Name: ferromagnetic bare metal stent for endothelial cell capture and retention
Uploaded: 2015-09-18T15:00:00
Description: Watch this Scientific Journal Video about Ferromagnetic Bare Metal Stent for Endothelial Cell Capture and Retention at JoVE.com

The authors thank Tyra Witt, Cheri Mueske, Brant Newman and Dr. Peter J. Psaltis, MBBS, PhD for their valuable contributions. This study was financially supported by European Regional Development Fund &#8211; FNUSA-ICRC (No. CZ.1.05/1.100/02.0123), American Heart Association Scientist Development Grant (AHA #06-35185N), National Institutes of Health (T32HL007111) and The Grainger Innovation Fund &#8211; Grainger Foundation.

Alle dyrestudier ble godkjent av Institutional Animal Care og Utnyttelse Committee (IACUC) ved Mayo Clinic. 1. Design og analyse av et 2 205 Stainless Steel Stent <ol><li> Utforme en bart metall stent ved hjelp av DAK <ol><li> Gjør en ekstrudert hul sylinder ved å velge på &#39;ekstrudert boss / base-funksjonen med veggtykkelse lik stent strut tykkelse. </li><li> Designe en stent mønster på en annen skisse plan tangentialt til den ekstruderte sylinder. Gjøre bredden av det flate møn...

<ol>
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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coronary+stents:+current+status.">Coronary stents: current status.</a> J Am Coll Cardiol. 56, 1-42 (2010).</li><li>Austin, D., 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=Drug-eluting+stents+versus+bare-metal+stents+for+off-label+indications:+a+propensity+score-matched+outcome+study.">Drug-eluting stents versus bare-metal stents for off-label indications: a propensity score-matched outcome study.</a> Circ Cardiovasc Interv. 1 (1), 45-52 (2008).</li><li>Polyak, 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=High+field+gradient+targeting+of+magnetic+nanoparticle-loaded+endothelial+cells+to+the+surfaces+of+steel+stents.">High field gradient targeting of magnetic nanoparticle-loaded endothelial cells to the surfaces of steel stents.</a> P Natl Acad Sci USA. 105 (2), 698-703 (2008).</li><li>Tassiopoulos, A. 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Vi utviklet et magnetisk stent som kan fungere som en bart metall stent og kan tiltrekke Spion-merket endotelceller. I tidligere studier med magnetiske stenter, har forskerne brukt nikkel belagt kommersielle stenter og spoler eller masker laget av magnetiske materialer på grunn av utilgjengelighet av en ferromagnetisk stent 5,10-14. Andre grupper har også brukt det paramagnetiske natur kommersielt tilgjengelige 304-grade rustfritt stål stenter for målretting nanopartikkel lastet endotelceller 3.</sup...

Patients implanted with vascular stents manufactured from thrombogenic materials like stainless steel, cobalt chromium, and platinum chromium &#8211; both bare metal stents (BMS) and drug eluting stents (DES) &#8211; need anti-platelet therapy to prevent thrombus formation. BMS heal rapidly, but are subject to late stage restenosis due to incomplete healing. DES require long term anti-platelet therapy due to delayed healing. Anti-platelet therapy administered to avoid thrombosis as a result of incomplete or delayed healing leads to increased bleeding risk and may not be suitable for certain patients1,2. An ideal stent will heal completely and quickly thus avoiding long-term anti-platelet therapy and late stage restenosis. This complete healing can only be achieved if the stent is rapidly coated with a monolayer of endothelial cells after implantation. Coating the stents with biocompatible materials such as gold or other biopolymers has been shown to improve thrombo-resistance, but none of these techniques achieved ideal blood compatibility as may be possible by coating with endothelial cells3,4.
A stent can be coated with endothelial cells post implantation by attracting circulating progenitor cells. This self-seeding technique can be achieved by utilizing ligands and antibodies. But this technique is limited by the low number of circulating endothelial progenitor cells. A promising strategy is to deliver cells directly to the stent immediately following implantation during a short period of blood flow occlusion3,5. This strategy requires a technique for rapidly capturing cells and retaining them on the stent even after restoring blood flow. We have developed a technique in which a magnetic stent is used to attract and retain magnetically-labeled endothelial cells delivered post implantation. To achieve this, a functional BMS with sufficient magnetic properties to capture and retain magnetically-labeled endothelial cells is required6.
In this paper, we discuss the methods for designing, manufacturing, and testing a 2205 stainless steel stent. The stents were designed using CAD and FEA. The manufactured stents were magnetized using a neodymium magnet and the retained magnetic field was measured using a magneto-resistance microsensor probe. We then tested the stents for magnetically-labeled cell capture in a culture dish during our in-vitro experiments. Finally, the stents were tested in-vivo by implanting magnetic and non-magnetic stents in 4 pigs and histologically analyzing the stented arteries.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>2205 Stainless steel</td>
			<td>Carpenter Technology Corporation</td>
			<td>N/A</td>
			<td>Round bar stock material</td>
		</tr>
		<tr>
			<td>Abaqus</td>
			<td>Dassault systems</td>
			<td>N/A</td>
			<td>Software</td>
		</tr>
		<tr>
			<td>Atropine</td>
			<td></td>
			<td></td>
			<td>Prescription drug.</td>
		</tr>
		<tr>
			<td>Clopidogrel</td>
			<td></td>
			<td></td>
			<td>Commercial name: Plavix. Prescription drug.</td>
		</tr>
		<tr>
			<td>CM-DiI</td>
			<td>Life Technologies</td>
			<td>V-22888</td>
			<td>Molecular Probes, Eugene, OR</td>
		</tr>
		<tr>
			<td>Endothelial growth medium-2</td>
			<td>Lonza</td>
			<td>CC-3162</td>
			<td></td>
		</tr>
		<tr>
			<td>Hand Held Crimping tool</td>
			<td>Blockwise engineering</td>
			<td>M1-RMC</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric acid (HCl)</td>
			<td>Sigma Aldrich</td>
			<td>MFCD00011324</td>
			<td>CAUTION: wear proptective equipment and handle under fume hood</td>
		</tr>
		<tr>
			<td>Isoflurane anesthesia</td>
			<td>Piramal Critical Care, Inc.&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>Sigma Aldrich</td>
			<td>MFCD00011674</td>
			<td></td>
		</tr>
		<tr>
			<td>NdFeB magnet 2&#34; Dia x 1&#34; thick</td>
			<td>Amazing magnets</td>
			<td>D1000P</td>
			<td>Axially magnetized disc magnet with poles on flat faces</td>
		</tr>
		<tr>
			<td>Over-The-Wire trifold balloon</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Any commercially available OTW trifold balloon can be used</td>
		</tr>
		<tr>
			<td>Phosphate buffered saline</td>
			<td>Life Technologies</td>
			<td>10010-023</td>
			<td>Commonly known as PBS</td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate (NaHCO3)</td>
			<td>Sigma Aldrich</td>
			<td>MFCD00003528</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium pentobarbital</td>
			<td>Zoetis</td>
			<td></td>
			<td>Commercial Name: Sleepaway (26%), FatalPlus, Beuthanasi.&#160; Controlled substance to be ordered only by licensed veternarian</td>
		</tr>
		<tr>
			<td>SolidWorks</td>
			<td>Dassault systems</td>
			<td>N/A</td>
			<td>Software</td>
		</tr>
		<tr>
			<td>SpinTJ-020 micro sensor</td>
			<td>MicroMagneitcs Sensible Solutions</td>
			<td>N/A</td>
			<td>Long probe STJ-020 microsensor</td>
		</tr>
		<tr>
			<td>SPION</td>
			<td>Mayo Clinic</td>
			<td>N/A</td>
			<td>Nanoparticles synthesized internally (Ref: Lee, S. J. et al. Nanoparticles of magnetic ferric oxides encapsulated with poly(D,L latide-co-glycolide) and their applications to magnetic resonance imaging contrast agent. J Magn Magn Mater 272, 2432-2433, doi:DOI 10.1016/j.jmmm.2003.12.416 (2004))</td>
		</tr>
		<tr>
			<td>Telazol</td>
			<td>Zoetis</td>
			<td></td>
			<td>Controlled substance to be ordered only by licensed veternarian</td>
		</tr>
		<tr>
			<td>Trypsin EDTA</td>
			<td>Life Technologies</td>
			<td>25200-056</td>
			<td>Gibco, Grand Island, NY</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Bayer Animal Health</td>
			<td></td>
			<td>Commercial name: Rompun. Controlled sunstance to be ordered only by a licensed veternarian</td>
		</tr>
	</tbody>
</table>

ferromagnetic bare metal stent for endothelial cell capture and retention

Rapid endotelialisering av kardiovaskulære stenter er nødvendig for å redusere stenttrombose og for å unngå platehemmende behandling som kan redusere blødningsrisiko. Muligheten for å bruke magnetiske krefter for å fange opp og holde på endotel-celler utvekst (EOC) merket med superparamagnetiske jernoksid nanopartikler (Spion) har blitt vist tidligere. Men denne teknikken krever utviklingen av en funksjonell mekanisk stent fra et magnetisk og biokompatibelt materiale, etterfulgt av in-vitro og in-vivo-test for å bevise hurtig endotelialisering. Vi utviklet et svakt ferromagnetisk stent fra 2 205 duplex rustfritt stål ved hjelp av dataassistert konstruksjon (DAK) og dens design ble videreutviklet ved hjelp av finite element analyse (FEA). Den endelige utforming av stenten oppviste en hovedstamme under bruddgrensen for materialet under mekanisk krymping og ekspansjon. Ett hundre stenter ble fremstilt, og en undergruppe av dem ble brukt for mekanisk testing, retained magnetiske feltmålinger, in-vitro celle fangst studier, og in-vivo implantasjon studier. Ti stenter ble testet for utplassering for å verifisere om de vedvarende krymping og utvidelse syklus uten feil. En annen 10 stenter ble magnetisert ved hjelp av en sterk neodymium magnet og deres beholdt magnetfelt ble målt. Stentene viste at det tilbakeholdt magnetisme var tilstrekkelig til å fange Spion merkede EOC i våre in vitro-studier. Spion-merket EOC fangst og oppbevaring ble bekreftet i store dyremodeller ved å implantere en magnetisert stent og en ikke-magnetisert kontroll stent i hver av 4 griser. Stentet arteriene ble Eksplanterte etter 7 dager og analysert histologisk. De svakt magnetiske stenter utviklet i denne studien var i stand til å tiltrekke seg og beholde Spion-merket endotelceller som kan fremme rask helbredelse.

<html> Iterativ stent design basert på FEA (figur 1) viste en stent som kan klemme og ekspandere med en hovedstamme på 20% som er mindre enn 30% bruddforlengelsen. Krympe- og ekspansjon test (figur 2) viste ingen tegn på brudd. Bilder av deformert stent viste god avtale med FEA beregnede deformasjoner og også mikros bildene viste ingen brudd (figur 3). Som forventet fra de tilbakeholdte magnetiske feltmålinger (figurene 4 og 5), ble Spion-merkede ce...

Watch this Scientific Journal Video about Ferromagnetic Bare Metal Stent for Endothelial Cell Capture and Retention at JoVE.com

Ferromagnetic Bare Metal Stent for Endothelial Cell Capture and Retention

Våre mål var å designe, produsere og teste ferromagnetiske stenter for endotelceller fangst. Ti stenter ble testet for brudd og 10 flere stenter ble testet for beholdt magnetisme. Til slutt ble 10 stenter testet in-vitro og 8 flere stenter ble implantert i 4 griser for å vise celle fangst og oppbevaring.

Ferromagnetisk Bare Metal Stent for endotelcelle Capture og oppbevaring

Rapid endothelialization of cardiovascular stents is needed to reduce stent thrombosis and to avoid anti-platelet therapy which can reduce bleeding risk. ...

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Corneal endothelial cell cultures have a tendency to undergo epithelial-to-mesenchymal transition (EMT) after loss of cell-to-cell contact. EMT is ...

Mitigation of Blood Borne Cell Attachment to Metal Implants through CD47-Derived Peptide Immobilization

Reduksjon av blodbårne celletilbehør til metallimplantater gjennom CD47-avledet peptidimmobilisering

The key complications associated with bare metal stents and drug eluting stents are in-stent restenosis and late stent thrombosis, respectively. Thus, ...

Segmenting Growth of Endothelial Cells in 6-Well Plates on an Orbital Shaker for Mechanobiological Studies

Segmentering av vekst av endotelceller i 6-brønnsplater på en orbital shaker for mekanobiologiske studier

Shear stress imposed on the arterial wall by the flow of blood affects endothelial cell morphology and function. Low magnitude, oscillatory and ...

A Microphysiological System to Study Leukocyte-Endothelial Cell Interaction during Inflammation

Et mikrofysiologisk system for å studere leukocytt-endotelial celleinteraksjon under betennelse

Leukocyte-endothelial cell interactions play an important role in inflammatory diseases such as sepsis. During inflammation, excessive migration of ...