2. 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.  ECM - main structural tissue of skin › Helps skin renew and generate › Provides signals to intercellular pathways  Engineered ECMs are known as scaffolds

4.  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

5.  Biocompatible polymer  Biodegradable at a rate that allows increased cell growth and stability  Easy to manipulate  Relatively low melting point - easy to use

6.  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

7.  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

8.  Natural polymer that exhibits biocompatible and biodegradable qualities  Cellular binding capabilities  Anti-bacterial properties  High viscosity which limits electrospinning

9.  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

10.  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

11.  Observing cells › Inverted light microscope  Analyze cell growth › Cell counts in cells per unit area (mm 2 ) › Means and standard deviations › ANOVA (Analysis of Variance) tests

12.  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)

13.  Analyze with SEM  Culture fibroblast cells and seed into meshes created  Determine cell density  Analyze with means, standard deviations, and ANOVA tests

14.  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

15. Date Acetic AcidStir/LevelHeat/LevelTimeResults 12/8/10.5 MolarYes /8No Approx. 1.5 hoursNot Dissolved 12/8/10GlacialYes/8No Approx. 1.5 hoursSlightly Dissolved 12/14/10GlacialYes/9No3 hours Almost Completely Dissolved 12/20/10GlacialYes/5 20 minutes Dissolved and then hardened 12/20/10GlacialYes/7Yes/22.5 hours Dissolved, hardened by next class 12/20/10GlacialYes/5No3 hours Dissolved, still liquid next class

16.  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.

18.  Background Research  Experimental Design  ISEF (International Science and Engineering Fair) Forms  Started solutions  Just began spinning

19.  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 21 st century. Progress in Polymer Science. doi: 10.1016/j.progpolymsci.2010.04.002