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  • Polymer Question

    Hello All,

    I am working with a polymer for tissue engineering applications and was
    wondering if any of the list subscribers would have insightful suggestions
    on how to breakdown the scaffold for subsequent biochemical analyses.

    The polymer I am using is a 5 mm in diameter and 3 mm in height
    fiber-reinforced, porous, resorbable scaffolds, composed of a matrix made
    from a random copolymer of 75:25 poly(D,L-lactide-co-glycolide) with a glass
    transition onset temperature (Tg) of 50 C, and reinforcing poly(glycolide)
    fibers with glass transition onset temperature Tg of 42 C. The porosity of
    the scaffold is between about 65% and 70% by volume.

    I am seeding chondrocytes onto these scaffolds and I am interested in
    measuring DNA content, Proteoglycan and Collagen (hydroxyproline) content,
    as well as isolate RNA for RT-PCR analysis. All of these assays need not be
    done on one sample, as I can dedicate certain numbers of scaffolds for each
    of the analyses.

    To do these analyses, I tried to digest the scaffold (after lyophilizing it)
    with 1 milliliter of papain solution (25 mg/mL papain solution in 2mM of
    L-cysteine and 2mM of EDTA with Phosphate Buffered Solution as the solvent
    base; pH 6.5) at 65C for 15 to 18 hours overnight. But this treatment
    seemed to do nothing on the scaffolds. Of course I could use a harsher
    solvent (like Soluene) that would dissolve the polymer but that would also
    dissolve the plastic plates I use for the assays I mentioned.

    I would really appreciate any input, and will be glad to share the answers I
    get with the rest of the group.

    Sincerely,

    Hani Awad, PhD
    Post Doctoral Research Associate
    Duke University Medical Center
    haniawad@duke.edu

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