Name: scaling of engineered vascular grafts using 3d printed guides and the ring stacking method
Uploaded: 2017-03-27T11:30:00
Description: Watch this Scientific Journal Video about Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method at JoVE.com

笔者想感谢我们的同胞林的实验室同事阿马尔奇什蒂和Bijal帕特尔他们与一些组织学和细胞培养的实物援助。资金来自韦恩州立大学纳米医学奖学金（CBP）提供的启动资金和心血管病研究所种子格兰特（MTL）。

1.细胞培养制备<ol><li>利用商业购买的人主动脉平滑肌细胞。 </li><li>保持的88.6％由平滑肌细胞的生长培养基中的细胞231培养基，0.1％每种重组人胰岛素（RH-胰岛素），重组人成纤维细胞生长因子（RH-FGF），重组人表皮生长因子（RH-FGF）的，和抗坏血酸;和5％每胎牛血清（FBS）和L-谷氨酰胺;和1％抗生素/抗真菌剂。 注意：每个生长因子，FBS和L-谷氨酰胺购买作为血管培养基上生长套件。 </li><li...

<ol>
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A., Lahti, M. T., Johnson, S. L., Tranquillo, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantation+of+completely+biological+engineered+grafts+following+decellularization+into+the+sheep+femoral+artery.">Implantation of completely biological engineered grafts following decellularization into the sheep femoral artery.</a> Tissue Eng Part A. 20 (11-12), 1726-1734 (2014).</li><li>Syedain, Z. H., Meier, L. A., Bjork, J. W., Lee, A., Tranquillo, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantable+arterial+grafts+from+human+fibroblasts+and+fibrin+using+a+multi-graft+pulsed+flow-stretch+bioreactor+with+noninvasive+strength+monitoring.">Implantable arterial grafts from human fibroblasts and fibrin using a multi-graft pulsed flow-stretch bioreactor with noninvasive strength monitoring.</a> Biomaterials. 32 (3), 714-722 (2011).</li><li>Meier, L. 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=Blood+outgrowth+endothelial+cells+alter+remodeling+of+completely+biological+engineered+grafts+implanted+into+the+sheep+femoral+artery.">Blood outgrowth endothelial cells alter remodeling of completely biological engineered grafts implanted into the sheep femoral artery.</a> J Cardiovasc Transl Res. 7 (2), 242-249 (2014).</li><li>Pinnock, C. B., Meier, E. M., Joshi, N. N., Wu, B., Lam, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Customizable+engineered+blood+vessels+using+3D+printed+inserts.">Customizable engineered blood vessels using 3D printed inserts.</a> Methods. S1046-2023 (15), 30184-30185 (2015).</li><li>Blakely, A. M., Manning, K. L., Tripathi, A., Morgan, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bio-Pick,+Place,and+Perfuse:+A+New+Instrument+for+Three-Dimensional+Tissue+Engineering.">Bio-Pick, Place,and Perfuse: A New Instrument for Three-Dimensional Tissue Engineering.</a> Tissue Eng Part C Methods. 21 (7), 737-746 (2015).</li><li>Gwyther, T. 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=Engineered+vascular+tissue+fabricated+from+aggregated+smooth+muscle+cells.">Engineered vascular tissue fabricated from aggregated smooth muscle cells.</a> Cells Tissues Organs. 194 (1), 13-24 (2011).</li><li>Fearon, W. 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午夜凶铃堆叠方法提出过电流血管组织工程技术构建多重优势。该RSM可以适应通过简单地定制职位和外壳尺寸创建任意大小的人类的船只。我们的方法允许对无聚合物完全人类细胞并迅速降解在身体的自然伤口愈合过程中发现的载体材料组成的工程化血管的发展。聚合物移植物已知会引起再狭窄在诊所和如果包含在工程化的移植物可能成为问题。细胞接种数目需要修改每个不同尺寸的组织环。细?...

在治疗冠状动脉疾病（CAD）的，患者自身的血管被收获作为用于旁路手术移植物材料。然而，通常情况下，危重病人没有可行的船只捐赠给自己，在他们这样做的情况下，供体部位造成相当大的额外伤害，并有感染的严重风险。 1个工程血管移植物可以填补这方面的需求。可扩展性是用于工程船舶最重要的，以满足广泛的患者容器的尺寸要求。然而，对于工程船本发明的方法不容易可扩展的，并且通常需要复杂的模具或聚合物支架的再制造。大多数工程化移植物或者使用被接种的成纤维细胞的血管平滑肌或血管内皮细胞的聚合物管状支架;或滚动围绕心轴的细胞片以形成组织管。在临床试验中的两个工程血管移植物是基于脱细胞化ð聚合物ECM平台。 2，3，4可用于血管修复使用聚合物移植物已经公知的有通畅的问题，这可能会出现与具有持续的聚合物存在下，接枝的长期应用的一个主要问题。管状模具已被用于制造完全细胞容器，5，6，7，8，9，10，11，12，13，该程序将需要定制模具额外的设计和刀具制造，以生产各种尺寸的船只。 本文所描述的方法包括一种新颖的技术，用于创建易于扩展工程化血管移植物使用定制的3D印刷插片和传统的培养板中。 14个细胞接种到具有中心柱和外壳的插入板。后控制管腔直径，并允许细胞单层自组装到组织的环。该环的外壳厚度控制，最终容器的因而壁厚。然后完成组织环被堆叠以形成管状，血管移植物。这种方法的优点，被称为“环堆积的方法，”是任何粘附细胞类型可以接种到板设置和组织环或需要的所期望的应用的任何尺寸的管子可以通过简单地修改引导插入物来产生。在组织的组织工程创建环比较技术仍然难以规模，15，16，要求模具的再制造为每个所需的尺寸。此外，血管移植物使用这种方法，可以制造由在2-3周D，几周更快相比其他工程血管。 6对于诊所，这个时间差可以使在恶化患者的治疗中的显著差异。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Human Aortic Smooth Muscle Cells&#160;</td>
			<td>ATCC</td>
			<td>PCS-100-012</td>
			<td>vascular smooth muscle cells</td>
		</tr>
		<tr>
			<td>Medium 231</td>
			<td>Gibco (Life Technologies&#160;</td>
			<td>M-231-500</td>
			<td>media specific to vascular smooth muscle cells</td>
		</tr>
		<tr>
			<td>Human Aortic Smooth Muscle Cell Growth Kit&#160;</td>
			<td>ATCC</td>
			<td>PSC-100-042</td>
			<td>growth factors for maintaining vascular smooth muscle cell viability</td>
		</tr>
		<tr>
			<td>Replicator Mini 3D printer&#160;</td>
			<td>MakerBot&#160;</td>
			<td>N/A</td>
			<td>3D printer</td>
		</tr>
		<tr>
			<td>Poly(lactic acid) 3D ink (PLA)</td>
			<td>MakerBot&#160;</td>
			<td>N/A</td>
			<td>3D printer filament</td>
		</tr>
		<tr>
			<td>Poly(dimethlysiloxane) (PDMS)</td>
			<td>Ellworth Adhesives&#160;</td>
			<td>3097358-1004</td>
			<td>polymer for gluing plate parts</td>
		</tr>
		<tr>
			<td>Fibrinogen</td>
			<td>Hyclone Labratories, Inc.</td>
			<td>SH30256.01</td>
			<td>fibrin gel component</td>
		</tr>
		<tr>
			<td>Thrombin&#160;</td>
			<td>Sigma Life Sciences</td>
			<td>F3879-5G</td>
			<td>fibrin gel component</td>
		</tr>
		<tr>
			<td>Tranforming Growth Factor-Beta 1&#160;</td>
			<td>PeproTech</td>
			<td>100-21</td>
			<td>growth factor for stimulating collagen production</td>
		</tr>
		<tr>
			<td>Hemocytometer&#160;</td>
			<td>Hausser Scientific Co.</td>
			<td>3200</td>
			<td>for cell counting</td>
		</tr>
		<tr>
			<td>Polycarbonate tubing&#160;</td>
			<td>US Plastics&#160;</td>
			<td>PCTUB1.750X1.625</td>
			<td>material for making tall, ring stacking plates</td>
		</tr>
		<tr>
			<td>Polycarbonate sheet&#160;</td>
			<td>Home Depot</td>
			<td>409497</td>
			<td>material for making tall, ring stacking plates</td>
		</tr>
		<tr>
			<td>Adhesive polymer solvent&#160;</td>
			<td>SCIGRIP</td>
			<td>10799</td>
			<td>material for making tall, ring stacking plates</td>
		</tr>
		<tr>
			<td>Instron&#160; 5940</td>
			<td>Instron</td>
			<td>N/A</td>
			<td>tensile testing machine</td>
		</tr>
		<tr>
			<td>U-Stretch</td>
			<td>Cell Scale</td>
			<td>N/A</td>
			<td>tensile testing machine</td>
		</tr>
		<tr>
			<td>Smooth Muscle Actin&#160;</td>
			<td>MA5-11547</td>
			<td>Thermo Fisher</td>
			<td>antibody</td>
		</tr>
		<tr>
			<td>Tropomyosin</td>
			<td>MA5-11783</td>
			<td>Thermo Fisher</td>
			<td>antibody</td>
		</tr>
	</tbody>
</table>

scaling of engineered vascular grafts using 3d printed guides and the ring stacking method

冠状动脉疾病仍然死亡的主要原因，影响数百万美国人。随着缺乏可用的自体血管移植，移植工程提供住院治疗的巨大潜力。然而，工程化血管移植物通常是不容易扩展，因此需要定制模具或聚合物管的制造，以定制到不同的尺寸，构成一个费时和昂贵的做法。人体动脉的范围中管腔直径约2.0-38毫米，在约0.5-2.5毫米的壁厚。我们创建了一个方法，称为“环堆积法”，其中所需要的细胞类型的组织的大小可变环，这里演示血管平滑肌细胞（SMC），可以通过中心的职位导游来控制管腔直径创建壳和外壳口授血管壁的厚度。然后，这些组织环堆叠以创建管状结构，模仿血管的自然形式。该船长度be。通过简单的叠加构成所需的长度需要环的数量量身定做。与我们的技术中，管状形式，类似于血管，组织可以容易地在各种尺寸和长度的制造符合临床和患者的需要。

这里演示是3点不同的工程化血管移植物的尺寸（ 图1）的制造，显示出环堆积方法（RSM）是可伸缩的。为了证明适用性，所述3种不同的容器尺寸选择关联于实际的人类血管尺寸的左前降支（小;内腔直径= 4 MM）17，降主动脉（中间体;内腔直径= 10毫米），升主动脉（大;管腔直径= 20mm）的18。壁厚为小环约500微米，两者的中间?...

Watch this Scientific Journal Video about Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method at JoVE.com

Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method

可扩展的工程血管会提高临床适用性。使用容易可观3D印刷指南，创建并堆叠成管状形式，形成血管移植血管平滑肌的环。移植物的大小可通过简单地改变三维印刷导向大小，以满足人冠状动脉的尺寸范围。

工程血管移植缩放使用3D打印指南和环堆叠方法

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The kynurenine pathway and the tryptophan catabolites called kynurenines have received increased attention for their involvement in immune regulation and ...