Name: melt electrospinning writing of three-dimensional poly(&#949;-caprolactone) scaffolds with controllable morphologies for tissue engineering applications
Uploaded: 2017-12-23T13:00:00
Description: Watch this Scientific Journal Video about Melt Electrospinning Writing of Three-dimensional PolyÃŽÂµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications at JoVE.c

Dette arbejde er blevet økonomisk støttet af Cooperative Research Center CRC for celle terapi fremstilling, den australske Rådet ARC Forskningscenter i tilsætningsstof Biomanufacturing og Institute for Advanced Study af det tekniske universitet i München. Denne forskning blev foretaget af den australske forskning Rådet industrielle Transformation Training Centre i tilsætningsstof Biomanufacturing http://www.additivebiomanufacturing.org (IC160100026). Besøg webstedet for artikler, bøger, tv eller radio programmer, elektroniske medier eller andre litterære værker relateret til projektet. Yderligere, forfatterne taknemmeligt anerkender Maria Flandes Iparraguirre til støtte i optagelserne, Philip Hubbard for voice over og Luise Grossmann til at filme og redigere.

1. materielle forberedelse
<ol>
	<li>Fyld 2 g af PCL i en 3 mL plastik sprøjte med en tragt og indsætte et stempel i den åbne ende.</li>
	<li>Læg sprøjten i en forvarmet ovn ved 65 ° C til 8 h. punkt spidsen opad så luftbobler til samlede tæt på åbningen.</li>
	<li>Skubbe stemplet med en tynd genstand til at frigive den fanget luft inden for det smeltede materiale.</li>
	<li>Lad det køle ned til stuetemperatur, der opnås, når polymeren ikke er gennemsigtig længere efter 10 minutter.</li>
	<li>Opbevar PCL ...

<ol>
	<li>Muerza-Cascante, M. L., Haylock, D., Hutmacher, D. W., Dalton, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melt+Electrospinning+and+Its+Technologization+in+Tissue+Engineering.">Melt Electrospinning and Its Technologization in Tissue Engineering.</a> Tissue Eng Part B Rev. 21 (2), 187-202 (2015).</li><li>Brown, T. D., Dalton, P. D., Hutmacher, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melt+electrospinning+today:+An+opportune+time+for+an+emerging+polymer+process.">Melt electrospinning today: An opportune time for an emerging polymer process.</a> Prog. in pol Sci. , (2015).</li><li>Brown, T. D., Dalton, P. D., Hutmacher, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+writing+by+way+of+melt+electrospinning.">Direct writing by way of melt electrospinning.</a> Adv Mater. 23 (47), 5651-5657 (2011).</li><li>Farrugia, B. L., 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=Dermal+fibroblast+infiltration+of+poly(&#949;-caprolactone)+scaffolds+fabricated+by+melt+electrospinning+in+a+direct+writing+mode.">Dermal fibroblast infiltration of poly(&#949;-caprolactone) scaffolds fabricated by melt electrospinning in a direct writing mode.</a> Biofabrication. 5 (2), 025001 (2013).</li><li>Brown, T. 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=Melt+electrospinning+of+poly+(&#949;-caprolactone)+scaffolds:+Phenomenological+observations+associated+with+collection+and+direct+writing.">Melt electrospinning of poly (&#949;-caprolactone) scaffolds: Phenomenological observations associated with collection and direct writing.</a> Mater. Sci. Eng. C. 45, 698-708 (2014).</li><li>Jungst, T., 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=Melt+electrospinning+onto+cylinders:+effects+of+rotational+velocity+and+collector+diameter+on+morphology+of+tubular+structures.">Melt electrospinning onto cylinders: effects of rotational velocity and collector diameter on morphology of tubular structures.</a> Polym Int. 64 (9), 1086-1095 (2015).</li><li>Brown, T. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melt+electrospinning.">Melt electrospinning.</a> Chem Asian J. 6 (1), 44-56 (2011).</li><li>Wei, C., Gang, T. Q., Chen, L. J., Zhao, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+condition+for+the+transformation+from+Taylor+cone+to+cone-jet.">Critical condition for the transformation from Taylor cone to cone-jet.</a> Chin. Phys. B. 23 (6), 064702 (2014).</li><li>Mikl, 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=Discrete+viscous+threads.">Discrete viscous threads.</a> ACM Trans. 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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimensional+Metrology+of+Cell-matrix+Interactions+in+3D+Microscale+Fibrous+Substrates.">Dimensional Metrology of Cell-matrix Interactions in 3D Microscale Fibrous Substrates.</a> Procedia CIRP. 65, 32-37 (2017).</li><li>Muerza-Cascante, M. L., 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=Endosteal-like+extracellular+matrix+expression+on+melt+electrospun+written+scaffolds.">Endosteal-like extracellular matrix expression on melt electrospun written scaffolds.</a> Acta Biomater. 52, 145-158 (2017).</li><li>Delalat, 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=3D+printed+lattices+as+an+activation+and+expansion+platform+for+T+cell+therapy.">3D printed lattices as an activation and expansion platform for T cell therapy.</a> Biomaterials. 140, 58-68 (2017).</li><li>Bas, O., 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=Biofabricated+soft+network+composites+for+cartilage+tissue+engineering.">Biofabricated soft network composites for cartilage tissue engineering.</a> Biofabrication. 9 (2), 025014 (2017).</li><li>Hansske, 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=Via+precise+interface+engineering+towards+bioinspired+composites+with+improved+3D+printing+processability+and+mechanical+properties.">Via precise interface engineering towards bioinspired composites with improved 3D printing processability and mechanical properties.</a> J. 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Integrere AM for at finde innovative løsninger på udfordringerne på det medicinske område præsenterer et nyt paradigme for 21st århundrede. Den såkaldte "Bio-fabrikation" er på fremmarch og innovationer i fabrikation teknologi aktiverer produktionen af meget avancerede arkitekturer for TE applikationer. Electrospinning af polymer smelter i en direkte skrivning mode (her MEW) ses som en af de mest lovende fremstilling kandidater til at opfylde behovene i Fællesskabets TE hvor bestilte strukturer af bio...

Fremstilling af tre-dimensionelle (3D) biokompatible strukturer for celler er et af de vigtigste bidrag af tilsætningsstoffet biomanufacturing til tissue engineering (TE), satsning hen til genoprette væv ved at anvende tilpassede biomaterialer, celler, biokemiske faktorer, eller en kombination af disse. Derfor, de vigtigste krav i stilladser for TE programmer omfatte: manufacturability fra biokompatible materialer, kontrollerbar morfologiske egenskaber for målrettede celle invasion og optimeret overflade egenskaber til forbedret celle interaktion 1.
MEW er en opløsningsmiddelfri fremstilling teknik, der kombinerer principperne for tilsætningsstof fremstillingsindustrien (ofte kaldet 3D udskrivning) og electrospinning til fremstilling af polymert masker med stærkt bestilte ultratynde fiber morfologier2. Det er en direkte skrivning tilgang og indskud præcist fibre ifølge forprogrammerede koder3, kaldet G-koder. Smelt electrospun konstruktioner er i øjeblikket udarbejdet ved hjælp af en flad4,5 eller en Dorn6,7 samler til at fabrikere porøse flad og rørformede stilladser, henholdsvis.
Denne teknik giver betydelige fordele til TE og regenerativ medicin (RM) samfund på grund af muligheden for at udskrive direkte medicinsk-grade polymerer, såsom poly(ε-caprolactone) (PCL), der præsenterer fremragende biokompatibilitet8. Andre fordele er muligheden for at tilpasse størrelsen og fordelingen af porøsitet, ved at deponere fibre i en meget organiseret måde at fremstille stilladser af høj overflade-til-volumen forhold. Før MEW kan udføres, kræver polymeren først anvendelsen af varme9. Én gang i en flydende tilstand, en anvendt lufttryk tvinger det til at flyde ud gennem en metallisk spindedyse, der er forbundet med en høj spændingskilde. Kraftudligningen mellem overfladespænding og tiltrækning af elektrostatisk opladet slipværktøjet til jordet opkøber fører til dannelsen af en Taylor kegle efterfulgt af udslyngning af en jet10.
Billeder og en skematisk tegning af in-house opbygge MEW enheden bruges til denne protokol er vist i figur 1. Det viser desuden principper ved hjælp af isolerende tape til at undgå elektrisk udladning mellem varmeelementer og elektrisk ladede messing del omkring spindedyse. Tilstrækkelig isolering ville føre til indre skader den implementerede hardware.
Afhængigt af justeringen af de tre systemparametre (temperatur, samling hastighed og lufttryk) muliggør MEW fabrikation af fibre med forskellige diametre, forklaret i afsnittet diskussion. I de fleste tilfælde dog vil finjustering og optimering af jet være påkrævet før en stabil jet vil blive bortvist. Visualisering af de elektrificerede omrejsende jet er en effektiv måde at kontrollere konsistens og ensartethed i processen. I sag ideel ligner flyvevejen et køreledningsanlæg kurve erhvervet som følge af en force balance styres af system parametre11. Yderligere, mikro - og makro-struktur af stilladser er afhængig af flyvevejen polymer jet12. En detaljeret tabel over forskellige afbøjning adfærd og foranstaltninger for optimering er givet i afsnittet diskussion.
I den foreliggende undersøgelse præsenterer vi en protokol, der beskriver fabrication trinnene til fremstilling af meget kontrolleret fibrøst stilladser MEW teknologi. I dette arbejde, medicinsk grade PCL (Molekylær vægt 95-140 kg/mol) blev brugt, da denne medicinsk grade PCL har forbedret renhed over tekniske kvalitet, og dens mekaniske og forarbejdning egenskaber er fremragende til MEW. Bred smelte forarbejdning vifte af PCL stammer fra dens lavt smeltepunkt (60 ° C) og høj termisk stabilitet. PCL er desuden en langsom sats bionedbrydeligt polymer, hvilket gør det til et fremragende materiale til mange tissue engineering programmer13.
For denne undersøgelse, temperatur og samler afstanden vil blive holdt konstant (65 ° C og 82 ° C for at sprøjte og spindedyse temperaturer (henholdsvis) og 12 mm for collector afstand); anvendt spænding, samler hastighed og lufttryk, dog skal varieres for at fremstille fibre med målrettede diametre. En detaljeret liste over offentliggjorte undersøgelser ved hjælp af MEW stilladser er fastsat i afsnittet resultater og afslører forskellige applikationer for felterne af TE og RM (tabel 1).

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Plastic syringe</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012072</td>
			<td>EFD BARREL O 3mL Clear 50</td>
		</tr>
		<tr>
			<td>Medical grade Poly (&#949;-caprolactone) (mPCL)</td>
			<td>Corbion Purac, The Netherlands</td>
			<td>PURASORB PC12</td>
			<td></td>
		</tr>
		<tr>
			<td>23 GA needle</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7018302</td>
			<td>#23GP .013 X .25 ORANGE 50 PC</td>
		</tr>
		<tr>
			<td>Plunger</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012166</td>
			<td>PISTON O 3mL WH WIPER 50</td>
		</tr>
		<tr>
			<td>Pressure adapter</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012059</td>
			<td>ADAPTER ASM O 3mL BL 1.8M</td>
		</tr>
		<tr>
			<td>Aluminium collector</td>
			<td>Action Aluminium, Australia</td>
			<td>SHP2</td>
			<td>Sheet 5005 H34</td>
		</tr>
		<tr>
			<td>Acrylic glass</td>
			<td>Mulford Plastics Pty Ltd</td>
			<td>ACC6-13094</td>
			<td></td>
		</tr>
		<tr>
			<td>Mach 3 software</td>
			<td>Art Soft</td>
			<td></td>
			<td>Purchased online</td>
		</tr>
		<tr>
			<td>Safety switch interlock</td>
			<td>RS components Pty Ltd</td>
			<td>12621330</td>
			<td></td>
		</tr>
		<tr>
			<td>High voltage generator</td>
			<td>EMCO High Voltage Co.</td>
			<td>DX250R</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature controller</td>
			<td>WATLOW</td>
			<td>PM9R1FJ</td>
			<td></td>
		</tr>
		<tr>
			<td>X and Y positioning slide</td>
			<td>VELMEX Inc.</td>
			<td>XN-10-0020-M011</td>
			<td></td>
		</tr>
	</tbody>
</table>

melt electrospinning writing of three-dimensional poly(&#949;-caprolactone) scaffolds with controllable morphologies for tissue engineering applications

Denne tutorial afspejler på de fundamentale principper og retningslinjer for electrospinning skriftligt med polymer smelter, et tilsætningsstof fremstillingsindustrien teknologi med stort potentiale for biomedicinske programmer. Teknikken letter direkte aflejring af biokompatible polymer fibre til at fabrikere velordnet stilladser i sub micron til mikro skala rækkevidde. Etablering af en stabil, viskoelastiske, polymer jet mellem en spindedyse og en solfanger er opnået ved hjælp af en anvendt spænding og kan være direkte-skrevet. En vigtig fordel af et typisk porøse stillads er et højt overflade-til-volumen forhold, som giver øget effektiv vedhæftning websteder for celle udlæg og vækst. Styre udskrivningen af finjustering systemparametrene giver høj reproducerbarhed i kvaliteten af de udskrevne stilladser. Det giver også en fleksibel fremstilling platform for brugerne til at skræddersy de morfologiske strukturer af stilladser til deres specifikke krav. Til dette formål præsenterer vi en protokol for at opnå forskellige fiber diameter ved hjælp af smelte electrospinning skrivning (MEW) med en guidet ændring af parametre, herunder flow sats, spænding og samling hastighed. Desuden demonstrere vi hvordan man optimere jet, diskutere ofte erfarne tekniske udfordringer, forklare fejlfindingsteknikker og fremvise en bred vifte af printbare stillads arkitekturer.

To forskellige metoder til indsamling er almindeligt anvendt i MEW, som er flad samling og dornen samling. De resulterende arkitekturer afhænger programmering af G-koden (tabel 2), som er udført af softwaren.
Flad samling Anvende flad samlere refererer til den mest almindelige metode og letter direkte aflejring af materiale henviser til de præ-programmerede G-code. 0/90 og 0/60 strukturer ...

Watch this Scientific Journal Video about Melt Electrospinning Writing of Three-dimensional PolyÃŽÂµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications at JoVE.c

Melt Electrospinning Writing of Three-dimensional PolyÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Âµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications

Denne protokol serverer en omfattende vejledning til at fabrikere stilladser via electrospinning med polymer smelter i en direkte skrivning-tilstand. Vi systematisk skitsere processen og definere de relevante parameterindstillinger for at opnå målrettede stillads arkitekturer.

Smelte Electrospinning skrivning af tre-dimensionelle Poly(ε-caprolactone) stilladser med kontrollerbare morfologier for Tissue Engineering applikationer

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

Research

JoVE Journal

Bioengineering

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Udarbejdelse af 3D Fibrin stilladser til Stem celledyrkningsapplikationer

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Selvrapportering Stilladser for 3-Dimensional Cell Culture

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Menneskelige bruskvæv Fabrication Brug Tre-dimensionel Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds

Electrospinning Growth Factor Releasing Mikrosfærer i Fiber Stilladser

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth.  This system incorporates mechanical, topographical, ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

Generation af en tre-dimensionel fuld tykkelse Skin Equivalent og Automated Anskydning

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Fabrikation af mekanisk Tunable og bioaktive Metal Stilladser til biomedicinske anvendelser

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

Engineering Tredimensionale epitelvæv indlejret i ekstracellulær matrix

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

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

Fabrikation af Inverted Kolloid Crystal Poly (ethylenglycol) Stillads: En Tre-dimensionel Cell Culture Platform for levervæv Engineering

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing ...

Culturing Mammalian Cells in Three-dimensional Peptide Scaffolds

Dyrkning pattedyrsceller i tre-dimensionelle peptid stilladser

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an ...