Engineering Multi-Functional Three- Dimensional Nervous Tissue Constructs Advisor: Dr. Bryan J. Pfister Presenter: Mevan Siriwardane, C 2 PRISM Research Fellow 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 Axon Stretch Growth Mechanical tension in nerves is natural during development in most species
Background Nerve Guidance Conduits for Treating Nerve Injury Hollow tubes used in support of nerve regeneration Nerve Guidance Conduits for Treating Nerve Injury Hollow tubes used in support of nerve regeneration Questions: What is tissue engineering?
2D Axon Stretch-Growth Building a Nerve Tissue Transplant: “Axon Stretch-Growth” 2-D Stretch-Growth
2D Axon Stretch-Growth
3D Axon Stretch-Growth Building a Nerve Tissue Transplant: “Axon Stretch-Growth” 3-D Stretch-Growth
3D Axon Stretch-Growth Design Considerations Sustaining stretch-grown axons in a gel Design Considerations Sustaining stretch-grown axons in a gel
3D Axon Stretch-Growth Axons crossing through mesh!
Axons also grow along the fibers of the nylon mesh!!! 3D Axon Stretch-Growth
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) 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)
Axon Outgrowth on 2D Rat Tail and Bovine Tendon Collagen
Morphology Differences b) DRG on Rat Tail Collagen Type I a) DRG on Bovine Tendon Collagen Type I
2D vs. 3D Axonal Outgrowth
Hypothesis The incorporation of uni-axial fibers to soft 3D collagen gels will provided a rigid framework for guiding directional outgrowth for regenerating axons
Axon Growth on Fibers a) DRG on Collagen Fibers b) DRG on PLLA Fibers
Axon Growth on Fibers PLLA Fiber Collagen Fiber
Axon Outgrowth Rate on Collagen Fibers
Collagen Bulk Fibers vs. Collagen-Coated PLLA Fibers
Future Directions a) DRG on Thick PLLA Fiber b) DRG on Thick Collagen Fiber c) DRG on Thin PLLA Fiber d) DRG on Thin Collagen Fiber Fiber Diameter and Molecular Orientation
Future Directions a) DRG on Thick PLLA vs. Collagen Fiber b) DRG on Thin PLLA vs. Collagen Fiber Preferential Growth
Future Directions 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) Multi-filament and Spacing
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 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
Questions? 3D Nervous Tissue Construct Fibers