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Engineering a TMJ Disc

Engineering a TMJ Disc. Danielle Lewis Louisiana Tech University REU, University of Louisiana at Lafayette Dr. David Mills Louisiana Tech University. What is a TMJ Disc?. Temporomandibular Joint Disc Located between the base of the skull and the lower jaw

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Engineering a TMJ Disc

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  1. Engineering a TMJ Disc Danielle Lewis Louisiana Tech University REU, University of Louisiana at Lafayette Dr. David Mills Louisiana Tech University

  2. What is a TMJ Disc? • Temporomandibular Joint Disc • Located between the base of the skull and the lower jaw • Allows for smooth opening and closing of the jaw

  3. TMDs: Temporomandibular Disorders • Improper disc placement hampers proper jaw movement, called disc displacements • 7-28% of adult population affected, mainly females between 18 and 25 • No known treatment for disorder • In severe cases discs are completely removed and patients can no longer move jaw

  4. http://www.jawjointpainrelief.com/what_is_TMJ.asp

  5. Current Obstacles in Engineering a TMJ disc • Recreating the intricate cellular structure • 3 regions: anterior band, intermediate zone, and posterior band • Anterior and posterior bands: interlaced collagen fiber bundles • Intermediate zone: aligned collagen fibers along with tiny fibrochondrocytes and small elastin fibers

  6. Approach of the Mills Lab:Electrospun Scaffolds • Electrospinning: • A high voltage is passed through a polymer solution inducing an electrostatic repulsion force • The polymer is pumped through an insulin syringe, the repulsion force results in the formation of a thin jet • This jet is directed toward a grounded collection plate, the solvent evaporates before hitting the collection plate and results in the formation of a polymer scaffold

  7. My Research Plan • Culture bovine fibrochondrocytes (FBCs) in 3 different gel environments – agarose, collagen, alginate • Treat cells with growth factors to observe their effect on proliferation, cell survival and protein expression • bFGF • TGF alpha and beta • CTGF • Characterize the extracellular matrix being produced by the FBCs

  8. Cell Isolations • Bovine FBCs were isolated from TMJ discs taken from cow skulls • Cells used in experiments were passage 6, slightly old but still exhibited the characteristic FBC shape

  9. Gels:Collagen and Agarose • FBCs were suspended in both collagen and agarose gels and allowed to grow for several days, 18 day group and 8 day group

  10. Fixing, Dehydration, and Paraffin Imbedding • Gels fixed in 2% paraformaldehyde • Gels were dehydrated by exposing them to a variety of ethanol solutions then infiltrated with pariffin to preserve them indefinitely • Gels were then imbedded in a paraffin block in preparation for creating slides • Next… Immunohistochemistry

  11. Results • Simple examination under phase contrast showed that FBCs thrived better in collagen gels, cells attached and displayed characteristic shape

  12. Is there any explanation for this observation? • Expected result • Collagen type I abundant in TMJ disc

  13. Conclusions • Bovine fibrochondrocytes appeared to prefer the collagen gel environment over the agarose gel environment

  14. Next Steps…. • Begin staining gels for extra-cellular matrix proteins • Hypothesis: FBCs grown in collagen gels will exhibit an increase in extra-cellular matrix proteins over those grown in agarose gels • Use this experiment as a control and continue by adding growth factors to FBCs in gel culture • Compare the amounts and types of extra-cellular matrix proteins found in gels, both with growth factors and without, to that found in actual disc and FBCs grown on electrospun scaffolds

  15. Thank you… • Dr. Mills, Kanthi, Skylar, Deepak, Stephanie, Paul • Dr. Jones • Louisiana Tech for lab facilities in Carson Taylor and the BME building • National Science Foundation

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