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Attenuating the inflammatory wound-healing response through bilayer, bifunctional HA materials

Attenuating the inflammatory wound-healing response through bilayer, bifunctional HA materials. Sarah Mayes Genzyme June 29, 2011. Post-surgical adhesion formation is an aberrant response to injury, often permanently tethering tissues that should remain separate.

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Attenuating the inflammatory wound-healing response through bilayer, bifunctional HA materials

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  1. Attenuating the inflammatory wound-healing response through bilayer, bifunctional HA materials • Sarah Mayes • Genzyme • June 29, 2011

  2. Post-surgical adhesion formation is an aberrant response to injury, often permanently tethering tissues that should remain separate

  3. Delivery of drugs or agents combined with reduced approximation of tissues can guide the wound-healing response by speeding the process Injury Response Delivery Remain Inflammation Recruitment Repair Normal Scar Time Injury Response Resolve Significant time reduction may eliminate unwanted scar formation Film Repair Normal Inflammation Normal

  4. Film synthesis 1. Syringe filter solution 2% Alginate / 1% GMHA / 4% Urea 2. Film-by-casting at 25°C and 70% Rh, closed to ambient 3. Add seed crystal to nucleate crystallization. Alginate Alginate Urea 4. Expose to UVA 1000 mW/cm2 for 30 seconds 8. Re-cast dehydrated film in GMHA solution at 25°C and 70% Rh, closed to ambient 5. Add 100 mM CaCl2 to film to crosslink Alginate in current configuration. H2O 7. Controlled dessication at 50% Rh and under force to remove H2O. 6. Rinse out urea crystals with methanol / ddiH2O. F Schmidt, Zawko, Mayes SCH 5337, 5884, 5885 CONFIDENTIAL

  5. Cryo-SEM images of film surface showing pore / fiber alignment Alginate Alginate / GMHA patterned GMHA patterned Alginate patterned

  6. Instron ASTM D638 (V) Mechanical Characterization of Film Method used to calculate modulus: Febriyenti, 2010

  7. Instron ASTM D638 (V) Mechanical Characterization of Film

  8. Instron ASTM D638 (V) Mechanical Characterization of Film Method used to calculate modulus: Febriyenti, 2010

  9. In vitro Cytotoxicity (with Vanessa) 4. At 24 hrs replace media with leaching media, 8 wells per film 2. Incubate 24 hours at 37 deg C 1. Place 1” x 1.5” film in 3 mL DMEM media 3. Seed 3,000 cell/well in 96 well plate with 200 uL of media for 6 hours 5. Seed between 20k and 4k cells for calibration, 2 wells per cell amount 6. After bringing to RT, add 1:1 media : cell titer glo for luminescence

  10. In vitro anti-adhesion study (with Vanessa and Jessie S.)

  11. in vivo Efficacy Study – Peritoneal Abrasion Model, N = 40

  12. Peritoneal model results No barrier Grade 3 adhesions Day 14 HA/Alginate barrier Grade 0 adhesions (no adhesions) Day 14

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