fabrication of an electrospun nanofibrous scaffold for use in the field of tissue engineering
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Fabrication of an electrospun nanofibrous scaffold for use in the field of tissue engineering. Tyler Crawford Shannon Daily. Purpose:. - PowerPoint PPT PresentationTRANSCRIPT
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To create a polycaprolactone mesh which enables cell activity and seeks to eventually provide an application in the field of tissue engineering toward biomimetic skin graft.
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ECM - main structural tissue of skin› Helps skin renew and generate› Provides signals to intercellular pathways
Engineered ECMs are known as scaffolds
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Ability to create scaffolds › Mimic the ECM (size and porosity)› High surface to volume ratio
Easy to vary mechanical and biological properties through changing materials
Flexible- allows cells to manipulate their environment
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Biocompatible polymer
Biodegradable at a rate that allows increased cell growth and stability
Easy to manipulate
Relatively low melting point - easy to use
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Clinically safe (FDA approval)
Proven to have potential for scaffolds in relation to tissue regeneration› Has created scaffolds w/ ideal conditions
High porosities Large amounts of surface areas
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Adding another biochemical can:› Increase stress resistance› Provide better adhesion of cells to the final scaffold› Increase the potential for cell proliferation
Biochemical should› Be a component of skin naturally› Must be able to be combined in a solution to be
electrospun
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Natural polymer that exhibits biocompatible and biodegradable qualities
Cellular binding capabilities
Anti-bacterial properties
High viscosity which limits electrospinning
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Create control meshes of pure PCL› Solution= PCL and acetic acid (solvent)› Electrospin
Starting parameters: 15 wt.% concentration, 20 cm from tip of syringe to collector plate, & 20 kV
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Vary voltage to create 9 meshes› 3 Voltages- 3 trials for each
20 kV 15 kV 25 kV
Examine mesh using Scanning Electron Microscope (SEM)
Culture fibroblast cells onto mesh
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Observing cells› Inverted light microscope
Analyze cell growth› Cell counts in cells per unit area (mm2)› Means and standard deviations› ANOVA (Analysis of Variance) tests
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Create solutions of PCL and chitosan Electrospin Vary concentration of chitosan to PCL
› .5% CHT› 1% CHT› 2% CHT
Total of 9 meshes (3 trials of each concentration)
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Analyze with SEM Culture fibroblast cells and seed into
meshes created Determine cell density Analyze with means, standard
deviations, and ANOVA tests
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Data obtained:› Fiber diameter
and pore diameter of mesh
› Cell density amounts
Analysis includes:› Means*› Standard
Deviations*› ANOVA tests
3 comparisons
*5-7 measurements/areas for these methods
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DateAcetic Acid Stir/Level Heat/Level Time Results
12/8/10 .5 Molar Yes /8 No
Approx. 1.5 hours Not Dissolved
12/8/10 Glacial Yes/8 No Approx. 1.5 hours Slightly Dissolved
12/14/10 Glacial Yes/9 No 3 hours Almost Completely Dissolved
12/20/10 Glacial Yes/5 Yes/520 minutes
Dissolved and then hardened
12/20/10 Glacial Yes/7 Yes/2 2.5 hours Dissolved, hardened by next class
12/20/10 Glacial Yes/5 No 3 hours Dissolved, still liquid next class
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15 wt.% solution created› 17 g. acetic acid, 3 g. PCL
Electrospun› 5 mL syringe with bevel tip› Flow rate: .02??
Mesh created within 2 hrs.
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Background Research Experimental Design ISEF (International Science and
Engineering Fair) Forms Started solutions Just began spinning
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Akhyari, P., Kamiya, H., Haverich, A., Karck, M., & Lichtenberg, A. (2008). Myocardial tissue engineering: The extracellular matrix. European Journal of Cardio-Thoracic Surgery, 34, 229-241. doi: 10.1016/j.ejcts.2008.03.062
Bhardwaj, N. & Kundu, S. C. (2010). Electrospinning: A fascinating fiber fabrication technique. Biotechnology Advances, 28, 325-347. doi: 10.1016/j.biotechadv.2010.01.004
Chong, E.J., Phan, T.T., Lim, I.J., Zhang, Y.Z., Bay, B.H., Ramakrishna, S., & Lim, C.T. (2007). Evaluation of electrospun PCL/gelatin nanofibrous scaffold for wound healing and layered dermal reconstitution. Acta Biomaterialia, 3, 321-330. doi: 10.1016/j.actbio.2007.01.002
Geng, X., Kwon, O-H., & Jang, J. (2005). Electrospinning of chitosan dissolved in concentrated acetic acid solution. Biomaterials, 26, 5427-5432.
Han, J., Branford-White, C.J., & Zhu, L.M. (2010). Preparation of poly(є-caprolactone)/poly(trimethylene carbonate) blend nanofibers by electrospinning. Carbohydrate Polymers, 79, 214-218. doi: 10.1016/j.carbpol.2009.07.052
Homayoni, H., Ravandi, S.A.H., & Valizadeh, M. (2009). Electrospinning of chitosan nanofibers: Processing optimization. Carbohydrate Polymers, 77, 656-661.
Lowery, J.L., Datta, N., & Rutledge, G.C. (2010). Effect of fiber diameter, pore size and seeding method on growth of human dermal fibroblasts in electrospun poly(є-caprolactone) fibrous mats. Biomaterials, 31, 491-504. doi: 10.1016/j.biomaterials.2009.09.072
Nisbet, D.R., Forsythe, J.S., Shen, W., Finkelstein, D.I., & Horne, M.K. (2009). A review of the cellular response on electrospun nanofibers for tissue engineering. Journal of Biomaterials Application, 24, 7-29.
Pham, Q.P., Sharama, V., & Mikos, A.G. (2006). Electrospinning of polymeric nanofibers for tissue engineering applications: A review. Tissue Engineering, 12,1197-1211.
Shevchenko, R.V., James, S.L., & James, S.E. (2010). A review of tissue-engineered skin bioconstructs available for skin reconstruction. Journal of the Royal Society Interface, 7, 229-258. doi: 10.1098/rsif.2009.0403
Sill, T.J., & von Recum, H.A. (2008). Electrospinning: Applications in drug delivery and tissue engineering. Biomaterials, 29, 1989-2006. doi: 10.1016/j.biomaterials.2008.01.011
Woodruff, M.A., & Hutmacher, D.W. (in press). The return of a forgotten polymer- Polycaprolactone in the 21st century. Progress in Polymer Science. doi: 10.1016/j.progpolymsci.2010.04.002
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