Name: procedure for human saphenous veins ex vivo perfusion and external reinforcement
Uploaded: 2014-10-01T15:00:00
Description: Watch this Scientific Journal Video about Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement at JoVE.com

Dette arbejde blev støttet af tilskud fra SNF [31003A-138528] har Octav og Marcella Botnar Fonden, Novartis Foundation og Emma Muschamp Foundation. Vi takker Martine Lambelet, og Jean-Christophe Stehle for deres fremragende teknisk bistand.

Den Etiske Komité Lausanne Universitet godkendte forsøg, som er i overensstemmelse med principperne i Helsinki-erklæringen fra 1975, som revideret i 1983 for brugen af ​​humane væv. 1. Menneskelige Store saphenavene Harvest <ol><li> Opnå overskydende segmenter af ikke-åreknuder humane saphenous vener fra patienter, der gennemgår underekstremitet bypass operation for iskæmi. I operationsstuen, desinficere hele benet med en jodopløsning og drapere patienten at eksponere benet fr...

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O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reprinted+article+&amp;#34;Pathophysiology+of+vein+graft+failure:+a+review&amp;#34;.">Reprinted article &amp;#34;Pathophysiology of vein graft failure: a review&amp;#34;.</a> European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery. 42, S19-S29 (2011).</li><li>Berard, X., 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=Role+of+hemodynamic+forces+in+the+ex+vivo+arterialization+of+human+saphenous+veins.">Role of hemodynamic forces in the ex vivo arterialization of human saphenous veins.</a> Journal of Vascular Surgery. 57, 1371-1382 (2013).</li><li>Vijayan, V., 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=Long-term+reduction+of+medial+and+intimal+thickening+in+porcine+saphenous+vein+grafts+with+a+polyglactin+biodegradable+external+sheath.">Long-term reduction of medial and intimal thickening in porcine saphenous vein grafts with a polyglactin biodegradable external sheath.</a> Journal of Vascular Surgery. 40, 1011-1019 (2004).</li><li>Jeremy, J. Y., 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=On+the+biology+of+saphenous+vein+grafts+fitted+with+external+synthetic+sheaths+and+stents.">On the biology of saphenous vein grafts fitted with external synthetic sheaths and stents.</a> Biomaterials. 28, 895-908 (2007).</li><li>Zilla, P., 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=Constrictive+external+nitinol+meshes+inhibit+vein+graft+intimal+hyperplasia+in+nonhuman+primates.">Constrictive external nitinol meshes inhibit vein graft intimal hyperplasia in nonhuman primates.</a> The Journal of Thoracic and Cardiovascular Surgery. 136, 717-725 (2008).</li><li>Zilla, P., 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=Utilization+of+shape+memory+in+external+vein-graft+meshes+allows+extreme+diameter+constriction+for+suppressing+intimal+hyperplasia:+a+non-human+primate+study.">Utilization of shape memory in external vein-graft meshes allows extreme diameter constriction for suppressing intimal hyperplasia: a non-human primate study.</a> Journal of Vascular Surgery. 49, 1532-1542 (2009).</li><li>Yeoman, M. S., 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=A+constitutive+model+for+the+warp-weft+coupled+non-linear+behavior+of+knitted+biomedical+textiles.">A constitutive model for the warp-weft coupled non-linear behavior of knitted biomedical textiles.</a> Biomaterials. 31, 8484-8493 (2010).</li><li>Longchamp, 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=The+use+of+external+mesh+reinforcement+to+reduce+intimal+hyperplasia+and+preserve+the+structure+of+human+saphenous+veins.">The use of external mesh reinforcement to reduce intimal hyperplasia and preserve the structure of human saphenous veins.</a> Biomaterials. 35, 2588-2599 (2014).</li><li>Saucy, F., 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=Ex+vivo+pulsatile+perfusion+of+human+saphenous+veins+induces+intimal+hyperplasia+and+increased+levels+of+the+plasminogen+activator+inhibitor+1.">Ex vivo pulsatile perfusion of human saphenous veins induces intimal hyperplasia and increased levels of the plasminogen activator inhibitor 1.</a> European Surgical Research. Europaische Chirurgische Forschung. Recherches Chirurgicales Europeennes. 45, 50-59 (2010).</li><li>Dubuis, C., 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=Atorvastatin-loaded+hydrogel+affects+the+smooth+muscle+cells+of+human+veins.">Atorvastatin-loaded hydrogel affects the smooth muscle cells of human veins.</a> The Journal of pharmacology and experimental. 347, 574-581 (2013).</li><li>Deglise, S., 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=Increased+connexin43+expression+in+human+saphenous+veins+in+culture+is+associated+with+intimal+hyperplasia.">Increased connexin43 expression in human saphenous veins in culture is associated with intimal hyperplasia.</a> Journal of Vascular Surgery. 41, 1043-1052 (2005).</li><li>Muto, A., Model, L., Ziegler, K., Eghbalieh, S. 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Denne undersøgelse afdækker en ex vivo vene perfusionssystem (EVPs) for at udføre omfattende hæmodynamiske studier i menneskelige årer. Dette system giver saphenous vener perfusion under definerede hæmodynamiske parametre i fravær af skærpende inflammatoriske og vækstfaktorer frigivet af cirkulerende celler in vivo. Således er det giver en bedre forståelse af de underliggende veje er involveret i kontrollen af IH i menneskelige vener podninger 5,11,12,15. <p class="jove_conten...

Kardiovaskulære sygdomme er den førende årsag til sygelighed og dødelighed i de vestlige lande 1. Trods fremskridt i endovaskulære behandlinger, bypass operation stadig grundpillen i moderne behandlingsformer, således over en halv million vene grafts udføres årligt i USA. Men på trods af årtiers forskning, 30-60% af lavere ekstremitet vene podninger mislykkes inden for de første år på grund af intimahyperplasi (IH) 2. Mekaniske kræfter, især lav forskydningsspænding (SS) og høj mur spænding, er omdrejningspunktet i indledningen og udviklingen af denne hyperplastisk respons 3,4. For at løse dette problem, blev en ex vivo vener perfusionssystem (EVPs), der genereres for at studere under strengt kontrollerede hæmodynamiske forhold (tryk og shear stress), adfærd menneskelige saphenous vener. I denne undersøgelse efter indsætning i det arterielle-lignende omsætning, højt tryk (middelværdi = 100 mm Hg) var tilstrækkeligt til at stimulere spredningsrelateredebejde og migrering af glatte muskelceller i det intimale lag (IH) 5. Mammale undersøgelser har foreslået brug af ekstern forstærkning som en effektiv metode til at støtte &quot;arterialized vene&quot;, og modvirke den akutte hæmodynamiske ændrer vene ansigter engang implanteret i en arteriel miljø. Redskabets forhindret over-udspiling, forøget forskydningsspænding og reduceret vægspænding og dermed IH 6-10. Men de underliggende mekanismer og dens anvendelse på de menneskelige vener at forbedre bypass åbenhed er ikke fuldt karakteriseret. Vores EVPs blev brugt til at sammenligne, i stand efterligne ændringer en vene ansigter engang indsat i en arteriel regime (høj forskydning stress og pres), adfærd menneskelige saphenous vener i fravær og nærvær af en ekstern makroporoes polyester rørformet mesh. Ved at forhindre patologisk remodeling og landbrugsredskaber masken fremlagde dokumentation for sin potentielle kliniske effektivitet 11 </sup&gt;. Denne undersøgelse 1) indfører en model af ex vivo humane saphenous vener perfusion under kontrolleret tryk og shear stress 2) viser, at ekstern makroporøs mesh polyester reducerer IH og indeholder afgørende information til dens potentielle kliniske anvendelse.

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			<td>Fisher Scientific</td>
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			<td>Idex Health &#38; Science GMBH</td>
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			<td>Sigma-Aldrich</td>
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			<td>BD Bioscence</td>
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			<td>BD Bioscence</td>
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			<td>Idex Health &#38; Science GMBH</td>
			<td>SM 895&#160;</td>
			<td>&#160;App-Nr 03736-00194</td>
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			<td height="21" style="height:21px;">Pump Head</td>
			<td>Idex Health &#38; Science GMBH</td>
			<td>MI0008&#160;</td>
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			<td height="21" style="height:21px;">Monitoring Kit TRANSPAC IV</td>
			<td>icumedical</td>
			<td>011-0J736-01</td>
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			<td>B. Braun Medical SA</td>
			<td>4612041-02</td>
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			<td height="21" style="height:21px;">Etibon 3-0 FS-2</td>
			<td>Ethicon- Johnson&#38;Johnson</td>
			<td>EH7346H</td>
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			<td height="21" style="height:21px;">Mesh ProVena 6-8mm</td>
			<td>B. Braun Medical SA</td>
			<td>1105012-14</td>
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			<td height="21" style="height:21px;">NaCl: Sodium Chlorure Solution perfusion 0.9% (100 ml)</td>
			<td>B. Braun Medical SA</td>
			<td>534534</td>
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			<td>Cole-Parmer Instrument Co</td>
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			<td>National Instruments</td>
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			<td>Transonic Systems Inc.</td>
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			<td>BD Connexta Luerlock</td>
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procedure for human saphenous veins ex vivo perfusion and external reinforcement

Grundpillen i moderne behandlinger for omfattende okklusiv arteriel sygdom er venøs bypass. Imidlertid er dens holdbarhed truet af intimahyperplasi (IH), der til sidst fører til tillukning af blodkar og organsvigt. Mekaniske kræfter, særlig lav forskydningsspænding og høj mur spænding, menes at indlede og opretholde disse cellulære og molekylære forandringer, men deres nøjagtige bidrag stadig at blive opklaret. For selektivt at vurdere betydningen af tryk og shear stress på biologi IH blev en ex vivo perfusionssystem (EVPs) oprettet for at perfundere segmenter af humane saphenous vener under arteriel regime (high shear stress og højt tryk). Yderligere tekniske nyskabelser tillod samtidig perfusion af to segmenter fra samme vene, en forstærket med en ekstern mesh. Vener blev høstet ved hjælp af en no-touch teknik og straks overført til laboratoriet til montage i EVPs. Et segment af frisk isolATED vene ikke blev perfuseret (kontrol, dag 0). De to andre segmenter blev perfunderet i op til 7 dage, idet én helt i læ med en 4 mm (diameter) ekstern mesh. Det tryk, flow hastighed og puls blev løbende overvåget og justeret for at efterligne de hæmodynamiske forhold i den femorale arterie. Ved afslutningen af ​​perfusionen blev vener afmonteres og bruges til histologiske og molekylære analyser. Under ex vivo-betingelser, højtryk perfusion (arterielt, middelværdi = 100 mm Hg) er tilstrækkelig til at generere IH og ombygning af menneskelige årer. Disse ændringer er reduceret i nærværelse af en ekstern mesh polyester.

Den EVPs giver et værdifuldt redskab til selvstændigt at vurdere de hæmodynamiske kræfter på menneskers saphenusvene podekviste remodeling og IH. Figur 1 viser perfusionskammeret og venen støtte. I figur 1A og B, venen støtte før (figur 1A) og efter (figur 1B) samling, henholdsvis, er afbildet. Det er sammensat af (fra top til bund) af en glat rør af rustfrit stål måler 9 cm, som tjener som en und...

Watch this Scientific Journal Video about Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement at JoVE.com

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

De mekanismer, der fører til udviklingen af ​​intimahyperplasi (IH) og venegraft svigt stadig dårligt forstået. Denne undersøgelse beskriver en ex vivo-system til at perfundere menneskelige vener under styret strømning og tryk. Desuden effektiviteten af ​​ekstern netforstærkning at begrænse udviklingen af ​​IH blev evalueret.

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Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

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