This research was supported by the Technology Innovation Program (Contract grant No. 0037915, WPM Biomedical Materials-Implant Materials) and Industrial Strategic Technology Development Program (Contract grant No. 10045329, Development of customized implant with porous structure for bone replacement), funded by the Ministry of Trade, industry &#38; Energy (MI, Korea), and BK21 PLUS SNU Materials Division for Educating Creative Global Leaders (Contract grant No. 21A20131912052).

多孔性金属足場の作製1。 <ol><li> 4初期多孔度（40、50、60、及び70）を有する多孔質チタン足場のために、表1に記載したように材料の適切な量を秤量した後、市販のTi粉末、カンフェン、及びKD-4を混合したTi-カンフェンスラリーを準備します。 500 mlのポリエチレン（PE）ボトルにスラリーを注ぎ、30 rpmでボールミルオーブン中で30分間55℃でボトルを回転させます。 </li><li> 60ミリメートルと60ミリメートルの高さの直径を有する円筒状のアルミニウム（Al）鋳型に、PEボトルからスラリーを注ぎます。対応するアルミニウムカバースリップで各アルミニウム金型を密閉し、10分間55℃で、30 rpmの速度でボールミルオーブン中で金型を回転させます。 <ol><li>その後、44°Cまでボールミルオーブンの温度を低下させ、かつ連続的に12時間、44℃の一定温度で、30 rpmの速度で金型を回転させます。 </li></ol></li><li>ボール - から金型を取り出しさらに、冷却プロセスのために1時間室温で金型を回転させた後、ミルオーブン。アルプランジャーを使用したAl金型から凝固したチタン/カンフェン緑体を削除します。 </li><li>手でゴム製の袋に入れて固化したチタン/カンフェン緑体を配置し、完全に文字列を使用して袋の口を結ぶことにより、ゴム袋を密封します。 （CIP）プレス機を冷間静水圧の水タンクにゴム袋を置き、10分間200 MPaでの静水圧を適用します。ゴム袋から圧縮されたグリーン体を削除します。 </li><li>手でアルミナるつぼに上のTi-カンフェン緑体を移し、凍結乾燥機でるつぼを配置します。凍結乾燥で素地にカンフェンフェーズを昇華する素地を - 24時間、40℃。 </li><li>その後、アルミナカバースリップでるつぼを閉じ、室温で（10 -6トル以下）の真空炉内で閉じたるつぼを配置します。その後、加熱Rで1300℃まで炉の温度を上昇させます5℃/分の食べて、2時間1300℃の温度を保持します。 </li><li>熱処理後、炉を完全に室温に冷却されるまで6-7時間炉内で焼結多孔質チタンを保ちます。 注：冷却プロセスの6時間の間に、400℃以上の炉の平均冷却速度は〜15℃/分で、400℃以下の炉の平均冷却速度は〜2℃/分です。 </li><li>必要に応じて、放電加工（EDM）から直径16mmの円盤状の試料に焼結多孔質チタンのブロックをカット27 注：アルミニウム金型の大きさに応じて、焼結多孔質チタンの大きさは、加工工程（ 図2A）を介して変更する必要があります。 </li><li>オートクレーブ内の多孔質チタンサンプルを有するガラスビーカーを置き、15分間121℃でサンプルを滅菌します。オートクレーブからサンプルを削除します。二回の70％エタノールで2回、次いで蒸留水を有する多孔質チタンサンプルを洗浄します。最後に、UV光の下でペトリ皿やクリーンベンチに室温で空気乾燥サンプルに多孔質チタンを残します。 </li></ol>生物活性剤と足場の2ディップコーティング<ol><li> 9ダルベッコのリン酸緩衝生理食塩水（DPBS、pHは7.4）の溶液10mlの中でMLは滅菌でGFPの1ミリリットルを混合することにより、クリーンベンチ内を100μg/ mlの1 mgの/から商業緑色蛍光タンパク質（GFP）を希釈表1に示すように、ポリスチレン（PS）チューブ。 </li><li> RTにおけるGFP溶液とPSチューブにチタンサンプルを配置し、クリーンベンチに置くことによって、GFP希釈溶液（100μg/ ml）の10 mlの滅菌高密度又は多孔質チタンを浸します。 </li><li>真空デシケーター中のPSチューブを置き、GFPソリューションをより効果的に多孔質チタンの細孔を貫通確保するために10分間デシケーターを避難。 </li><li>ピンセットを使用してPSチューブから多孔質チタンを削除します。直径10cm PEでGFP-コーティングされた多孔質チタンを配置トライディッシュとクリーンベンチに室温で空気乾燥O / N。 </li><li>ガラスビーカー内のダルベッコのリン酸緩衝生理食塩水（DPBS）10mlで二回、多孔質チタンをすすぎ、クリーンベンチにRTでピンセットと空気乾燥を使用した直径10cmのペトリ皿に多孔質チタンを移動します。 </li></ol>多孔質足場の3緻密化<ol><li>円筒状の鋼製ダイに様々な高さと、GFP-コーティングされた多孔質チタンのサンプルを配置し、スチールダイ（ 図3A）の上部と下部の穴にパンチのセットを挿入します。 </li><li> 0.05〜0.1秒の中間ひずみ速度でプレス機を使用したサンプル（ 図3A）のz方向の室温での鋼金型内に多孔質チタンを圧縮-1、表2に示す所定の適用株に対する圧力を保持アンロードする前に1分間。 </li><li>鋼ダイから緻密化のTiサンプルを削除します。 DPBS 10mlで二回緻密化サンプルを洗いますクリーンベンチ上室温でビーカーや空気乾燥O / Nインチ</li></ol> GFP-コーティングされた足場の4放出試験<ol><li>標本の3種類を浸し（GFP-コーティングされた密度の高いチタン（ステップ後2）、GFPは、コーティングされた多孔質のTi（ステップ1と2）およびGFP-コーティングされた高密度化された多孔質のTi（ステップ1-3）後の後）5ミリリットルのDPBS中（pH値クリーンベンチ上で37℃で10ミリリットル滅菌PSチューブに含まれている7.4）ソリューション。 </li><li> GFPコーティングされたサンプルと吸引アウト各PS管からのすべてのDPBS溶液を1、2、3、5、8、12の予め定められた回数に応じて、ピペットを使用して、新しい5 mLのDPBS溶液（pH7.4）で補充し、 15、22および29日浸漬後。 </li><li>共焦点レーザー走査型分光法（CLSM）を用いて、浸漬（0日目）前22日浸漬後のGFPでコーティングされたサンプルの蛍光像を撮ります。 </li><li>使用してセクション4.2で各PSチューブから引き出された5ミリリットルのDPBS溶液の総から1ミリリットル溶液中で放出されたGFPの蛍光シグナル強度を測定します215ナノメートルの波長でUV分光法。強度値は、標準曲線を用いて、GFP溶液の濃度に変換します。 注：測定の前に、0 NG / mlの濃度範囲でGFP溶液の蛍光シグナル強度を測定することにより、GFP溶液の標準曲線を描く - を10μg/ mlです。 </li></ol>傾斜多孔質チタンの足場の5作製<ol><li> 1.7ステップにステップ1.1を繰り返して、焼結多孔質のTiのブロックを生成します。 </li><li>マシンEDMにより所定の構造設計（例えば、 図5aおよび図5d）に応じて焼結多孔質チタンブロック。 </li><li>ダイの直径よりも小さい0.1ミリメートル〜多孔質チタンの直径はスチール製金型に高さ分布と機械加工のTiサンプルを置き、スチールダイの上部と下部の穴にパンチのセットを挿入します。 </li><li>ステップ3.2および3.3を実行します。 </li></ol> 6.気孔率ミーTiの足場のasurement <ol><li>チタン足場の質量（M s）を測定します。 </li><li>チタン足場の長さ、幅と高さを測定することによって見かけの体積（V s）を計算します。 </li><li>以下の式を用いて気孔率を計算します。 <img alt="式（1）" src="/files/ftp_upload/53279/53279eq1.jpg" /> ここでPは、全気孔率の割合で、ρTiはチタンの理論密度であり、m のS / V Sはサンプルの測定された密度です。 注：マイクロCTイメージングは​​、マイクロコンピュータ断層撮影スキャナを使用して実施された後のTi試料の気孔率を直接マイクロCT画像から取得することができます。 </li></ol>

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Porous+titanium+(Ti)+scaffolds+by+freezing+TiH2/camphene+slurries.">Porous titanium (Ti) scaffolds by freezing TiH2/camphene slurries.</a> Mater. Lett. 62 (30), 4506-4508 (2008).</li><li>Yook, S. W., Kim, H. E., Koh, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+porous+titanium+scaffolds+with+high+compressive+strength+using+camphene-based+freeze+casting.">Fabrication of porous titanium scaffolds with high compressive strength using camphene-based freeze casting.</a> Mater. Lett. 63 (17), 1502-1504 (2009).</li></ol>

The authors declare that they have no competing financial interests.

バイオメタルシステムは、広く、生物医学的用途に使用されているが、特に、荷重支持材料として、高い剛性と低い金属の生物活性は、主要な課題とみなされてきました。本研究では、新しい金属系、生体模倣機械的特性ならびに持続的な放出挙動を持つ生理活性表面を有する緻密化多孔性金属足場の製造方法を確立しました。当社の製造方法の主要な利点は、1を含む）に、我々はすでに開...

金属の生体材料が広く、それらの優れた機械的強度や弾力性、1-3の耐荷重インプラント、内部固定デバイスとして使用されてきたが、それらは2つの重要な課題が伴う：1）機械的な不一致を金属は生体組織よりもはるかに硬いので、望ましくない損害を引き起こします周囲組織としばしばしばしば異物反応（例えば、炎症または血栓症）を引き起こす、生物学的組織と悪い界面をもたらす。4-6多孔性金属足場は改善、構造において骨成長を促進するために提案されている2）低い生物活性に骨-インプラント接触応力遮蔽効果は、それらの低減剛性を抑制しつつ7-9また、種々の表面改質は、金属インプラントの生物学的活性を増強するために適用されています。このような改変は、生物活性分子（ 例えば、成長 FACで金属表面を被覆含まター）または薬物 （例えば、バンコマイシン、テトラサイクリン）。10-12しかし、このような多孔性金属足場の減少、機械的特性等の問題が、剛性が低下し、生物活性コーティング層の迅速な放出は未解決のまま。13-16 具体的には、チタン（Ti）およびTi合金は、その優れた機械的特性、化学的安定性、および良好な生体適合性の中で最も人気のあるバイオメタルシステムの一つです。13,17-19彼らの泡状のアプリケーションが3Dので、関心の高まりをもを集めています多孔質のネットワークは、骨様の機械的特性に加えて、骨の内部成長を促進する。20-22努力は、ポリマースポンジの複製、金属粒子の焼結、ラピッドプロトタイピング（RP）方式を含む新しい製造技術を開発することにより、機械的特性を改善するためになされている、および毛穴の様々な機能を制御するために、スペースホルダー法（例えば、細孔画分、形状、大きさ、分布、および接続）と材料特性（ 例えば 、金属相と不純物）。23-25 ​​近年、水ベースの金属スラリーの凍結鋳造は、よく整列細孔と機械的に強化されたTi形態を製造するために、かなりの注目を集めています凝固時に一方向の氷のデンドライト成長を利用した構造。しかし、水と金属粉末の接触による酸素汚染は、Ti足場の脆化を最小限にするために、特別な注意が必要です。14,15 したがって、我々は、生物活性と機械的に同調可能な多孔性のTi足場を製造に向けた新たなアプローチを開発しました。25足場は最初に50％以上の気孔率を有する多孔質構造を有しています。製造された多孔質足場は、生物活性分子でコーティングされ、その後、最終的な多孔性、機械的特性および薬物放出挙動は、アプリによって制御された時に機械的なプレスを用いて圧縮しました。ED株。緻密化した多孔質チタンインプラントは、骨（3-20万気圧）。2ので、コーティング層のと同等の剛性が低いのにもかかわらず、良好な強度と低い多孔性を示してきた、緻密化多孔質チタンの生物活性が大幅に改善されました。また、ために高密度化プロセスにより誘導されたユニークなフラットな細孔構造の、コーティングされた生物活性分子は徐々に長期間その有効性を維持し、足場から放出されることが見られました。 本研究では、生物医学的用途における潜在的な使用のために緻密化した多孔質チタンの足場を製造するために、当社の確立された方法を紹介しました。プロトコルは、金属スラリーと多孔質足場の緻密化とダイナミック凍結鋳造が含まれています。まず、 図1（a）に示すように 、動的凍結鋳造法が導入された良好な延性を有する多孔質チタン足場を作製しました。 Ti粉末を液体カンフェン中に分散させました。次に、温度を下げることにより、液相は、Ti粉末ネ​​ットワークと固体カンフェン結晶間の相分離をもたらす、固化させました。その後、凝固したTi-カンフェングリーン体は、Ti粉末を連続チタン支柱と縮合され、カンフェン相が完全に多孔質構造を得るために除去された焼結しました。得られた多孔質足場を有するコーティング及び緻密化プロセスが緻密化し、初期空隙率の度合いを変化させること、を使用しました。コー​​ティング層およびその放出挙動は、GFPでコーティングされた高密度のチタンと比較して緻密化して多孔質とすることなく、チタンコーティングを採用して可視化し、緑色蛍光タンパク質（GFP）を用いて定量しました。最後に、二つの異なる多孔質構造を有する傾斜機能チタン足場が提案され、多孔質足場の内側及び外側部分の緻密化の度合いを変えることによって実証されました。

<table>
	<tbody>
		<tr>
			<td>Titanium powder</td>
			<td>Alfa Aesar</td>
			<td>#42624</td>
			<td>-325 mesh, 99.5% (metals basis)</td>
		</tr>
		<tr>
			<td>Camphene</td>
			<td>SigmaAldrich</td>
			<td>#456055</td>
			<td>95%, C10H16</td>
		</tr>
		<tr>
			<td>KD-4</td>
			<td>Croda</td>
			<td>&#173;</td>
			<td>Hypermer, polymeric dispersant</td>
		</tr>
		<tr>
			<td>Phosphate Buffer Solution (PBS)</td>
			<td>Welgene</td>
			<td>ML 008-01</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Green Fluorescent Protein (GFP)</td>
			<td>Genoss Co.</td>
			<td>-</td>
			<td>&#62;98% purity, 1mg/ml</td>
		</tr>
		<tr>
			<td>Ball mill oven</td>
			<td>SAMHENUG ENERGY</td>
			<td>SH-BDO150</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Freeze dryer</td>
			<td>Ilshin Lab.</td>
			<td>PVTFD50A</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Cold isostatic pressing (CIP) machine</td>
			<td>SONGWON SYSTEMS</td>
			<td>CIP 42260</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Vaccum furnace</td>
			<td>JEONG MIN INDUSTRIAL</td>
			<td>JM-HP20</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>electical chaege machine</td>
			<td>FANUC robocut</td>
			<td>0iB</td>
			<td>External use</td>
		</tr>
		<tr>
			<td>Press machine</td>
			<td>CG&#38;S</td>
			<td>AJP-200</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Confocal laser scanning spectroscopy (CLSM)</td>
			<td>Olympus</td>
			<td>FluoView FV1000</td>
			<td>External use</td>
		</tr>
	</tbody>
</table>

fabrication of mechanically tunable and bioactive metal scaffolds for biomedical applications

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness and low bioactivity of metals. In this study, we have developed a new method towards fabricating a new type of bioactive and mechanically reliable porous metal scaffolds-densified porous Ti scaffolds. The method consists of two fabrication processes, 1) the fabrication of porous Ti scaffolds by dynamic freeze casting, and 2) coating and densification of the porous scaffolds. The dynamic freeze casting method to fabricate porous Ti scaffolds allowed the densification of porous scaffolds by minimizing the chemical contamination and structural defects. The densification process is distinctive for three reasons. First, the densification process is simple, because it requires a control of only one parameter (degree of densification). Second, it is effective, as it achieves mechanical enhancement and sustainable release of biomolecules from porous scaffolds. Third, it has broad applications, as it is also applicable to the fabrication of functionally graded porous scaffolds by spatially varied strain during densification.

多孔質チタン足場を製造するために使用される製造プロセスは、 図1Aに示されています。 Ti粉末を12時間44℃で容器の連続的な回転によってカンフェン中に均一に分散保持され、液体カンフェンが完全に固化している間、比較的重いTi粉末のいずれかの堆積物を最小限にすることができます。その結果、均質のTi-カンフェングリーン体は、図1（b）に示すように 、動的...

Watch this Scientific Journal Video about Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications at JoVE.com

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

バイオメディカル応用のための機械的に調整可能な生物活性金属スキャフォールドの作製

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

fabrication-mechanically-tunable-bioactive-metal-scaffolds-for

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Mechanical Engineering

Bioengineering

Unit 1

Transformations of Stress and Strain

SCIENTISTS IN ACTION

10.6K ビュー.  Korea Institute of Industrial Technology. Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

ビデオ: Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Microfabricated Platforms for Mechanically Dynamic Cell Culture

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or fundamental cell biology studies is limited by current bioreactor technologies, which cannot simultaneously apply a variety of mechanical stimuli to cultured cells. In order to address this issue, we have developed a series of microfabricated platforms designed to screen for the effects of mechanical stimuli in a high-throughput format. In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and further demonstrate how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

機械的にダイナミックな細胞培養のための微細加工プラットフォーム

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or ...

生物工学

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells inside of 3D biomaterial scaffolds provides a way to accurately mimic these microenvironments, providing an advantage over traditional 2D culture methods using polystyrene as well as a method for engineering replacement tissues 2. While 2D tissue culture polystrene has been used for the majority of cell culture experiments, 3D biomaterial scaffolds can more closely replicate the microenvironments found in vivo by enabling more accurate establishment of cell polarity in the environment and possessing biochemical and mechanical properties similar to soft tissue.3 A variety of naturally derived and synthetic biomaterial scaffolds have been investigated as 3D environments for supporting stem cell growth. While synthetic scaffolds can be synthesized to have a greater range of mechanical and chemical properties and often have greater reproducibility, natural biomaterials are often composed of proteins and polysaccharides found in the extracelluar matrix and as a result contain binding sites for cell adhesion and readily support cell culture. Fibrin scaffolds, produced by polymerizing the protein fibrinogen obtained from plasma, have been widely investigated for a variety of tissue engineering applications both in vitro and in vivo 4. Such scaffolds can be modified using a variety of methods to incorporate controlled release systems for delivering therapeutic factors 5. Previous work has shown that such scaffolds can be used to successfully culture embryonic stem cells and this scaffold-based culture system can be used to screen the effects of various growth factors on the differentiation of the stem cells seeded inside 6,7.
 This protocol details the process of polymerizing fibrin scaffolds from fibrinogen solutions using the enzymatic activity of thrombin. The process takes 2 days to complete, including an overnight dialysis step for the fibrinogen solution to remove citrates that inhibit polymerization. These detailed methods rely on fibrinogen concentrations determined to be optimal for embryonic and induced pluripotent stem cell culture. Other groups have further investigated fibrin scaffolds for a wide range of cell types and applications - demonstrating the versatility of this approach 8-12.

This work details the preparation of 3D fibrin scaffolds for culturing and differentiating plutipotent stem cells. Such scaffolds can be used to screen the effects of various biological compounds on stem cell behavior as well as modified to contain drug delivery systems.

幹細胞培養アプリケーションのための3Dフィブリン足場の調製

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

Designing Silk-silk Protein Alloy Materials for Biomedical Applications

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, nanomedicine, tissue regeneration, and drug delivery. Designing materials based on the molecular-scale interactions between these proteins will help generate new multifunctional protein alloy biomaterials with tunable properties. Such alloy material systems also provide advantages in comparison to traditional synthetic polymers due to the materials biodegradability, biocompatibility, and tenability in the body. This article used the protein blends of wild tussah silk (Antheraea pernyi) and domestic mulberry silk (Bombyx mori) as an example to provide useful protocols regarding these topics, including how to predict protein-protein interactions by computational methods, how to produce protein alloy solutions, how to verify alloy systems by thermal analysis, and how to fabricate variable alloy materials including optical materials with diffraction gratings, electric materials with circuits coatings, and pharmaceutical materials for drug release and delivery. These methods can provide important information for designing the next generation multifunctional biomaterials based on different protein alloys.

Blending is an efficient approach to generate biomaterials with a broad range of properties and combined features. By predicting the molecular interactions between different natural silk proteins, new silk-silk protein alloy platforms with tunable mechanical resiliency, electrical response, optical transparency, chemical processability, biodegradability, or thermal stability can be designed.

バイオメディカル応用のためのシルクシルクプロテイン合金材料設計

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, ...

Fabrication of a Bioactive, PCL-based "Self-fitting" Shape Memory Polymer Scaffold

Tissue engineering has been explored as an alternative strategy for the treatment of critical-sized cranio-maxillofacial (CMF) bone defects. Essential to the success of this approach is a scaffold that is able to conformally fit within an irregular defect while also having the requisite biodegradability, pore interconnectivity and bioactivity. By nature of their shape recovery and fixity properties, shape memory polymer (SMP) scaffolds could achieve defect &#8220;self-fitting.&#8221; In this way, following exposure to warm saline (~60 &#186;C), the SMP scaffold would become malleable, permitting it to be hand-pressed into an irregular defect. Subsequent cooling (~37 &#186;C) would return the scaffold to its relatively rigid state within the defect. To meet these requirements, this protocol describes the preparation of SMP scaffolds prepared via the photochemical cure of biodegradable polycaprolactone diacrylate (PCL-DA) using a solvent-casting particulate-leaching (SCPL) method. A fused salt template is utilized to achieve pore interconnectivity. To realize bioactivity, a polydopamine coating is applied to the surface of the scaffold pore walls. Characterization of self-fitting and shape memory behaviors, pore interconnectivity and in vitro bioactivity are also described.

Fabrication of a Bioactive, PCL-based &quot;Self-fitting&quot; Shape Memory Polymer Scaffold

Scaffolds capable of fitting within cranio-maxillofacial (CMF) bone defects while exhibiting osteoconductivity and bioactivity are of interest. This protocol describes the preparation of a shape memory scaffold based on polycaprolactone diacrylate (PCL-DA) using a solvent-casting particulate-leaching (SCPL) method employing a fused salt template and application of a bioactive polydopamine coating.

生理活性、PCLベースの「セルフフィット」形状記憶ポリマー足場の作製

Tissue engineering has been explored as an alternative strategy for the treatment of critical-sized cranio-maxillofacial (CMF) bone defects. Essential to ...

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial applications. Here we describe how electrospinning or electrospraying a polymer mixture containing a biodegradable, biocompatible aliphatic polyester (e.g., polycaprolactone and poly(lactide-co-glycolide)), as the major component, doped with a hydrophobic copolymer composed of the polyester and a stearate-modified poly(glycerol carbonate) affords a superhydrophobic biomaterial. The fabrication techniques of electrospinning or electrospraying provide the enhanced surface roughness and porosity on and within the fibers or the particles, respectively. The use of a low surface energy copolymer dopant that blends with the polyester and can be stably electrospun or electrosprayed affords these superhydrophobic materials. Important parameters such as fiber size, copolymer dopant composition and/or concentration, and their effects on wettability are discussed. This combination of polymer chemistry and process engineering affords a versatile approach to develop application-specific materials using scalable techniques, which are likely generalizable to a wider class of polymers for a variety of applications.

Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.

バイオメディカル応用のための超疎水性高分子材料を製造します

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial ...

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions.

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

水試料中の微量金属分析のためのダイポールアシスト固相抽出マイクロチップの作製

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest regarding the use of bioinspired nanoscale hydroxyapatite (nHA). However, biological apatite is known to be calcium-deficient and carbonate-substituted with a nanoscale platelet-like morphology. Bioinspired nHA has the potential to stimulate optimal bone tissue regeneration due to its similarity to bone and tooth enamel mineral. Many of the methods currently used to fabricate nHA both in the laboratory and commercially, involve lengthy processes and complex equipment. Therefore, the aim of this study was to develop a rapid and reliable method to prepare high quality bioinspired nHA. The rapid mixing method developed was based upon an acid-base reaction involving calcium hydroxide and phosphoric acid. Briefly, a phosphoric acid solution was poured into a calcium hydroxide solution followed by stirring, washing and drying stages. Part of the batch was sintered at 1,000 &#176;C for 2 h in order to investigate the products&#39; high temperature stability. X-ray diffraction analysis showed the successful formation of HA, which showed thermal decomposition to &#946;-tricalcium phosphate after high temperature processing, which is typical for calcium-deficient HA. Fourier transform infrared spectroscopy showed the presence of carbonate groups in the precipitated product. The nHA particles had a low aspect ratio with approximate dimensions of 50 x 30 nm, close to the dimensions of biological apatite. The material was also calcium deficient with a Ca:P molar ratio of 1.63, which like biological apatite is lower than the stoichiometric HA ratio of 1.67. This new method is therefore a reliable and far more convenient process for the manufacture of bioinspired nHA, overcoming the need for lengthy titrations and complex equipment. The resulting bioinspired HA product is suitable for use in a wide variety of medical and consumer health applications.

This paper describes a novel method for the rapid manufacture of high quality bioinspired nanoscale hydroxyapatite. This biomaterial is of great significance in the manufacture of a wide range of innovative medical devices for clinical applications in orthopedics, craniofacial surgery and dentistry.

バイオメディカル応用のためのバイオインスパイアードナノハイドロキシアパタイトの急速なミックス準備

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest ...

Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications

There are many biological stimuli that can influence cell behavior and stem cell differentiation. General cell culture approaches rely on soluble factors within the medium to control cell behavior. However, soluble additions cannot mimic certain signaling motifs, such as matrix-bound growth factors, cell-cell signaling, and spatial biochemical cues, which are common influences on cells. Furthermore, biophysical properties of the matrix, such as substrate stiffness, play important roles in cell fate, which is not easily manipulated using conventional cell culturing practices. In this method, we describe a straightforward protocol to provide patterned bioactive proteins on synthetic polyethylene glycol (PEG) hydrogels using photochemistry. This platform allows for the independent control of substrate stiffness and spatial biochemical cues. These hydrogels can achieve a large range of physiologically relevant stiffness values. Additionally, the surfaces of these hydrogels can be photopatterned with bioactive peptides or proteins via thiol-ene click chemistry reactions. These methods have been optimized to retain protein function after surface immobilization. This is a versatile protocol that can be applied to any protein or peptide of interest to create a variety of patterns. Finally, cells seeded onto the surfaces of these bioactive hydrogels can be monitored over time as they respond to spatially specific signals.

In this method, we use photopolymerization and click chemistry techniques to create protein or peptide patterns on the surface of polyethylene glycol (PEG) hydrogels, providing immobilized bioactive signals for the study of cellular responses in vitro.

生理活性タンパク質や生物学的応用の光化学ハイドロゲルのペプチドをパターニング

There are many biological stimuli that can influence cell behavior and stem cell differentiation. General cell culture approaches rely on soluble factors ...

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 ...

Fabrication and Characterization of Griffithsin-modified Fiber Scaffolds for Prevention of Sexually Transmitted Infections

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to prevent and treat sexually transmitted infections (STIs). Moreover, many EF technologies focus on encapsulating the active agent, relative to utilizing the surface to impart biofunctionality. Here we describe a method to fabricate and surface-modify poly(lactic-co-glycolic) acid (PLGA) electrospun fibers, with the potent antiviral lectin Griffithsin (GRFT). PLGA is an FDA-approved polymer that has been widely used in drug delivery due to its outstanding chemical and biocompatible properties. GRFT is a natural, potent, and safe lectin that possesses broad activity against numerous viruses including human immunodeficiency virus type 1 (HIV-1). When combined, GRFT-modified fibers have demonstrated potent inactivation of HIV-1 in vitro. This manuscript describes the methods to fabricate and characterize GRFT-modified EFs. First, PLGA is electrospun to create a fiber scaffold. Fibers are subsequently surface-modified with GRFT using 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS)chemistry. Scanning electron microscopy (SEM) was used to assess the size and morphology of surface-modified formulations. Additionally, a gp120 or hemagglutinin (HA)-based ELISA may be used to quantify the amount of GRFT conjugated to, as well as GRFT desorption from the fiber surface. This protocol can be more widely applied to fabricate fibers that are surface-modified with a variety of different proteins.

This manuscript describes the procedure to fabricate and characterize Griffithsin-modified poly(lactic-co-glycolic acid) electrospun fibers that demonstrate potent adhesive and antiviral activity against human immunodeficiency virus type 1 infection in vitro. Methods used to synthesize, surface-modify, and characterize the resulting morphology, conjugation, and desorption of Griffithsin from surface-modified fibers are described.

作製と評価 Griffithsin 変更繊維足場の防止のため性的感染

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to ...