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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. 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 l.jpg

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 l.jpg
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

Tmds temporomandibular disorders l.jpg
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

Current obstacles in engineering a tmj disc l.jpg
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

    • 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

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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

    • TGF alpha and beta

    • 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

Results l.jpg

  • 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

Conclusions l.jpg

  • 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

  • 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