この研究はMKにファリス家族イノベーション賞とNIH 1R01NR015674-01によってサポートされていました。著者らは、電磁発電システムだけでなく、彼の技術相談を提供するためのジョセップNayfach（Qteris、Inc）をお願いいたします。著者らはまた、彼女の技術的なアシスタントのフアンヤン（LCI＆化学物理学際プログラム、ケント州立大学）を感謝します。

注：磁場応答ゼラチンミクロゲルを製造する全体的なプロセスは、 図1Aに示されています。 1.準備溶液及び懸濁液<ol><li>リン酸緩衝生理食塩水2mlにゲニピン20mgを溶解することにより架橋剤ゲニピン溶液（w / vで1％）を調製（1×PBS、pH7.4）で。渦2時間50 O Cの水浴中で溶液と場所は完全に溶液に溶解します。 </li><li>ポリ（エチレングリコール）、ポリ20mgの（エチレングリコール） -ポリ（プロピレングリコール）を溶解することにより、界面活性剤溶液を調製し（M W = 2,900ダ; L64と称する）100の濃度になるようにPBS 200mlにppmです。 </li><li> PBS 0.43 ml中のゼラチンの64.5 mgを溶解することにより15％（w / v）のゼラチン溶液を調製します。渦ソリューションと、それは解決策が流体になるゾル相に達するまで37 O℃で水浴上に置きます。目を確実にするために3回 - その後、ゼラチン溶液2をボルテックスサンプルの電子の均一性。 </li><li> P（NIPAM- 共同 -AAm）モデル薬物と/のMNP溶液（BSA）の調製： <ol><li> PBSの0.43ミリリットル中に親水性のMNPの10.75ミリグラムを分散した後、3％の濃度を作るためにMNPサスペンションのpの12.9ミリグラム（NIPAM- 共同 -AAm）を溶解（w / v）の。 Pの濃度増加（NIPAM- 共 -AAm）はミクロゲルの増大した脱膨潤挙動を達成することができます。 </li><li>使用テキサスレッド共役ウシ血清アルブミン（TR-BSA; M ワット 〜66 kDaの）モデル薬物としては。 P（NIPAM- 共同 -AAm）/のMNPの混合物でTR-BSA 0.5mgのを溶かします。 </li></ol></li><li>その後、ゼラチン溶液（0.43ミリリットル）へのpの混合物（NIPAM- 共同 -AAm）/のMNP（0.43ミリリットル）を加えることにより、ゼラチン/ P（NIPAM- 共同 -AAm）/のMNP / BSA溶液（0.86ミリリットル）の混合物を準備し、徹底的に均質な混合物を作るためにそれらをボルテックス。このように、ポリマーおよびMNPの濃度は、最終混合物中の初期濃度の半分になります。 </li></ol><ol><li>清潔で滅菌ビーカーにシリコーンオイル[ポリジメチルシロキサン（粘度350センチストークス）]の15ミリリットルを注ぎます。 </li><li>すぐにシリコーン油にゼラチン/ P（NIPAM- 共 -AAm）/のMNP / BSA溶液（0.86 ml）を予め調製した水性混合物を追加して、900で磁気攪拌棒で攪拌することにより油相中の水性混合物を乳化30分間、30°Cの時rpmで。 </li></ol> 3.ゲル化し、水溶液にマイクロ液滴の譲渡<ol><li> 50ミリリットルチューブにビーカーからエマルション（〜16ミリリットル）を転送します。 </li><li>油中の微小液滴のゲル化のために4℃で10分間チューブをクールダウン。 </li><li> 50ミリリットルまで（4 O Cで ）準備L64溶液でチューブを記入し、精力的にチューブを振ります。また、L64の界面活性剤の一部がミクロゲル内であることが可能であってもよいです。 </li><li> 4 O℃で2300×gで20分間チューブを遠心</li><li> Regularlyは、チューブの側のゲル粒子のペレットが存在するかどうかをチェック。粒子が見られない場合は、同じ速度および温度でさらに20分間遠心します。慎重にチューブの内壁に形成されたペレットを乱すことなく、上清を除去するために進んでください。 </li><li>繰り返し手順（3.5）から（3.3）を1回以上。各時間は、ミクロゲル懸濁液中の任意の油滴の混入を避けるために、新しいチューブに試料を移します。このステップの後、界面活性剤または油滴が試料懸濁液中に存在しないことを確認してください。しかし、反復分離工程は、最初の材料の損失につながる可能性があります。 </li></ol>ミクロゲルの4.共有結合架橋<ol><li>ゲル粒子のペレットに（セクション1で調製）ゲニピン溶液2 mlを加え、その溶液をボルテックスしてよく混ぜます。 </li><li>迅速デの間に共有結合架橋反応を開始するために23°Cの水浴中で懸濁液のチューブを移しますIRED架橋時間 （例えば、5 - 。120分）。 </li><li>架橋後、直ちに、ゲニピン液を捨て、PBSでマイクロゲルを再懸濁し、2300×gで（4 O℃）で20分間チューブを遠心分離することにより、任意の過度の架橋剤を除去します。必要に応じて、慎重にピペットチップで離れて形成されたペレットを破ります。ゲニピンは依然として溶液中に残っている場合は、この洗浄工程を3回まで繰り返すことができます。 </li><li>血球計数をカウントすることにより、上清を廃棄し、所望の濃度でPBS中のミクロゲルを再懸濁（ 例えば 、5×10 6ミクロゲル/ミリリットル）。 </li><li>顕微鏡観察のために、スライドガラスとカバーガラスとの間の空間にミクロゲル懸濁液をロードし、エポキシ樹脂とカバースリップの境界をシールします。 </li></ol>薬物放出をトリガするための交番磁界の5.アプリケーション<ol><li>水性媒体中のミクロゲルの所望の濃度にしてチューブを配置磁気コイルのチャンバー。必要であれば、AMFの適用中にメディアの温度変化を監視するために、チューブ内に光ファイバ温度プローブを挿入します。 </li><li>高周波（&gt; 100 kHzの）はAMF定義された電界強度（&gt; 5 kAの/メートル）で、指定した期間に適用されます。 AMF、遠心分離機の適用後2,273×gで（4 O℃）で20分間、試料管およびTR-BSAの量を定量するために上清を回収することは、分光光度法を使用して周囲の媒体へのミクロゲルから放出。テキサスレッドの励起および発光波長はそれぞれ、584 nmおよび612 nmです。 </li></ol>

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The authors have nothing to disclose.

ここで説明する技術は、磁気熱誘発型薬物放出のためのナノ粒子ミクロゲルハイブリッドの使用に関する概念の証拠を示しています。これは、物理的にゲニピンで架橋マイクロ三次元ゼラチンネットワーク内のMNP及びp（NIPAM- 共 -AAm）鎖を捕捉することによって達成されました。磁場応答プラットフォームは次に、モデル薬物、BSAの放出を引き起こしリモート印加AMFに応答してミクロゲ...

内因性または外因性のいずれかの刺激に応答してしっかりと制御された薬物送達を可能にする刺激応答性薬物送達系（ 例えば 、温度またはpH）は、広範囲の薬物送達のためのスマートソフト装置の新しいタイプとして研究されてきました。マイクロヒドロゲルは広く、彼らが制御可能かつ持続可能な薬物放出プロファイルと同様に調整可能な化学的および機械的性質1-3を与えるという点で、薬物送達プラットフォームとして使用されています。具体的には、コロイド状ミクロゲルは、低侵襲的方法4の局所組織に外部刺激及び適切な注射によるもの、それらの迅速な応答性薬物送達のためのビヒクルとして多くの利点を示します。ポリ（N-イソプロピルアクリルアミド）（PNIPAM）又はその共重合体が広く、ゼラチン、キトサン、アルギン酸塩、酸、またはヒアルロン酸5,6-含む生分解性/生体適合性ポリマーでPNIPAMをグラフトした熱応答性マイクロゲルの合成に採用されています、ここで、その下限臨界溶液温度（LCST）でPNIPAMの相転移特性が薬物放出7のトリガとして使用することができます。我々は最近、3次元ゼラチンネットワーク8内の微量成分としてポリ（N-isopropylacrylamide- コ -アクリルアミド）[P（NIPAM- 共同 -AAm）]チェーンを組み込むことにより、生分解性、ゼラチン系熱応答性マイクロゲルの製造を実証しました。ゼラチン/ P（NIPAM- 共 -AAm）ミクロゲルは正にウシ血清アルブミン（BSA）の放出に相関温度上昇に同調可能な脱膨潤を示しました。 過去数年の間に、オンデマンドファッション9,10に薬物の放出を誘発することができる磁気応答性薬物送達プラットフォームを開発する努力が増加しています。磁気応答性薬物送達プラットフォームの合成のための基本原理は、（超常磁性ナノ粒子の特性を利用しますMNP）は、温度感受性薬物の放出を誘発磁界を高周波交流（AMF）を受信したときに熱を発生させます。これは、このシステムは、深部組織に標的化することができるという点で、将来の臨床応用のための有望な非侵襲的なリモート制御された薬物放出を可能にし、温熱治療と磁気共鳴イメージング装置10〜12と組み合わせることができます。このようなプラットフォームは、（1）のMNP / PNIPAMハイブリッドミクロゲル粒子13-15をし、（2）肉眼ヒドロゲル足場を固定化組み込んだのMNP 16-18。 PNIPAMベースのミクロゲルプラットフォームは磁気熱刺激に細かく調整可能な体積相転移の応答性を実証しました。しかしながら、それらは依然として製造が複雑で洗練された技術に依存して、高含量のPNIPAMポリマーの使用は、それらのin vivo適用を制限することができる、細胞19に潜在的に細胞毒性であることができます。巨視的足場は相対性を示しますLY遅い外部刺激に応答し、コロイド状マイクロゲルに比べて侵襲的な外科的移植を必要とします。 油中水型乳化はサブミリまたはマイクロメートルサイズのゲル粒子20を製造するための標準的な方法でした。エマルションの水 - 油界面で、ミクロゲル粒子は、機械的せん断力下での水の液滴の表面エネルギーの最小化のために球形を形成します。この方法は、簡単な製造手順水性球状ゲル滴の大量生産を可能にし、正常な薬物送達用途21-23ゼラチンベースのミクロゲルを製造するために採用されています。 ここでは、油中水型乳化法を用いて、薬物送達用途のためmagnetothermally応答ゼラチンベースのミクロゲルを合成するための容易な方法を提示します。これは、物理的に、酸化鉄のMNP及びP（NIPAM- COを組み込むことによって達成されました-AAmと）共有結合磁界（AMF）アプリケーションシステムを高周波交流と連携して、天然由来の架橋剤ゲニピンにより架橋されている球状のマイクロスケールゼラチンネットワーク内の微量成分としてチェーン。

<table border="0" cellpadding="0" cellspacing="0" width="706">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Gelatin</td>
			<td style="width:204px;">Sigma-Aldrich, MO, USA</td>
			<td style="width:119px;">G2500</td>
			<td style="width:167px;">Gelatin type A, porcine skin</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">poly(N-isopropylacrylamide-co-acrylamide)&#160;</td>
			<td style="width:204px;">Sigma-Aldrich, MO, USA</td>
			<td>738727</td>
			<td>MW=20,000, LCST=34-38 oC</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Silicon oil</td>
			<td style="width:204px;">Sigma-Aldrich, MO, USA</td>
			<td>378372</td>
			<td>Viscosity 350 cSt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pluoronic L64</td>
			<td style="width:204px;">Sigma-Aldrich, MO, USA</td>
			<td style="width:119px;">435449</td>
			<td>100 ppm poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">genipin</td>
			<td style="width:204px;">TimTec LLC, DE, USA</td>
			<td>ST080860</td>
			<td>Mw = 226.23;&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Magnetic nanoparticles (MNPs)</td>
			<td style="width:204px;">Micromod Inc, Germany</td>
			<td>79-00-102</td>
			<td>nanomag-D-spio, 100 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">TR-BSA</td>
			<td style="width:204px;">Life Technologies, NY USA</td>
			<td>A23017</td>
			<td>Albumin from Bovine Serum (BSA), Texas Red conjugate</td>
		</tr>
	</tbody>
</table>

alternating magnetic field-responsive hybrid gelatin microgels for controlled drug release

厳密に制御、オンデマンドの薬物送達を可能にする磁気応答性ナノ/マイクロ工学生体材料は、生物医学的応用のためのスマートソフトのデバイスの新しいタイプとして開発されてきました。磁気応答性薬物送達システムの数は、概念研究のインビトロでの証拠またはインビボ前臨床用途におけるいずれかを介して有効性を実証しているが、臨床設定におけるそれらの使用は、依然として、その不十分な生体適合性または生分解性によって制限されます。さらに、既存のプラットフォームの多くは、彼らの捏造のための洗練された技術に依存しています。我々は最近、物理的に3次元ゼラチンネットワーク内の微量成分としてポリ（N-isopropylacrylamide- コ -アクリルアミド）鎖を捕捉することによって、生分解性、ゼラチン系熱応答性マイクロゲルの製造を実証しました。本研究では、磁気トンを可能にする生分解性の薬物放出プラットフォームを製造する容易な方法を提示hermally薬物放出を引き起こしました。これは、交流磁場印加装置と連動して、超常磁性酸化鉄ナノ粒子およびゼラチンベースコロイドミクロゲル中の熱応答性のポリマーを組み込むことによって達成されました。

プロトコルが正しく実行された場合、製造されたマイクロゲルは、5ミクロン（ 図1BおよびC）〜20μmの間の範囲の直径でよく特徴付け球状の形態およびコロイド分散性を示すべきです。蛍光のMNPや蛍光BSAのいずれかがのMNPまたは薬物（この研究では、BSA）が正しくミクロゲル（ 図1D）内にカプセル化されているかどうかを確認するために使用?...

Watch this Scientific Journal Video about Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release at JoVE.com

Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release

制御された薬物放出のための磁場応答性ハイブリッドゼラチンマイクロゲルを交互に

我々は、磁気熱応答性である生分解性ゼラチン系薬物放出プラットフォームを製造する容易な方法を提示します。これは、交流磁界印加装置と連動して、ゲニピンで架橋球状ゼラチンマイクロネットワーク内の超常磁性酸化鉄ナノ粒子とポリ（N-isopropylacrylamide- コ -アクリルアミド）を組み込むことによって達成されました。

Magnetically-responsive nano/micro-engineered biomaterials that enable a tightly controlled, on-demand drug delivery have been developed as new types of ...

alternating-magnetic-field-responsive-hybrid-gelatin-microgels-for

Research

JoVE Journal

Bioengineering

9.7K ビュー.  Kent State University. 我々は、磁気熱応答性である生分解性ゼラチン系薬物放出プラットフォームを製造する容易な方法を提示します。これは、交流磁界印加装置と連動して、ゲニピンで架橋球状ゼラチンマイクロネットワーク内の超常磁性酸化鉄ナノ粒子とポリ（N-isopropylacrylamide- コ -アクリルアミド）を組み込むことによって達成されました。 

ビデオ: Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release

Magnetically-Assisted Remote Controlled Microcatheter Tip Deflection under Magnetic Resonance Imaging

X-ray fluoroscopy-guided endovascular procedures have several significant limitations, including difficult catheter navigation and use of ionizing radiation, which can potentially be overcome using a magnetically steerable catheter under MR guidance.
	The main goal of this work is to develop a microcatheter whose tip can be remotely controlled using the magnetic field of the MR scanner. This protocol aims to describe the procedures for applying current to the microcoil-tipped microcatheter to produce consistent and controllable deflections.
	A microcoil was fabricated using laser lathe lithography onto a polyimide-tipped endovascular catheter. In vitro testing was performed in a waterbath and vessel phantom under the guidance of a 1.5-T MR system using steady-state free precession (SSFP) sequencing. Various amounts of current were applied to the coils of the microcatheter to produce measureable tip deflections and navigate in vascular phantoms.
	The development of this device provides a platform for future testing and opportunity to revolutionize the endovascular interventional MRI environment.

Current applied to an endovascular microcatheter with microcoil tip made by laser lathe lithography can achieve controllable deflections under magnetic resonance (MR) guidance, which may improve speed and efficacy of navigation of vasculature during various endovascular procedures.

磁気共鳴イメージング下磁気アシストリモート制御マイクロカテーテルの先端偏向

X-ray fluoroscopy-guided  endovascular procedures have several significant limitations, including  difficult catheter navigation and use of ionizing ...

生物工学

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 several advantages over traditional microsphere fabrication techniques. Contrary to emulsion, suspension, and dispersion techniques, microgels formed by precipitation are of uniform shape and size, i.e.&#160;low polydispersity index, without the use of organic solvents or stabilizers. The mild conditions of the precipitation reaction, customizable properties of the microgels, and low viscosity for injections make them applicable for in vivo purposes. Unlike other fabrication techniques, microgel characteristics can be modified by changing the starting polymer molecular weight. Increasing the starting PEG molecular weight increased microgel diameter and swelling ratio. Further modifications are suggested such as encapsulating molecules during microgel crosslinking. Simple adaptations to the PEG microgel building blocks are explored for future applications of microgels as drug delivery vehicles and tissue engineering scaffolds.

This work describes the formation of poly(ethylene glycol) (PEG) microgels via a photopolymerized precipitation reaction. Increasing the PEG molecular weight increased microgel diameter and swelling ratio. Simple adaptations to the PEG microgel precipitation reaction are explored for future applications of microgels as drug delivery vehicles and tissue engineering scaffolds.

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

Hybrid &#181;CT-FMT imaging and image analysis

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to assess the biodistribution of novel probes and to assess disease progression using established molecular probes or reporter genes. The combination with an anatomical modality, e.g., micro computed tomography (&#181;CT), is beneficial for image analysis and for fluorescence reconstruction. We describe a protocol for multimodal &#181;CT-FMT imaging including the image processing steps necessary to extract quantitative measurements. After preparing the mice and performing the imaging, the multimodal data sets are registered. Subsequently, an improved fluorescence reconstruction is performed, which takes into account the shape of the mouse. For quantitative analysis, organ segmentations are generated based on the anatomical data using our interactive segmentation tool. Finally, the biodistribution curves are generated using a batch-processing feature. We show the applicability of the method by assessing the biodistribution of a well-known probe that binds to bones and joints.

Hybrid  &#181;CT-FMT imaging and image analysis

We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (&#181;CT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.

ハイブリッドμCT-FMT撮像と画像解析

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to ...

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

磁気共鳴画像誘導による感熱リポソームからのシスプラチンのトリガリリースのためのカスタム設計されたレーザーベースの加熱装置

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Composite Scaffolds of Interfacial Polyelectrolyte Fibers for Temporally Controlled Release of Biomolecules

Various scaffolds used in tissue engineering require a controlled biochemical environment to mimic the physiological cell niche. Interfacial polyelectrolyte complexation (IPC) fibers can be used for controlled delivery of various biological agents such as small molecule drugs, cells, proteins and growth factors. The simplicity of the methodology in making IPC fibers gives flexibility in its application for controlled biomolecule delivery. Here, we describe a method of incorporating IPC fibers into two different polymeric scaffolds, hydrophilic polysaccharide and hydrophobic polycaprolactone, to create a multi-component composite scaffold. We showed that IPC fibers can be easily embedded into these polymeric structures, enhancing the capability for sustained release and improved preservation of biomolecules. We also created a composite polymeric scaffold with topographical cues and sustained biochemical release that can have synergistic effects on cell behavior. Composite polymeric scaffolds with IPC fibers represent a novel and simple method of recreating the cellular niche.

Scaffolds for tissue engineering need to recapitulate the complex biochemical and biophysical microenvironment of the cellular niche. Here, we show the use of interfacial polyelectrolyte complexation fibers as a platform to create composite, multi-component polymeric scaffolds with sustained biochemical release.

生体分子の時間的に制御された放出のための界面高分子電解質繊維の複合足場

Various scaffolds used in tissue engineering require a controlled biochemical environment to mimic the physiological cell niche. Interfacial ...

Magnetic and Thermal-sensitive Poly(N-isopropylacrylamide)-based Microgels for Magnetically Triggered Controlled Release

Magnetically and thermally sensitive poly(N-isopropylacrylamide) (PNIPAAm)/Fe3O4-NH2 microgels with the encapsulated anti-cancer drug curcumin (Cur) were designed and fabricated for magnetically triggered release. PNIPAAm-based magnetic microgels with a spherical structure were produced via a temperature-induced emulsion followed with physical-crosslinking by mixing PNIPAAm, polyethylenimine (PEI), and Fe3O4-NH2 magnetic nanoparticles. Because of their dispersity, the Fe3O4-NH2 nanoparticles were embedded inside the polymer matrix. The amine groups exposed on the Fe3O4-NH2 and PEI surface supported the spherical structure by physically crosslinking with the amide groups of the PNIPAAm. The hydrophobic anti-cancer drug curcumin can be dispersed in water after encapsulation into the microgels. The microgels were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), and UV-Vis spectral analysis. Furthermore, magnetically triggered release was studied under an external high frequency magnetic field (HFMF). A significant &#34;burst release&#34; of curcumin was observed after applying the HFMF to the microgels due to the magnetic inductive heating (hyperthermia) effect. This manuscript describes the magnetically triggered controlled release of Cur-PNIPAAm/Fe3O4-NH2 encapsulated curcumin, which can be potentially applied for tumor therapy.

Magnetic and Thermal-sensitive PolyN-isopropylacrylamide-based Microgels for Magnetically Triggered Controlled Release

This manuscript describes the preparation of magnetic and thermal-sensitive microgels via a temperature-induced emulsion without chemical reaction. These sensitive microgels were synthesized by mixing poly(N-isopropylacrylamide) (PNIPAAm), polyethylenimine (PEI) and Fe3O4-NH2 nanoparticles for the potential use in magnetically and thermally triggered drug-release.

磁気および感熱性ポリ（<em&gt; N</em&gt; - イソプロピルアクリルアミド）ベースのミクロゲル

Magnetically and thermally sensitive poly(N-isopropylacrylamide) (PNIPAAm)/Fe3O4-NH2 microgels with the encapsulated anti-cancer drug curcumin (Cur) were ...

Repressing Gene Transcription by Redirecting Cellular Machinery with Chemical Epigenetic Modifiers

Regulation of chromatin compaction is an important process that governs gene expression in higher eukaryotes. Although chromatin compaction and gene expression regulation are commonly disrupted in many diseases, a locus-specific, endogenous, and reversible method to study and control these mechanisms of action has been lacking. To address this issue, we have developed and characterized novel gene-regulating bifunctional molecules. One component of the bifunctional molecule binds to a DNA-protein anchor so that it will be recruited to an allele-specific locus. The other component engages endogenous cellular chromatin-modifying machinery, recruiting these proteins to a gene of interest. These small molecules, called chemical epigenetic modifiers (CEMs), are capable of controlling gene expression and the chromatin environment in a dose-dependent and reversible manner. Here, we detail a CEM approach and its application to decrease gene expression and histone tail acetylation at a Green Fluorescent Protein (GFP) reporter located at the Oct4 locus in mouse embryonic stem cells (mESCs). We characterize the lead CEM (CEM23) using fluorescent microscopy, flow cytometry, and chromatin immunoprecipitation (ChIP), followed by a quantitative polymerase chain reaction (qPCR). While the power of this system is demonstrated at the Oct4 locus, conceptually, the CEM technology is modular and can be applied in other cell types and at other genomic loci.

Regulation of the chromatin environment is an essential process required for proper gene expression. Here, we describe a method for controlling gene expression through the recruitment of chromatin-modifying machinery in a gene-specific and reversible manner.

化学のエピジェネティックな修飾剤の細胞機械装置をリダイレクトすることによって遺伝子の転写を抑制

Regulation of chromatin compaction is an important process that governs gene expression in higher eukaryotes. Although chromatin compaction and gene ...

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

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of experimental conditions while minimizing potential process variability. Applications of this approach include: clone screening, temperature and pH shifts,&#160;media and supplement optimization. Furthermore, the small reactor volumes are conducive to large Design of Experiments that investigate a wide range of conditions. This allows upstream processes to be significantly optimized before scale-up where experimentation is more limited in scope due to time and economic constraints. Automated microscale bioreactor systems offer various advantages over traditional small scale cell culture units, such as shake flasks or spinner flasks. However, during pilot scale process development&#160;significant care must be taken to ensure that these advantages are realized. When run with care, the system can enable high level automation, can be programmed to run DOE&#39;s with a higher number of variables&#160;and can reduce sampling time when integrated with a nutrient analyzer or cell counter. Integration of the expert-derived heuristics presented here, with current automated microscale bioreactor experiments can minimize common pitfalls that hinder meaningful results. In the extreme, failure to adhere to the principles laid out here can lead to equipment damage that requires expensive repairs. Furthermore, the microbioreactor systems have small culture volumes&#160;making characterization of cell culture conditions difficult. The number and amount of samples taken in-process in batch mode culture is limited as operating volumes cannot fall below 10 mL. This method will discuss the benefits and drawbacks of microscale bioreactor systems.

A detailed protocol for the concurrent operation of 48 parallel cell cultures under varied conditions in a microbioreactor system is presented. Cell culture process, harvest and subsequent antibody titer analysis are described.

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 ...

Wet-spinning-based Molding Process of Gelatin for Tissue Regeneration

This article presents an inexpensive method to fabricate gelatin, as a natural polymer, into monofilament fibers or other appropriate forms. Through the wet spinning method, gelatin fibers are produced by smooth extrusion in a suitable coagulation medium. To increase the functional surface of these gelatin fibers and their ability to mimic the features of tissues, gelatin can be molded into a tube form by referring to this concept. Examined by in vitro and in vivo tests, the gelatin tubes demonstrate a great potential for application in tissue engineering. Acting as a suitable filling gap material, gelatin tubes can be used to substitute the tissue in the damaged area (e.g., in the nervous or cardiovascular system), as well as to promote regeneration by providing a direct replacement of stem cells and neural circuitry. This protocol provides a detailed procedure for creating a biomaterial based on a natural polymer, and its implementation is expected to greatly benefit the development of correlative natural polymers, which help to realize tissue regeneration strategies.

We developed and describe a protocol based on the wet spinning concept, for the construction of gelatin-based biomaterials used for the application of tissue engineering.

組織再生用ゼラチンのウェット ベース回転成形プロセス

This article presents an inexpensive method to fabricate gelatin, as a natural polymer, into monofilament fibers or other appropriate forms. Through the ...

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

Primary clarification is an essential step in a biomanufacturing process for the initial removal of cells from therapeutic products within the harvested cell culture fluid. While traditional methods like centrifugation or filtration are widely implemented for cell removal, the equipment for these processes have large footprints and operation can involve contamination risks and filter fouling. Additionally, traditional methods may not be ideal for continuous bioprocessing schemes for primary clarification. Thus, an alternate application using acoustic (sound) waves was investigated to continuously separate cells from the cell culture fluid. Presented in this study is a detailed protocol for using a bench-scale acoustic wave separator (AWS) for the primary separation of culture fluid containing a monoclonal IgG1 antibody from a CHO cell bioreactor harvest. Representative data are presented from the AWS and demonstrate how to achieve effective cell clarification and product recovery. Finally, potential applications for AWS in continuous bioprocessing are discussed. Overall, this study provides a practical and general protocol for the implementation of AWS in primary clarification for CHO cell cultures and further describes its application potential in continuous bioprocessing.

Presented here is a protocol for the primary clarification of CHO cell culture using an acoustic separator. This protocol can be used for the primary clarification of shake flask cultures or bioreactor harvests and has the potential application for continuous clarification of the cell bleed material during perfusion bioreactor operations.

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 ...

制御された薬物放出のための磁場応答性ハイブリッドゼラチンマイクロゲルを交互に

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