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Junior Research

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

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Junior Research

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

  2. Purpose: To create a nanofibrous mesh consisting of polycaprolactone and another biological polymer which enables cell activity and seeks to eventually provide an application in the field of tissue engineering toward a biomimetic skin graft.

  3. Main Qualities to Replicate in the Creation of an Artificial Skin Graft • Protection from infection • Prevent fluid/heat loss • Ability to support and maintain tissue growth • Skin properties • Friction & elasticity • For easy movement and manipulation

  4. The Extracellular Matrix (ECM) • ECM - main structural tissue of skin • Helps skin renew and generate • Provides signals to intercellular pathways • Main components • Glycoproteins (such as collagen) • Proteoglycans • Hyaluronic Acid • Engineered ECMs are known as scaffolds

  5. Electrospinning • Ability to create scaffolds • mimic the ECM in size and porosity • Have high surface to volume ratio • More space for cells to attach and grow • Increases biocompatibility • Easy to vary mechanical and biological properties through changing materials • Flexible- allows cells to manipulate their environment

  6. Polycaprolactone (PCL) • Biocompatible polymer • Biodegradable at a slow enough rate to allow increased cell growth and stability • Easy to manipulate • Relatively low melting point- easy to use • 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. Additional Biochemical Material • Much research has shown that 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. Potential Biochemical Polymers • Collagen • Advantages • biodegradable and biocompatible • plays important role in tissue formation • Disadvantages • Very expensive • complex handling properties • Gelatin • Advantages • naturally derived from collagen, similar properties • Cost efficient and easy to manipulate • Disadvantages • can provoke inflammatory response • Poor electrospinnability unless combined with specific solvents

  9. Potential Polymers continued… • Hyaluronic Acid • Advantages: • Excellent biocompatibility and biodegradability • Main component of ECM • Disadvantages •  High viscosity, surface tension, and water retention make it difficult to form uniform sized fibers • Elastin • Advantages • Provides elasticity to skin- essential for this skin quality • Disadvantages •  highly insoluble • Potential health risk • Fibrinogen • Advantages • Essential for wound healing • Promotes cell migration and cellular interaction • Disadvantages •  difficult to control matrix properties

  10. Potential Polymers Continued: • Alginate • Advantages • Good for health reasons (low toxicity, immunogenic) • Low cost • Disadvantages • Poor spinnability (possibly be fixed with addition of a synthetic polymer) • Chitosan • Advantages • natural polymer, biocompatible and biodegradable • Cellular binding capabilities • Accelerates wound healing • Anti-bacterial properties • Disadvantages • high viscosity limits spinnability • Fibers can swell in aqueous solution- need to be cross linked to maintain structural qualities

  11. Experimental Design Progress: Procedure • Create solutions of PCL and other polymer varying the concentrations • Spin these solutions creating nanofilament meshes • Analyze meshes for fiber and pore qualities using scanning electron microscope • Culture fibroblast cells and seed into meshes created

  12. Experimental Design Progress:Data and Analysis • Data obtained will include: • Fiber diameter and pore diameter of the mesh • Concentration of the chemical • Amount of cell activity throughout mesh • Analysis will include: • For what concentration of chemical did the most cell activity occur

  13. References 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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