1. Scott Zawko 2012.11.19 Data Blitz

2. Bioresorption and Efficacy Study

3. Sodium displacement of calcium Na + Ca 2+ Na + Ca 2+

4. Calcium chelation Citrate EDTA Na + Ca 2+ Na + Ca 2+ Chelator

5. Surgical procedure

6. Rat cecal abrasion studies DateFilmsChelatorOutcome July 2012 Templated Non-templated EDTA or Citrate 0.2 mL 100 mM 34 of 35 films did not degrade

7. Chelator treatment promotes degradation in rat carcass

8. Rat cecal abrasion studies DateFilmsChelatorOutcome July 2012 Templated Non-templated EDTA or Citrate 0.2 mL 100 mM 34 of 35 films did not degrade Oct 2012 Templated Non-templated Citrate 0.5 mL 300 or 400 mM 5 mg/mL HA 4 of 38 films did not degrade

9. Group Film Found? Yes No Totals Film8210 Film, 300 mM citrate18 9 Film, 400 mM citrate1910 Film, templated8210 Film, templated, 300 mM citrate18 9 Film, templated, 400 mM citrate1910 Film FoundChelatorNo ChelatorTotals Yes04 (11%)16 (80%)20 No34 (89%)04 (20%)38 Totals382058 p < 0.000001

10. Grade scale for adhesion severity

11. Group Adhesion Grade p 01232 or 3 Control20088-- SepraFilm501450.22 Film, no chelator300770.74 Film, 300 mM citrate303360.66 Film, 400 mM citrate600440.09 Film, templated, no chelator501450.22 Film, templated, 300 mM citrate502240.15 Film, templated, 400 mM citrate201781.00 Incidence of adhesions

12. Swelling measurements

13. Swelling formulas

14. Prelim swelling measurements Trial #BlottingWet Weight (mg) 1none204 2none178 3none244 4none197 5blot on glove125 6blot on glove128 7blot on glove134 8more blotting106 9more blotting108 10more blotting101

15. Prelim swelling measurements Trial #Weight (mg) 1108 2103 3107 4106 5109 6105 7103 8105 9103 10100 11101 12108 Average105 Stdev2.9

16. Wet Wt (mg) Thickness (um) Film #123AvgStdev Dry Wt (mg) SR% WCMinMax 15152 0.6104.2813060 21281311251283.0224.88370220 3535251521.0104.2815090 4108110 1091.2194.88350260 5565955572.1114.2814070 6566051564.5104.6825060 787 83862.3154.78260120 887 0.0164.48280120 91151081131123.6204.68260130 1057 56570.6104.78240100 111071051081071.5194.68260170 1259 56581.7104.8835080 Average4.582 Stdev0.250.8 Swelling data – HA/Alg Film

17. SR weakly correlated with thickness Pearson’s Correlation Coefficient, R = 0.58

18. HA increases film swelling

20. Film thickness measurements

21. 21 Film thickness Thickness (um) 200 0 100 2.7 in Thickness (um) 200 0 100

22. Leveling is very important Top Surface Bottom Surface Shelf

23. Films 1234567 200 0 100 Thickness (μm) Thickness (um) 200 0 100 Re-leveled 2.7 in Not-leveled

24. Leveling is difficult 2.7 in 10 µm θ

25. Doctor blade for film casting

27. Solution castingDoctor blading Cast in a moldSpread on a flat surface without sides Use a solution with low viscosityUse a solution with high viscosity Variables: concentration, volumeVariables: concentration, gap height Requires high precision machined molds (Cost: $100's) Requires a doctor blade (Cost: $100's) Leveling is absolutely necessaryLeveling is not necessary Film thickness is non-uniformFilm thickness is uniform Contrast two techniques

28. Doctor Blade Films Gap height: 750 – 1000 μm750 μm 1500 μm 1000 μm 1.7 in 3.5 in

29. Doctor Blade Films Gap height (µm) 750 1000 1500 750-1000 Films 1234567 200 0 100 Thickness (μm)

30. Film # Thickness (μm) MinMaxRange 110210200 230150120 340140100 440120080 530140110 630150120 730150120 750-100050090040 075050070020 100050100050 150090170080 Doctor Blade Films