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

Forfatterne vil gerne takke vores kollega Lam lab kolleger Ammar Chishti og Bijal Patel for deres venlige hjælp med nogle af histologi og cellekultur. Finansieringen blev leveret af Wayne State University Nanomedicin Fellowship (CBP), Start-Up fonde og Cardiovascular Research Institute Seed Grant (MTL).

1. Cell Culture Fremstilling <ol><li> Udnytte humane aorta glatmuskelceller købes kommercielt. </li><li> Opretholde celler i glatte muskelceller vækstmedier sammensat af 88,6% 231 medier, 0,1% hver af rekombinant human insulin (rH-insulin), rekombinant human fibroblastvækstfaktor (rH-FGF), rekombinant human epidermal vækstfaktor (rH-FGF), og ascorbinsyre; og 5% hver af føtalt bovint serum (FBS) og L-glutamin; og 1% antibiotisk / antimykotisk. Bemærk: Hver vækstfaktor, FBS og L-glutamin købes som en vask...

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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. 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The Ring Stacking Method præsenterer flere fordele i forhold til de nuværende vaskulære manipuleret væv konstruere teknikker. RSM kan tilpasses at skabe menneskelige fartøjer af enhver størrelse ved blot at tilpasse post- og ydre shell dimensioner. Vores fremgangsmåde muliggør udvikling af polymer-fri manipulerede fartøjer udelukkende består af humane celler og hurtigt nedbrydelige bæremateriale findes i kroppens naturlige sårheling proces. Polymer transplantater er kendt for at forårsage restenose i klinik...

I behandling af koronararteriesygdom (CAD), er en patientens egne blodkar høstes som implantatmateriale til bypass-kirurgi. Men oftentimes, syge patienter ikke har levedygtige fartøjer til at donere til sig selv, og i tilfælde, hvor de gør, donor websted forårsager betydelig yderligere skade og har en alvorlig risiko for infektion. 1 Udviklet vaskulære transplantater kunne fylde dette behov. Skalerbarhed er af allerstørste betydning for engineering fartøjer for at opfylde den brede vifte af patientens krav fartøj størrelse. Men de nuværende metoder til tekniske skibe er ikke let skalerbar, og kræver typisk reproduceres af komplekse forme eller polymerstativer. Mest manipuleret transplantater enten anvende en polymer rørformet stillads, der podes med vaskulære fibroblaster, glatte muskelceller eller endothelceller; eller rulle en celle ark omkring en dorn for at skabe et væv rør. To manipuleret kargrafter i kliniske forsøg er baseret på en decellularized polymer-ECM platform. 2, 3, 4 Polymer transplantater til rådighed til brug i vaskulær reparation er allerede kendt for at have problemer med åbenhed, som kan opstå som et stort problem med langsigtet anvendelse af et transplantat med en polymer til stede. Rørformede forme er blevet anvendt til at fabrikere helt cellulære fartøjer, 5, 6, 7, 8, 9, 10, 11, 12, 13, hvilke procedurer ville kræve ekstra design og værktøjsfremstilling for brugerdefinerede forme for at fremstille fartøjer i forskellige størrelser . Den heri beskrevne fremgangsmåde omfatter en hidtil ukendt teknik til at skabe let skalerbar manipuleret vaskulærtransplantater bruger tilpasses 3D trykte indsatser og traditionelle kultur plader. 14 Celler podes i plader med indsatse af en central post og ydre skal. De efterfølgende kontrol lumen diameter og tillader cellemonolaget til selv samle ind i en ring af væv. Den ydre skal kontrollen tykkelse af ringen, og således vægtykkelse af den endelige beholder. Tilbagelagt væv ringe derpå stablet til dannelse af en rørformet, vaskulært transplantat. Fordelen ved denne metode, betegnet &quot;Ring Stacking Method&quot;, er, at for vedhængende celletype kan podes ind i pladen opsætning og væv ringe eller rør af enhver størrelse nødvendig for den ønskede anvendelse kan genereres ved tilpasse guide skær. Komparative teknikker i vævsmanipulering Oprettelse ringe af væv er fortsat svært at skala, 15, 16 trænger genfremstilling af forme til hver ønsket størrelse. Derudover vaskulære transplantater fremstillet ved hjælp af denne metode kan producered i 2-3 uger, flere uger hurtigere i forhold til andre manipuleret fartøjer. 6 For klinikken, kan denne gang uoverensstemmelse gøre en væsentlig forskel i behandlingen af en forværret patient.

<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

Koronararteriesygdom fortsat en førende dødsårsag, der påvirker millioner af amerikanere. Med manglen på autologe vaskulære transplantater rådighed, manipuleret transplantater tilbyde et stort potentiale for patientbehandlingen. Men manipuleret vaskulære transplantater er generelt ikke let skalerbar, kræver fremstilling af brugerdefinerede forme eller polymer rør med henblik på at tilpasse til forskellige størrelser, som udgør en tidskrævende og bekostelig praksis. Humane arterier varierer i lumen diameter fra ca. 2,0 til 38 mm og i vægtykkelse fra ca. 0,5-2,5 mm. Vi har skabt en metode, kaldet den &quot;Ring Stacking Method&quot;, hvor variable størrelse ringe af væv af den ønskede celletype, demonstrerede her med vaskulære glatte muskelceller (SMC&#39;er), kan oprettes ved hjælp styrene på center stillinger for at kontrollere lumen diameter og ydre skaller at diktere godstykkelsen. Disse væv ringe er derefter stablet at skabe en rørformet konstruktion, der efterligner den naturlige form af et blodkar. Fartøjet Længden kan be skræddersyet ved blot at stable det antal ring, der kræves for at udgøre længden nødvendig. Med vores teknik, kan væv af rørformede typer, svarende til et blodkar, let fremstilles i forskellige dimensioner og længder for at opfylde behovene i klinikken og patienten.

Demonstreret her er fabrikation af 3 forskellige manipuleret vaskulære graft størrelser (figur 1), der viser, at Ring Stacking Method (RSM) er skalerbar. For at bevise anvendelighed, de 3 forskellige skibsstørrelser valgt korrelat til den faktiske menneskelige fartøj størrelse til venstre forreste nedadgående arterie (lille, lumen diameter = 4 mm) 17, faldende aorta (mellemprodukt lumen diameter = 10 mm) og aorta ascendens (stor; lumendiamet...

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

Skalerbar manipulerede blodkar ville forbedre klinisk anvendelighed. Brug af let betragtelige 3D-trykte vejledninger, blev ringe af vaskulær glat muskulatur skabt og stablet til en rørformet form, og danner et vaskulært transplantat. Transplantater kan være dimensioneret til at opfylde området af humane koronararterie dimensioner ved blot at ændre 3D-trykte vejledning størrelse.

Skalering af Engineered kargrafter Brug 3D Trykt Guides og Ring Stacking Method

Coronary artery disease remains a leading cause of death, affecting millions of Americans. With the lack of autologous vascular grafts available, ...

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Bioengineering

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Surgical Technique for the Implantation of Tissue Engineered Vascular Grafts and Subsequent In Vivo Monitoring

Surgical Technique for the Implantation of Tissue Engineered Vascular Grafts and Subsequent In Vivo Monitoring

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The development of Tissue Engineered Vessels (TEVs) is advanced by the ability to routinely and effectively implant TEVs (4-5 mm in diameter) into a large ...

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