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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

Fabrication of an electrospunnanofibrous scaffold for use in the field of tissue engineering

By: Shannon Daily & Tyler Crawford

purpose
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.
progress since last meeting
Progress since last meeting
  • Spun 8 more meshes
  • Visited Janelia Farm and used SEM
  • Details on cell procedure
  • Began cell work
electrospun meshes
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
electrospun meshes1
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
electrospun meshes2
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
electrospun meshes3
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
chitosan solution
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
conclusion from sem
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
cell work
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
cell work1
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
to be worked on
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
bibliography
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