この作品は、NIH K08 EB003996とアメリカ研究財団助成2573の麻痺退役軍人によってサポートされていました。

1。ポリマーを選択してください<ol><li>ユーザの仕様に基づいて、ポリマー（例えば、ポリ- L -乳酸（PLLA）、ポリカプロラクトン（PCL）、ポリスチレン（PS）またはナイロン）（例えば、生分解性、熱可塑性または架橋性）とそのポリマーの溶剤を選択してください。あなたの選択に基づいて適切な保護具を選択してください。 </li><li>アプリケーションに基づいて、基板（例えば、ガラス、プラスチック、金属またはシリコンウェハ）を選択します。 </li></ol> 2。コレクタを選択します。 <ol><li>あなたの仕様に基づいてコレクタの形状を選択します。ランダムな繊維は、静止プレート上で収集することができます。整列繊維は、急速に回転する車輪、ドラムまたはロッド、またはパラレルプレート上に収集することができます。 </li><li>コレクタは、導電性でなければならず、それはまた、隣接するオブジェクト、テーブルトップ、等を接地せずに接地することができるような方法で、そのアクセルから絶縁しておく必要があります</li></ol> 3。経験的に1およその臨界絡み合い濃度<ol><li>いくつかの候補のポリマーの濃度（例えば、4、10、15、20、30重量％）を準備してから開始する（解決策は、粘性の高い液体ではなくゲルでなければならない）フロー濃度を選択してください。 </li><li>エレクトロスピニング装置2,3,4,5を設定する（図1を参照） <ol><li>シリンジポンプをロードし、先端からワイプソリューションのいずれかのビーズが、即座に交換されるようにポンプ速度を設定します。 </li><li>グランドコレクタと導体プレート（そのような注射器の先端の突出を介してアルミ箔等の導電性材料の小さい正方形）に高電圧線をクリッピング。 </li><li>車輪が回転し始める。 </li><li>電源装置はワークステーションの電源を入れる前にゼロに設定されていることを確認します。 </li></ol></li><li>ストリームを観察<ol><li>徐々に電圧をランプアップし、針先での解のビーズを見る。 </li><li>長いと安定したストリームを取得するために電圧を調整する。安定した流れが得られない場合、ポリマー溶液の濃度を調整する。例えば、表1を参照してください。 </li></ol></li></ol> 4。トラブルシューティング - ストリーム： <ol><li>ストリームの表示に問題<ol><li>濃いマットの背景を使用し、視聴者とストリームの間に単方向の光源（例えば、懐中電灯など）（図2参照）に置きます。 </li></ol></li><li>シリンジの先端から滴り落ちる<ol><li>ポリマー溶液は、車輪にない魅力でまっすぐ下方に滴下されている場合は、導体板は、針の先端との接触を作っているし、コレクターが地面との接触をしていることを確認してください。 </li><li>シリンジ先端のポリマー溶液の液滴は、ホイールの方向に傾いてもストリームを形成していない場合、電圧を増加させる。ストリームの品質が安定した流れが見えるようになるまでの距離と電圧を変えることによって調整することができます。 4％のPLLAの解と8x8 cmの導体板に対応する電圧で、推奨距離については、図3を参照してください。 </li></ol></li><li>シリンジの先端に大きなグロブ<ol><li>ポリマー溶液は、非導電性の棒に接続されたペーパータオルでグロブ離れて針、スワイプの先端で収集し、硬化を開始したとき。 </li></ol></li><li>振動または&#39;振る&#39;ストリーム ストリームが急速に上下に振るとされている場合、電圧を下げるか、注射器の先端とホイールの間の距離を増加させる。ストリームは、WAGポリマーのより高い濃度を使用するか、遅い蒸発で少し溶剤を追加し続けている場合。 </li><li>ショートまたは不連続のストリーム 高回転速度でホイールをセットして観察可能な接触することは一定した流れは均一とアラインメントの最高品質が得られます。ストリームは、溶媒の蒸発、およびストリームの長さと安定性を向上される電圧を調整し、より遅いの追加、ポリマー溶液を増やし、短いと不連続である場合。 </li></ol> 5。トラブルシューティング-繊維形態6,7,8（図4を参照） <ol><li>ビーズ ビーズが繊維に発見されている場合、ポリマー溶液を増加させ、導体板は、針で連続的な接触をしていると、接地ワイヤブラシはホイールで連続的な接触をしていることを確認してください。 </li><li>リボンと出血繊維 繊維のリボンとして形成されているか一緒に出血しているときは、ポリマーのより高い濃度または蒸発率が高い（より揮発性）を持つ溶剤を使用してください。 </li><li>渦巻き図形または波状繊維 繊維は波や渦巻き様を形成している場合、車輪の速度を増やすか、コレクターから、さらに針先を移動する。また、導体板とコレクタが振動していないことを確認してください。 </li><li>気孔率9 毛穴が必要な場合は、急速に蒸発溶剤を使用してください。毛穴が望まれていない場合は、主要な溶剤よりも揮発性である共溶媒を少量添加してみてください。 </li><li>アライメント10 コレクターはMOVのとき低回転時や安静時のING、アライメントの品質が悪いです。車輪の速度を増加させることによりアライメントを増やします。 </li></ol> 6。代表的な結果： 典型的な繊維の結果の描写については、図4を参照してください。 <img src="/files/ftp_upload/2494/2494fig1.jpg" alt="図1" /> 図1典型的なエレクトロのセットアップ。ポリマー溶液（青）はシリンジポンプ（オレンジ）から分配される。高電圧直流電源（緑）を理由配向ナノファイバーが収集され、その上に急速に回転するホイール​​のコレクタ（グレー）。シリンジとコレクタの間にポリマージェットは安定したストリーミングセグメントと急速に振動ホイップセグメントで構成されています。 <img src="/files/ftp_upload/2494/2494fig2.jpg" alt="図2" /> 図2ストリーミングジェットは、シリンジの先端から出る可視であり、ホイップ、ジェットが見られるように小さすぎる。 PLLAの臨界絡み合い濃度の近似<table border="1"><tbody><tr><td> PLLA（％重量/ V） </td><td> 観測 </td><td> 濃度調整 </td></tr><tr><td> 0.5 </td><td>ドリッピングありません。なくストリームを</td><td>増加する</td></tr><tr><td> 2.0 </td><td>小さなグロブ唾吐きありません。なくストリームを</td><td>少しIncrese </td></tr><tr><td> 4.0 </td><td>安定したストリーム</td><td>良好な</td></tr><tr><td> 6.0 </td><td>大グロブやビーズを吐き</td><td>わずかに減少</td></tr><tr><td> 12.0 </td><td>先端部に凝集しない;ないストリーム</td><td>減少する</td></tr></tbody></table> 表1の例では、PLLAの臨界絡み合い濃度の近似を描いた。いろいろなポリマー濃度が試されており、安定したストリームになるまで観測された結果のストリーミングジェットが得られる。 <img src="/files/ftp_upload/2494/2494fig3.jpg" alt="図3" /> 図3シリンジの先端とコレクタ間の距離は、安定したストリーミングジェットを得るために印加電圧とバランスを取る必要があります。過剰な印加電圧はあまり揃え繊維でその結果を形成するために、振動や&quot;振る&quot;ジェットが発生します。電圧が低すぎると、何ジェットが形成されず、ソリューションは、唯一のシリンジの先端から滴下される。紫色の網掛け領域は、上記の安定したストリーミングジェットはシリンジ - コレクタ間距離の関数としてPLLAのために得ることができる以上の電圧範囲を表します。 <img src="/files/ftp_upload/2494/2494fig4.jpg" alt="図4" /> 図4。エレクトロ繊維は玉が付く（）、リボン（B）、渦巻き様（C）、多孔質のグロブ（D）、良好なアライメント（E）と貧しいアライメント（F）を含めて、形態の様々な展示ができます。

<ol>
	<li>Shenoy, S. L., Bates, W. D., Frisch, H. L., Wnek, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+chain+entanglements+on+fiber+formation+during+electrospinning+of+polymer+solutions:+good+solvent,+non-specific+polymer-polymer+interaction+limit.">Role of chain entanglements on fiber formation during electrospinning of polymer solutions: good solvent, non-specific polymer-polymer interaction limit.</a> Polymer. 46, 3372-3384 (2005).</li><li>Gertz, C. C., Leach, M. K., Birrel, L. K., Martin, D. C., Feldman, E. L., Corey, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+neuritogenesis+and+maturation+of+primary+spinal+motor+neurons+in+response+to+nanofibers.">Accelerated neuritogenesis and maturation of primary spinal motor neurons in response to nanofibers.</a> Dev. Neurobiol. 70, 589-603 (2010).</li><li>Lin, D. Y., Johnson, M. A., Vohden, R. A., Chen, D., Martin, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tailored+nanofiber+morphologies+using+modulated+electrospinning+for+biomedical+applications.">Tailored nanofiber morphologies using modulated electrospinning for biomedical applications.</a> Mat. Res. Soc. Symp. Proc. 736, D3.8.1-D3.8.6 (2003).</li><li>Corey, J. M., Gertz, C. C., Wang, B. S., Birrell, L. K., Johnson, S. L., Martin, D. C., Feldman, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+design+of+electrospun+PLLA+nanofiber+scaffolds+compatible+with+serum-free+growth+of+primary+motor+and+sensory+neurons.">The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons.</a> Acta. Biomater. 4, 863-875 (2008).</li><li>Corey, J. M., Lin, D. Y., Mycek, K. B., Chen, Q., Samuel, S., Feldman, E. L., Martin, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aligned+electrospun+nanofibers+specify+the+direction+of+dorsal+root+ganglia+neurite+growth.">Aligned electrospun nanofibers specify the direction of dorsal root ganglia neurite growth.</a> J. Biomed. Mater. Res. A. 83, 636-645 (2007).</li><li>Tan, S. -. H., Kotaki, M., Ramakrishna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+parameter+study+for+ultra-fine+fiber+fabrication+via+electrospinning+process.">Systematic parameter study for ultra-fine fiber fabrication via electrospinning process.</a> Polymer. 46, 6128-6134 (2005).</li><li>Yang, F., Murugan, R., Wang, S., Ramakrishna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning+of+nano/micro+scale+poly(L-lactic+acid)+aligned+fibers+and+their+potential+in+neural+tissue+engineering.">Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neural tissue engineering.</a> Biomaterials. 26, 2603-2610 (2005).</li><li>Li, W., Laurencin, C. T., Caterson, E. J., Tuan, R. S., Ko, F. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospun+nanofibrous+structure:+A+novel+scaffold+for+tissue+engineering.">Electrospun nanofibrous structure: A novel scaffold for tissue engineering.</a> J. Biomed. Mater. Res. A. 60, 613-621 (2002).</li><li>Kim, C. H., Jung, Y. H., Kim, H. Y., Lee, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+collector+temperature+on+the+porous+structure+of+electrospun+fibers.">Effect of collector temperature on the porous structure of electrospun fibers.</a> Macromol. Res. 14, 59-65 (2006).</li><li>Wang, H. B., Mullins, M. E., Cregg, J. M., Hurtado, A., Oudega, M., Trombley, M. T., Gilbert, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Creation+of+highly+aligned+electrospun+poly-L-lactic+acid+fibers+for+nerve+regeneration+applications.">Creation of highly aligned electrospun poly-L-lactic acid fibers for nerve regeneration applications.</a> J. Neural Eng. 6, (2009).</li></ol>

利害の衝突は宣言されません。

注：例の大部分は、ポリ- L -乳酸（PLLA）ナノファイバーをエレクトロスピニングを扱うここで紹介。 PLLAは、我々の研究室で最も一般的にスピンポリマーであるため、これは単純です。しかし、我々はまた、成功した他のポリマー（例えば、PLGA、PCL、PS）electrospinとここに示す手法は、高分子量のポリマー溶液半ばまでの大部分に容易に適用可能であることを信じるようにこれらのメソッドを使用している。

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>High voltage DC power supply</td>
<td>&nbsp;</td>
<td>Gamma High Voltage</td>
<td>ES40P-5W</td>
<td>&nbsp;</td>
<tr>
<td>Syringe pump</td>
<td>&nbsp;</td>
<td>KD Scientific</td>
<td>KDS100</td>
<td>&nbsp;</td>
<tr>
<td>Aluminum foil</td>
<td>&nbsp;</td>
<td>Reynolds</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Blunt metal tips, 23ga</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>13-850-102 </td>
<td>&nbsp;</td>
<tr>
<td>Polypropylene syringe</td>
<td>&nbsp;</td>
<td>BD</td>
<td>309585</td>
<td>&nbsp;</td>
<tr>
<td>Rotating or stationary collector</td>
<td>&nbsp;</td>
<td>Custom built</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Various alligator clips and wires</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Dimethylformamide</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>AC11622-0010</td>
<td>&nbsp;</td>
<tr>
<td>Chloroform</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>AC42355-0040 </td>
<td>&nbsp;</td>
<tr>
<td>PLLA</td>
<td>&nbsp;</td>
<td>Boehringer Ingelheim</td>
<td>Resomer L210</td>
<td>&nbsp;</td>
<tr>
<td>PLGA 85:15</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>43471</td>
<td>&nbsp;</td>
<tr>
<td>Carbon tape</td>
<td>&nbsp;</td>
<td>Ted Pella</td>
<td>13073-1</td>
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electrospinning fundamentals: optimizing solution and apparatus parameters

エレクトロスピニングナノファイバーの足場は、成熟を ​​加速成長を向上させ、in vitroでの細胞の遊走を誘導することが示されている。エレクトロスピニングは、帯電したポリマーのジェットが接地コレクタで収集されるプロセスです。整列ナノファイバーで急速に回転するコレクタの結果をランダムに配向した繊維マットで固定コレクターの結果ながら。ポリマージェットは、適用の静電荷は溶液の表面張力を克服した時に形成される。所与のポリマーの最小濃度がある、重要なエンタングルメントの濃度は、安定したジェットを達成することができないとはナノファイバーが形成されませんこれを下回る、と呼ば - ナノ粒子を達成することができるが（エレクトロ）。安定したジェットは、2つのドメイン、ストリーミングセグメントとホイップセグメントがあります。ホイップジェットは通常肉眼では見えないですが、ストリーミングの部分は、多くの場合、適切な照明条件の下で表示されます。ストリームの長さ、太さ、一貫性と動きを観察することが形成されるナノファイバーの配置と形態を予測するのに便利です。短期、不均一な、一貫性のない、および/または振動ストリームは、飛散ビーズ、そして渦巻き図形または波状のパターン、貧しいファイバーアライメントを含め、さまざまな問題の指標となる。ストリームは、このように生産されている繊維のアラインメントと形態を最適化する、溶液の組成とエレクトロスピニング装置の構成を調整することによって最適化することができます。このプロトコルでは、我々は経験的に、ポリマー溶液の臨界絡み合い濃度を近似し、エレクトロスピニングプロセスを最適化し、基本的なエレクトロスピニング装置を設定する手順を提示する。さらに、我々はいくつかの一般的な問題とトラブルシューティングのテクニックについて説明します。

Watch this Scientific Journal Video about Electrospinning Fundamentals: Optimizing Solution and Apparatus Parameters at JoVE.com

Electrospinning Fundamentals: Optimizing Solution and Apparatus Parameters

エレクトロスピニング技術は、組織工学やその他のアプリケーションのためのナノ繊維足場の様々を作成することができます。ここでは、所望の形態と配置に繊維を得るためにエレクトロソリューションと装置のパラメータを最適化する手順を説明します。一般的な問題とトラブルシューティングのテクニックも紹介されています。

エレクトロスピニングの基礎：最適化ソリューションと装置のパラメータ

Electrospun nanofiber scaffolds have been shown to accelerate the maturation, improve the growth, and direct the migration of cells in vitro. ...

electrospinning-fundamentals-optimizing-solution-apparatus

Research

JoVE Journal

Bioengineering

64.6K ビュー.  University of Michigan. エレクトロスピニング技術は、組織工学やその他のアプリケーションのためのナノ繊維足場の様々を作成することができます。ここでは、所望の形態と配置に繊維を得るためにエレクトロソリューションと装置のパラメータを最適化する手順を説明します。一般的な問題とトラブルシューティングのテクニックも紹介されています。

ビデオ: Electrospinning Fundamentals: Optimizing Solution and Apparatus Parameters

Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study

Throughout native artery, collagen and elastin play an important role, providing a mechanical backbone, preventing vessel rupture, and promoting recovery under pulsatile deformations. The goal of this study was to mimic the structure of native artery by fabricating a multi-layered electrospun conduit composed of poly(caprolactone) (PCL) with the addition of elastin and collagen with blends of 45-45-10, 55-35-10, and 65-25-10 PCL-ELAS-COL to demonstrate mechanical properties indicative of native arterial tissue, while remaining conducive to tissue regeneration. Whole grafts and individual layers were analyzed using uniaxial tensile testing, dynamic compliance, suture retention, and burst strength. Compliance results revealed that changes to the middle/medial layer changed overall graft behavior with whole graft compliance values ranging from 0.8 - 2.8 % / 100 mmHg, while uniaxial results demonstrated an average modulus range of 2.0 - 11.8 MPa. Both modulus and compliance data displayed values within the range of native artery. Mathematical modeling was implemented to show how changes in layer stiffness affect the overall circumferential wall stress, and as a design aid to achieve the best mechanical combination of materials. Overall, the results indicated that a graft can be designed to mimic a tri-layered structure by altering layer properties.

The aim of this study was to mimic the native three layered architecture of the arterial wall. To accomplish this, electrospinning was employed with the use of a 3-1 (input-output) nozzle and blends of polycaprolactone, elastin, and collagen.

ポリカプロラクトン、エラスチン、コラーゲンを使用してネイティブ動脈アーキテクチャを模倣するためにトライレイヤーエレクトロスピニング：予備的研究

Throughout native artery, collagen and elastin play an important role, providing a mechanical backbone, preventing vessel rupture, and promoting recovery ...

生物工学

Quantitatively Measuring In situ Flows using a Self-Contained Underwater Velocimetry Apparatus (SCUVA)

The ability to directly measure velocity fields in a fluid environment is necessary to provide empirical data for studies in fields as diverse as oceanography, ecology, biology, and fluid mechanics. Field measurements introduce practical challenges such as environmental conditions, animal availability, and the need for field-compatible measurement techniques. To avoid these challenges, scientists typically use controlled laboratory environments to study animal-fluid interactions. However, it is reasonable to question whether one can extrapolate natural behavior (i.e., that which occurs in the field) from laboratory measurements. Therefore, in situ quantitative flow measurements are needed to accurately describe animal swimming in their natural environment.
We designed a self-contained, portable device that operates independent of any connection to the surface, and can provide quantitative measurements of the flow field surrounding an animal. This apparatus, a self-contained underwater velocimetry apparatus (SCUVA), can be operated by a single scuba diver in depths up to 40 m. Due to the added complexity inherent of field conditions, additional considerations and preparation are required when compared to laboratory measurements. These considerations include, but are not limited to, operator motion, predicting position of swimming targets, available natural suspended particulate, and orientation of SCUVA relative to the flow of interest. The following protocol is intended to address these common field challenges and to maximize measurement success.

Quantitatively Measuring In situ Flows using a Self-Contained Underwater Velocimetry Apparatus SCUVA

This protocol provides instructions on how to use a self-contained underwater velocimetry apparatus (SCUVA), which is designed for quantification of in situ animal-generated flows. In addition, this protocol addresses challenges posed by field conditions, and includes operator motion, predicting position of animals, and orientation of SCUVA.

定量的に測定その場で自己完結型水中速度計測装置（SCUVA）を使用してフロー

The ability to directly measure velocity fields in a fluid environment is necessary to provide empirical data for studies in fields as diverse as ...

Small and Wide Angle X-Ray Scattering Studies of Biological Macromolecules in Solution

In this paper, Small and Wide Angle X-ray Scattering (SWAXS) analysis of macromolecules is demonstrated through experimentation. SWAXS is a technique where X-rays are elastically scattered by an inhomogeneous sample in the nm-range at small angles (typically 0.1 - 5&deg;) and wide angles (typically &gt; 5&deg;). This technique provides information about the shape, size, and distribution of macromolecules, characteristic distances of partially ordered materials, pore sizes, and surface-to-volume ratio. Small Angle X-ray Scattering (SAXS) is capable of delivering structural information of macromolecules between 1 and 200 nm, whereas Wide Angle X-ray Scattering (WAXS) can resolve even smaller Bragg spacing of samples between 0.33 nm and 0.49 nm based on the specific system setup and detector. The spacing is determined from Bragg's law and is dependent on the wavelength and incident angle.
In a SWAXS experiment, the materials can be solid or liquid and may contain solid, liquid or gaseous domains (so-called particles) of the same or another material in any combination. SWAXS applications are very broad and include colloids of all types: metals, composites, cement, oil, polymers, plastics, proteins, foods, and pharmaceuticals. For solid samples, the thickness is limited to approximately 5 mm.
Usage of a lab-based SWAXS instrument is detailed in this paper. With the available software (e.g., GNOM-ATSAS 2.3 package by D. Svergun EMBL-Hamburg and EasySWAXS software) for the SWAXS system, an experiment can be conducted to determine certain parameters of interest for the given sample. One example of a biological macromolecule experiment is the analysis of 2 wt% lysozyme in a water-based aqueous buffer which can be chosen and prepared through numerous methods. The preparation of the sample follows the guidelines below in the Preparation of the Sample section. Through SWAXS experimentation, important structural parameters of lysozyme, e.g. the radius of gyration, can be analyzed.

The demonstration of the small and wide angle X-ray scattering (SWAXS) procedure has become instrumental in the study of biological macromolecules. Through the use of the instrumentation and procedures of specific angle methods and preparation, the experimental data from the SWAXS displays the atomic and nano-scale characterization of macromolecules.

溶液中の生体高分子の小型·広角X線散乱による研究

In this paper, Small and Wide Angle X-ray Scattering (SWAXS) analysis of macromolecules is demonstrated through experimentation. SWAXS is a technique ...

Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth. This system incorporates mechanical, topographical, adhesive and chemical signals. Mechanical properties are controlled by the type of material used to fabricate the electrospun fibers. In this protocol we use 30% methacrylated Hyaluronic Acid (HA), which has a tensile modulus of ~500 Pa, to produce a soft fibrous scaffold. Electrospinning on to a rotating mandrel produces aligned fibers to create a topographical cue. Adhesion is achieved by coating the scaffold with fibronectin. The primary challenge addressed herein is providing a chemical signal throughout the depth of the scaffold for extended periods. This procedure describes fabricating&#160;poly(lactic-co-glycolic acid) (PLGA) microspheres that contain Nerve Growth Factor (NGF) and directly impregnating the scaffold with these microspheres during the electrospinning process. Due to the harsh production environment, including high sheer forces and electrical charges, protein viability is measured after production. The system provides protein release for over 60 days and has been shown to promote primary nerve cell growth.

This protocol combines electrospinning and microspheres to develop tissue engineered scaffolds to direct neurons. Nerve growth factor was encapsulated within PLGA microspheres and electrospun into Hyaluronic Acid (HA) fibrous scaffolds. The protein bioactivity was tested by seeding the scaffolds with primary chick Dorsal Root Ganglia and culturing for 4-6 days.

繊維状足場に微小球を離すエレクトロ成長因子

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth.  This system incorporates mechanical, topographical, ...

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as Aniridia or Steven Johnson Syndrome as well as by external factors such as chemical burns or radiation. Current treatments are (i) the use of corneal grafts and (ii) the use of stem cell expanded in the laboratory and delivered on carriers (e.g., amniotic membrane); these treatments are relatively successful but unfortunately they can fail after 3-5 years. There is a need to design and manufacture new corneal biomaterial devices able to mimic in detail the physiological environment where stem cells reside in the cornea. Limbal stem cells are located in the limbus (circular area between cornea and sclera) in specific niches known as the Palisades of Vogt. In this work we have developed a new platform technology which combines two cutting-edge manufacturing techniques (microstereolithography and electrospinning) for the fabrication of corneal membranes that mimic to a certain extent the limbus. Our membranes contain artificial micropockets which aim to provide cells with protection as the Palisades of Vogt do in the eye.

We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.

角膜再生のためのニッチを装備膜を製造するためのマイクロ光とエレクトロスピニングの組み合わせ

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as ...

Fabricating Degradable Thermoresponsive Hydrogels on Multiple Length Scales via Reactive Extrusion, Microfluidics, Self-assembly, and Electrospinning

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), their ultimate clinical use has been hampered by the lack of biologically-relevant degradation observed for most smart materials. This is particularly true for temperature-responsive hydrogels, which are almost uniformly based on polymers that are functionally non-degradable (e.g., poly(N-isopropylacrylamide) (PNIPAM) or poly(oligoethylene glycol methacrylate) (POEGMA)). As such, to effectively translate the potential of thermoresponsive hydrogels to the challenges of remote-controlled or metabolism-regulated drug delivery, cell scaffolds with tunable cell-material interactions, theranostic materials with the potential for both imaging and drug delivery, and other such applications, a method is required to render the hydrogels (if not fully degradable) at least capable of renal clearance following the required lifetime of the material. To that end, this protocol describes the preparation of hydrolytically-degradable hydrazone-crosslinked hydrogels on multiple length scales based on the reaction between hydrazide and aldehyde-functionalized PNIPAM or POEGMA oligomers with molecular weights below the renal filtration limit. Specifically, methods to fabricate degradable thermoresponsive bulk hydrogels (using a double barrel syringe technique), hydrogel particles (on both the microscale through the use of a microfluidics platform facilitating simultaneous mixing and emulsification of the precursor polymers and the nanoscale through the use of a thermally-driven self-assembly and cross-linking method), and hydrogel nanofibers (using a reactive electrospinning strategy) are described. In each case, hydrogels with temperature-responsive properties similar to those achieved via conventional free radical cross-linking processes can be achieved, but the hydrazone cross-linked network can be degraded over time to re-form the oligomeric precursor polymers and enable clearance. As such, we anticipate these methods (which may be generically applied to any synthetic water-soluble polymer, not just smart materials) will enable easier translation of synthetic smart materials to clinical applications.

Protocols are described for the fabrication of degradable thermoresponsive hydrogels based on hydrazone cross-linking of polymeric oligomers on the bulk scale, microscale, and nanoscale, the latter for preparation of both gel nanoparticles and nanofibers.

自己組織化反応押出、マイクロフルイディクス, 経由で複数の長さスケールでの分解性感温ハイドロゲルを加工・ エレクトロスピニング

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.

Melt Electrospinning Writing of Three-dimensional Poly(&amp;#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

組織工学的応用の制御可能な形態の三次元 Poly(ε-caprolactone) 足場のエレクトロスピニング記述を溶かす

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

High-resolution Patterning Using Two Modes of Electrohydrodynamic Jet: Drop on Demand and Near-field Electrospinning

Electrohydrodynamic (EHD) jet printing has drawn attention in various fields because it can be used as a high-resolution and low-cost direct patterning tool. EHD printing uses a fluidic supplier to maintain the extruded meniscus by pushing the ink out of the nozzle tip. The electric field is then used to pull the meniscus down to the substrate to produce high-resolution patterns. Two modes of EHD printing have been used for fine patterning: continuous near-field electrospinning (NFES) and dot-based drop-on-demand (DOD) EHD printing. According to the printing modes, the requirements for the printing equipment and ink viscosity will differ. Even though two different modes can be implemented with a single EHD printer, the realization methods significantly differ in terms of ink, fluidic system, and driving voltage. Consequently, without a proper understanding of the jetting requirements and limitations, it is difficult to obtain the desired results. The purpose of this paper is to present a guideline so that inexperienced researchers can reduce the trial and error efforts to use the EHD jet for their specific research and development purposes. To demonstrate the fine-patterning implementation, we use Ag nanoparticle ink for the conductive patterning in the protocol. In addition, we also present the generalized printing guidelines that can be used for other types of ink for various fine-patterning applications.

Here, we present a protocol to produce high-resolution conductive patterns using electrohydrodynamic (EHD) jet printing. The protocol includes two modes of EHD jet printing: the continuous near-field electrospinning (NFES) and the dot-based drop-on-demand (DOD) EHD printing.

パターン化する電気流体力学的ジェットの 2 つのモードを使用して高解像度: 需要と近傍電界紡糸ドロップ

Electrohydrodynamic (EHD) jet printing has drawn attention in various fields because it can be used as a high-resolution and low-cost direct patterning ...

Apparatus for Harvesting Tissue Microcolumns

This manuscript describes the production process for a laboratory apparatus, made from off-the-shelf components, that can be used to collect microcolumns of full-thickness skin tissue. The small size of the microcolumns allows donor sites to heal quickly without causing donor site scarring, while harvesting full-thickness tissue enables the incorporation of all cellular and extracellular components of skin tissue, including those associated with deeper dermal regions and the adnexal skin structures, which have yet to be successfully reproduced using conventional tissue engineering techniques. The microcolumns can be applied directly into skin wounds to augment healing, or they can be used as the autologous cell/tissue source for other tissue engineering approaches. The harvesting needles are made by modifying standard hypodermic needles, and they can be used alone for harvesting small amounts of tissue or coupled with a simple suction-based collection system (also made from commonly available laboratory supplies) for high-volume harvesting to facilitate studies in large animal models.

Here we describe a protocol for producing harvesting needles that can be used to collect full-thickness skin tissue without causing donor site scarring. The needles can be combined with a simple collection system to achieve high-volume harvesting.

配向柱状組織を採取するための装置

This manuscript describes the production process for a laboratory apparatus, made from off-the-shelf components, that can be used to collect microcolumns ...

Three-Dimensional Echocardiographic Method for the Visualization and Assessment of Specific Parameters of the Pulmonary Veins

The dimensions of the pulmonary veins are important parameters when planning pulmonary vein isolation (PVI), especially with the cryoballoon ablation technique. Acknowledging the dimensions and anatomical variations of the pulmonary veins (PVs) may improve the outcome of the intervention. Conventional 2D transoesophageal echocardiography can only provide limited data about the dimensions of the PVs; however, 3D echocardiography can further evaluate relevant diameters and areas of the PVs, as well as their spatial relationship to surrounding structures. In previous literature data, parameters influencing the success rate of PVI have already been identified. These are the left lateral ridge, the intervenous ridge, the ostial area of the PVs and the ovality index of the ostium. Proper imaging of the PVs by 3D echocardiography is a technically challenging method. One crucial step is the collection of images. Three individual transducer positions are necessary to visualize the important structures; these are the left lateral ridge, the ostium of the PVs and the intervenous ridge of the left and right PVs. Next, 3D images are acquired and saved as digital loops. These datasets are cropped, which result in the en face views displaying spatial relationships. This step can also be employed to determine the anatomical variations of the PVs. Finally, multiplanar reconstructions are created to measure each individual parameter of the PVs.
Optimal quality and orientation of the acquired images are paramount for the appropriate assessment of PV anatomy. In the present work, we examined the 3D visibility of the PVs and the suitability of the above method in 80 patients. The aim was to provide a detailed outline of the essential steps and potential pitfalls of PV visualization and assessment with 3D echocardiography.

The dimensions of the pulmonary veins (PV) are important parameters when planning pulmonary vein isolation. 2D transoesophageal echocardiography can only provide limited data about the PVs; however, 3D echocardiography can evaluate relevant diameters and areas of the PVs, as well as their spatial relationship to surrounding structures.

肺静脈の特異的パラメータの可視化と評価のための3次元心エコー法

The dimensions of the pulmonary veins are important parameters when planning pulmonary vein isolation (PVI), especially with the cryoballoon ablation ...