Name: Postproduction Processing of Electrospun Fibres for Tissue Engineering (Video) | JoVE Journal
Uploaded: 2012-08-09T13:00:00.000+00:00
Duration: 15 min 52 s
Description: Learn postproduction processing of electrospun biodegradable scaffolds for tissue engineering applications. This method enables researchers to create sterilizable, mechanically robust multilayered constructs for clinical use.

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We thank BBSRC for funding a PhD for Mr. Frazer Bye.

<ol><li>Canton, I., McKean, R., Charnley, M., Blackwood, K., Fiorica, C., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+an+Ibuprofen-releasing+biodegradable+PLA%2FPGA+electrospun+scaffold+for+tissue+regeneration%5BTitle%5D)+AND+%22Biotechnology+and+bioengineering%22%5BJournal%5D)">Development of an Ibuprofen-releasing biodegradable PLA/PGA electrospun scaffold for tissue regeneration.</a> Biotechnology and bioengineering. 105, 396-408 (2010).</li>
<li>Blackwood, K., McKean, R., Canton, I., Freeman, C., Franklin, K., Cole, A., Brook, I., Farthing, P., Rimmer, S., Haycock, J., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+biodegradable+electrospun+scaffolds+for+dermal+replacement%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Development of biodegradable electrospun scaffolds for dermal replacement.</a> Biomaterials. 29, 3091-3104 (2008).</li>
<li>Yang, F., Maurugan, R., Wang, S., Ramakrishna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrospinning+of+nano%2Fmicro+scale+poly%28L-lactic+acid%29+aligned+fibers+and+their+potential+in+neural+tissue+engineering%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neural tissue engineering.</a> Biomaterials. 26, 2603-2610 (2005).</li>
<li>Sittichokechaiwut, A., Edwards, J. H., Scutt, A. M., Reilly, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Short+bouts+of+mechanical+loading+are+as+effective+as+dexamethasone+at+inducing+matrix+production+by+human+bone+marrow+mesenchymal+stem+cell%5BTitle%5D)+AND+%22Eur.+Cell+Mater%22%5BJournal%5D)">Short bouts of mechanical loading are as effective as dexamethasone at inducing matrix production by human bone marrow mesenchymal stem cell.</a> Eur. Cell Mater. 20, 45-57 (2010).</li>
<li>Sill, T. J., von Recum, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrospinning%3A+applications+in+drug+delivery+and+tissue+engineering%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Electrospinning: applications in drug delivery and tissue engineering.</a> Biomaterials. 29 (13), 1989-2006 (2008).</li>
<li>Deitzel, J., Kleinmeyer, J., Harris, D., Beck Tan, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effect+of+processing+variables+on+the+morphology+of+electrospun+nanofibers+and+textiles%5BTitle%5D)+AND+%22Polymer%22%5BJournal%5D)">The effect of processing variables on the morphology of electrospun nanofibers and textiles.</a> Polymer. 42, 261-272 (2001).</li>
<li>Fridrikh, S., Yu, J., Brenner, M., Rutledge, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Controlling+the+fiber+diameter+during+electrospinning%5BTitle%5D)+AND+%22Physical+review+letters%22%5BJournal%5D)">Controlling the fiber diameter during electrospinning.</a> Physical review letters. 90, 1-4 (2003).</li>
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<li>Tong, H. -W., Wang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrospinning+of+poly%28hydroxybutyrate-co-hydroxyvalerate%29+fibrous+tissue+engineering+scaffolds+in+two+different+electric+fields%5BTitle%5D)+AND+%22Polymer+Engineering+%26+Science%22%5BJournal%5D)">Electrospinning of poly(hydroxybutyrate-co-hydroxyvalerate) fibrous tissue engineering scaffolds in two different electric fields.</a> Polymer Engineering & Science. 51 (7), 1325-1338 (2011).</li>
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No conflicts of interest declared.

Electrospinning is a very popular technique for producing scaffolds for tissue engineering.14, 15, 16 While it is relatively simple to produce basic electrospun scaffolds for experimental use the technique is also complex and multifaceted with many variables.6 There are many studies describing how the electrospinning parameters determine the scaffold produced. In this study the focus is on the considerable challenges post production to make scaffolds of appropriate architectures and mechanical pr...

<table><tbody><tr><td>Poly lactic-co-glycolic acid</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Poly lactic acid</td><td>Sigma Aldrich</td><td>81273</td><td>Inherent viscosity ~2.0dl/g</td></tr><tr><td>Poly &amp;#949;-caprolactone</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Poly hydroxybutyrate-co-hydroxyvalerate 12:1</td><td>Goodfellow</td><td>578-446-59</td><td>PHB88/PHV12</td></tr><tr><td>Dichloromethane</td><td>Sigma Aldrich or Fisher</td><td>270997 or D/1850/17</td><td>&amp;gt;99.8% contains 50-150ppm amylene stabiliser</td></tr><tr><td>50 multi coloured balloons</td><td>Wilkinson&amp;#8217;s Hardware Stores Ltd.</td><td>0105790</td><td></td></tr><tr><td>Goat anti-rabbit IgG (FC):FITC</td><td>AbDserotec</td><td>STAR121F</td><td></td></tr><tr><td>Rabbit anti-human alpha elastin</td><td>AbDserotec</td><td>4060-1060</td><td></td></tr><tr><td>Screw Cap GL45 PP 2 Port, pk/2</td><td>SLS</td><td>1129750</td><td></td></tr><tr><td>4&amp;#8242;,6-Diamidino-2-phenylindole dihydrochloride</td><td>Sigma Aldrich</td><td>32670</td><td></td></tr><tr><td>CellTracker green CMFDA</td><td>Invitrogen</td><td>C7025</td><td></td></tr><tr><td>CellTracker red CMTX</td><td>Invitrogen</td><td>C34552</td><td></td></tr></tbody></table>

postproduction processing of electrospun fibres for tissue engineering

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Electrospinning is a commonly used and versatile method to produce scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo undergo biaxial distension to varying extents such as skin, bladder, pelvic floor and even the hard palate as children grow. In producing scaffolds for these purposes there is a need to develop scaffolds of appropriate biomechanical properties (whether achieved without or with cells) and which are sterile for clinical use. The focus of this paper is not how to establish basic electrospinning parameters (as there is extensive literature on electrospinning) but on how to modify spun scaffolds post production to make them fit for tissue engineering purposes - here thickness, mechanical properties and sterilisation (required for clinical use) are considered and we also describe how cells can be cultured on scaffolds and subjected to biaxial strain to condition them for specific applications.
Electrospinning tends to produce thin sheets; as the electrospinning collector becomes coated with insulating fibres it becomes a poor conductor such that fibres no longer deposit on it. Hence we describe approaches to produce thicker structures by heat or vapour annealing increasing the strength of scaffolds but not necessarily the elasticity. Sequential spinning of scaffolds of different polymers to achieve complex scaffolds is also described. Sterilisation methodologies can adversely affect strength and elasticity of scaffolds. We compare three methods for their effects on the biomechanical properties on electrospun scaffolds of poly lactic-co-glycolic acid (PLGA).
Imaging of cells on scaffolds and assessment of production of extracellular matrix (ECM) proteins by cells on scaffolds is described. Culturing cells on scaffolds in vitro can improve scaffold strength and elasticity but the tissue engineering literature shows that cells often fail to produce appropriate ECM when cultured under static conditions. There are few commercial systems available that allow one to culture cells on scaffolds under dynamic conditioning regimes - one example is the Bose Electroforce 3100 which can be used to exert a conditioning programme on cells in scaffolds held using mechanical grips within a media filled chamber.4 An approach to a budget cell culture bioreactor for controlled distortion in 2 dimensions is described. We show that cells can be induced to produce elastin under these conditions. Finally assessment of the biomechanical properties of processed scaffolds cultured with or without cells is described.

Electrospun scaffolds can be processed post production for tissue engineering applications. Here we describe methods for spinning complex scaffolds (by consecutive spinning), for making thicker scaffolds (by multi-layering using heat or vapour annealing), for achieving sterility (aseptic production or sterilisation post production) and for achieving appropriate biomechanical properties.

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

The aim of this procedure is to produce electro spun biodegradable scaffolds that can be sterilized and made thick enough and of appropriate mechanical strength for clinical tissue engineering. This is accomplished by first electro spinning biodegradable polymers into primary architectures, either random or reliant and orientated or sequentially layered. The second step is to apply post spinning modifications to the scaffolds by putting several layers together and sterilizing them.Next cells are introduced to the scaffolds and their performance during and after culture is assessed. The final step is to subject the cells to an exercise regime to mimic the physiological stress the cells will encounter in the body in order to prepare them for transplantation. Ultimately, we can assess the response of cells to exercise by measuring the extracellular matrix they produce in response to dynamic tension.This technique is versatile and flexible, allowing a large variety of scaffolds to be made with properties specifically modified for the required application. For example, scaffolds can be designed to treat difficult conditions where tissue repairs needed, such as replacement of extensive s

postproduction-processing-of-electrospun-fibres-for-tissue-engineering

Research

JoVE Journal

Bioengineering

The process of electrospinning polymers for tissue engineering and cell culture is addressed in this article.  Specifically, the electrospinning of photoreactive macromers with additional processing capabilities of photopatterning and multi-polymer electrospinning is described.

Electrospinning Fibrous Polymer Scaffolds for Tissue Engineering and Cell Culture

The aim of this study was to mimic the native three layered architecture of the arterial wall.  To accomplish this, electrospinning was employed with the use of a 3-1 (input-output) nozzle and blends of polycaprolactone, elastin, and collagen.

Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study

Electrospinning techniques can create a variety of nanofibrous scaffolds for tissue engineering or other applications.  We describe here a procedure to optimize the parameters of the electrospinning solution and apparatus to obtain fibers with the desired morphology and alignment.  Common problems and troubleshooting techniques are also presented.

Electrospinning Fundamentals: Optimizing Solution and Apparatus Parameters

This protocol combines electrospinning and microspheres to develop tissue engineered scaffolds to direct neurons. Nerve growth factor was encapsulated within PLGA microspheres and electrospun into Hyaluronic Acid (HA) fibrous scaffolds.  The protein bioactivity was tested by seeding the scaffolds with primary chick Dorsal Root Ganglia and culturing for 4-6 days.

Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds

Synthesis and fabrication of electrospun long fibers spanning a larger deposit area via a newly designed collector from a novel biodegradable polymer named poly(glycerol-dodecanoate) (PGD) was reported. The fibers were able to support the growth of cells derived from mouse pluripotent stem cells.

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells

Here, we present a protocol to fabricate electrospun nanofiber scaffolds with gradated organization of fibers and explore their applications in regulating cell morphology/orientation. Gradients with regard to physical and chemical properties of the nanofiber scaffolds offer a wide variety of applications in the biomedical field.

Electrospun Nanofiber Scaffolds with Gradations in Fiber Organization

Advancements in biomaterial technologies enable the development of three-dimensional multi-cell-type constructs. We have developed electrospinning protocols to produce three individual scaffolds to culture the main structural cells of the airway to provide a 3D in vitro model of the airway bronchiole wall.

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

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

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

Learn postproduction processing of electrospun biodegradable scaffolds for tissue engineering applications. This method enables researchers to create sterilizable, mechanically robust multilayered constructs for clinical use.

Postproduction Processing of Electrospun Fibres for Tissue Engineering

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