著者はニューヨーク·首都圏·リサーチ·アライアンス助成の支援を承認したいと思います。

1。Bioprintingプラットフォームの確立プリンタ修飾は、HPデスクジェット500サーマルインクジェットプリンタおよびHP 51626Aブラックインクカートリッジに基づいていた。 <ol><li>プリンタの上部のプラスチックカバーを取り外し、慎重にカバーからコントロールパネルを取り外す。 </li><li>プリンタ頂部と基部との間に3ケーブルの接続を外します。ベースからプリンタのトップ部分を取り外します。 </li><li>プリンタのトップ部には、インクカートリッジの下に右側に小さなプラスチックとゴム付属品（プリントヘッドクリーニングシステム）を外します。 </li><li>スプリングを用紙トレイのベースを取り外します。 </li><li>プラスチック、紙送りバーをカバーする金属製のプレートを取り外します。 </li><li>ハンドのこぎり又は他の切断ツールを使用して中央給電車輪位置におけるプラスチック、紙送りバーを切り落とした。 </li><li>前のステップの後に公開される2給紙ホイールを取り外します。ホイールプラスチックは非常に難しいと電子であるSAWは参考になります。 </li><li>缶詰の空気とエタノールワイプを使用してほこりやごみを清掃してください。 </li><li>ベースにプリンタ頂部を取り付けます。 </li><li> UVは、使用する前に、層流フード内で少なくとも2時間変性プリンタを滅菌する。 </li><li>ハンドソーや他の切削工具を使用して、HPの51626Aのインクカートリッジのキャップを切り取ります。 </li><li>インクを空にし、軟骨の下の井戸貯水池をカバーしてフィルタを削除します。 </li><li>水道水を使用して徹底的にカートリッジを洗浄します。 </li><li> Ultrasonicate残留インクを除去するために10分間、脱イオン（DI）水中のカートリッジ。 </li><li>すべてのインクが削除されていることを確認するカートリッジを調べます。滅菌脱イオン水に続いて殺菌のため70％エタノールで徹底的にカートリッジを、すすぎ又はスプレー。 </li><li>同時光重合能力を提供するために、印刷プラットフォームの上に長波長紫外線ランプを設定する。 </li><li> UV光を用いて印刷プラットフォームでのUV強度を測定メートル。印刷対象の強度が4-8の間MW / cm 2と （プリンタプラットフォームへのランプから約25 cm）で ​​すので、UVランプとプリンタプラットフォーム間の距離を調整します。 </li></ol> 2。Bioink準備<ol><li>単層の軟骨細胞の拡大<ol><li>ダルベッコでの細胞増殖のための各T175組織培養フラスコにプレート500万ヒト軟骨細胞は10％ウシ血清および1×ペニシリン - ストレプトマイシン - グルタミン（PSG）を補足したイーグル培地（DMEM）を修正しました。 5％CO 2を含む加湿空気で37℃で培養細胞。フラスコに85％コンフルエントになるまで、3日ごとに培養液を変更します。同じ継代から細胞を使用してください。 </li></ol></li><li> 10％w / vの最終濃度にPBSでPEGDAを溶解0.05％w / vの最終濃度になるように光開始剤I-2959を追加解決策を滅菌ろ過する。 </li><li> 5×10 6細胞で調製PEGDA液中で培養したヒト軟骨細胞を一時停止/ミリリットル。 </li></ol> 3。軟骨組織印刷<ol><li>プリンタとノートパソコンの電源をオンにします。 </li><li> Microsoft WordやAdobe Photoshopのを使用して、直径4ミリメートルと黒丸の印刷パターンを作成します。 <ol><li>パターンの位置を調整して、プラスチック金型に正確に印刷されることを確認してください。 </li><li>足場の所望の厚さに達するのに必要な印刷部数を計算する。高さ4ミリメートルの場合は、220プリントが必要な足場を作成する必要があります。 </li></ol></li><li>インクカートリッジにbioinkをロードします。印刷中に直接UV暴露から保護するためにアルミホイルでカートリッジを覆う。 </li><li>プリンタに印刷コマンドを送信します。プリンタが印刷を開始したときに、用紙センサーを引き出します。全体の印刷プロセスは、直径4ミリメートル、高さ4ミリメートルと足場未満4分を取る必要があります。 </li><li>転写は、24ウェルプレートに新軟骨を印刷し、各ウェルに1.5ミリリットルの培養培地を加える。 </li></ol><ol><li>暗所で15分間室温での生/死生存率/細胞毒性作用溶液で印刷新軟骨を培養する。 </li><li>半分に細胞を含んだハイドロゲルをカットし、切削領域の蛍光画像を撮影。 </li><li> 5ランダムに撮影した画像で盲目観察者（緑）、死者（赤）細胞生存数える。生細胞および死細胞の総数によって生細胞数で割ることによって細胞生存率を計算する。 </li></ol>

<ol>
	<li>Cui, X., Boland, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+microvasculature+fabrication+using+thermal+inkjet+printing+technology.">Human microvasculature fabrication using thermal inkjet printing technology.</a> Biomaterials. 30, 6221-6227 (2009).</li><li>Cui, X., Breitenkamp, K., Finn, M. G., Lotz, M., D'Lima, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+human+cartilage+repair+using+three-dimensional+bioprinting+technology.">Direct human cartilage repair using three-dimensional bioprinting technology.</a> Tissue Eng Part A. 18, 1304-1312 (2012).</li><li>Cui, X., Breitenkamp, K., Lotz, M., D'Lima, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+action+of+fibroblast+growth+factor-2+and+transforming+growth+factor-beta1+enhances+bioprinted+human+neocartilage+formation.">Synergistic action of fibroblast growth factor-2 and transforming growth factor-beta1 enhances bioprinted human neocartilage formation.</a> Biotechnol. Bioeng. 109, 2357-2368 (2012).</li><li>Cui, X., Breitenkamp, K., Finn, M. G., Lotz, M., Colwell, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+human+cartilage+repair+using+thermal+inkjet+printing+technology.">Direct human cartilage repair using thermal inkjet printing technology.</a> Osteoarthritis and Cartilage. 19, (2011).</li><li>Cui, X., Boland, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+deposition+of+human+microvascular+endothelial+cells+and+biomaterials+for+human+microvasculature+fabrication+using+inkjet+printing.">Simultaneous deposition of human microvascular endothelial cells and biomaterials for human microvasculature fabrication using inkjet printing.</a> NIP24/digital Fabrication 2008: 24th International Conference on Digital Printing Technologies, Technical Program and Proceedings. 24, 480-483 (2008).</li><li>Cui, X., Dean, D., Ruggeri, Z. M., Boland, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+damage+evaluation+of+thermal+inkjet+printed+Chinese+hamster+ovary+cells.">Cell damage evaluation of thermal inkjet printed Chinese hamster ovary cells.</a> Biotechnol. Bioeng. 106, 963-969 (2010).</li><li>Cui, X., Hasegawa, A., Lotz, M., D'Lima, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structured+three-dimensional+co-culture+of+mesenchymal+stem+cells+with+meniscus+cells+promotes+meniscal+phenotype+without+hypertrophy.">Structured three-dimensional co-culture of mesenchymal stem cells with meniscus cells promotes meniscal phenotype without hypertrophy.</a> Biotechnol. 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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=Hydrogel+properties+influence+ECM+production+by+chondrocytes+photoencapsulated+in+poly(ethylene+glycol)+hydrogels.">Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels.</a> Journal of Biomedical Materials Research. 59, 63-72 (2002).</li><li>Elisseeff, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoencapsulation+of+chondrocytes+in+poly(ethylene+oxide)-based+semi-interpenetrating+networks.">Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks.</a> Journal of Biomedical Materials Research. 51, 164-171 (2000).</li><li>Buskirk, W. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+A+High-Resolution+Thermal+Inkjet+Printhead.">Development of A High-Resolution Thermal Inkjet Printhead.</a> Hewlett-Packard Journal. 39, 55-61 (1988).</li><li>Harmon, J. P., Widder, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+the+Printhead+Into+the+HP+Deskjet+Printer.">Integrating the Printhead Into the HP Deskjet Printer.</a> Hewlett-Packard Journal. 39, 62-66 (1988).</li><li>Kim, T. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+model+for+cartilage+tissue+engineering+to+regenerate+the+zonal+organization+of+articular+cartilage.">Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage.</a> Osteoarthritis and Cartilage. 11, 653-664 (2003).</li><li>Sharma, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Designing+zonal+organization+into+tissue-engineered+cartilage.">Designing zonal organization into tissue-engineered cartilage.</a> Tissue Engineering. 13, 405-414 (2007).</li><li>Cohen, D. L., Lipton, J. I., Bonassar, L. J., Lipson, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Additive+manufacturing+for+in+situ+repair+of+osteochondral+defects.">Additive manufacturing for in situ repair of osteochondral defects.</a> Biofabrication. 2, (2010).</li></ol>

著者らは、この研究には経済的利害関係を持っていません。

同時光重合能力を持つこの3D bioprintingシステムは、細胞のアルギン酸ハイドロゲル16を押し出し、注射器を用いて骨軟骨欠損のその場での印刷において最高の以前に報告された方法よりもかなり大きい印刷解像度を提供します。高い印刷解像度は、解剖学的軟骨帯状組織を復元するために軟骨組織工学のために特に重要です。層ごとの組み立て中の同時光重合は、3D建設?...

サーマルインクジェット印刷に基づくBioprintingは、組織工学や再生医療の分野で最も有望な実現技術の一つです。デジタル制御および高スループットのプリントヘッドを有する細胞、足場、および成長因子は、正確に迅速に所望の2次元（2D）および3次元（3D）の位置に堆積させることができる。多くの成功したアプリケーションは、組織工学および再生医療1-9に、この技術を用いて達成されてきた。本論文では、bioprintingプラットフォームが変更されたヒューレット·パッカード社（HP）DeskJet 500サーマルインクジェットプリンタと同時光重合系に設立されました。ポリ（エチレングリコール）（PEG）から配合する合成ヒドロゲルは、軟骨細胞の生存能力を維持し、軟骨ECM産生を促進10,11の容量を示している。また、光架橋PEGは同時polymeに最適です低粘度、水への溶解性が高いです。3次元bioprinting中にrization。本論文では、ポリ（エチレン）グリコールジアクリレート（PEGDA、MW 3400）に懸濁させ、人間の軟骨細胞を精密に3次元解像度1,400 dpiので新軟骨の層ごとに構築するために印刷した。 3D足場における細胞の堆積の均一な分布は、優れた機械的性質および拡張ECM産生に軟骨組織を生成した、観察された。対照的に、マニュアル製造において細胞は、培養後の2,3不均一な軟骨形成をもたらし代わりに遅い足場重合によるそれらの最初に堆積位置のゲルの底部に溜まった。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">HP Deskjet 500 thermal inkjet printer</td>
			<td style="width:221px;">Hewlett-Packard</td>
			<td style="width:195px;">C2106a</td>
			<td style="width:433px;">Discontinued. Purchased refurbished from internet vendor.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">HP black ink cartridge</td>
			<td style="width:221px;">Hewlett-Packard</td>
			<td style="width:195px;">51626a</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Ultraviolet lamp</td>
			<td style="width:221px;">UVP</td>
			<td style="width:195px;">B-100AP</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">UV light meter</td>
			<td style="width:221px;">General Tools</td>
			<td style="width:195px;">UV513AB</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Zeiss LSM 510 laser scanning microscope</td>
			<td style="width:221px;">Carl Zeiss</td>
			<td style="width:195px;">LSM 510</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Dulbeccos Modified Eagles Medium (DMEM)</td>
			<td style="width:221px;">Mediatech</td>
			<td style="width:195px;">10-013</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Penicillin-streptomycin-glutamine (PSG)</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">10378-016</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Accutase cell dissociation reagent</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">A11105-01</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Phosphate buffered saline (PBS)</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">10010-023</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Live/Dead viability/cytotoxicity Kit</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">L-3224</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Poly(ethylene glycol) diacrylate (PEGDA)</td>
			<td style="width:221px;">Glycosan Biosystems</td>
			<td style="width:195px;">GS700</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Irgacure 2959</td>
			<td style="width:221px;">Ciba Specialty Chemicals</td>
			<td style="width:195px;">I-2959</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Human articular chondrocytes</td>
			<td style="width:221px;">Lonza</td>
			<td style="width:195px;">CC-2550</td>
			<td style="width:433px;"></td>
		</tr>
	</tbody>
</table>

human cartilage tissue fabrication using three-dimensional inkjet printing technology

サーマルインクジェット印刷に基づいていますBioprintingは、組織工学や再生医療の分野で最も魅力的な実現技術の一つです。急速にデジタルコントロール細胞、足場、および成長因子が正確に所望の2次元（2D）に付着させることができるとともに、三次元（3D）の位置。そのため、この技術は、ネイティブの解剖学的構造を模倣組織を作製するための理想的なアプローチです。天然の帯状組織、細胞外マトリックス組成物（ECM）、および機械的特性を有する軟骨を操作するために、我々は3次元軟骨組織工学のための可能な同時重合を有する市販のインクジェットプリンタを用いbioprintingプラットフォームを開発した。ポリ（エチレングリコール）ジアクリレート（PEGDA）に懸濁したヒト軟骨細胞を、レイヤーバイレイヤーアセンブリを介して3D新軟骨構築のために印刷した。印刷された細胞は、surroでサポートされて、元の堆積の位置に固定し、同時光重合に足場をunding。印刷された組織の機械的特性は、天然の軟骨と同様であった。長いUV露光を必要とする従来の組織の製造と比較して、光重合と同時印刷細胞の生存率は有意に高かった。印刷された新軟骨は、遺伝子発現と一致した優れたグリコサミノグリカン（GAG）およびコラーゲン型II産生を実証した。そのため、このプラットフォームは、解剖学的、組織工学のための正確な細胞の分布や配置に最適です。

修飾されたサーマルインクジェットプリンタは、高スループット、優れた細胞生存率及び足場における細胞の堆積のために可能であった。同時光重合感光生体材料との組み合わせは、この技術は、最初に堆積場所に細胞および他の印刷された物質を固定することができます。修飾されたサーマルインクジェットプリンタの特性に応じて、2D印刷解像度が130 plの単一のインク滴の体積の300解像?...

Watch this Scientific Journal Video about Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology at JoVE.com

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

このホワイトペーパーで説明する方法は、同時UV重合とbioprinterに市販のインクジェットプリンターを変換する方法を示しています。プリンタは、細胞と生体材料との三次元組織構造を構築することが可能である。ここに実証研究では、3Dの新軟骨を構築した。

三次元インクジェット印刷技術を用いて、ヒト軟骨組織の作製

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

human-cartilage-tissue-fabrication-using-three-dimensional-inkjet

Research

JoVE Journal

Bioengineering

15.6K ビュー.  Rensselaer Polytechnic Institute. このホワイトペーパーで説明する方法は、同時UV重合とbioprinterに市販のインクジェットプリンターを変換する方法を示しています。プリンタは、細胞と生体材料との三次元組織構造を構築することが可能である。ここに実証研究では、3Dの新軟骨を構築した。 

ビデオ: Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Planar and Three-Dimensional Printing of Conductive Inks

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical devices1-3. In this paper, we focus on one- (1D), two- (2D), and three-dimensional (3D) printing of conductive metallic inks in the form of flexible, stretchable, and spanning microelectrodes.
Direct-write assembly4,5 is a 1-to-3D printing technique that enables the fabrication of features ranging from simple lines to complex structures by the deposition of concentrated inks through fine nozzles (~0.1 - 250 &mu;m). This printing method consists of a computer-controlled 3-axis translation stage, an ink reservoir and nozzle, and 10x telescopic lens for visualization. Unlike inkjet printing, a droplet-based process, direct-write assembly involves the extrusion of ink filaments either in- or out-of-plane. The printed filaments typically conform to the nozzle size. Hence, microscale features (&lt; 1 &mu;m) can be patterned and assembled into larger arrays and multidimensional architectures.
In this paper, we first synthesize a highly concentrated silver nanoparticle ink for planar and 3D printing via direct-write assembly. Next, a standard protocol for printing microelectrodes in multidimensional motifs is demonstrated. Finally, applications of printed microelectrodes for electrically small antennas, solar cells, and light-emitting diodes are highlighted.

Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.

導電性インクの平面および三次元印刷

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

生物工学

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

三次元の全層皮膚等価物と自動傷つけ処理の発生

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Three-dimensional Biomimetic Technology: Novel Biorubber Creates Defined Micro- and Macro-scale Architectures in Collagen Hydrogels

Tissue scaffolds play a crucial role in the tissue regeneration process. The ideal scaffold must fulfill several requirements such as having proper composition, targeted modulus, and well-defined architectural features. Biomaterials that recapitulate the intrinsic architecture of in vivo tissue are vital for studying diseases as well as to facilitate the regeneration of lost and malformed soft tissue. A novel biofabrication technique was developed which combines state of the art imaging, three-dimensional (3D) printing, and selective enzymatic activity to create a new generation of biomaterials for research and clinical application. The developed material, Bovine Serum Albumin rubber, is reaction injected into a mold that upholds specific geometrical features. This sacrificial material allows the adequate transfer of architectural features to a natural scaffold material. The prototype consists of a 3D collagen scaffold with 4 and 3 mm channels that represent a branched architecture. This paper emphasizes the use of this biofabrication technique for the generation of natural constructs. This protocol utilizes a computer-aided software (CAD) to manufacture a solid mold which will be reaction injected with BSA rubber followed by the enzymatic digestion of the rubber, leaving its architectural features within the scaffold material.

An innovative biofabrication technique was developed to engineer three-dimensional constructs that resemble the architectural features, components, and mechanical properties of in vivo tissue. This technique features a newly developed sacrificial material, BSA rubber, which transfers detailed spatial features, reproducing the in vivo architectures of a wide variety of tissues.

三次元バイオミメティック・テクノロジー：小説Biorubberはコラーゲンハイドロゲルで定義されたマイクロ・マクロスケールのアーキテクチャを作成します。

Tissue scaffolds play a crucial role in the tissue regeneration process. The ideal scaffold must fulfill several requirements such as having proper ...

Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing drugs targeted at the liver, requires a platform in which cells receive proper biochemical and mechanical cues. Recent liver tissue engineering systems have employed three-dimensional (3D) scaffolds composed of synthetic or natural hydrogels, given their high water retention and their ability to provide the mechanical stimuli needed by the cells. There has been growing interest in the inverted colloidal crystal (ICC) scaffold, a recent development, which allows high spatial organization, homotypic and heterotypic cell interaction, as well as cell-extracellular matrix (ECM) interaction. Herein, we describe a protocol to fabricate the ICC scaffold using poly (ethylene glycol) diacrylate (PEGDA) and the particle leaching method. Briefly, a lattice is made from microsphere particles, after which a pre-polymer solution is added, properly polymerized, and the particles are then removed, or leached, using an organic solvent (e.g., tetrahydrofuran). The dissolution of the lattice results in a highly porous scaffold with controlled pore sizes and interconnectivities that allow media to reach cells more easily. This unique structure allows high surface area for the cells to adhere to as well as easy communication between pores, and the ability to coat the PEGDA ICC scaffold with proteins also shows a marked effect on cell performance. We analyze the morphology of the scaffold as well as the hepatocarcinoma cell (Huh-7.5) behavior in terms of viability and function to explore the effect of ICC structure and ECM coatings. Overall, this paper provides a detailed protocol of an emerging scaffold that has wide applications in tissue engineering, especially liver tissue engineering.

Fabrication of Inverted Colloidal Crystal Polyethylene glycol Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

This manuscript presents a detailed protocol for the fabrication of an emerging three-dimensional hepatocyte culture platform, the inverted colloidal crystal scaffold, and the concomitant techniques to assess hepatocyte behavior. The size-controllable pores, interconnectivity and ability to conjugate extracellular matrix proteins to the poly(ethylene glycol) (PEG) scaffold enhance Huh-7.5 cell performance.

肝組織工学のための三次元細胞培養プラットフォーム：逆コロイド結晶ポリ（エチレングリコール）足場の作製

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and directed within a layer of paper. Moreover, stacking multiple layers of patterned paper creates sophisticated three-dimensional microfluidic networks that can support the development of analytical and bioanalytical assays. Paper-based microfluidic devices are inexpensive, portable, easy to use, and require no external equipment to operate. As a result, they hold great promise as a platform for point-of-care diagnostics. In order to properly evaluate the utility and analytical performance of paper-based devices, suitable methods must be developed to ensure their manufacture is reproducible and at a scale that is appropriate for laboratory settings. In this manuscript, a method to fabricate a general device architecture that can be used for paper-based immunoassays is described. We use a form of additive manufacturing (multi-layer lamination) to prepare devices that comprise multiple layers of patterned paper and patterned adhesive. In addition to demonstrating the proper use of these three-dimensional paper-based microfluidic devices with an immunoassay for human chorionic gonadotropin (hCG), errors in the manufacturing process that may result in device failures are discussed. We expect this approach to manufacturing paper-based devices will find broad utility in the development of analytical applications designed specifically for limited-resource settings.

We detail a method to fabricate three-dimensional paper-based microfluidic devices for use in the development of immunoassays. Our approach to device assembly is a type of multilayer, additive manufacturing. We demonstrate a sandwich immunoassay to provide representative results for these types of paper-based devices.

イムノアッセイのための3次元紙ベースのマイクロ流体デバイスの作製

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

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 ...

Culturing Mammalian Cells in Three-dimensional Peptide Scaffolds

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an environment that closely mimics the structural features of non-polarized tissue. Furthermore, the particular intrinsic nanofiber structure of the scaffold makes it transparent to visual light, which allows for easy visualization of the sample under microscopy. This advantage was largely used to study cell migration, organization, proliferation, and differentiation and thus any development of their particular cellular function by staining with specific dyes or probes. Furthermore, in this work, we describe the good performance of this system to easily study the redifferentiation of expanded human articular chondrocytes into cartilaginous tissue. Cells were encapsulated into self-assembling peptide scaffolds and cultured under specific conditions to promote chondrogenesis. Three-dimensional cultures showed good viability during the 4 weeks of the experiment. As expected, samples cultured with chondrogenic inducers (compared to non-induced controls) stained strongly positive for toluidine blue (which stains glycosaminoglycans (GAGs) that are highly present in cartilage extracellular matrix) and expressed specific molecular markers, including collagen type I, II and X, according to Western Blot analysis. This protocol is easy to perform and can be used at research laboratories, industries and for educational purposes in laboratory courses.

Here, we present a protocol to obtain 3D-culture systems in self-assembling peptide scaffolds to promote the differentiation of dedifferentiated human articular chondrocytes into cartilage-like tissue.

三次元ペプチド足場で哺乳動物の細胞を培養

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an ...

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic experimentation is a stable fluid handling system capable of accurately providing an inlet flow rate or inlet pressure. Here, we have developed a syringe pump system capable of controlling and regulating the inlet fluid pressure delivered to a microfluidic device. This system was designed using low-cost materials and additive manufacturing principles, leveraging three-dimensional (3D) printing of thermoplastic materials and off-the-shelf components whenever possible. This system is composed of three main components: a syringe pump, a pressure transducer, and a programmable microcontroller. Within this paper, we detail a set of protocols for fabricating, assembling, and programming this syringe pump system. Furthermore, we have included representative results that demonstrate high-fidelity, feedback control of inlet pressure using this system. We expect this protocol will allow researchers to fabricate low-cost syringe pump systems, lowering the entry barrier for the use of microfluidics in biomedical, chemical, and materials research.

Here we present a protocol to construct a pressure-controlled syringe pump to be used in microfluidic applications. This syringe pump is made from an&#160;additively manufactured body, off-the-shelf hardware, and open-source electronics. The resulting system is low-cost, straightforward to build, and delivers well-regulated fluid flow to enable rapid microfluidic research.

印刷作成する熱可塑性材料の三次元マイクロ流体用フィードバック制御を用いたシリンジ ポンプを自動

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...

Fabrication of Decellularized Cartilage-derived Matrix Scaffolds

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage research has focused on the development of regenerative scaffolds. This article describes the development of scaffolds that are completely derived from natural cartilage extracellular matrix, coming from an equine donor. Potential applications of the scaffolds include producing allografts for cartilage repair, serving as a scaffold for osteochondral tissue engineering, and providing in vitro models to study tissue formation. By decellularizing the tissue, the donor cells are removed, but many of the natural bioactive cues are thought to be retained. The main advantage of using such a natural scaffold in comparison to a synthetically produced scaffold is that no further functionalization of polymers is required to drive osteochondral tissue regeneration. The cartilage-derived matrix scaffolds can be used for bone and cartilage tissue regeneration in both in vivo and in vitro settings.

Decellularized cartilage-derived scaffolds can be used as a scaffold to guide cartilage repair and as a means to regenerate osteochondral tissue. This paper describes the decellularization process in detail and provides suggestions to use these scaffolds in in vitro settings.

脱軟骨由来のマトリックス足場の作製

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage ...

Pancreatic Tissue-Derived Extracellular Matrix Bioink for Printing 3D Cell-Laden Pancreatic Tissue Constructs

The transplantation of pancreatic islets is a promising treatment for patients who suffer from type 1 diabetes accompanied by hypoglycemia and secondary complications. However, islet transplantation still has several limitations such as the low viability of transplanted islets due to poor islet engraftment and hostile environments. In addition, the insulin-producing cells differentiated from human pluripotent stem cells have limited ability to secrete sufficient hormones that can regulate the blood glucose level; therefore, improving the maturation by culturing cells with proper microenvironmental cues is strongly required. In this article, we elucidate protocols for preparing a pancreatic tissue-derived decellularized extracellular matrix (pdECM) bioink to provide a beneficial microenvironment that can increase glucose sensitivity of pancreatic islets, followed by describing the processes for generating 3D pancreatic tissue constructs using a microextrusion-based bioprinting technique.

Decellularized extracellular matrix (dECM) can provide suitable microenvironmental cues to recapitulate the inherent functions of target tissues in an engineered construct. This article elucidates the protocols for the decellularization of pancreatic tissue, evaluation of pancreatic tissue-derived dECM bioink, and generation of 3D pancreatic tissue constructs using a bioprinting technique.

3D細胞-細胞-膵組織コンストラクトを印刷するための膵組織由来細胞外マトリックスバイオインク

The transplantation of pancreatic islets is a promising treatment for patients who suffer from type 1 diabetes accompanied by hypoglycemia and secondary ...