Name: comprehensive evaluation of the effectiveness and safety of placenta-targeted drug delivery using three complementary methods
Uploaded: 2018-09-10T14:30:00
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この作品は、国立自然科学財団 (81771617) と自然科学財団の広東省 (2016A030313178); x. f. に与えられるからの補助金によって支えられました。深セン基礎研究基金 (JCYJ20170413165233512) 場合であって; に与えられるからの助成金ユーニス · ケネディ · シュライバー国立衛生研究所の子と受賞番号 R01HD088549 の下で健康の国民の協会の人間の開発 (内容は著者の責任と、必ずしも公式健康の国民の協会の意見) に N.N.

すべてのマウス実験徹底プロトコル (SIAT-IRB-160520-YYS-FXJ-A0232) 動物のケアおよび使用委員会の深セン機関の高度な技術、中国科学院によって承認しました。
1. 胎盤コンドロイチン硫酸 A をターゲットとした脂質高分子ナノ粒子の合成
<ol><li>MTX と ICG ロード脂質高分子ナノ粒子を合成 (MNPs と INPs それぞれ) と plCSA BP 共役ナノ粒子 (plCSA MNPs と plCSA INPs) の説明に従って詳細他...

<ol>
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本稿では、3 法システム plCSA BP 導かれるナノ粒子が胎盤への薬の配達の対象化の効率的なツールであるかどうかを決定するための概要を説明します。生体内でイメージ投射赤外線蛍光 ICG 信号をモニターできるの使用確認 plCSA 跪く使用の胎盤のターゲット特異性 plCSA BP 共役ナノ粒子が MTX をだけに実現効率的にできることを示した我々 HFUS と高速液体クロマトグラフィー、胎児が胎盤?...

胎盤を介する妊娠合併症、子癇前症、妊娠の損失、胎盤早期剥離、小さな胎 (SGA) などが一般的なへと導く実質的な胎児と母体の罹患率と死亡率1,2、4、5の障害妊娠の治療の効果があると証明された3、および非常に少数の薬。妊娠中より選択と安全の胎盤をターゲットとした薬物送達のための戦略の開発、現代の薬物療法でやりがいのままです。
近年、いくつかのレポートが焦点子宮組織への薬のターゲットを絞った配信ペプチドや抗体でコーティング ナノ胎盤向けツールとして。抗上皮成長因子受容体 (EGFR)6抗体、ペプチド腫瘍ホーミング (CGKRK および iRGD)7、胎盤標的ペプチド8、胎盤血管標的ペプチド9および抗体が含まれます、オキシトシン受容体10。
ここでは、胎盤の11へのナノ粒子およびそれらの薬物のペイロードのターゲット配信用合成胎盤コンドロイチン硫酸 A 結合ペプチド (plCSA BP) することができますを示します。PlCSA BP 導かれるナノ粒子は、報告された子宮胎盤絨毛を対象にできるので、ターゲット設定方法に補完されます。
非侵襲的な方法として生体内イメージングは、マウス12、胎盤特異的遺伝子の発現を監視する使用されています、インドシアニン グリーン (ICG) 蛍光イメージング システム13を用いたナノ粒子の追跡に用いられています。 14,15。したがって、我々 は静脈内妊娠マウスにおける蛍光イメージャ plCSA INP の分布を可視化する (plCSA-INPs) ICG 搭載 plCSA BP 共役系ナノ粒子を注入しました。我々 は、静脈内メトトレキサート (MTX) を注入-妊娠マウスに plCSA NPs を読み込まれます。別の非侵襲的高周波超音波 (HFUS)、リアルタイム イメージング ツール16,17は、マウス胎子及び胎盤の開発を監視するために使用されました。最後に、胎盤と胎児の MTX 分布を定量化するのに高速液体クロマトグラフィー (HPLC) を使用します。
このプロトコルでは 3 方式 plCSA BP 誘導ナノキャリアによる胎盤標的薬配信の効率を評価するために使用詳細に述べる.

<table border="0" cellpadding="0" cellspacing="0" style="width:785px;" width="786">
	<tbody>
		<tr>
			<td>CD-1 mice</td>
			<td>Beijing Vital River</td>
			<td>201</td>
			<td>Female (8-12 week)</td>
		</tr>
		<tr>
			<td>Insulin syringe</td>
			<td>BD</td>
			<td>328421</td>
			<td>for IV injection</td>
		</tr>
		<tr>
			<td>Ethanol absolute</td>
			<td>Sinopharm Chemical</td>
			<td>10009218</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>Soybean lecithin</td>
			<td>Avanti Polar Lipids</td>
			<td>441601</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>DSPE-PEG-COOH</td>
			<td>Avanti Polar Lipids</td>
			<td>880125</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>PLGA</td>
			<td>Sigma-Aldrich</td>
			<td>719897</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>Ultrasonic processor</td>
			<td>Sonics</td>
			<td>VCX130</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>Methotrexate (MTX)</td>
			<td>Sigma-Aldrich</td>
			<td>V900324</td>
			<td>for nanoparticles synthesis</td>
		</tr>
		<tr>
			<td>Indocyanine green (ICG)</td>
			<td>Sigma-Aldrich</td>
			<td>1340009</td>
			<td>for in vivo imaging</td>
		</tr>
		<tr>
			<td>phosphate-buffered saline (PBS)</td>
			<td>Hyclone</td>
			<td>SH30028.01</td>
			<td></td>
		</tr>
		<tr>
			<td>IVIS spectrum instrument</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td>for in vivo imaging</td>
		</tr>
		<tr>
			<td>Ultrasound transmission gel</td>
			<td>Guanggong</td>
			<td>ZC4252418</td>
			<td>for ultrasound imaging</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Lunan Pharmaceutical</td>
			<td>I0040</td>
			<td>for maintain the anesthesia</td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Nair</td>
			<td>TMG001</td>
			<td>for removing fur</td>
		</tr>
		<tr>
			<td>40 MHz transducer</td>
			<td>VisualSonics</td>
			<td>MS550S</td>
			<td>for ultrasound imaging</td>
		</tr>
		<tr>
			<td>High-frequency ultrasound imaging system</td>
			<td>VisualSonics</td>
			<td>Vevo2100</td>
			<td>for ultrasound imaging</td>
		</tr>
		<tr>
			<td>Avertin</td>
			<td>Sigma-Aldrich</td>
			<td>T48402</td>
			<td>for anesthesia</td>
		</tr>
		<tr>
			<td>Syringe pump</td>
			<td>Mindray</td>
			<td>SK-500III</td>
			<td>forcardiac perfusion</td>
		</tr>
		<tr>
			<td>0.9% saline solution</td>
			<td>Meilunbio</td>
			<td>MA0083</td>
			<td>forcardiac perfusion</td>
		</tr>
		<tr>
			<td>1.5 mL Polypropylene tubes</td>
			<td>AXYGEN</td>
			<td>MCT-150-C</td>
			<td></td>
		</tr>
		<tr>
			<td>-80 &#176;C freezer</td>
			<td>Thermo Fisher Scientific</td>
			<td>88600V</td>
			<td></td>
		</tr>
		<tr>
			<td>Centriguge</td>
			<td>Cence</td>
			<td>H1650R</td>
			<td></td>
		</tr>
		<tr>
			<td>Perchloric acid</td>
			<td>Sigma-Aldrich</td>
			<td>311421</td>
			<td>for precipitating protein</td>
		</tr>
		<tr>
			<td>Homogenizer</td>
			<td>SCIENTZ</td>
			<td>SCIENTZ-48</td>
			<td>for homogenizing tissue</td>
		</tr>
		<tr>
			<td>Syringe filter (0.45 &#956;m)</td>
			<td>Millipore</td>
			<td>SLHV033RS01</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium hydroxide</td>
			<td>Sinopharm Chemical</td>
			<td>10019763</td>
			<td>for solving MTX</td>
		</tr>
		<tr>
			<td>HPLC vials</td>
			<td>Waters</td>
			<td>670650620</td>
			<td>for HPLC</td>
		</tr>
		<tr>
			<td>Potassium phosphate dibasic</td>
			<td>Sinopharm Chemical</td>
			<td>20032117</td>
			<td>for HPLC</td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>JKchemical</td>
			<td>932537</td>
			<td>for HPLC</td>
		</tr>
		<tr>
			<td>C18 column</td>
			<td>Waters</td>
			<td>186003966</td>
			<td>for HPLC</td>
		</tr>
		<tr>
			<td>HPLC system</td>
			<td>Shimadzu</td>
			<td></td>
			<td>for HPLC</td>
		</tr>
	</tbody>
</table>

comprehensive evaluation of the effectiveness and safety of placenta-targeted drug delivery using three complementary methods

現在は、胎盤の妊娠合併症は、効果的な治療法が存在しないと胎児と母体の副作用を最小限に抑えながら胎盤を薬のターゲットを絞った配信のための戦略を開発、やりがいのままです。対象となるナノ粒子のキャリアは、胎盤疾患を治療するために新しい機会を提供します。我々 は最近、合成胎盤コンドロイチン硫酸 A 結合ペプチド (plCSA BP) を使用して、胎盤に薬を提供するナノ粒子をガイドできる示した。このプロトコルで述べる plCSA BP の組み合わせで使用される 3 つの別々 のメソッドを採用するによって胎盤への薬物送達の効率を評価するためのシステム:生体内イメージング、高周波超音波 (HFUS)、および高パフォーマンス液体クロマトグラフィー (HPLC)。生体内でを使用してイメージング、plCSA BP 導かれるナノ粒子, 可視化された、生きている動物の胎盤で HFUS と HPLC 実証 plCSA BP 共役ナノ粒子効率的かつ具体的に配信されるメトトレキサート胎盤。したがって、これらのメソッドの組み合わせは、胎盤を薬のターゲットを絞った配信といくつかの妊娠合併症の新たな治療戦略の開発のための効果的なツールとして使用できます。

本稿では、MTX (plCSA MNPs) または ICG (plCSA INPs) 搭載の plCSA BP 共役ナノ粒子静脈内妊娠マウスに注入しました。In vivoイメージング plCSA INP 注入後 30 分子宮の強い ICG 信号を明らかにしました。INPs は主に肝臓と脾臓領域 (図 1 a) にローカライズされました。PlCSA INP 注射後 48 時間の妊娠マウスが犠牲になった、ICG 信号しかない信号との間の胎盤...

Watch this Scientific Journal Video about Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods at JoVE.com

Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods

安全性と胎盤をターゲットとした薬剤投与の有効性を評価する 3 つの方法を利用するシステムについて述べる:生体内のイメージ投射ナノ粒子蓄積、胎盤と胎児の開発を監視する高周波超音波を監視するには、および高速液体クロマトグラフィー組織への薬物送達を定量化します。

3 つの補完的な手法を使用して胎盤をターゲットとした薬剤投与の安全性の総合評価

No effective treatments currently exist for placenta-associated pregnancy complications, and developing strategies for the targeted delivery of drugs to ...

Research

JoVE Journal

Bioengineering

Self-reporting Scaffolds for 3-Dimensional Cell Culture

3次元細胞培養のための自己申告足場

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

携帯の障壁を通過する薬物送達システムの輸送を評価するためのモデルと方法

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. ...

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

三層のための3つのユニークな環境を提供するために、エレクトロスピニング法を適応させます<em&gt;インビトロ</em&gt;気道壁のモデル

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to ...

Manufacture and Drug Delivery Applications of Silk Nanoparticles

シルクナノ粒子の製造および薬物送達アプリケーション

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

Automated Contraction Analysis of Human Engineered Heart Tissue for Cardiac Drug Safety Screening

心臓医薬品安全性スクリーニングのための人間エンジニア心臓組織の自動収縮分析

Cardiac tissue engineering describes techniques to constitute three dimensional force-generating engineered tissues. For the implementation of these ...

Use of High-Throughput Automated Microbioreactor System for Production of Model IgG1 in CHO Cells

高スループットの自動モデル IgG1 CHO 細胞での生産のための Microbioreactor システム

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of ...

Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System

自動化された微生物叢システムを用いた中国ハムスター卵巣細胞からのモノクローナル抗体の精製と分析

Monoclonal antibodies (mAbs) are one of the most popular and well-characterized biological products manufactured today. Most commonly produced using ...

Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates

ケラチン生成細胞増殖の 2 次元と 3 次元の評価タイプ I コラーゲン基板

Type I collagen, useful as a substrate for cell culture, exists in two forms: the two-dimensional, non-fibrous form and three-dimensional, fibril form. ...

Preparing Protein Producing Synthetic Cells using Cell Free Bacterial Extracts, Liposomes and Emulsion Transfer

細胞を含まない細菌抽出物、リポソーム、エマルジョン移動を用いた合成細胞の調製

The bottom-up assembly approach for construction of synthetic cells is an effective tool for isolating and investigating cellular processes in a cell ...

Primary Clarification of CHO Harvested Cell Culture Fluid using an Acoustic Separator

音響セパレータを用いたCHO採取細胞培養液の一次解明

Primary clarification is an essential step in a biomanufacturing process for the initial removal of cells from therapeutic products within the harvested ...