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 PolyÎµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications at JoVE.c

Эта работа финансовую поддержку КПР центр совместных исследований для клетки терапии производства, Австралийский центр дуги Совета исследований в Институте перспективных исследований в техническом университете Мюнхена и добавка Biomanufacturing. Это исследование было проведено, Австралийский исследовательский совет промышленного преобразования учебный центр в Biomanufacturing добавка http://www.additivebiomanufacturing.org (IC160100026). Пожалуйста, посетите сайт для статей, книг, программ телевидения или Радио, электронные средства массовой информации или любые другие литературные произведения, связанные с проектом. Кроме того авторы с благодарностью признаем Мария Фландес ИПАРРАГИРРЕ для поддержки в съемки, Филипп Хаббард для голоса и Luise Гроссман для съемки и редактирования.

1. Подготовка
<ol>
	<li>Заполните 2 g PCL в пластиковый шприц 3 мл с воронкой и вставьте поршень в открытый конец.</li>
	<li>Поместите шприц в предварительно разогретой духовке при температуре 65 ° C для 8 h. точку кончик вверх разрешить пузырьков воздуха в совокупных закрыть для открытия.</li>
	<li>Толкат...

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	<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. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+Fabrication+of+Tubular+Scaffolds+via+Direct+Writing+in+a+Melt+Electrospinning+Mode.">Design and Fabrication of Tubular Scaffolds via Direct Writing in a Melt Electrospinning Mode.</a> Biointerphases. 7 (1), 1-16 (2012).</li><li>Woodruff, M. A., 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=The+return+of+a+forgotten+polymer-Polycaprolactone+in+the+21st+century.">The return of a forgotten polymer-Polycaprolactone in the 21st century.</a> Prog. in pol Sci. 35 (10), 1217-1256 (2010).</li><li>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.">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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Rapid Commun. 37 (1), 93-99 (2016).</li><li>Wagner, 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=A+validated+preclinical+animal+model+for+primary+bone+tumor+research.">A validated preclinical animal model for primary bone tumor research.</a> JBJS. 98 (11), 916-925 (2016).</li><li>Baldwin, J., 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=Periosteum+tissue+engineering+in+an+orthotopic+in+vivo+platform.">Periosteum tissue engineering in an orthotopic in vivo platform.</a> Biomaterials. 121, 193-204 (2017).</li><li>Tourlomousis, F., Chang, R. 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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Mater. Eng. 298 (5), 504-520 (2013).</li><li>Wunner, F. 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=.">.</a> Comprehensive Biomaterials II. , 217-235 (2017).</li><li>Loh, Q. L., Choong, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Scaffolds+for+Tissue+Engineering+Applications:+Role+of+Porosity+and+Pore+Size.">Three-Dimensional Scaffolds for Tissue Engineering Applications: Role of Porosity and Pore Size.</a> Tissue Eng Part B Rev. 19 (6), 485-502 (2013).</li><li>Ristovski, 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=Improved+fabrication+of+melt+electrospun+tissue+engineering+scaffolds+using+direct+writing+and+advanced+electric+field+control.">Improved fabrication of melt electrospun tissue engineering scaffolds using direct writing and advanced electric field control.</a> Biointerphases. 10 (1), 011006 (2015).</li><li>Wei, C., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+fabrication+of+high-resolution+three-dimensional+polymeric+scaffolds+using+electrohydrodynamic+hot+jet+plotting.">Direct fabrication of high-resolution three-dimensional polymeric scaffolds using electrohydrodynamic hot jet plotting.</a> J Micromech Microeng. 23 (2), 025017 (2013).</li><li>Hacker, 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=Electrospinning+of+polymer+melt+:+steps+towards+an+upscaled+multi-jet+process.">Electrospinning of polymer melt : steps towards an upscaled multi-jet process.</a> , 71-76 (2009).</li><li>Nayak, R., Padhye, R., Kyratzis, I. 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F., Luxenhofer, R., Groll, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+definition+fibrous+poly(2-ethyl-2-oxazoline)+scaffolds+through+melt+electrospinning+writing.">High definition fibrous poly(2-ethyl-2-oxazoline) scaffolds through melt electrospinning writing.</a> Polymer. 55 (20), 5017-5023 (2014).</li><li>Chen, 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=Additive+Manufacturing+of+a+Photo-Cross-Linkable+Polymer+via+Direct+Melt+Electrospinning+Writing+for+Producing+High+Strength+Structures.">Additive Manufacturing of a Photo-Cross-Linkable Polymer via Direct Melt Electrospinning Writing for Producing High Strength Structures.</a> Biomacromolecules. 17 (1), 208-214 (2016).</li><li>Hochleitner, G., 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=Additive+manufacturing+of+scaffolds+with+sub-micron+filaments+via+melt+electrospinning+writing.">Additive manufacturing of scaffolds with sub-micron filaments via melt electrospinning writing.</a> Biofabrication. 7 (3), 035002 (2015).</li></ol>

Интеграция утра для того, чтобы найти новаторские решения проблем в области медицины представляет новую парадигму в веке 21Сент . Так называемые поля «Био изготовление» находится на подъеме и инновации в технологии изготовления позволяют производство сложных архитектур для TE пр?...

Производство трехмерных (3D) биосовместимых структур для клеток является одним из основных вкладов добавка biomanufacturing ткани инженерия (TE), направленных на восстановление тканей путем применения настраиваемого биоматериалов, клетки, биохимические факторы, или их сочетание. Таким образом, основные требования подмости для TE приложений включают в себя: технологичность из биосовместимых материалов, контролируемый морфологических свойств для целевой ячейки вторжения и оптимизированные свойства поверхности для расширения взаимодействия клеток 1.
МЭВ является техника производства растворителей, которая сочетает в себе принципы аддитивного производства (часто называемые 3D печать) и electrospinning для производства полимерных сеток с высоко упорядоченную ультратонких волокон морфологии2. Это прямой написания подход и точно депозиты волокон согласно предварительно запрограммированных кодов3, называется G-кодов. Расплава electrospun конструкции в настоящее время готовятся с использованием плоский4,5 или7 коллекционер оправки6,для изготовления пористых плоские и трубчатых лесов, соответственно.
Эта техника предлагает значительные преимущества для TE и регенеративной медицины (RM) сообщества благодаря возможности непосредственно печатать медико класса полимеров, таких как poly(ε-caprolactone) (PCL), который представляет отличная биосовместимость8. Другими преимуществами являются возможность настраивать размер и распределение пористости, сдав волокна образом высоко организованной изготовить подмости высоких коэффициентов поверхности к объему. Прежде чем могут быть выполнены МэВ, полимер сначала требует применения тепла9. Однажды в состоянии жидкости, давление прикладной воздуха заставляет его вытекать через металлический прядильная, который подключен к источнику высокого напряжения. Силу баланс между поверхностное натяжение и привлечение электростатически заряженной капли сборщику обоснованных приводит к образованию Тейлор конуса, следуют выброса струи10.
На рисунке 1показаны изображения и схема, чертеж устройства МЭВ внутреннего построения, используемого для этого протокола. Он дополнительно демонстрирует принципы использования изолента во избежание электрического разряда между нагревательными элементами и электрически заряженных латуни вокруг прядильная. Недостаточная изоляция приведет к внутреннего повреждения оборудования реализована.
В зависимости от регулировки трех системных параметров (температуры, коллекции скорости и давления воздуха) МэВ позволяет изготовления волокон с различными диаметрами, описано в разделе обсуждения. В большинстве случаев Однако, тонкая настройка и оптимизация струи потребуется прежде чем стабильной jet будет извлечена. Визуализация электрифицированных путешествия jet является эффективным способом для проверки согласованности и единообразия процесса. В идеальном случае траектории полета напоминает катенарными кривую, приобретенных в результате баланс силы, контролируемые параметры системы11. Кроме того микро - и макро структура лесов зависит от траектории полета полимер струи12. Подробная таблица мер по оптимизации и различные отклонения поведения приводится в разделе обсуждения.
В настоящем исследовании мы представляем протокол, который описывает шаги производства для изготовления высоко контролируемых волокнистых подмостей, с использованием технологии МэВ. В этой работе, медицинский класс PCL (молекулярный вес 95-140 кг/моль) был использован, как этот медицинский класс PCL улучшилось чистоты за технический сорт, и его механические свойства и свойства обработки превосходны для МЭВ. Широкий расплава, обработки диапазона PCL исходит от его низкой температурой плавления (60 ° C) и высокой термостойкостью. Кроме того PCL является медленно скорость биоразлагаемые полимерные, что делает его отличным материалом для многих тканей, инженерные приложения13.
Для этого исследования, температура и коллекционер расстояние будет оставаться постоянной (65 ° C и 82 ° C для шприца и прядильная температур (соответственно) и 12 мм для расстояния коллектор); применяется напряжения, коллекционер скорости и давления воздуха, однако, будут изменяться для изготовления волокон с целенаправленных диаметров. Подробный перечень опубликованных исследований с использованием МЭВ подмостей приводится в разделе результаты и раскрывает различные приложения для полей TE и RM (Таблица 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

Этот учебник отражает основополагающие принципы и руководящие принципы для electrospinning, написание с полимерными расплавов, аддитивного производства технологии с большим потенциалом для биомедицинских приложений. Техника облегчает прямое осаждение биосовместимых полимерных волокон для изготовления упорядоченной леса в субмикронных для микро шкалы. Создание стабильной, вязкоупругих полимерных jet между прядильная и коллекционер достигается с помощью приложенного напряжения и может быть написано прямой. Значительное преимущество типичный пористых эшафот — высокий коэффициент поверхности к объему, который обеспечивает увеличение эффективной адгезии сайты для клеток привязанность и роста. Управление процессом печати доводкой параметров системы позволяет высокую воспроизводимость качества печатной подмостей. Он также предоставляет гибкие производственные платформы для пользователей адаптировать морфологических структур подмости с их конкретными требованиями. Для этой цели мы представляем протокол для получения различных волокно диаметров с использованием расплава electrospinning написания (МэВ) с гидом поправки параметров, включая скорость, напряжения и коллекции скорость потока. Кроме того мы показали, как оптимизировать струи, обсудить часто опытные технические проблемы, объяснить неполадок и демонстрируют широкий спектр печати эшафот архитектур.

Два различных метода коллекции обычно используются в МэВ, которые являются оправки и плоские коллекцию. Результате архитектуры зависит от программирования G-код (Таблица 2), который выполняется программное обеспечение.
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Watch this Scientific Journal Video about Melt Electrospinning Writing of Three-dimensional PolyÎµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications at JoVE.c

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

Этот протокол служит в качестве всеобъемлющего руководства изготовить подмости через electrospinning полимером тает в режиме прямой записи. Мы систематически изложить процесс и определить соответствующие параметры для достижения целевых эшафот архитектуры.

Расплава Electrospinning написания трехмерных Poly(ε-caprolactone) леса с контролируемым морфологии тканей инженерных приложений

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

This video tutorial reflects on electrospinning with polymer melts in a direct writing mode and provides necessary guidelines to fabricate scaffolds with well ordered architectures. Electrospinning refers to a polymer processing technique where a viscous polymer is extruded through an orifice while the application of an electrical field provokes the the rise of a jet. Electrostatic forces accelerate the fiber towards an oppositely charged or grounded collector.In order to reach a required level of viscosity, polymers were traditionally liquified in solvents. These evaporate during the flight phase, which, in combination with the electrical field, provokes strong buckling of the fibers. This results in polymeric scaffolds whose morphological properties are challenging to control, characterize, and reproduce.This experiment demonstrates electrospinning with polymer melts. Instead of dissolving, the required level of viscosity is reached by the application of heat to virgin polymers. The lack of solvents improves the macromolecular entanglement of the polymer and results in a more stable fiber flight path.The micron scale fibers can be accurately deposited according to the movement of the collector. Electrospinning with polymer melts in a direct writing mode allows for the fabrication of well ordered scaffold architectures, down to the micron and even nano scale. We predict that the technology will play a key role in the future of tissue engineering and regenerative medicine.The technology bridges a long-awaited gap between conventional fiber-forming processes and 3D-printing principles and opens up new avenues for designing the next generation of scaffolds. Micro electrospinning writing facilitates the direct processing of medical-grade plastics, such polycaprolactone or PCL. Because of the small diameters and highly organized structures and porosity, melt electrospun PCL scaffolds are ideal structures for this as in vitro and in vivo.One major prerequisite for achieving an effective pyrolysis with valuable results is a highly functional soft-and hardware. Therefore, we engineered a fully integrated system with automated perimeter control. This facilitates a reliable and specifically reproducible production of scaffolds to sufficiently meet the requirements of the users.Melt electrospin writing involves manufacturing scaffolds with different architectures. The following tutorial describes material preparation, scaffold production, fiber diameter adjustment, and optimization of the jets. Fill two grams of PCL in a three milliliter plastic syringe and push the plunger in gently.Place the syringe in a preheated oven for eight hours at 65 degrees Celsius. Carefully squeeze the plunger until all the trapped air bubbles are removed. Connect a 23ga needle and attach it to the air pressure system.Press it down until the needle tip emerges from the brass pot. Install the collector on the stage. Mount and clean it with 70%ethanol.Determine speed and direction of the collector movement by programming the G-code to vary size, spatial fiber distance, and number of layers of the scaffold. For tubular scaffolds, select rotational speed, translational speed, and its repetitions to define the thickness of the scaffold. A detailed guideline to individually program G-codes for flat or tubular scaffolds can be found in the manuscript.Open the software Mach3 and upload the generated G-code. There are five adjustable system parameters which significantly determine the quality of the resulting scaffold. These are air pressure, voltage, collection speed, temperature, and working distance and should be adjusted in the following order.Place a 12-millimeter high 3D-printed plastic block between needle and collector to adjust the working distance. Switch on both heaters and set to 65 degrees for the top and 82 degrees for the bottom part. Initiate the air pressure and wait until the molten polymer is extruded from the needle.Before starting the process, ensure that all grounding cables are connected firmly. Close the front door of the enclosure which connects the interlock. Increase the voltage until the extruded melt transforms into a Taylor cone and ejects a molten jet towards the collector.Allow the polymer melt to be extruded on the still collector plate for five minutes to stabilize the jet. Electrospinning with polymer melts allows for the fabrication of different fiber diameters. This can be achieved via specifically varying the system parameters pressure, voltage, and speed, whilst maintaining distance and temperature on a constant level.Small-diameter fibers between three and 10 micron can be printed via applying a relatively low pressure, small to medium electrical field, and high collection speeds. Based on the experience in our lab, we can print four micrometer diameter fibers with the following values:0.9 bar, eight kilovolt, and 1, 700 millimeters per minute. Medium-sized diameters between 10 and 20 micron can be printed via applying relatively low pressure level, medium to high voltage, and collection speeds in the medium range.Based on the experience in our lab, we can print 13-micrometer diameter fibers with the following values:1.5 bar, 11 kilovolt, and 1, 200 millimeters per minute. Fibers in the range of 20 to 30 micron or more require large polymer extrusion rates alongside the application of comparably high voltages and the lowest possible collection speeds. Based on the experience in our lab, we can print 25-micrometer diameter fibers with the following values:2.6 bar, 12 kilovolts, and 700 millimeters per minute.In all three cases however, fine tuning and optimization will be required to obtain stable results with homogenous diameters. In melt electrospinning, only a perfectly balanced equilibrium between the forces determining the flow of the polymer mass and the forces attracting the jet towards the collector will eventually lead to consistent scaffold morphologies. A well balanced equilibrium is achieved once the flight path of the fiber resembles a catenary curve over a long period of time.However, three different deviating behaviors are known. First, an unbalanced distribution between delivered mass to the Taylor cone and respective drag forces on the fibers results in a unstable electrified jet. Here the Taylor cone is constantly overfed with polymer and releases its mass in frequent time periods.Frequently changing angles are visible and result in extremely differing diameters. In order to restabilize the process, decrease the flow rate to minimize polymer delivery and increase the speed and voltage to amplify pulling forces on the fiber. Secondly, the fiber travels down and perpendicularly deposits on the collector.Hereby the amount of acceleration forces surpasses the polymer supply in the Taylor cone. Slight increases of pressure and lowering the voltage resolves the situation, or altering the collection speed. Thirdly, the fiber lags behind and resembles a straight line under a high angle.Hereby relatively thick fibers are produced and dragged. This leads to increased inaccuracies of the deposited fibers. A decrease in collection speed will solve the problem.However, slight increases in the electrical field strength are needed. In melt electrospinning, there are two methods of collection:flat collectors and mandrel collectors. Applying flat collectors refers to the most common method.090 and 060 degree structures of different sizes are widely reported in literature. The capability of directly depositing molten fibers on the collector also facilitates the production of non-linear structures, such as repetitive circular structures. Tubular architectures can be achieved with cylindrical collectors which rotate as well as traverse parallel to the needle.Through fine tuning the decode of both the rotational as well as the translational speed, the orientation of the fibers can be modified. Higher rotational speeds than translational speeds lead to radially orientated pores and vice versa. The diameter of the tube depends on the diameter of the implemented mandrel.A detailed list of different scaffold architectures and their proposed applications in published studies can be found in the manuscript. Due to the highly sensitive electrical field and its specific dependency on the distance between the two electrodes, it is of great importance to regularly level both the collecting stage and the bottom side of the print head. If the distance between the collector and the head is set too close, the electrical field significantly increases until a spark rises between the components.Turn down the voltage immediately on the electrical control box, or open the door. Before every start of a process, double check all the cable connections and confirm that they are not loose. Any needle damage potentially leads to deformed Taylor cones and thus irregularities within the printing.Therefore, needles where the tip was bent accidentally must be disposed of. We hope that this video assists in the establishment of a stable electrified jet and that you become convinced, as we are, that melt electrospinning writing will play a significant role in the field of tissue engineering and regenerative medicine.

Research

JoVE Journal

Bioengineering

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Эластомерные PGS Строительные леса в Артериальная тканевой инженерии

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

Подготовка 3D фибрина лесов для стволовых Приложения культуре клеток

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

Строительные леса Self-отчетности для 3-Dimensional культуре клеток

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

Человека хрящевой ткани Изготовление с использованием трехмерных струйных технологии печати

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

Электропрядения фактор роста Освобождение микросфер в волокнистых Строительные леса

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

Генерация трехмерной всю толщину кожи Эквивалент техники и автоматизированных Ранение

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

Изготовление механически перестраиваемых и биологически активных металлов строительные леса для биомедицинских применений

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

Инженерная Трехмерные эпителиальной ткани Встроенные в внеклеточной матрицы

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

Изготовление Перевернутый коллоидный кристалл поли (этиленгликоль) строительные леса: трехмерная Cell Culture платформа для печени тканевой инженерии

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

Культивирование клеток млекопитающих в трехмерной пептид подмости

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