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Fabrication of an electrospun nanofibrous scaffold for use in the field of tissue engineering

Fabrication of an electrospun nanofibrous scaffold for use in the field of tissue engineering. By: Shannon Daily & Tyler Crawford. Purpose.

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Fabrication of an electrospun nanofibrous scaffold for use in the field of tissue engineering

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  1. Fabrication of an electrospunnanofibrous scaffold for use in the field of tissue engineering By: Shannon Daily & Tyler Crawford

  2. Purpose • 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.

  3. Progress since last meeting • Spun 8 more meshes • Visited Janelia Farm and used SEM • Details on cell procedure • Began cell work

  4. Electrospun Meshes • Mesh 5 & 6 • Spun on 3/1 • 15 kV • 15 cm from tip of pipette to collector plate • Pasteur pipette used • Glass slide down middle

  5. Electrospun Meshes • Mesh 7 & 8 • Spun on 3/11 • 15 wt.% solution from 3/9 • Mesh 7 • 20 kV • 15 cm • No glass slide • Mesh 8 • 20 kV • 20 cm • Glass slide

  6. Electrospun Meshes • Mesh 9 & 10 • Spun on 3/17 • 10 wt.% solution from 3/15 • Mesh 9 • 15 kV • 15 cm • Glass slide • Mesh 10 • 20 kV • 15 cm • Glass slide

  7. Electrospun Meshes • Mesh 11 & 12 • Spun on 3/23 • 20 wt.% solution from 3/21 • Mesh 11 • 15 kV • 15 cm • Glass slide • Mesh 12 • 20 kV • 15 cm • Glass slide

  8. Chitosan Solution • Created PCL/chitosan solution to begin electrospinning • 15 wt.% PCL, 1 wt.% chitosan • 3 g PCL, .2 g chitosan, 16.8 g acetic acid

  9. SEM Images Mesh 4

  10. SEM Images Mesh 4

  11. SEM Images Mesh 6

  12. SEM Images Mesh 6

  13. SEM Images Mesh 6

  14. Conclusion from SEM • Beading is still a problem • Sections of fibers and sections of beading • Ideas to fix • Increase the voltage • Decrease the concentration • Increase the distance • Comparison of fiber size • Mesh 1: 185 nm vs. Mesh 4&6: ~300 nm

  15. Cell Work • Meetings with Anu, Estelle, and Ms. Curley • Determined basic procedure • Centrifuge thawed cells • Add media to pellet of cells & resuspend • Transfer to flask (in our case, tissue culture plates) • Allow cells to grow until about plate covered about 80% • Split to new containers

  16. Cell Work • Checked to see if mesh dissolves in media • Placed small piece of mesh in 500 µL of media • Let sit for 2 days- checking periodically • Mesh did not dissolve meaning cell growth is possible

  17. To be worked on: • Use SEM on meshes created since last time (7-12) • Different SEM? • Cell work • Begin with Mesh 4 (best fibers imaged) • Continue with other meshes • Begin spinning PCL/chitosan solutions

  18. Bibliography 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. ActaBiomaterialia, 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 chitosannanofibers: 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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