Name: synthesis of biocompatible liquid crystal elastomer foams as cell scaffolds for 3d spatial cell cultures
Uploaded: 2017-04-11T15:00:00
Description: Watch this Scientific Journal Video about Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures at JoVE.com

Die Autoren möchten sich Kent State University (kollaborativen Forschungsförderung und Unterstützung für die Regenerative Medizin Initiative an der Kent State - ReMedIKS) danken für die finanzielle Unterstützung dieses Projekts.

HINWEIS: Die folgenden Schritte für die 3D LCE schaumartige Zubereitung des 3-armige Sternblockcopolymer verwendet , ist in Abbildung 1 dargestellt. Für Kernspinresonanz (NMR) Charakterisierung, die Spektren in deuteriertem Chloroform (CDCl 3) bei Raumtemperatur auf einem Bruker DMX - 400-MHz - Instrumente und intern referenzierten Restpeaks bei 7,26 aufgezeichnet. Fourier-Transformations-Infrarot (FT-IR) Spektren aufgenommen werden, einen Bruker Vector 33 FT-IR-Spektrometer u...

<ol>
	<li>Khor, E., Lim, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantable+applications+of+chitin+and+chitosan.">Implantable applications of chitin and chitosan.</a> Biomaterials. 24 (13), 2339-2349 (2003).</li><li>Chung, H. J., Park, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+engineered+and+drug+releasing+pre-fabricated+scaffolds+for+tissue+engineering.">Surface engineered and drug releasing pre-fabricated scaffolds for tissue engineering.</a> Adv. Drug Deliv. Rev. 59 (4-5), 249-262 (2007).</li><li>Yakacki, C. M., Gall, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shape-Memory+Polymers+for+Biomedical+Applications.">Shape-Memory Polymers for Biomedical Applications.</a> Shape-Memory Polymers. 226, 147-175 (2010).</li><li>Agrawal, 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=Stimuli-responsive+liquid+crystal+elastomers+for+dynamic+cell+culture.">Stimuli-responsive liquid crystal elastomers for dynamic cell culture.</a> J. of Mat. Res. 30 (4), 453-462 (2015).</li><li>Agrawal, A., Yun, T. H., Pesek, S. L., Chapman, W. G., Verduzco, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shape-responsive+liquid+crystal+elastomer+bilayers.">Shape-responsive liquid crystal elastomer bilayers.</a> Soft Matter. 10 (9), 1411-1415 (2014).</li><li>Sharma, 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=Biodegradable+and+Porous+Liquid+Crystal+Elastomer+Scaffolds+for+Spatial+Cell+Cultures.">Biodegradable and Porous Liquid Crystal Elastomer Scaffolds for Spatial Cell Cultures.</a> Macromol. Biosci. 15 (2), 200-214 (2015).</li><li>Yakacki, C. M., 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=Tailorable+and+programmable+liquid-crystalline+elastomers+using+a+two-stage+thiol-acrylate+reaction.">Tailorable and programmable liquid-crystalline elastomers using a two-stage thiol-acrylate reaction.</a> RSC Adv. 5 (25), 18997-19001 (2015).</li><li>deGennes, P. G., Hebert, M., Kant, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+muscles+based+on+nematic+gels.">Artificial muscles based on nematic gels.</a> Macromolecular Symposia. 113, 39-49 (1997).</li><li>Fleischmann, E. -. K., Zentel, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liquid-Crystalline+Ordering+as+a+Concept+in+Materials+Science:+From+Semiconductors+to+Stimuli-Responsive+Devices.">Liquid-Crystalline Ordering as a Concept in Materials Science: From Semiconductors to Stimuli-Responsive Devices.</a> Angew. Chem. Int. Ed. 52 (34), 8810-8827 (2013).</li><li>Finkelmann, H., Kim, S. T., Munoz, A., Palffy-Muhoray, P., Taheri, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tunable+mirrorless+lasing+in+cholesteric+liquid+crystalline+elastomers.">Tunable mirrorless lasing in cholesteric liquid crystalline elastomers.</a> Adv. Mater. 13 (14), 1069-1072 (2001).</li><li>Artal, 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=SHG+characterization+of+different+polar+materials+obtained+by+in+situ+photopolymerization.">SHG characterization of different polar materials obtained by in situ photopolymerization.</a> Macromolecules. 34 (12), 4244-4255 (2001).</li><li>Camacho-Lopez, M., Finkelmann, H., Palffy-Muhoray, P., Shelley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+liquid-crystal+elastomer+swims+into+the+dark.">Fast liquid-crystal elastomer swims into the dark.</a> Nat. Mater. 3 (5), 307-310 (2004).</li><li>Yamada, M., 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=Photomobile+polymer+materials:+Towards+light-driven+plastic+motors.">Photomobile polymer materials: Towards light-driven plastic motors.</a> Angew. Chem. Int. Ed. 47 (27), 4986-4988 (2008).</li><li>Ohm, C., Brehmer, M., Zentel, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liquid+Crystalline+Elastomers+as+Actuators+and+Sensors.">Liquid Crystalline Elastomers as Actuators and Sensors.</a> Adv. Mater. 22 (31), 3366-3387 (2010).</li><li>Fleischmann, E. -. K., 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=One-piece+micropumps+from+liquid+crystalline+core-shell+particles.">One-piece micropumps from liquid crystalline core-shell particles.</a> Nat. Commun. 3, (2012).</li><li>Herzer, N., 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=Printable+Optical+Sensors+Based+on+H-Bonded+Supramolecular+Cholesteric+Liquid+Crystal+Networks.">Printable Optical Sensors Based on H-Bonded Supramolecular Cholesteric Liquid Crystal Networks.</a> J. Am. Chem. Soc. 134 (18), 7608-7611 (2012).</li><li>Lockwood, N. 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=Thermotropic+liquid+crystals+as+substrates+for+imaging+the+reorganization+of+matrigel+by+human+embryonic+stem+cells.">Thermotropic liquid crystals as substrates for imaging the reorganization of matrigel by human embryonic stem cells.</a> Adv. Funct. Mater. 16 (5), 618-624 (2006).</li><li>Sharma, 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=Effects+of+structural+variations+on+the+cellular+response+and+mechanical+properties+of+biocompatible,+biodegradable,+and+porous+smectic+liquid+crystal+elastomers.">Effects of structural variations on the cellular response and mechanical properties of biocompatible, biodegradable, and porous smectic liquid crystal elastomers.</a> Macromol. Biosci. , (2016).</li><li>Bera, 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=Liquid+Crystal+Elastomer+Microspheres+as+Three-Dimensional+Cell+Scaffolds+Supporting+the+Attachment+and+Proliferation+of+Myoblasts.">Liquid Crystal Elastomer Microspheres as Three-Dimensional Cell Scaffolds Supporting the Attachment and Proliferation of Myoblasts.</a> ACS Appl. Mater. Interfaces. 7 (26), 14528-14535 (2015).</li><li>Bera, T., Malcuit, C., Clements, R. J., Hegmann, E. <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+Surfactant+during+Microemulsion+Photopolymerization+for+the+Creation+of+Three-Dimensional+Liquid+Crystal+Elastomer+Microsphere+Spatial+Cell+Scaffolds.">Role of Surfactant during Microemulsion Photopolymerization for the Creation of Three-Dimensional Liquid Crystal Elastomer Microsphere Spatial Cell Scaffolds.</a> Front. Mater. 3 (31), (2016).</li><li>McKee, C. T., Last, J. A., Russell, P., Murphy, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Indentation+Versus+Tensile+Measurements+of+Young's+Modulus+for+Soft+Biological+Tissues.">Indentation Versus Tensile Measurements of Young's Modulus for Soft Biological Tissues.</a> Tissue Eng. Part B Rev. 17 (3), 155-164 (2011).</li><li>Kung, C. -. 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=Preparation+and+characterization+of+three+dimensional+graphene+foam+supported+platinum-ruthenium+bimetallic+nanocatalysts+for+hydrogen+peroxide+based+electrochemical+biosensors.">Preparation and characterization of three dimensional graphene foam supported platinum-ruthenium bimetallic nanocatalysts for hydrogen peroxide based electrochemical biosensors.</a> Biosens. Bioelectron. 52, 1-7 (2014).</li><li>Amsden, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Curable,+biodegradable+elastomers:+emerging+biomaterials+for+drug+delivery+and+tissue+engineering.">Curable, biodegradable elastomers: emerging biomaterials for drug delivery and tissue engineering.</a> Soft Matter. 3 (11), 1335-1348 (2007).</li><li>Sinturel, C., Vayer, M., Morris, M., Hillmyer, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Solvent+Vapor+Annealing+of+Block+Polymer+Thin+Films.">Solvent Vapor Annealing of Block Polymer Thin Films.</a> Macromolecules. 46 (14), 5399-5415 (2013).</li><li>Riboldi, S. 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=Skeletal+myogenesis+on+highly+orientated+microfibrous+polyesterurethane+scaffolds.">Skeletal myogenesis on highly orientated microfibrous polyesterurethane scaffolds.</a> J. Biomed. Mater. Res. A. 84 (4), 1094-1101 (2008).</li><li>Chung, S., Moghe, A. K., Montero, G. A., Kim, S. H., King, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanofibrous+scaffolds+electrospun+from+elastomeric+biodegradable+poly(L-lactide-co-epsilon-caprolactone)+copolymer.">Nanofibrous scaffolds electrospun from elastomeric biodegradable poly(L-lactide-co-epsilon-caprolactone) copolymer.</a> Biomed. Mater. 4 (1), 9 (2009).</li><li>Gao, Y. 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=Biocompatible+3D+Liquid+Crystal+Elastomer+Cell+Scaffolds+and+Foams+with+Primary+and+Secondary+Porous+Architecture.">Biocompatible 3D Liquid Crystal Elastomer Cell Scaffolds and Foams with Primary and Secondary Porous Architecture.</a> ACS Macro Lett. 5 (1), 14-19 (2016).</li><li>Lenoir, 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=Ring-opening+polymerization+of+alpha-chloro-is+an+element+of-caprolactone+and+chemical+modification+of+poly(alpha-chloro-is+an+element+of-caprolactone)+by+atom+transfer+radical+processes.">Ring-opening polymerization of alpha-chloro-is an element of-caprolactone and chemical modification of poly(alpha-chloro-is an element of-caprolactone) by atom transfer radical processes.</a> Macromolecules. 37 (11), 4055-4061 (2004).</li><li>Younes, H. M., Bravo-Grimaldo, E., Amsden, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis,+characterization+and+in+vitro+degradation+of+a+biodegradable+elastomer.">Synthesis, characterization and in vitro degradation of a biodegradable elastomer.</a> Biomaterials. 25 (22), 5261-5269 (2004).</li><li>Donaldson, T., Henderson, P. A., Achard, M. F., Imrie, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chiral+liquid+crystal+tetramers.">Chiral liquid crystal tetramers.</a> J. Mater. Chem. 21 (29), 10935-10941 (2011).</li><li>Palmgren, R., Karlsson, S., Albertsson, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+degradable+crosslinked+polymers+based+on+1,5-dioxepan-2-one+and+crosslinker+of+bis-epsilon-caprolactone+type.">Synthesis of degradable crosslinked polymers based on 1,5-dioxepan-2-one and crosslinker of bis-epsilon-caprolactone type.</a> J. Pol. Sci. A Polym. Chem. 35 (9), 1635-1649 (1997).</li></ol>

Flüssigkristalline Elastomere als biokompatible Zellgerüste untersucht aufgrund ihrer Reize Ansprechbarkeit kürzlich. Sie haben sich als ideale Plattformen als Zellgerüste sein. Jedoch ist ein wichtiger Faktor im Auge zu behalten bei der Vorbereitung und ein neues LCE Gerüstes Gestaltung ist Porosität. Die Einarbeitung von auslaugbaren Feststoffen oder Gas 23 führt nicht immer in homogener Porosität oder vollständig miteinander verbundene Poren. Die Verwendung einer Metallschablone, die ...

Es gibt mehrere Beispiele von biologischen und biologisch verträglichen synthetischen Materialien zur Anwendung , in Zellstudien und für die Geweberegeneration darauf abzielen, die Zellanheftung und Proliferation 1, 2, 3, 4, 5. Es gibt einige Beispiele von biokompatiblen Materialien gewesen, bekannt als Flüssigkristall Elastomere (LCEs), die Molekular auf äußere Reize mit anisotropen reagieren könnten Bestellung 6, 7. LCEs sind Stimuli reagierende Materialien , die die mechanischen und elastischen Eigenschaften von Elastomeren , die mit den optischen Funktionen und molekularen Anordnung von Flüssigkristallen 8, 9 verbinden. LCEs können Änderungen in der Form, mechanische Verformung, elastisches Verhalten auf, und die optischen Eigenschaften in Reaktion auf externe stimUli (dh., Wärme, Druck, Licht, etc.) 10, 11, 12, 13, 14, 15, 16. Frühere Studien haben gezeigt , dass Flüssigkristallen (LCs) 4 , das Wachstum und die Orientierung von Zellen abtasten kann, 17. Es ist möglich, dann davon ausgehen, dass LCEs für biologisch und medizinisch relevante Anwendungen geeignet sein kann, einschließlich Zellgerüsten und Ausrichtung. Wir haben bereits berichtet , die Herstellung von smektischen biokompatibles, biologisch abbaubarem, gussgeformt, und dünnen Filme von LCEs einen „Swiss-Käse - Typ“ poröse Morphologie 6, 18. Wir haben auch nematische biokompatiblen LCEs mit globuläre Morphologie als Gerüste für das Zellwachstum 19 hergestellt &lt;sup&gt; 20. Unsere Arbeit wurde bei Abstimmung der mechanischen Eigenschaften der Materialien richtet 21 diejenigen des Gewebes von Interesse zu entsprechen. Außerdem konzentrieren sich diese Studien auf Elastomer-Zell-Wechselwirkungen zu verstehen, sowie zelluläre Reaktion, wenn die Elastomere auf äußere Reize unterliegen. Die wichtigsten Herausforderungen waren teilweise die Porosität des LCEs maßzuschneidern für die Zellanheftung und Permeation durch die Elastomermatrix und zum besseren Massentransport zu ermöglichen. Die Porosität dieser Dünnschichten 6 für Zellpermeation durch die Masse der Matrix erlaubt, aber nicht alle Poren vollständig miteinander verbunden waren oder hatten ein regelmäßigeres (homogen) Größe Pore. Wir berichten dann auf biokompatiblen nematischen LCE Elastomere mit globulären Morphologien. Diese nematische Elastomere für die Anheftung und Proliferation von Zellen erlaubt, aber die Porengröße lag im Bereich von 10 bis 30 nur um, was verhindert, oder die Verwendung von dieser beschränktElastomere mit einer größeren Vielfalt von Zelllinien 19, 20. Frühere Arbeiten von Kung et al. im Zusammenhang mit der Bildung von Graphen Schäumen einer „Opfer“ Metallschablone zeigte , dass der erhaltene Graphitschaum eine sehr regelmäßige poröse Morphologie durch die gewählte Metallschablone 22 diktiert hatte. Diese Methode bietet die volle Kontrolle über die Porosität und Porengröße. Zur gleichen Zeit, die Geschmeidigkeit und Flexibilität der Metallschablone ermöglichen die Bildung von verschiedener Vorlage Schaumherstellung vor prägt. Andere Techniken, wie beispielsweise Material Auslaugung 23, Gas Templat 24 oder elektrogesponnenen Fasern 25, 26 bieten auch das Potential für die Herstellung von porösen Materialien, aber sie sind zeitaufwendig , und in einigen Fällen wird die Porengröße begrenzt auf nur wenige Mikrometer. Schaum-ähnlichen 3D LCEs vorbereitet Metallschablonen für eine höhere Zellenlast erlauben verwendet; eine verbesserte Proliferationsrate; Cokultivieren; und last but not least, eine bessere Massentransportmanagement (dh Nährstoffe, Gase und Abfall) , um sicherzustellen , vollständige Gewebeentwicklung 27. Schaumartige 3D LCEs scheint auch Zellausrichtung zu verbessern; Dies ist wahrscheinlich in Bezug auf die LC-Anhänger Abfühlen Zellwachstum und die Zellorientierung. Die Anwesenheit von LC-Einheiten innerhalb des LCE erscheint Zellenausrichtung mit Bezug auf Zellenposition innerhalb des LCE Gerüstes zu verbessern. Zellen auszurichten innerhalb der Streben des LCE, während keine klare Orientierung beobachtet wird , wo die Verstrebungen miteinander verbinden (junctions) 27. Insgesamt ist unsere LCE Zellgerüstplattform als Zellträgermedium bietet die Möglichkeit zur Abstimmung der Elastomer Morphologie und elastische Eigenschaften und insbesondere die Ausrichtung der (individuellen) Zelltypen zu lenken, eine geordnete, räumlichen Anordnungen o zu schaffenf Zellen ähnlich wie lebende Systeme. Abgesehen von einem Gerüst der Lage erhalt Bereitstellung und langfristiges Zellwachstum und -proliferation zu leiten, LCEs erlaubt auch für dynamische Experimente, in denen Zellorientierung und Wechselwirkungen können im laufenden Betrieb geändert werden.

<table border="0" cellpadding="0" cellspacing="0" width="723">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:291px;">1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane</td>
			<td style="width:148px;">Alfa Aesar</td>
			<td style="width:132px;">L16606</td>
			<td style="width:153px;">Silanizing agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-bis(4-hydroxy-cyclohexyl)propane</td>
			<td>TCI</td>
			<td><a href="http://www.tcichemicals.com/eshop/en/us/commodity/B0928/">B0928</a></td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-chlorohexanone&#160;</td>
			<td>Alfa Aesar</td>
			<td>A18613</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-heptanone&#160;</td>
			<td>Sigma Aldrich</td>
			<td>W254401</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-propanol&#160;</td>
			<td>Sigma Aldrich</td>
			<td>278475</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3-chloroperbenzoic acid, m-CPBA</td>
			<td>Sigma Aldrich</td>
			<td>273031</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4-dimethylaminopyridine</td>
			<td>Alfa Aesar</td>
			<td>A13016</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4&#39;,6-diamidino-2-phenylindole, DAPI&#160;</td>
			<td>Invitrogen</td>
			<td>D1306</td>
			<td>Nuclear Stain</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-hexynoic acid&#160;</td>
			<td>Alfa Aesar</td>
			<td>B25132-06</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Acetic acid</td>
			<td>VWR</td>
			<td>36289</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Acetone</td>
			<td>Sigma Aldrich</td>
			<td>34850</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alcohol 200 proof ACS Grade&#160;</td>
			<td>VWR</td>
			<td>71001-866</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Benzene</td>
			<td>Alfa Aesar</td>
			<td>AA33290</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#949;-caprolactone&#160;</td>
			<td>Alfa Aesar</td>
			<td>A10299-0E</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Chloroform</td>
			<td>VWR</td>
			<td>BDH1109</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cholesterol</td>
			<td>Sigma Aldrich</td>
			<td>C8503</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Chromium(VI) oxide</td>
			<td>Sigma Aldrich</td>
			<td>232653</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Copper (I) iodide</td>
			<td>Strem Chemicals</td>
			<td>100211-060</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D,L-Lactide&#160;</td>
			<td>Alfa Aesar</td>
			<td>L09026</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dichloromethane</td>
			<td>Sigma Aldrich</td>
			<td>320269</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Diethyl ether&#160;</td>
			<td>Emd Millipore</td>
			<td>EX0190</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N-Dimethylformamide</td>
			<td>Sigma Aldrich</td>
			<td>270547</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dulbecco&#8217;s modified Eagle medium, DEME&#160;</td>
			<td>CORNING Cellgo</td>
			<td>10-013</td>
			<td>Cell Media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Alfa Aesar</td>
			<td>33361</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Formaldehyde&#160;</td>
			<td>SIGMA Life Science</td>
			<td>F8775</td>
			<td>Fixative</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal bovine serum, FBS&#160;</td>
			<td>HyClone</td>
			<td>SH30071.01</td>
			<td>Media Component</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Filter paper, Grade 415, qualitative, crepe</td>
			<td>VWR</td>
			<td>28320</td>
			<td>Filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glycerol</td>
			<td>Sigma Aldrich</td>
			<td>G5516</td>
			<td>Central node (3-arm)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hexamethylene diisocyanate, HDI</td>
			<td>Sigma Aldrich</td>
			<td>52649</td>
			<td>Crosslinker</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Iron(III) chloride&#160;</td>
			<td>Alfa Aesar</td>
			<td>12357</td>
			<td>Etching agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isopropyl alcohol</td>
			<td>VWR</td>
			<td>BDH1133</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Methanol</td>
			<td>Alfa Aesar</td>
			<td>L13255</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N&#39;-dicyclohexylcarbodiimide</td>
			<td>Aldrich</td>
			<td>D80002</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">N,N-Dimethylformamide</td>
			<td>Sigma Aldrich</td>
			<td>270547</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nickel metal template</td>
			<td>American Elements</td>
			<td>Ni-860</td>
			<td>Foam template</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Neuroblastomas cells (SH-SY5Y)</td>
			<td>ATCC</td>
			<td>CRL-2266</td>
			<td>Cell line</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Penicillin streptomycin&#160;</td>
			<td>Thermo SCIENTIFIC</td>
			<td>15140122</td>
			<td>Antibiotics</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polyethylene glycol 2000, PEG</td>
			<td>Alfa Aesar</td>
			<td>B22181</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium azide&#160;</td>
			<td>VWR</td>
			<td>97064-646</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium bicarbonate</td>
			<td>AMRESCO</td>
			<td>865</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium chloride</td>
			<td>BDH</td>
			<td>BDH9286</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium phosphate dibasic heptahydrate</td>
			<td>Fisher Scientific</td>
			<td>S-374</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium phosphate monobasic monohydrate</td>
			<td>Sigma Aldrich</td>
			<td>S9638</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium sulfate</td>
			<td>Sigma Aldrich</td>
			<td>239313</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tetrahydrofuran</td>
			<td>Alfa Aesar</td>
			<td>41819</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thiosulfate de sodium</td>
			<td>AMRESCO</td>
			<td>393</td>
			<td>Drying salt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tin(II) 2-ethylhexanoate</td>
			<td>Aldrich</td>
			<td>S3252</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Toluene&#160;</td>
			<td>Alfa Aesar</td>
			<td>22903</td>
			<td>Solvent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triethylamine</td>
			<td>Sigma Aldrich</td>
			<td>471283</td>
			<td>Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypsin&#160;</td>
			<td>HyClone</td>
			<td>SH30042.01</td>
			<td>Cell Detachment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olympus FV1000</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of biocompatible liquid crystal elastomer foams as cell scaffolds for 3d spatial cell cultures

Hier präsentieren wir eine Schritt-für-Schritt Herstellung eines 3D, biologisch abbaubaren, schaumartige Zellgerüst. Diese Gerüste wurden hergestellt durch vernetzende Stern-Block-Copolymere mit Cholesterin-Einheiten als Seitenketten-Seitengruppen, die sich in der smektischen A (SmA) Flüssigkristall Elastomere (LCEs). Schaumartige Gerüste, hergestellt Metall Vorlagen verfügen miteinander verbundene Mikrokanäle, so dass sie geeignet als 3D-Zellkulturgerüste. Die kombinierten Eigenschaften der regelmäßigen Struktur des Metallschaums und des Elastomer Ergebnis in einem 3D - Zell Gerüst , das nicht nur eine höhere Zellproliferation im Vergleich zu herkömmlichen porösen Templat Filme, sondern auch eine bessere Verwaltung der Massentransport (dh Nährstoffe, Gase, Abfall fördert usw.). Die Art der Metallschablone ermöglicht die einfache Bearbeitung von Schaumformen (dh Rollen oder Folien) und zur Herstellung von Gerüsten unterschiedlicher Porengrößen für unterschiedliche Zellstudien unter Beibehaltung die interconnected poröse Natur der Vorlage. Der Ätzprozess wirkt sich nicht auf die Chemie der Elastomere, ihre biokompatibel und biologisch abbaubar Natur zu bewahren. Wir zeigen, dass diese smektische LCEs, wenn sie für ausgedehnte Zeiträume gewachsen, die Untersuchung von klinisch relevanten und komplexen Gewebekonstrukte ermöglichen, während das Wachstum und die Vermehrung von Zellen zu fördern.

Dieser Bericht zeigt die Herstellungsverfahren eines porösen 3D LCE als Gerüst für die Zellkultur einer Nickel-Metall-Vorlage. Der erhaltene 3D LCE zeigt ein Komplex miteinander verbundenes Kanalnetzwerk , das ebenso wie weitere geeigneten Massentransport 27 für eine einfache Zellinfiltration ermöglicht. Es wurde festgestellt, dass die Zellen der Lage sind, in vollem Umfang das miteinander verbundene Kanalnetzwerk eindringen und ebenfalls innerhalb des LCE au...

Watch this Scientific Journal Video about Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures at JoVE.com

Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures

Diese Studie stellt eine Methode zur Herstellung von 3D, biologisch abbaubare, schaumartige Gerüste Zelle basierend auf biokompatible Seitenkettenflüssigkristall Elastomere (LCEs). Die konfokale Mikroskopie Experimente zeigen, dass schaumartige LCEs ermöglichen Zellanheftung, Proliferation und die spontane Ausrichtung der C2C12s Myoblasten.

Synthese von biokompatiblen Liquid Crystal Elastomerschäume als Zellgerüste für 3D Spatial Zellkulturen

Here, we present a step-by-step preparation of a 3D, biodegradable, foam-like cell scaffold. These scaffolds were prepared by cross-linking star block ...

The goal of this procedure is to prepare biodegradable, three dimensional foam-like cell scaffolds based on biocompatible side chain liquid crystal elastomers. This method can help answer key questions in the liquid crystal and biomedical fields, such as the effects of liquid crystal elastomer properties on cell proliferation and alignment. The main advantage of this method of two dimensional cell scaffolds is it permits the study of spatial cell-cell interactions which is rarely possible in 2D macro environments.Generally, those new to this method will struggle with the tactile compression tests. We first had the idea for this method after having to change the media in hundreds of Petri dishes. We wondered if liquid crystal elastomers could support muscle cells on a 3D network, to eliminate the use of so many Petri dishes.Visual demonstration of this method is critical, as some steps require careful manipulation of chemicals. Further, shaping the metal foam template may be challenging. First, fill a 20 milliliter ampoule with a 2%by volume solution of PFOTES in toluene.Stir the solution in the ampoule for 24 hours to silanize the ampoule interior. Rinse the silanized ampoule with isopropyl alcohol and dry it at 140 degrees celsius for 30 minutes. Place 3.64 grams of distilled epsilon-caprolactone, 0.5 grams of alpha-chloro-epsilon-caprolactone, and 0.25 milliliters of glycerol in the dry ampoule.Vortex the mixture for one minute. Then, add 4.90 grams of DL-lactide to the mixture, and purge the ampoule atmosphere with nitrogen gas for one minute. Cover the ampoule opening with aluminum foil, and heat the mixture at 120 degrees Celsius for about two hours to melt the DL-lactide.Vortex the mixture to disrupt any unmelted solids, and then, add 66 microliters of 10-2-ethylhexanoate to the ampoule, and vortex again. Close the ampoule with aluminum foil and heat the mixture at 120 degrees Celsius for 10 minutes to remelt the DL-lactide. Once the DL-lactide has melted again, vigorously vortex the mixture, and purge the ampoule atmosphere with nitrogen gas.Seal the ampoule with a rubber septum. Insert a needle connected to a vacuum line through the septum, and start the vacuum. Flame-seal the neck of the ampoule, being careful not to melt the rubber stopper.Once the neck is sealed, heat the reaction mixture at 140 degrees Celsius for 48 hours. Then, allow the mixture to cool to room temperature. Break open the ampoule and dissolve the viscous reaction mixture in 10 milliliters of dichloromethane.Transfer the mixture to a separatory funnel. Place a flask containing 100 milliliters of methanol in a dry ice and acetone bath to cool down the methanol to approximately minus 78 degrees Celsius. Once the methanol has cooled down, secure the separatory funnel over the flask.Add the reaction mixture to the cold methanol at a rate of two drops every second. Collect the resulting white precipitate on filter paper. Dry the precipitate in a vacuum oven between 50 and 60 degrees Celsius to obtain the three-arm alpha-chloro SBC product.To prepare the alpha cholesteryl three-arm SBC, the pendant chlorine atom is replaced by an azide. A click reaction with cholesteryl 5-hexynoate results in a pendant cholesteryl as the liquid crystal moiety. To begin preparing the liquid crystal elastomer scaffold, combine 0.75 grams of alpha-cholesteryl three-arm SBC with 0.25 milliliters of HDI and 0.24 milliliters of distilled epsilon-caprolactone.Add to this 60 microliters of 10-2-ethylhexanoate and vortex the mixture. Then, cut out a one centimeter by four centimeter piece of nickel metal foam. Roll the nickel foam into a cylinder one centimeter in diameter and one centimeter tall to form a template for the liquid crystal elastomer foam scaffold.Place the template in a glass vial or aluminum foil enclosure, and pour the liquid crystal elastomer mixture over the template until it is completely covered. Allow the template to sit in the liquid crystal elastomer mixture for two minutes, and then, remove the excess with a pipette. Heat the mixture and template at 80 degrees Celsius overnight.Then, peel away the aluminum foil, or break the glass. Use a razor blade to remove excess liquid crystal elastomer to expose the nickel metal template. Place the foam in a flask and add 70 milliliters of a saturated aqueous solution of iron three chloride.Stir the foam in the solution for three days at room temperature to dissolve the nickel template. Every 24 hours, stir the foam in deionized water for 30 minutes, and then, resume stirring in a fresh iron three chloride solution. After the second day of stirring, perform a tactile compression test on the liquid crystal elastomer foam.Resistance to tactile compression indicates that the nickel template is still present in the foam. After the third day, all nickel template has been eliminated, and the resulting foam appears very soft and easy to fully compress. The nickel template must be completely eliminated.It is important to thoroughly rinse the liquid crystal elastomer foam in iron chloride until it's soft to the touch when a tactile compression test is performed. The liquid crystal elastomer foam scaffold is now ready to be characterized and used. Once the liquid crystal elastomer foam is soft, rinse it with 70%ethanol to sterilize it.To begin the seeding procedure, wash again the liquid crystal elastomer foam scaffolds twice in one milliliter of 70%ethanol to sterilize the elastomer surfaces. Then, irradiate the scaffolds with UV light for 10 minutes. Wash the scaffolds with another one milliliter portion of 70%ethanol.Rinse the scaffolds with one milliliter each of sterile water and phosphate-buffered saline. Load the sterilized liquid crystal elastomer scaffolds into 24-well culture plates. Prepare and count a suspension of the cells of interest in the appropriate cell growth medium with penicillin and streptomycin.Dilute the cell suspension to 1.5 times 10 to the fifth cells per 100 microliters using growth medium. Deposit a drop of diluted cell suspension on top of each liquid crystal elastomer scaffold. Incubate the seeded liquid crystal elastomer scaffolds at 37 degrees Celsius in a 5%CO2 atmosphere for two hours.Add another 0.5 milliliters of growth medium to each scaffold, and continue incubation. Every 48 hours, wash the seeded scaffolds with one milliliter of PBS, and add fresh growth medium. Continue incubating the scaffolds until the cells are ready for microscopy.These smetic LCs enable the study of complex tissue constructs while promoting cell growth and proliferation. In order to maintain cell viability, foams must be sterilized and cleanliness of cultures maintained at all times. To prepare for microscopy, fix the cells on the scaffolds with a solution of 4%paraformaldehyde in PBS for 15 minutes.Soak the samples three times in three milliliters of PBS for five minutes. Place one scaffold sample in an Eppendorf tube. Stain the sample with a solution of 0.1%DAPI in 500 microliters of PBS for 10 minutes.Soak the sample twice in one milliliter of PBS for five minutes. And then, immediately image the sample with confocal microscopy. Acquire image stacks spanning the sample and analyze the data in an image processing program.A three-arm SBC with liquid crystal moieties was cast on a nickel foam template with HDI as the cross-linker. The nickel template was removed by etching to obtain the liquid crystal elastomer foam. Compression deformation testing was periodically performed to monitor the etching process.Once the nickel had been completely dissolved, the compression deformation testing showed a 70%reduction in size when compressed. Upon releasing compression, the liquid crystal elastomer foam consistently recovered its original size and shape. This behavior is attributed to the liquid crystal moieties, as similar elastomer foams lacking the cholesterol epsilon-caprolactone moiety did not recover from compression.SCM of the internal foam morphology showed an interconnected network of hollow foam struts. The regular morphology was attributed to the structure of the nickel foam, indicating that pore size and overall shape can be controlled by selection of an appropriate metal template. The foam was seeded with neuroblastoma cells, which attached to the walls of the network within two days.30 days after seeding, confocal microscopy revealed that the cells extended over the liquid crystal elastomer network, and had formed multiple layers dispersed throughout the elastomer foam. Elongation of cell nuclei was observed, and was correlated to cell alignment. Once mastered, this technique can be done within three to four weeks if performed properly.After watching this video, you should have a good understanding of how to design and prepare liquid crystal elastomer cell scaffolds with specific pore sizes and morphologies. While attempting this procedure, it's important to fully characterize each product and to monitor cell viability and expansion. Further, proper staining technique is important to easily acquire confocal microscopy images.Following this procedure, liquid crystal elastomers can also be exposed to external stimuli, such as stress, or magnetic and electric fields, to answer additional questions about how the anisotropic molecular ordering of the liquid crystal elastomers affects cell response. The implications of this technique apply to the study of disease processes, as it provides a platform to dynamically study the effects of external agents on simulated disease activity, and subsequent repair processes. Though this smectic liquid crystal elastomer foam can provide insight into cell-cell interactions, they can also be used with other materials, such as semiconductor or metal nanostructures.After its development, this technique paved the way for researchers in biomedical fields to design long-term experimental platforms mirroring living systems.

Research

JoVE Journal

Bioengineering

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

Alginat-Mikrokapseln als 3D-Plattform für die Vermehrung und Differenzierung von humanen embryonalen Stammzellen (hES) auf verschiedenen Linien

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

Forschung

Biotechnik

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Erstellung von 3D-Fibrin-Gerüste für Stem Cell Culture Anwendungen

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

Selbstberichts Gerüste für 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 ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Herstellung von mechanisch Tunable und Bioactive Metallgerüste für biomedizinische Anwendungen

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

Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release

Magnetisches Wechselfeld-Responsive Hybrid Gelatine Mikrogele der gezielten Medikamentenapplikation

Magnetically-responsive nano/micro-engineered biomaterials that enable a tightly controlled, on-demand drug delivery have been developed as new types of ...

Preparation of Monodomain Liquid Crystal Elastomers and Liquid Crystal Elastomer Nanocomposites

Herstellung von Monodomäne Liquid Crystal Elastomere und Liquid Crystal Elastomer Nanokomposite

LCEs are shape-responsive materials with fully reversible shape change and potential applications in medicine, tissue engineering, artificial muscles, and ...

Micropatterning and Assembly of 3D Microvessels

Mikrostrukturierung und Montage von 3D-Micro-Ader

In vitro platforms to study endothelial cells and vascular biology are largely limited to 2D endothelial cell culture, flow chambers with polymer or glass ...

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

Die Herstellung von extrazellulären Matrix-derived Foams und Mikroträgern als gewebespezifische Zellkultur und Delivery-Plattformen

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol

Ein Mikrocontroller betrieben Gerät zur Erzeugung von flüssige Extrakte aus herkömmlichen Zigarette Rauch und elektronische Zigarette Aerosol

Electronic cigarettes are the most popular tobacco product among middle and high schoolers and are the most popular alternative tobacco product among ...

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients

3D-Analyse von multizellulären Reaktionen auf tschoattraktive Verläufe

Various limitations of 2D cell culture systems have sparked interest in 3D cell culture and analysis platforms, which would better mimic the spatial and ...