この作品は、国立科学財団グラントNo.によってサポートされています。CBET-1236351およびGWバイオミメティクス・バイオインスピレーション工学センター(COBRE)。

この研究では、壁に取り付けられたプロレート半球形、 すなわち モデルの声帯状ポリープは、5:1収縮比の吸引型風洞の試験部床に配置される。不安定な3次元流の分離と壁圧負荷への影響を、オイルフロービジュアライゼーション、壁圧測定、粒子画像のvelocimetryを使用して調査します。不安定な圧力測定は、圧抵抗型圧力センサを備えた16チャンネルの走査型圧力トランスデューサを使用して取得します。圧力センサは670 Hzの周波数応答を持っています。ステンレス鋼管から形成された静圧タップは、表面圧測定を容易にし、スキャン圧力装置に短く配管するために、モデルボーカルフォールドポリープの上流および下流にフラッシュマウントされています。油流の視覚化と表面圧力の測定は、油がファウリングを引き起こす圧力タップに流れ込むため、同時に取得することはできません。
次のセクションでは、壁に取り付けられたプロレートヘミスフェロイドの周囲のオイルフィルムの可視化と表面圧力測定を設定および取得するためのプロトコルを提供します。位相平均化および時間分解粒子画像の経過測定が取得されているが、PIV取得はこのプロトコルには含まれない。著者らはラッフェルらの参照を示唆している。PIV実験セットアップ、データ取得、およびデータ処理の深い理解のための36とエイドリアンとウェスターウィール39。
1. プロトゥービランスを生成する(すなわち、モデルポリープ)
<ol><li>目的のジオメトリを使用して、3 次元コンピュータ支援設計 (CAD) モデルを構築します。長さ5.08cm、幅2.54cm、高さ1.27cmのプロレート半球形としてモデルの声帯ポリープを生成します。モデルのボーカルフォールドポリープの底部に厚さ0.64cmの2.54センチの正方形のベースをマウントします。このベースは、モデルをテスト 断面の床に固定するために使用されます。</li>
	<li>3D CAD モデルをステレオリソグラフィ(STL)ファイルとして書き出します。STL ファイル形式では、モデルサーフェスが一連の三角形として生成されます。モデルのポリップ上のサーフェスを滑らかにするために適切な解像度を選択します。600 ドット/インチ以上の解像度を推奨します。</li>
	<li>STLファイルを適切なソフトウェアにアップロードし、高解像度の3次元プリンタまたは高速プロトタイプを使用してSTLファイルを印刷し、ビルドレイヤー解像度を20μm以上にします。</li>
	<li>風洞試験部は、 図1に示すように取り外し可能な底板を備えた約30.48cm x 30.48 cm x 121.92 cmです。2.54 cmの正方形の穴を約0.85 cmの深さの風洞試験セクションの床の取り外し可能な版にミルし、テストのためのモデルのボーカルフォールドポリープを取り付ける。穴はテストセクションの幅の中央に配置し、テストのために必要な下流の位置に配置する必要があります。</li>
</ol>2. オイルフロー可視化の準備
<ol><li>試験部を準備するために、風洞内部の試験部表面を白い粘着紙で覆う。慎重に配置し、テストセクションの床が接着剤の紙に気泡や折り目によるバンプがないことを確認するために、粘着紙を滑らかにします。テストセクションの壁に取り付けるモデルのポリープアンカーのために、テストセクションの床の正方形の穴の上にある粘着紙に穴を開けます。</li>
	<li>突起(モデルボーカルフォールドポリープ)をアンカー位置に挿入して、テストの準備をします。 図 1を参照してください。</li>
	<li>風洞試験部の上に高解像度カメラを取り付けます。モデルのポリープと周囲のテストセクション領域を含む、選択した視野に合ったカメラの焦点を合わせます。テスト用のカメラ取得パラメータを設定します。ビデオ設定は、オイルフロー ビジュアライゼーションの一時的な部分をキャプチャするために使用する必要があります。</li>
	<li>ベビーオイル、コピートナーパウダー、灯油を7:1:2の比率で組み合わせることで、フロービジュアライゼーションオイル混合物を準備します。例えば:35mlベビーオイル、5mlのコピートナーパウダー、10ml灯油を組み合わせます。ベビーオイルとトナーパウダーを容器に入れ、トナーが完全に溶けるまでかき混ぜます。その後、灯油を加え、よく混ぜます。</li>
	<li>テストセクション表面に簡単に適用できるようにスプレーボトルに混合物を転送します。</li>
</ol>3. オイルフロー可視化測定
<ol><li>オイル混合物の各アプリケーションの前にテストセクション表面をきれいにし、乾燥させます。</li>
	<li>油混合物で満たされたスプレーボトルを使用して、関心のあるセクションに薄い、均一な流体の層をスプレーします。薄い、さらには油の混合物層は適切なオイルフィルム視覚化イメージを作り出す上で重要である。</li>
	<li>カメラで画像またはビデオの取得を開始します。風洞の電源が入る前にカメラの取り込みを開始し、最初の過渡油混合運動をキャプチャします。</li>
	<li>吸引風洞を希望の速度に設定します。油混合物は試験セクション表面に沿って流れ始める。</li>
	<li>油の混合物が流れ止めて定常状態に達したら(すなわち 、パターンは静止している)、または所望の時間が経過したら、カメラの記録を停止し、風洞をパワーダウンする。 注: ビデオ 1 は、定常状態に達し、皮膚摩擦パターンが静止するまで流れる油混合物を表示します。ビデオでは、フローは左から右に移動しています。</li>
</ol>4. 表面圧力測定準備
<ol><li>ステンレス鋼管(0.16 cm外径0.16cm、長さ2.54cm)をテストセクションの床に取り付けるための穴を掘削して、静圧タップを構築することで、テストセクションの床面(取り外し可能なプレート)を準備します。プロレート半球体のアンカー位置の中線から始まり、スパン方向に8.89 cm、下流22.86cmの22.86 cmの下流に1.27 cmのスパンワイズグリッド間隔と2.54 cmの下流グリッド間隔を持つグリッド上の穴をドリルします( 図1を参照)。ステンレス鋼の管は適用範囲の高い管を取り付けるための一端に膨らみを備え、取付けのための反対側にまっすぐである。 注: 静圧タップは、圧力取得位置のより細かいグリッドの間隔を近い間隔で配置できます。</li>
	<li>テストの準備のためにテストセクションの床に所望の構成で壁取り付けプロレート半球体(すなわち モデルボーカルフォールドポリープ)のアンカー位置を囲む管を取り付けます。チューブは、テストセクションの床とフラッシュマウントする必要があります。</li>
	<li>取り付けられたステンレス鋼の管から取り付けられたステンレス鋼の管からスキャン圧力トランスデューサー測定ポートに短い柔軟な管(長さ6.35 cm、0.159 cm内径0.159cm、0.475 cmの透明なポリ塩化ビニルチューブ)を取り付けます。走査圧力トランスデューサには16個の圧力ポートがあります。</li>
</ol>5. 表面圧力測定の獲得
<ol><li>スキャン圧力トランスデューサをコンピュータに接続し、スキャン圧力トランスデューサソフトウェアを使用して取得パラメータを設定します。取得ソフトウェアを設定して、データ取得の目的の期間に 500 Hz でデータを取得します。 注:データは、低発振周波数での小さな圧力変動のために、スキャン圧力トランスデューサの最大サンプリングレート、500 Hzで取得されました。</li>
	<li>吸引風洞を希望の速度に設定します。</li>
	<li>圧力測定の取得を開始します。圧力測定は、任意の所望の流れの診断技術(例えば PIV、レーザードップラー麻酔、ホットワイヤー解剖 学など)と同時に取得することができる。</li>
</ol>

<ol>
	<li>Hirano, M., Kakita, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cover-body theory of vocal fold vibration. Speech science--recent advances. , 1-46 (1985).</li><li>Gray, S. D., Titze, I. R., Alipour, F., Hammond, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanical+and+histologic+observations+of+vocal+fold+fibrous+proteins.">Biomechanical and histologic observations of vocal fold fibrous proteins.</a> Ann. Otol. Rhinol. Laryngol. 109 (1), 77-85 (2000).</li><li>Titze, I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physics+of+small-amplitude+oscillation+of+the+vocal+fold.">The physics of small-amplitude oscillation of the vocal fold.</a> J. Acoustic. Soc. Am. 83 (4), 1536-1552 (1988).</li><li>Boessenecker, A., Berry, D. A., Lohscheller, J., Eysholdt, U., D&#246;llinger, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mucosal+wave+properties+of+a+human+vocal+fold.">Mucosal wave properties of a human vocal fold.</a> Acta Acustica. 93 (5), 815-823 (2007).</li><li>Shaw, H. S., Deliyski, 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=Mucosal+wave:+a+normophonic+study+across+visualization+techniques.">Mucosal wave: a normophonic study across visualization techniques.</a> J. Voice. 22 (1), 23-33 (2008).</li><li>Krausert, C. R., Olszewski, A. E., Taylor, L. N., McMurray, J. S., Dailey, S. H., Jiang, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mucosal+wave+measurement+and+visualization+techniques.">Mucosal wave measurement and visualization techniques.</a> J. Voice. 25 (4), 395-405 (2010).</li><li>Fant, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustic+Theory+of+Speech+Production.">Acoustic Theory of Speech Production.</a> Mouton and Co. N. V.: The Hague. , 15-79 (1960).</li><li>Wegel, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theory+of+vibration+of+the+larynx.">Theory of vibration of the larynx.</a> J. Acoustic. Soc. Am. 1, 1-21 (1930).</li><li>Den Berg, J. V. a. n., Zantema, J. T., Doornenbal, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+air+resistance+and+the+Bernoulli+effect+of+the+human+larynx.">On the air resistance and the Bernoulli effect of the human larynx.</a> J. Acoustic. Soc. Am. 29 (5), 626-631 (1957).</li><li>Scherer, R. C., Shinwari, D., De Witt, K. J., Zhang, C., Kucinschi, B. R., Afjeh, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraglottal+pressure+profiles+for+a+symmetric+and+oblique+glottis+with+a+divergence+angle+of+10+degrees.">Intraglottal pressure profiles for a symmetric and oblique glottis with a divergence angle of 10 degrees.</a> TJ. Acoustic. Soc. Am. 109 (4), 1616-1630 (2001).</li><li>Thomson, S. L., Mongeau, L., Frankel, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerodynamic+transfer+of+energy+to+the+vocal+folds.">Aerodynamic transfer of energy to the vocal folds.</a> TJ. Acoustic. Soc. Am. 118 (3), 1689-1700 (2005).</li><li>Erath, B. D., Plesniak, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+investigation+of+asymmetric+flow+features+in+a+scaled-up+driven+model+of+the+human+vocal+folds.">An investigation of asymmetric flow features in a scaled-up driven model of the human vocal folds.</a> Exp. Fluids. 49 (1), 131-146 (2010).</li><li>Petrovi&#263;-Lazi&#263;, M., Kosanovi&#263;, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustic+analysis+findings+in+patients+with+vocal+fold+polyp.">Acoustic analysis findings in patients with vocal fold polyp.</a> Acta Med. Saliniana. 38 (2), 63-66 (2009).</li><li>Erath, B. D., Plesniak, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+laryngeal+flow+fields+induced+by+a+model+vocal+fold+polyp.">Three-dimensional laryngeal flow fields induced by a model vocal fold polyp.</a> Int. J. Heat Fluid Flow. 35, 93-101 (2012).</li><li>Acarlar, M. S., Smith, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+study+of+hairpin+vortices+in+a+laminar+boundary+layer.+Part+1.+Hairpin+vortices+generated+by+a+hemisphere+protuberance.">A study of hairpin vortices in a laminar boundary layer. Part 1. Hairpin vortices generated by a hemisphere protuberance.</a> J. Fluid Mech. 175, 1-41 (1987).</li><li>Kawanisi, K., Maghrebi, M. F., Yokosi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+instantaneous+3-D+analysis+of+turbulent+flow+in+the+wake+of+a+hemisphere.">An instantaneous 3-D analysis of turbulent flow in the wake of a hemisphere.</a> Boundary-Layer Meteorol. 64, 1-14 (1992).</li><li>Savory, E., Toy, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemisphere+and+hemisphere-cylinders+in+turbulent+boundary+layers.">Hemisphere and hemisphere-cylinders in turbulent boundary layers.</a> J. Wind Eng. Ind. Aerodyn. 23, 345-364 (1986).</li><li>Tamai, N., Asaeda, T., Tanaka, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vortex+structures+around+a+hemispheric+hump.">Vortex structures around a hemispheric hump.</a> Boundary-Layer Meteorol. 39, 301-314 (1987).</li><li>Savory, E., Toy, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+separated+shear+layers+associated+with+hemispherical+bodies+in+turbulent+boyndary+layers.">The separated shear layers associated with hemispherical bodies in turbulent boyndary layers.</a> J. Wind Eng. Ind. Aerodyn. 28, 291-300 (1988).</li><li>Ogawa, T., Nakayama, M., Murayama, S., Sasaki, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+wind+pressures+on+basic+structures+with+curved+surfaces+and+their+response+in+turbulent+flow.">Characteristics of wind pressures on basic structures with curved surfaces and their response in turbulent flow.</a> J. Wind Eng. Ind. Aerodyn. 38, 427-438 (1991).</li><li>Manhart, M., Wengle, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-eddy+simulation+of+turbulent+bounday+layer+flow+over+a+hemisphere.">Large-eddy simulation of turbulent bounday layer flow over a hemisphere.</a> Direct and Large-Eddy Simulation I: Selected papers from the First ERCOFTAC Workshop on Direct and Large-Eddy Simulation. , 299-301 (1994).</li><li>Manhart, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vortex+shedding+from+a+hemisphere+in+a+turbulent+boundary+layer.">Vortex shedding from a hemisphere in a turbulent boundary layer.</a> Theor. Comp. Fluid Dyn. 12, 1-28 (1998).</li><li>Meroney, R. N., Letchford, C. W., Sarkar, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+numerical+and+wind+tunnel+simulation+of+wind+loads+on+smooth,+rough+and+dual+domes+immersed+in+a+boundary+layer.">Comparison of numerical and wind tunnel simulation of wind loads on smooth, rough and dual domes immersed in a boundary layer.</a> Wind Struct. 5 (2-4), 347-358 (2002).</li><li>Hunt, J. C. R., Abell, C. J., Peterka, J. A., Woo, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinematical+studies+of+the+flows+around+free+or+surface-mounted+obstacles;+applying+topology+to+flow+visualization.">Kinematical studies of the flows around free or surface-mounted obstacles; applying topology to flow visualization.</a> J. Fluid Mech. 86 (01), 179 (2006).</li><li>Legendre, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignes+de+courant+d'un+ecoulement+permanent:+decollement+et+separation.">Lignes de courant d'un ecoulement permanent: decollement et separation.</a> La Recherche A&#233;rospatiale. 6, 327-335 (1977).</li><li>Merzkirch, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+of+Surface+Flow.">Visualization of Surface Flow.</a> Flow Visual. , 82-89 (1987).</li><li>Simpson, R. L., Long, C. H. H., Byun, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+of+vortical+separation+from+an+axisymmetric+hill.">Study of vortical separation from an axisymmetric hill.</a> Int. J. Heat Fluid Flow. 23 (5), 582-591 (2002).</li><li>Byun, G., Simpson, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface-pressure+fluctuations+from+separated+flow+over+an+axisymmetric+bump.">Surface-pressure fluctuations from separated flow over an axisymmetric bump.</a> Am. Inst. Aeronaut. Astronaut. J. 48 (10), 2397-2405 (2010).</li><li>Martinuzzi, R., Tropea, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+flow+around+surface-mounted,+prismatic+obstacles+placed+in+a+fully+developed+channel+flow.">The flow around surface-mounted, prismatic obstacles placed in a fully developed channel flow.</a> J. Fluids Eng. 115, 85-92 (1993).</li><li>R&#246;diger, T., Knauss, H., Gaisbauer, U., Kr&#228;mer, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pressure+and+heat+flux+measurements+on+the+surface+of+a+low-aspect-ratio+circular+cylinder+mounted+on+a+ground+plate.">Pressure and heat flux measurements on the surface of a low-aspect-ratio circular cylinder mounted on a ground plate.</a> New Results Num. Exp. Fluid Mech. VI. , 121-128 (2007).</li><li>Martinuzzi, R., AbuOmar, M., Savory, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scaling+of+the+wall+pressure+field+around+surface-mounted+pyramids+and+other+bluff+bodies.">Scaling of the wall pressure field around surface-mounted pyramids and other bluff bodies.</a> J. Fluids Eng. 129, 1147-1156 (2007).</li><li>Taniguchi, S., Sakamoto, H., Kiya, M., Arie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-averaged+aerodynamic+forces+acting+on+a+hemisphere+immersed+in+a+turbulent+boundary.">Time-averaged aerodynamic forces acting on a hemisphere immersed in a turbulent boundary.</a> J. Wind Eng. Indust. Aerodyn. 9, 257-273 (1982).</li><li>Triep, M., Br&#252;cker, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+nature+of+the+glottal+jet.">Three-dimensional nature of the glottal jet.</a> The Journal of the Acoustic. Soc. Am. 127 (3), 1537-1547 (2010).</li><li>Khosla, S., Murugappan, S., Paniello, R., Ying, J., Gutmark, 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+vortices+in+voice+production:+normal+versus+asymmetric+tension.">Role of vortices in voice production: normal versus asymmetric tension.</a> Laryngoscope. 119 (1), 216-221 (2009).</li><li>Drechsel, J. S., Thomson, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+supraglottal+structures+on+the+glottal+jet+exiting+a+two-layer+synthetic,+self-oscillating+vocal+fold+model.">Influence of supraglottal structures on the glottal jet exiting a two-layer synthetic, self-oscillating vocal fold model.</a> The Journal of the Acoustical Society of America. 123 (6), 4434-4445 (2008).</li><li>Raffel, M., Willert, C., Kompenhans, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Particle Image Velocimetry: A Practical Guide. , (1998).</li><li>Neubauer, J., Zhang, Z., Miraghaie, R., Berry, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+structures+of+the+near+field+flow+in+a+self-oscillating+physical+model+of+the+vocal+folds.">Coherent structures of the near field flow in a self-oscillating physical model of the vocal folds.</a> J. Acoustic. Soc. Am. 121 (2), 1102-1118 (2007).</li><li>Krebs, F., Silva, F., Sciamarella, D., Artana, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+three-dimensional+study+of+the+glottal+jet.">A three-dimensional study of the glottal jet.</a> Exp. Fluids. 52 (5), 1133-1147 (2011).</li><li>Adrian, R. J., Westerweel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Particle image velocimetry. 30, (2010).</li><li>Tobak, M., Peake, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topology+of+three-dimensional+separated+flows.">Topology of three-dimensional separated flows.</a> Ann. Rev. Fluid Mech. 14, 61-85 (1982).</li><li>Zhang, Y., Jiang, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymmetric+Spatiotemporal+Chaos+Induced+by+a+Polypoid+Mass+in+the+Excised+Canine+Larynx.">Asymmetric Spatiotemporal Chaos Induced by a Polypoid Mass in the Excised Canine Larynx.</a> Chaos. 18, 43102 (2008).</li><li>D&#233;lery, J. M., Jean, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delery+Toward+the+elucidation+of+three-dimensional+separation.">Delery Toward the elucidation of three-dimensional separation.</a> Ann. Rev. Fluid Mech. 33, 129-154 (2001).</li></ol>

著者らは開示するものは何もない。

幾何学的突起からの渦構造の形成と伝播を理解し、その後の声帯力学的負荷に対する影響を理解することは、声帯ポリープおよび結節の治療を進めるために洞察とモデルを提供する必要がある。この実験におけるモデルポリープによって引き起こされる空力負荷の変動は、ポリープ13,41の患者に観察される不規則な声の折り目のダイナミクスに寄与すると予想される。今後の作業には...

ボーカルフォールドは、ボーカル気道を横切って伸びる組織の2つのバンドです。音声音声は、クリティカルな肺圧が達成されると生成され、付加された声帯を通して空気を強制する。声の折り目は、組織の多くの層で構成され、多くの場合、簡略化された2層のボディカバーシステム1によって表されます。カバー層の大部分を構成する細胞外マトリックスは、コラーゲンとエラスチン繊維で構成され、非線形応力-ひずみ特性を提供し、声帯1,2の適切な運動に重要である。空気力学的な力は声のひだの組織にエネルギーを与え、自己持続振動3を励起する。声が揺れるにつれて、それらの間の開口部は、グロティスと呼ばれ、収束性から均一に移行し、サイクル4,6を閉じて繰り返す前に発散通路に移行する時間的に変化するオリフィスを形成する。通常の音声に対する振動の周波数は、通常、男性と女性でそれぞれ100〜220Hzに及び、グロティス7を通過する脈動性流れ場を作り出す。通常の音声のための流体構造エネルギー交換プロセスは、広範囲に研究されています8-12;しかし、いくつかの病理に対するこのプロセスの中断は十分に理解されていない。声帯の病理的状態は、そのダイナミクスの劇的な変化をもたらし、音声を生成する能力に影響を与える可能性があります。
ポリープと結節は、声帯の内側表面に形成される幾何学的な異常である。これらの異常は、患者の13を通信する能力に影響を与える可能性があります。それにもかかわらず、最近になって、ポリープのような幾何学的な突起に起因する流動界の破壊が14と考えられていた。その研究は、「正常な」流体構造の音声交換プロセスが劇的に変化し、フローフィールドの改変がポリープおよび結節患者の声質の深刻な劣化の最も可能性の高い理由であることを示した。脈動流におけるポリープからの三次元流れ分離によって生じる流れ構造の包括的な理解は確立されていない。ポリープからの渦構造の生成と伝播、および声帯力学的な動きを駆動する空気力学的負荷へのその後の影響は、患者のポリープの外科的改善を進めるために必要な重要な要素である。
安定した流れにおける壁に取り付けられた半球体からの流れの分離は15-23で調査されているが、驚くべきことに、拍動性または不安定な流れ条件に従う壁の半球状からの不安定な三次元流の分離に関する情報はほとんどない。アカラーとスミス15 の精通した研究は、層状境界層内の壁に取り付けられた半球体を横切る安定した流れによって生成された三次元のコヒーレント構造の分析を提供した。アカラーとスミスは、2種類の渦構造を特定した。立っている馬蹄渦は、半球状突起の上流に形成され、両側の突起の下流に拡張された。さらに、ヘアピン渦は、壁から定期的にヘミスフェロイドをウェイクに取り付けた。ヘアピン渦の複雑な動きと進行を調査し、詳細に説明した。
滑らかに輪郭を描いた軸対称丘の上の流れは、表面静圧測定と表面油の視覚化の両方が乱流せん断流内のバンプの上および下流に取得された研究が以前に研究されました。オイルフィルム技術は、皮膚摩擦線、高速および低速度領域、および表面流れの中の分離およびアタッチメントポイントの可視化を可能にし、壁に取り付けられた物体のウェイクを調査するのに有用である。この技術のために、目的の表面は、油基剤と微粉末顔料(ランプ ブラック、グラファイト粉末、または二酸化チタン)混合物の薄膜でコーティングされています。所望の流れ条件では、摩擦力が表面に沿って油を動かすと、顔料粉末がストリークに沈着する。臨界点または特異点は、せん断応力が0又は2以上の成分の平均速度がゼロである場所、サドルポイントまたは節点24~26として得られた皮膚摩擦線パターンから分類することができる。
丘の幾何学では、分離によって引き起こされる特異点は上流で見つかっていない。これは、半球形突起で発生する不利な圧力勾配を生成しなかったバンプの滑らかに上昇する輪郭に起因した。その結果、その後の隆起の頂点まで流れが加速することが判明し、ヘアピン渦27,28の形成から予想されるように、バンプ中心線を過ぎて間もなく発達した不安定なサドル焦点分離点が発達した。異なる壁に取り付けられた幾何学を用いた同様の実験技術を用いた研究では、マルティンッツィとトロペア29 によって行われた安定した流れの表面に取り付けられた立方体の周りの油膜視覚化は、物体の上流に2つの明確な皮膚摩擦線を表示した。第1の皮膚摩擦線は、不利な圧力勾配に起因する一次分離線に対応し、第2の皮膚摩擦線は馬蹄渦の時間平均位置を示した。物体の上流で行われた表面圧力測定は、馬蹄渦線に沿った局所最小および一次分離と馬蹄渦線の間の局所圧力最大値を示した。同様の上流分離線は、円筒、ピラミッド、円錐29-31など、他の表面に取り付けられたジオメトリと形成されます。壁に取り付けられたオブジェクトの下流のサーフェスビジュアライゼーションは、通常、オブジェクト30の後ろの再循環領域によって引き起こされる2つのフォチを表示する。2つの渦は、病巣位置で生成され、壁に取り付けられたヘミスフェロイド32の後に見られる「アーチ型」またはヘアピン渦に対応する。
粒子画像のヴェロシメトリー(PIV)は、合成声の折り目モデル33-35の下流流を研究するために以前に使用されてきた。PIVは、画像が時空間流体力学36を捕捉するために平面内でトレーサー粒子の動きを流れる非侵襲的な視覚化技術である。振動する声の折り目の下流を形成する三次元の一貫した構造は、Neubauerらによって研究されてきた 。37;渦の発生と対流とジェットフラッピングが観察された。最近、クレブス ら38 は立体PIVを用いて眼内ジェットの三次元性を調べ、その結果、眼内ジェット軸切り替えを実証した。エラスとプレスニアック14 は、モデルボーカルフォールドポリープが7.5倍スケールアップされた動的に駆動されたボーカルフォールドモデルの内側表面に及ぼす影響を調査した。ポリープの下流に再循環領域が形成され、噴射ダイナミクスはフォナトリーサイクル全体を通して影響を受けた。エラスとプレスニアック14による駆動声帯ポリープ研究を阻止する以前の研究では、内側の声帯折りポリープまたは結節によって誘発される流体ダイナミクスを探求していない。
ボーカルフォールド移動壁の追加の複雑さ、誘導圧力勾配、限られた幾何学的体積および他の複雑さを含む前に、安定した、拍動性の流れ場内のモデルのポリープの流体動的効果を理解することが重要です。現在の作業は、安定したフロー条件と不安定なフロー条件の両方で、下流壁の流れ構造のシグネチャに焦点を当てています。突起から流される渦構造と下流壁との相互作用は、これらの相互作用が生物学的応答を引き出すので、声帯ポリープの調査と他の生物学的考察にとって大きな関心を持つ。

<table border="0" cellpadding="0" cellspacing="0" width="699">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">Rapid Prototyper</td>
			<td style="width:139px;">Objet</td>
			<td style="width:119px;">Objet24</td>
			<td style="width:227px;">Tray Size (X&#215;Y&#215;Z): 240 &#215; 200 &#215; 150 mm 
			Build layer thickness =&#160; 28 &#181;m&#160; 
			Accuracy = 0.1 mm 
			Build Resolution: X-axis: 600 dpi, Y-axis: 600 dpi, Z-axis: 900 dpi</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Rapid Prototyper Model Material</td>
			<td style="width:139px;">Objet</td>
			<td style="width:119px;">VeroWhite Plus Fullcure 835</td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Rapid Prototyper Support Material</td>
			<td style="width:139px;">Objet</td>
			<td style="width:119px;">FullCure 705 Support</td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Copy Toner</td>
			<td style="width:139px;">Xerox</td>
			<td style="width:119px;"></td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Kerosene</td>
			<td style="width:139px;">Sunnyside</td>
			<td style="width:119px;"></td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Baby Oil</td>
			<td style="width:139px;">Johnson&#39;s</td>
			<td style="width:119px;"></td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Adhesive Paper</td>
			<td style="width:139px;">Con-Tact Brand</td>
			<td style="width:119px;"></td>
			<td style="width:227px;">White adhesive covering</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tygon Tubing</td>
			<td style="width:139px;">Tygon</td>
			<td style="width:119px;">PVC Tubing</td>
			<td style="width:227px;">1/16&#34; ID, 3/16&#34;OD</td>
		</tr>
		<tr height="151">
			<td height="151" style="height:151px;width:216px;">Pressure Scanner (16 channel)</td>
			<td style="width:139px;">Scanivalve</td>
			<td style="width:119px;">DSA3217</td>
			<td style="width:227px;">Used for gas pressure measurements 
			Pressure range = +/- 5&#34; H2O 
			Full scale accuracy = +/-0.3% full scale accuracy.&#160; 
			Maximum scan rate = 500 Hz/channel</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stainless Steel Tubulations</td>
			<td style="width:139px;">Scanivalve</td>
			<td style="width:119px;">TUBN-063-1.0</td>
			<td style="width:227px;">0.063&#34; Diameter and 1&#34; Length</td>
		</tr>
	</tbody>
</table>

investigating the three-dimensional flow separation induced by a model vocal fold polyp

通常の音声のための流体構造エネルギー交換プロセスは広範囲に研究されているが、病理学的状態については十分に理解されていない。声帯の内側表面に形成される幾何学的な異常であるポリープと結節は、声の折り畳みダイナミクスを破壊し、したがって、患者の通信能力に壊滅的な結果をもたらす可能性があります。当研究室では、 インビトロ 駆動のボーカルフォールドモデルの内側表面に位置するモデルポリープの調査の中で、粒子画像のvelocimetry(PIV)測定が報告されており、このような幾何学的異常が気球ジェットの挙動を著しく破壊することを示しています。このフローフィールド調整は、ポリープ患者における声質の著しい劣化の可能性が高い理由である。声帯ポリープのような幾何学的突起からの渦構造の形成と伝播、および声の折り目のダイナミクスを駆動する空力負荷への結果の影響のより完全な理解は、この病理学的状態の治療を進める上で必要である。本研究は、クロスフローにおける2:1アスペクト比を有する壁に取り付けられたプロレート半球体、 すなわち モデルのボーカルフォールドポリープによって誘導される3次元流の分離に関する、オイルフィルム可視化技術を用いた。不安定な3次元流の分離と壁圧荷重の影響は、皮膚摩擦線の可視化と壁圧測定を使用して調べます。

7.5倍スケールアップ動的に駆動されたボーカルフォールドモデルを使用した以前の研究は、幾何学的突起、モデルボーカルフォールドポリープの存在が、フォナトリーサイクルを通じて眼球ジェットの正常なダイナミクスを破壊することを実証しました。前回の駆動ボーカルフォールドモデルスタディの代表的な結果を 、図2 と ビデオ2に表示します。ビデオは、収?...

Watch this Scientific Journal Video about Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp at JoVE.com

Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp

声帯ポリープは、声の折り目のダイナミクスを破壊することができ、したがって、患者の通信能力に壊滅的な結果をもたらすことができます。壁面取り付けモデルポリープによって誘導される3次元流れ分離と壁圧負荷への影響を、粒子画像の軌跡、皮膚摩擦線の可視化、および壁圧測定を用いて調べる。

モデルの声帯ポリープによる三次元流分離の調査

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. ...

investigating-three-dimensional-flow-separation-induced-model-vocal

Research

JoVE Journal

Bioengineering

8.4K ビュー.  The George Washington University. 声帯ポリープは、声の折り目のダイナミクスを破壊することができ、したがって、患者の通信能力に壊滅的な結果をもたらすことができます。壁面取り付けモデルポリープによって誘導される3次元流れ分離と壁圧負荷への影響を、粒子画像の軌跡、皮膚摩擦線の可視化、および壁圧測定を用いて調べる。

ビデオ: Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells behave differently when they are grown within a 3D extra-cellular matrix and also interact with other cells (1-5). Mouse models have been broadly utilized to study tissue morphogenesis in vivo. However mouse and human skin have significant differences in cellular architecture and physiology, which makes it difficult to extrapolate mouse studies to humans. Since melanocytes in mouse skin are mostly localized in hair follicles, they have distinct biological properties from those of humans, which locate primarily at the basal layer of the epidermis. The recent development of 3D human skin reconstruct models has enabled the field to investigate cell-matrix and cell-cell interactions between different cell types. The reconstructs consist of a "dermis" with fibroblasts embedded in a collagen I matrix, an "epidermis", which is comprised of stratified, differentiated keratinocytes and a functional basement membrane, which separates epidermis from dermis. Collagen provides scaffolding, nutrient delivery, and potential for cell-to-cell interaction. The 3D skin models incorporating melanocytic cells recapitulate natural features of melanocyte homeostasis and melanoma progression in human skin. As in vivo, melanocytes in reconstructed skin are localized at the basement membrane interspersed with basal layer keratinocytes. Melanoma cells exhibit the same characteristics reflecting the original tumor stage (RGP, VGP and metastatic melanoma cells) in vivo. Recently, dermal stem cells have been identified in the human dermis (6). These multi-potent stem cells can migrate to the epidermis and differentiate to melanocytes.

In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.

三次元ヒト皮膚は、モデルを再構築：ノーマルスキンとメラノーマ進行を勉強するツールを

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

生物工学

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 ...

Synthetic, Multi-Layer, Self-Oscillating Vocal Fold Model Fabrication

Sound for the human voice is produced via flow-induced vocal fold vibration. The vocal folds consist of several layers of tissue, each with differing material properties 1. Normal voice production relies on healthy tissue and vocal folds, and occurs as a result of complex coupling between aerodynamic, structural dynamic, and acoustic physical phenomena. Voice disorders affect up to 7.5 million annually in the United States alone 2 and often result in significant financial, social, and other quality-of-life difficulties. Understanding the physics of voice production has the potential to significantly benefit voice care, including clinical prevention, diagnosis, and treatment of voice disorders.
Existing methods for studying voice production include in vivo experimentation using human and animal subjects, in vitro experimentation using excised larynges and synthetic models, and computational modeling. Owing to hazardous and difficult instrument access, in vivo experiments are severely limited in scope. Excised larynx experiments have the benefit of anatomical and some physiological realism, but parametric studies involving geometric and material property variables are limited. Further, they are typically only able to be vibrated for relatively short periods of time (typically on the order of minutes).
Overcoming some of the limitations of excised larynx experiments, synthetic vocal fold models are emerging as a complementary tool for studying voice production. Synthetic models can be fabricated with systematic changes to geometry and material properties, allowing for the study of healthy and unhealthy human phonatory aerodynamics, structural dynamics, and acoustics. For example, they have been used to study left-right vocal fold asymmetry 3,4, clinical instrument development 5, laryngeal aerodynamics 6-9, vocal fold contact pressure 10, and subglottal acoustics 11 (a more comprehensive list can be found in Kniesburges et al. 12)
Existing synthetic vocal fold models, however, have either been homogenous (one-layer models) or have been fabricated using two materials of differing stiffness (two-layer models). This approach does not allow for representation of the actual multi-layer structure of the human vocal folds 1 that plays a central role in governing vocal fold flow-induced vibratory response. Consequently, one- and two-layer synthetic vocal fold models have exhibited disadvantages 3,6,8 such as higher onset pressures than what are typical for human phonation (onset pressure is the minimum lung pressure required to initiate vibration), unnaturally large inferior-superior motion, and lack of a "mucosal wave" (a vertically-traveling wave that is characteristic of healthy human vocal fold vibration).
In this paper, fabrication of a model with multiple layers of differing material properties is described. The model layers simulate the multi-layer structure of the human vocal folds, including epithelium, superficial lamina propria (SLP), intermediate and deep lamina propria (i.e., ligament; a fiber is included for anterior-posterior stiffness), and muscle (i.e., body) layers 1. Results are included that show that the model exhibits improved vibratory characteristics over prior one- and two-layer synthetic models, including onset pressure closer to human onset pressure, reduced inferior-superior motion, and evidence of a mucosal wave.

The methodology for fabricating synthetic vocal fold models is described. The models are life-sized and mimic the multi-layer structure of the human vocal folds. Results show the models to self-oscillate at pressures comparable to lung pressure and demonstrate flow-induced vibratory responses that are similar to those of human vocal folds.

合成、マルチレイヤ、自己発振声帯モデルの作製

Sound for the human voice is produced via flow-induced vocal fold vibration. The vocal folds consist of several layers of tissue, each with differing ...

Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling in the tumor microenvironment 1. Previously, research on the tumor microenvironment has focused primarily on tumor-stromal interactions 1-2. However, the tumor microenvironment also includes a variety of biophysical forces, whose effects remain poorly understood. These forces are biomechanical consequences of tumor growth that lead to changes in gene expression, cell division, differentiation and invasion3. Matrix density 4, stiffness 5-6, and structure 6-7, interstitial fluid pressure 8, and interstitial fluid flow 8 are all altered during cancer progression.
Interstitial fluid flow in particular is higher in tumors compared to normal tissues 8-10. The estimated interstitial fluid flow velocities were measured and found to be in the range of 0.1-3 &mu;m s-1, depending on tumor size and differentiation 9, 11. This is due to elevated interstitial fluid pressure caused by tumor-induced angiogenesis and increased vascular permeability 12. Interstitial fluid flow has been shown to increase invasion of cancer cells 13-14, vascular fibroblasts and smooth muscle cells 15. This invasion may be due to autologous chemotactic gradients created around cells in 3-D 16 or increased matrix metalloproteinase (MMP) expression 15, chemokine secretion and cell adhesion molecule expression 17. However, the mechanism by which cells sense fluid flow is not well understood. In addition to altering tumor cell behavior, interstitial fluid flow modulates the activity of other cells in the tumor microenvironment. It is associated with (a) driving differentiation of fibroblasts into tumor-promoting myofibroblasts 18, (b) transporting of antigens and other soluble factors to lymph nodes 19, and (c) modulating lymphatic endothelial cell morphogenesis 20.
The technique presented here imposes interstitial fluid flow on cells in vitro and quantifies its effects on invasion (Figure 1). This method has been published in multiple studies to measure the effects of fluid flow on stromal and cancer cell invasion 13-15, 17. By changing the matrix composition, cell type, and cell concentration, this method can be applied to other diseases and physiological systems to study the effects of interstitial flow on cellular processes such as invasion, differentiation, proliferation, and gene expression.

Interstitial fluid flow is elevated in solid tumors and can modulate tumor cell invasion. Here we describe a technique to apply interstitial fluid flow to cells embedded in a matrix and then measure its effects on cell invasion. This technique can be easily adapted to study other systems.

腫瘍細胞の浸潤を間質液流れに及ぼす影響を測定するための三次元細胞培養モデル

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling ...

A Novel Three-dimensional Flow Chamber Device to Study Chemokine-directed Extravasation of Cells Circulating under Physiological Flow Conditions

Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell homing and tumor metastasis. The three-dimensional flow chamber device (hereafter the 3D device) is a novel in vitro technology that recreates physiological shear stress and allows each step of the cell extravasation cascade to be quantified. The 3D device consists of an upper compartment in which the cells of interest circulate under shear stress, and a lower compartment of static wells that contain the chemoattractants of interest. The two compartments are separated by porous inserts coated with a monolayer of endothelial cells (EC). An optional second insert with microenvironmental cells of interest can be placed immediately beneath the EC layer. A gas exchange unit allows the optimal CO2 tension to be maintained and provides an access point to add or withdraw cells or compounds during the experiment. The test cells circulate in the upper compartment at the desired shear stress (flow rate) controlled by a peristaltic pump. At the end of the experiment, the circulating and migrated cells are collected for further analyses. The 3D device can be used to examine cell rolling on and adhesion to EC under shear stress, transmigration in response to chemokine gradients, resistance to shear stress, cluster formation, and cell survival. In addition, the optional second insert allows the effects of crosstalk between EC and microenvironmental cells to be examined. The translational applications of the 3D device include testing of drug candidates that target cell migration and predicting the in vivo behavior of cells after intravenous injection. Thus, the novel 3D device is a versatile and inexpensive tool to study the molecular mechanisms that mediate cellular extravasation.

The three-dimensional flow chamber device is a novel in vitro technology for the quantitative and step-wise evaluation of the extravasation cascade of cells circulating under conditions of physiological shear stress. The device therefore fills a critical need for basic, preclinical, and clinical studies of cell migration.

生理フローの状況の下で循環細胞のケモカイン指向溢出を研究する新規三次元フローチャンバーデバイス

Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

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 ...

Construction and Characterization of a Novel Vocal Fold Bioreactor

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 &#181;m. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

A novel vocal fold bioreactor capable of delivering physiologically relevant, vibratory stimulation to cultured cells is constructed and characterized. This dynamic culture device, when combined with a fibrous poly(&#949;-caprolactone) scaffold, creates a vocal fold-mimetic environment that modulates the behaviors of mesenchymal stem cells.

新規声帯バイオリアクターの構築とキャラクタリゼーション

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To ...

Analysis of Cell Migration within a Three-dimensional Collagen Matrix

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic development, wound healing, and immune responses. However, cell migration is also a key mechanism in cancer enabling these cancer cells to detach from the primary tumor to start metastatic spreading. Within the past years various cell migration assays have been developed to analyze the migratory behavior of different cell types. Because the locomotory behavior of cells markedly differs between a two-dimensional (2D) and three-dimensional (3D) environment it can be assumed that the analysis of the migration of cells that are embedded within a 3D environment would yield in more significant cell migration data. The advantage of the described 3D collagen matrix migration assay is that cells are embedded within a physiological 3D network of collagen fibers representing the major component of the extracellular matrix. Due to time-lapse video microscopy real cell migration is measured allowing the determination of several migration parameters as well as their alterations in response to pro-migratory factors or inhibitors. Various cell types could be analyzed using this technique, including lymphocytes/leukocytes, stem cells, and tumor cells. Likewise, also cell clusters or spheroids could be embedded within the collagen matrix concomitant with analysis of the emigration of single cells from the cell cluster/ spheroid into the collagen lattice. We conclude that the 3D collagen matrix migration assay is a versatile method to analyze the migration of cells within a physiological-like 3D environment.

Cell migration is a biological phenomenon that is involved in a plethora of physiological, such as wound healing and immune responses, and pathophysiological processes, like cancer. The 3D-collagen matrix migration assay is a versatile tool to analyze the migratory properties of different cell types within in a 3D physiological-like environment.

三次元コラーゲンマトリックス内の細胞移動の解析

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic ...

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 ...

Studying Protein Function and the Role of Altered Protein Expression by Antibody Interference and Three-dimensional Reconstructions

A strict management of protein expression is not only essential to every organism alive, but also an important strategy to investigate protein functions in cellular models. Therefore, recent research invented different tools to target protein expression in mammalian cell lines or even animal models, including RNA and antibody interference. While the first strategy has gathered much attention during the past two decades, peptides &#160;mediating a translocation of antibody cargos across cellular membranes and into cells, obtained much less interest. In this publication, we provide a detailed protocol how to utilize a peptide carrier named Chariot in human embryonic kidney cells as well as in primary hippocampal neurons to perform antibody interference experiments and further illustrate the application of three-dimensional reconstructions in analyzing protein function. Our findings suggest that Chariot is, probably due to its nuclear localization signal, particularly well-suited to target proteins residing in the soma and the nucleus. Remarkably, when applying Chariot to primary hippocampal cultures, the reagent turned out to be surprisingly well accepted by dissociated neurons.

Controlling protein expression is not only essential to every organism alive, but also an important strategy to investigate protein functions in cellular models. The protocol presented shows the application of antibody interference in mammalian cells including primary hippocampal neurons and demonstrates the use of three-dimensional reconstructions in studying protein function.

タンパク質機能および抗体干渉によるタンパク質発現の変化の役割と三次元再構成を学びます

A strict management of protein expression is not only essential to every organism alive, but also an important strategy to investigate protein functions ...