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Engineering Multi-Functional Three-Dimensional Nervous Tissue Constructs

Engineering Multi-Functional Three-Dimensional Nervous Tissue Constructs. Advisor: Dr. Bryan J. Pfister Presenter: Mevan Siriwardane, C 2 PRISM Research Fellow. Development and Growth. Axon Stretch Growth Mechanical tension in nerves is natural during development in most species.

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Engineering Multi-Functional Three-Dimensional Nervous Tissue Constructs

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  1. Engineering Multi-Functional Three-Dimensional Nervous Tissue Constructs Advisor: Dr. Bryan J. Pfister Presenter: Mevan Siriwardane, C2PRISM Research Fellow

  2. Development and Growth • Axon Stretch Growth • Mechanical tension in nerves is natural during development in most species

  3. Background • Nerve Guidance Conduits for Treating Nerve Injury • Hollow tubes used in support of nerve regeneration Questions: What is tissue engineering?

  4. 2D Axon Stretch-Growth • Building a Nerve Tissue Transplant: “Axon Stretch-Growth” 2-D Stretch-Growth

  5. 2D Axon Stretch-Growth

  6. 2D Axon Stretch-Growth

  7. 3D Axon Stretch-Growth • Building a Nerve Tissue Transplant: “Axon Stretch-Growth” 3-D Stretch-Growth

  8. 3D Axon Stretch-Growth • Design Considerations • Sustaining stretch-grown axons in a gel

  9. 3D Axon Stretch-Growth • Axons crossing through mesh!

  10. 3D Axon Stretch-Growth • Axons also grow along the fibers of the nylon mesh!!!

  11. Axon Outgrowth on 2D and 3D Collagen Substrates • Evaluation of Candidate Materials for Nerve Tissue Construct • Rat Tail Collagen Type I • Bovine Tendon Collagen Type I • Study Axon Growth Behavior • 2D coating on dishes (Collagen) • 3D Hydrogels (Collagen)

  12. Axon Outgrowth on 2D Rat Tail and Bovine Tendon Collagen

  13. Axon Outgrowth on 2D Rat Tail and Bovine Tendon Collagen • Morphology Differences a) DRG on Bovine Tendon Collagen Type I b) DRG on Rat Tail Collagen Type I

  14. 2D vs. 3D Axonal Outgrowth

  15. Hypothesis • The incorporation of uni-axial fibers to soft 3D collagen gels will provided a rigid framework for guiding directional outgrowth for regenerating axons

  16. Axon Growth on Fibers a) DRG on Collagen Fibers b) DRG on PLLA Fibers

  17. Axon Growth on Fibers PLLA Fiber Collagen Fiber

  18. Axon Outgrowth Rate on Collagen Fibers

  19. Collagen Bulk Fibers vs. Collagen-Coated PLLA Fibers

  20. Future Directions Fiber Diameter and Molecular Orientation a) DRG on Thick PLLA Fiber b) DRG on Thick Collagen Fiber c) DRG on Thin PLLA Fiber d) DRG on Thin Collagen Fiber

  21. Future Directions Preferential Growth a) DRG on Thick PLLA vs. Collagen Fiber b) DRG on Thin PLLA vs. Collagen Fiber

  22. Future Directions Multi-filament and Spacing a) DRG on Multi-filament Thin PLLA Fibers b) DRG on Multi-filament Thin Collagen Fibers c) DRG on Multi-filament Thin PLLA Fibers (Spaced) d) DRG on Multi-filament Thin Collagen Fibers (Spaced)

  23. Future Directions • Protein coatings on collagen fibers • Comparison of rat tail and bovine tendon collagen hydrogels on 3D axon outgrowth • Integration of living axon stretch-grown cultures into hydrogel matrix with fiber-reinforced composites • Clinical evaluation of construct using in vivo rat models

  24. Questions? 3D Nervous Tissue Construct Fibers Fibers

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