微泡（的LOM）被设计用于提供治疗剂量的氧，这可能保留终末器官的氧输送和防止在气道阻塞或低氧血症7期血流动力学不稳定。设计用于静脉内气体输送乳剂需要比那些用于超声造影剂或靶向药物递送不同的设计特点。首先，因为身体消耗大量的氧气（〜200毫升/分钟）的，的LOM必须生产和注射大规模。这需要在制造过程是有效的。第二，在制造过程中应该是闭环，以避免氮气污染过的LOM的曝光（它应该充满100％的氧气）到环境空气中。第三，因为的LOM的目的是静脉内气体输送，LOM中的气体馏分应最大化，认识到通过乳液粘度7所施加的限制。最后，与任何静脉注射，精确地控制粒度分布是必要的，避免微血管梗阻8。 有用于制造微泡几个成熟的方法。超声处理采用强度高，低频超声应用于乳液，它包括表面活性剂，如两亲性的磷脂，以气体的顶部空间，以产生微泡7,9的存在下的空气-液体界面。这个过程是可控的，通过改变超声音的频率，功率和脉冲持续时间，并将得到的粒度分布可以根据产生特定尺寸分布的微泡，虽然超声处理在临床上使用的微泡的制备很少使用。合并是表面活性剂和气体的封闭系统，该系统也难于扩大，以适应大容量的2的强烈机械搅拌。液滴型微流控使微气泡大小分布10-13的精确控制。虽然传统上难以扩大规模，多通道，高速微流体已经描述了增加微泡的生产效率13。使用上述任何一种方法制造的微泡可能需要制造后尺寸减小过程中，如离心分馏14,15和微泡浮选16,17。 另一种既定方法的高度稳定微泡的制造是剪切homogeniz振动性6，这可能会导致对微泡的表面18上的稳定六边形磷脂图案。在此基础上的概念，我们描述了在线高剪切均质机来创建自组装的LOM 19日注册成立。在这个过程中，利用匀化器快速旋转接近双细网眼emulsor屏幕叶片，产生高的机械和液压剪用于创建微泡。脂质乳剂通过本系统的串行浓度产生越来越浓的气体馏分，可进一步通过离心浓缩。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="42">
			<td height="42" style="height:43px;width:239px;">Name of Material/ Equipment</td>
			<td style="width:141px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:405px;">Comments/Description</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)</td>
			<td style="width:141px;">Avanti Polar Lipids</td>
			<td style="width:119px;">770365</td>
			<td>Alternate product: non-GMP from NOF America (Coatsome MC-8080)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">Cholesterol</td>
			<td style="width:141px;">Sigma Aldrich</td>
			<td style="width:119px;">C75209</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">Plasma-Lyte A</td>
			<td style="width:141px;">VWR</td>
			<td style="width:119px;">80089-818</td>
			<td>Alternatively can use NaCl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">Glass collection vessel</td>
			<td style="width:141px;">Specialty Glass, Inc.</td>
			<td style="width:119px;">Custom</td>
			<td>Contact: Pam Zurbrick - 281-595-2210</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">Gas composition (oxygen) monitor</td>
			<td style="width:141px;">Precision Medical</td>
			<td style="width:119px;">PM5900L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">Sarns 8000 roller pump</td>
			<td style="width:141px;">Calicut Medical</td>
			<td style="width:119px;">16407</td>
			<td>Part of a modular perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">BIOtherm Heat Exchanger</td>
			<td style="width:141px;">Medtronic</td>
			<td style="width:119px;">ECMOtherm-II</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">Verso laboratory in-line mixer</td>
			<td style="width:141px;">Silverson Machines, Inc</td>
			<td style="width:119px;">TH-IL-102-VERSO</td>
			<td>Use multistage workheads and front-end extension with T piece</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">T-piece for Silverson Verso inlet port</td>
			<td style="width:141px;">Process Innovations</td>
			<td style="width:119px;">Custom</td>
			<td>Contact: Brian Leavitt - 508-423-2266</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">L5M-A laboratory mixer</td>
			<td style="width:141px;">Silverson Machines, Inc</td>
			<td style="width:119px;">NC0136483</td>
			<td>Use mesh emulsor screen (fine)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">Rochester-Ochsner toothed forceps</td>
			<td style="width:141px;">Fisher Scientific</td>
			<td style="width:119px;">13-812-18</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">140 mL syringe</td>
			<td style="width:141px;">Kendall Healthcare Monoject&#160;</td>
			<td style="width:119px;">8881114030</td>
			<td>Ensure there is a luer lock.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:239px;">IX71 Inverted light microscope</td>
			<td style="width:141px;">Olympus</td>
			<td style="width:119px;">IX71</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:239px;">Retiga-2000R microscope camera</td>
			<td style="width:141px;">QImaging</td>
			<td style="width:119px;">RET-2000R-F-M-12</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:239px;">Accusizer 780A Autodilution</td>
			<td style="width:141px;">PSS-NICOMP Particle Sizing Systems</td>
			<td style="width:119px;">Out of production</td>
			<td></td>
		</tr>
	</tbody>
</table>

manufacture of concentrated, lipid-based oxygen microbubble emulsions by high shear homogenization and serial concentration

充气微泡已经开发作为超声造影和药物递送剂。微泡可以通过使用超声处理，机械搅拌，微流体装置，或均化处理的表面活性剂来制备。最近，基于脂质的氧气微泡（的LOM）已经被设计在医疗紧急静脉输送氧气，扭转危及生命的低氧血症，防止后续的脏器损伤，心脏骤停而死亡。我们提出使用闭环高剪切均化器的方法按比例扩大生产的高度氧化的微泡。该过程可产生2升浓缩的LOM（90体积％）中90分钟。产生气泡的平均直径为〜2μm时，以及流变更新与该血液时，稀释至60％（体积）是一致的。该技术生产的高容量和高纯度氧的LOM，这表明该技术可用于转化研究实验室有用。

高剪切均质使效率（ 即在一个下午）生产足够的LOM对动物的研究，也不需要专业技术。一旦熟练，高达2升浓缩的LOM的可以在90分钟来制造。 微气泡的大小和形态，光镜和光遮蔽评估。时的LOM的10微升样品是可视化，球形的LOM中所指出的，以及脂质碎片的相对贫乏（图3A）。这一点尤其是当GMP产物被使用。时的LOM的相同的样品，通过减光评估的平均粒径为2.62...

Watch this Scientific Journal Video about Manufacture of Concentrated, Lipid-based Oxygen Microbubble Emulsions by High Shear Homogenization and Serial Concentration at JoVE.com

Manufacture of Concentrated, Lipid-based Oxygen Microbubble Emulsions by High Shear Homogenization and Serial Concentration

我们描述了大容量的基于脂质的氧气微气泡（的LOM），使用高剪切均化和串行浓度设计用于静脉内的氧气输送的制造方法。

制造浓缩，脂质为基础的氧气微泡的乳液由高剪切均质化和串行集中

Gas-filled microbubbles have been developed as ultrasound contrast and drug delivery agents. Microbubbles can be produced by processing surfactants using ...

Research

JoVE Journal

Bioengineering

Introduction to the Ultrasound Targeted Microbubble Destruction Technique

有针对性的超声微泡销毁技术简介

In UTMD, bioactive molecules, such as negatively charged plasmid DNA vectors encoding a gene of interest, are added to the cationic shells of lipid ...

科研

生物工程

Characteristics of Precipitation-formed Polyethylene Glycol Microgels Are Controlled by Molecular Weight of Reactants

This work describes the formation of poly(ethylene glycol) (PEG) microgels via a photopolymerized precipitation reaction. Precipitation reactions offer ...

Generation of Shear Adhesion Map Using SynVivo Synthetic Microvascular Networks

的剪切粘合地图使用SynVivo合成微血管网络代

Cell/particle adhesion assays are critical to understanding the biochemical interactions involved in disease pathophysiology and have important ...

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

受体动力学通过单分子荧光显微镜的高分辨率时空分析

Single-molecule microscopy is emerging as a powerful approach to analyze the behavior of signaling molecules, in particular concerning those aspect (e.g., ...

Synthesis of Gold Nanoparticle Integrated Photo-responsive Liposomes and Measurement of Their Microbubble Cavitation upon Pulse Laser Excitation

在脉冲激光激发金纳米粒子集成光电响应脂质体及其微泡空化的测量的合成

Photo-responsive nanoparticles (NPs) have received considerable attention because of their potential in providing spatial, temporal, and dosage control ...

Manufacture and Drug Delivery Applications of Silk Nanoparticles

纳米丝绸制造和药物输送应用

Silk is a promising biopolymer for biomedical and pharmaceutical applications due to its outstanding mechanical properties, biocompatibility and ...

Quantifying Intermembrane Distances with Serial Image Dilations

用序列图像量化膜间隙距离伸缩

A recently-described extracellular nanodomain, termed the perinexus, has been implicated in ephaptic coupling, which is an alternative mechanism for ...

Catalytic Scavenging of Plant Reactive Oxygen Species In Vivo by Anionic Cerium Oxide Nanoparticles

Catalytic Scavenging of Plant Reactive Oxygen Species In Vivo by Anionic Cerium Oxide Nanoparticles

阴离子氧化铈纳米粒子催化清除体内植物活性氧的研究

Reactive oxygen species (ROS) accumulation is a hallmark of plant abiotic stress response. ROS play&#160;a dual role in plants by acting as signaling ...

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate (LAL) Assays

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate LAL Assays

用利穆鲁布莱巴细胞裂解液 (lal) 法检测纳米配方中的内毒素

When present in pharmaceutical products, a Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause ...

Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy

使用荧光显微镜在剪切流下描述单分子构象变化

Single-molecule behavior under mechanical perturbation has been characterized widely to understand many biological processes. However, methods such as ...