Name: fabrication of biologically derived injectable materials for myocardial tissue engineering
Uploaded: 2010-12-20T15:00:00
Description: Watch this Scientific Journal Video about Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering at JoVE.com

この研究は、助成金番号1 - DP2 - OD004309 - 01を介して、NIHのディレクターの新イノベーター賞プログラム、医学研究のためのNIHロードマップの一部が部分的にサポートされていました。 SBS - N。大学院研究フェローシップのためにNSFを感謝したいと思います。

1。前処理組織の準備<ol class="lalpha"><li>このプロトコルを使用する前に、一つは既に選択肢の組織を脱細胞されている必要があります。この例では、新鮮なブタと人間の心膜の試料は脱イオン（DI）水と硫酸ドデシルナトリウム（SDS）に低張と高張リンスを使用して脱細胞されています。 </li><li>特に、最初の30分間のDI水にブタ心膜を洗浄し、リン酸塩で1％SDS中で継続的にかき混ぜる生理食?...

<ol>
	<li>Seif-Naraghi, S. B., Salvatore, M. A., Magoffin-Schup, P. J., Hu, D. P., Christman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+characterization+of+an+injectable+pericardial+matrix+gel:+A+potentially+autologous+scaffold+for+cardiac+tissue+engineering.">Design and characterization of an injectable pericardial matrix gel: A potentially autologous scaffold for cardiac tissue engineering.</a> Tissue Engineering. , (2009).</li><li>Freytes, D. O., Martin, J., Velankar, S. S., Lee, A. S., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+rheological+characterization+of+a+gel+form+of+the+porcine+urinary+bladder+matrix.">Preparation and rheological characterization of a gel form of the porcine urinary bladder matrix.</a> Biomaterials. 29, 1630-1630 (2008).</li><li>Gilbert, T. W., Sellaro, T. L., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decellularization+of+tissues+and+organs.">Decellularization of tissues and organs.</a> Biomaterials. 27, 3675-3675 (2006).</li><li>Liao, J., Joyce, E. M., Sacks, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+decellularization+on+the+mechanical+and+structural+properties+of+the+porcine+aortic+valve+leaflet.">Effects of decellularization on the mechanical and structural properties of the porcine aortic valve leaflet.</a> Biomaterials. 29, 1065-1065 (2008).</li><li>Singelyn, J. M., DeQuach, J. A., Seif-Naraghi, S. B., Littlefield, R. B., Schup-Magoffin, P. J., Christman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Naturally+derived+myocardial+matrix+as+an+injectable+scaffold+for+cardiac+tissue+engineering.">Naturally derived myocardial matrix as an injectable scaffold for cardiac tissue engineering.</a> Biomaterials. 30, 5409-5409 (2009).</li><li>Christman, K. L., Vardanian, A. J., Fang, Q., Sievers, R. E., Fok, H. H., Lee, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injectable+fibrin+scaffold+improves+cell+transplant+survival,+reduces+infarct+expansion,+and+induces+neovasculature+formation+in+ischemic+myocardium.">Injectable fibrin scaffold improves cell transplant survival, reduces infarct expansion, and induces neovasculature formation in ischemic myocardium.</a> J Am Coll Cardiol. 44, 654-654 (2004).</li><li>Christman, K. L., Fok, H. H., Sievers, R. E., Fang, Q., Lee, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibrin+glue+alone+and+skeletal+myoblasts+in+a+fibrin+scaffold+preserve+cardiac+function+after+myocardial+infarction.">Fibrin glue alone and skeletal myoblasts in a fibrin scaffold preserve cardiac function after myocardial infarction.</a> Tissue Eng. 10, 403-410 (2004).</li><li>Huang, N. F., Sievers, R. E., Park, J. S., Fang, Q., Li, S., Lee, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rodent+model+of+myocardial+infarction+for+testing+the+efficacy+of+cells+and+polymers+for+myocardial+reconstruction.">A rodent model of myocardial infarction for testing the efficacy of cells and polymers for myocardial reconstruction.</a> Nat Protoc. (1), 1596-1609 (2006).</li><li>Ott, H. C., Matthiesen, T. S., Goh, S. K., Black, L. D., Kren, S. M., Netoff, T. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion-decellularized+matrix:+using+nature's+platform+to+engineer+a+bioartificial+heart.">Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart.</a> Nat Med. 14, 213-221 (2008).</li><li>Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+extracellular+matrix+as+a+biologic+scaffold+material.">The extracellular matrix as a biologic scaffold material.</a> Biomaterials. 28, 3587-3593 (2007).</li></ol>

このメソッドは、心筋組織工学のための生物学的に誘導された、注射足場の世代を可能にする。これらのメソッドが最初に製造するため、心筋マトリックスゲルの in vivo試験で開発され、心膜マトリックスのゲルをここに提示されたが、このプロトコルは、組織が ​​適切に脱細胞することができます提供されて、どんな組織で使用するために適応させることができる。細胞化?...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Reagents:</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Pepsin</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>p6887-1G</td>
<td>Lyophilized </td>
</tr>
<tr>
<td>Biotin</td>
<td>&nbsp;</td>
<td>Thermo Scientific</td>
<td>21217</td>
<td>&nbsp;</td>
<tr>
<td>Neutravidin-HRP</td>
<td>&nbsp;</td>
<td>Thomas Scientific</td>
<td>21130</td>
<td>&nbsp;</td>
<tr>
<td>Equipment: </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Wiley Mini Mill</td>
<td>&nbsp;</td>
<td>Thomas Scientific</td>
<td>3383L10</td>
<td>&nbsp;</td>
<tr>
<td>Labconco Lyophilizer</td>
<td>&nbsp;</td>
<td>Labconco, Inc</td>
<td>7670520</td>
<td>&nbsp;</td>
<tr>
<td>Surgical supplies:</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Betadine</td>
<td>&nbsp;</td>
<td>Purdue Products, L.P.</td>
<td>67618-154-16</td>
<td>&nbsp;</td>
<tr>
<td>Lactated Ringers Solution</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>003966</td>
<td>&nbsp;</td>
<tr>
<td>KY Jelly</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>28658</td>
<td>&nbsp;</td>
<tr>
<td>Lidocaine, 2%</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>17767</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine hydrochloride</td>
<td>&nbsp;</td>
<td>Reckitt Benckiser Healthcare (UK) Ltd.</td>
<td>12496-0757-1</td>
<td>&nbsp;</td>
<tr>
<td>Artificial tear ointment</td>
<td>&nbsp;</td>
<td>Fisher </td>
<td>NC9860843</td>
<td>&nbsp;</td>
<tr>
<td>Triple antibiotic ointment</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>19082795</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>60307-120-25</td>
<td>&nbsp;</td>
<tr>
<td>Otoscope </td>
<td>&nbsp;</td>
<td>MWI</td>
<td>008699</td>
<td>&nbsp;</td>
<tr>
<td>Stop cock</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>006245</td>
<td>&nbsp;</td>
<tr>
<td>3-0 Vicrile suture</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>J327H</td>
<td>&nbsp;</td>
<tr>
<td>5-0 Proline suture</td>
<td>&nbsp;</td>
<td>MWI</td>
<td>s-1173</td>
<td>&nbsp;</td>
<tr>
<td>Reverse cutting (RC) needle</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>8684G</td>
<td>&nbsp;</td>
<tr>
<td>Microhemostats </td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>13013-14</td>
<td>&nbsp;</td>
<tr>
<td>Rat tooth microforceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11084-07</td>
<td>&nbsp;</td>
<tr>
<td>No. 10 scalpel</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>10110-01</td>
<td>&nbsp;</td>
<tr>
<td>Blunt scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14108-09</td>
<td>&nbsp;</td>
<tr>
<td>Sharp, curved scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14085-08</td>
<td>&nbsp;</td>
<tr>
<td>Large, serrated forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>1106-12</td>
<td>&nbsp;</td>
<tr>
<td>PE160 suction tubing</td>
<td>&nbsp;</td>
<td>BD</td>
<td>427430 </td>
<td>&nbsp;</td>
<tr>
<td>Clippers </td>
<td>&nbsp;</td>
<td>MWI</td>
<td>21608</td>
<td>&nbsp;</td>
<tr>
<td>Skin staples/stapler </td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>PRR35</td>
<td>&nbsp;</td>
<tr>
<td>General supplies:</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Stir plates</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>0.1 M HCl</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1 M NaOH</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>10x PBS</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1x PBS</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>70% Ethanol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>0.1 mL syringes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>10 mL syringe</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Q-tips</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Surgical glue</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Surgical drape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Towel clamps</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Small hand-held vacuum </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

fabrication of biologically derived injectable materials for myocardial tissue engineering

このプロトコルは、心筋組織の工学的応用のための注射細胞外マトリックス（ECM）ゲルの調製のためのメソッドを提供します。簡単に言えば、脱細胞組織は、凍結粉砕、酵素的に消化し、生理的pHにもたらされる。凍結乾燥は、小さなミルで微粉末に粉砕することができる乾燥したECMで、その結果、組織からすべての水分を取り除きます。粉砕後、ECMの粉末を注射マトリックスを形成するために、ペプシンで消化する。 pH7.4に調整した後、液体マトリックス材料は、心筋内に注入することができます。前の特性評価試験の結果は、脱細胞心膜と心筋組織から生成行列のゲルは多様なタンパク質、ペプチドおよびグリコサミノグリカンを含むネイティブなECM成分を、保持することが示されている。組織工学のためのこの物質の使用を考えると、in vivoでの特性評価に特に有用であり、ここ、左心室に壁内注射を行う方法（LV）自由壁がで行列のゲルへのホストの応答を分析する手段として提示され小動物モデル。胸腔へのアクセスは、ダイヤフラムを介して得ていると注射は少しLV自由壁の頂部の上に作られています。生物由来スキャフォールドは、注射の前にビオチン標識によって視覚化し、ホースラディッシュペルオキシダーゼ結合ニュートラビジンで組織切片を染色し、ジアミノベンジジン（DAB）染色を介して可視化することができます。注入領域の解析も組織学的および免疫組織化学的染色を行うことができます。この方法では、以前に調べた心膜と心筋のマトリックスのゲルは、繊維状、多孔質ネットワークを形成し、注入の領域内の血管形成を促進することが示された。

Watch this Scientific Journal Video about Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering at JoVE.com

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

脱細胞組織から注射マトリックスのゲルを調製し、ラット心筋にそれを注入するための方法<em&gt; in vivoで</em&gt;説明して​​います。

心筋組織工学のための生物由来注射用材料の作製

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

Research

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生体力学的解析と組織工学のための細胞由来の組織のリングを製作する監督セルラーセルフアセンブリ

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do ...

Postproduction Processing of Electrospun Fibres for Tissue Engineering

組織工学のためのエレクトロ繊維のポスト処理

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

Capillary Force Lithography for Cardiac Tissue Engineering

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Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

エンジニアリング3D細胞化血管組織再生のためのコラーゲンゲル

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Design and Construction of Artificial Extracellular Matrix (aECM) Proteins from Escherichia coli for Skin Tissue Engineering

Design and Construction of Artificial Extracellular Matrix aECM Proteins from Escherichia coli for Skin Tissue Engineering

人工細胞外マトリックス（AECM）タンパク質からの設計と構築<em&gt;エシェリヒア·コリ</em&gt;皮膚組織工学のための

Recombinant technology is a versatile platform to create novel artificial proteins with tunable properties. For the last decade, many artificial proteins ...

Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer (SALB) Method

Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer SALB Method

溶剤アシスト脂質二重（SALB）法による生体膜の作製

In order to mimic cell membranes, the supported lipid bilayer (SLB) is an attractive platform which enables in vitro investigation of membrane-related ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

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

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

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 Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

組織特異的細胞培養および配信プラットフォームとして、細胞外マトリックス由来のフォームおよびマイクロキャリアの作製

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering

組織工学のための血管新生単位としてマウスの脂肪組織由来の微小血管フラグメントの単離

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several ...