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

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  • Engineering a TMJ Disc Danielle Lewis Louisiana Tech University REU, University of Louisiana at Lafayette Dr. David Mills Louisiana Tech University
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  • What is a TMJ Disc? Temporomandibul ar Joint Disc Located between the base of the skull and the lower jaw Allows for smooth opening and closing of the jaw
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  • 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
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  • http://www.jawjointpainrelief.com/what_is_TMJ.asp
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  • 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
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  • Approach of the Mills Lab: Electrospun Scaffolds Electrospinning: A high voltage is passed through a polymer solution inducing an electrostatic repulsion force 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 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 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
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  • 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 bFGF TGF alpha and beta TGF alpha and beta CTGF CTGF Characterize the extracellular matrix being produced by the FBCs
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  • 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
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  • 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
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  • 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
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  • Results Simple examination under phase contrast showed that FBCs thrived better in collagen gels, cells attached and displayed characteristic shape
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  • Is there any explanation for this observation? Expected result Collagen type I abundant in TMJ disc
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  • Conclusions Bovine fibrochondrocytes appeared to prefer the collagen gel environment over the agarose gel environment
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  • 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 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
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  • 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