1. Thomas Crouzier, 2015 Engineering mucins for biomedical applications Thomas Crouzier biopolymersforlife.org WBC 19 th of May 2016 Page 1

2. Thomas Crouzier, 2015 Page 2 Research on extracellular matrix Biomedical products Cosmetics Hyaluronic acid

3. Thomas Crouzier, 2015 Page 3 Collage n Research on extracellular matrix Biomedical products Cosmetics

4. Thomas Crouzier, 2015 Page 4 Chitosan and Chitin Agriculture Biomedical products Cosmetics

5. Thomas Crouzier, 2015 Mucins Page 5

6. Thomas Crouzier, 2015 Author unknown Page 6 Mucins

7. Thomas Crouzier, 2015 Page 7 From our infographic “Mucus, it’s snot what you think”: https://figshare.com/articles/Mucus_it_s_snot_what_you_think_/1277545

8. Thomas Crouzier, 2015 Mucins cover our wet epithelium Page 8

9. Thomas Crouzier, 2015 Tomas Sandberg, Hans Blom, Karin D. Caldwell. J Biomed Mater Res A. 2009 Dec;91(3):762-72. Mucins’ chemical structure Page 9

10. Thomas Crouzier, 2015 Mucins’ interesting properties Hydration Lubrication Selective filter Physical gels Intermolecular interactions Cell repulsion Page 10

11. Thomas Crouzier, 2015 In house from pig stomachs Commercially available mucin (Sigma-Aldrich) Other animal or plant sources: Mucin extraction Page 11

12. Thomas Crouzier, 2015 Mucin coatings Mucin thin films Mucin hydrogel 12 nm µm mm Page 12 Assembling mucins into materials Crouzier et al. Biomacromolecules 14, 3010–3016 (2013). Co et al. Adv. Mater. Interfaces 2, (2015). Polak et al. Biomacromolecules 15, 3093–3098 (2014). Crouzier et al. Biomacromolecules 13, 3401–3408 (2012). Duffy et al. Acta Biomater. 20, 51–59 (2015).

13. Thomas Crouzier, 2015 3D mucin hydrogels for drug delivery Page 13

14. Thomas Crouzier, 2015 Drug delivery in soft tissue engineering. 2011, Vol. 8(9) p1175-1188 Controlled drug release Page 14

15. Thomas Crouzier, 2015 Hydrogels for drug delivery Cortisone release from hyaluronic acid hydrogel J. Control. Release, 13 (1990), pp. 33–41 Page 15

16. Thomas Crouzier, 2015 Mucins are sticky molecules Mucins bind molecules that are hydrophobic, thiol-bearing, positively charged Page 16

17. Thomas Crouzier, 2015 Methacrylation of mucins Photo-crosslinked with Irgacure 2959 + UV light Page 17

18. Thomas Crouzier, 2015 Scanning electron microscopy 5 µm 1 µm 4 mm Page 18

19. Thomas Crouzier, 2015 Model drugs Paclitaxel (Hydrophobic anti-cancer) Polymyxin B (Positively charged antibiotic ) Page 19

20. Thomas Crouzier, 2015 Polymyxin B (hydrophilic, positively charged) Sustained release from BSM-MA gels Page 20

21. Thomas Crouzier, 2015 Hydrophilic positively charged Sustained release from BSM-MA gels Page 21

22. Thomas Crouzier, 2015 Paclitaxel (hydrophobic) Hydrophilic positively charged Sustained release from BSM-MA gels Page 22

23. Thomas Crouzier, 2015Page 23 Drug release from BSM-MA gels Lieleg, O. & Ribbeck, K. Biological hydrogels as selective diffusion barriers. Trends Cell Biol. 21, 543–551 (2011). Size filtering Interaction filtering

24. Thomas Crouzier, 2015 Control molecules Dextrans (Neutral, high MW) Polyglutamic acid (negatively charged, high MW) Page 24

25. Thomas Crouzier, 2015 Hydrophobic drug Hydrophilic drug Polyglutamic acid (hydrophilic) Sustained release from BSM-MA gels Page 25

26. Thomas Crouzier, 2015 Sustained release from BSM-MA gels Dextran (5 kDa) Hydrophilic drug (1.3 kDa) PGA (25 kDa) Hydrophobic drug (0.8 kDa) Page 26

27. Thomas Crouzier, 2015Page 27 Dextran release from BSM-MA gels Lieleg, O. & Ribbeck, K. Biological hydrogels as selective diffusion barriers. Trends Cell Biol. 21, 543–551 (2011). Size filtering Interaction filtering

28. Thomas Crouzier, 2015 Dextran release from BSM-MA gels Page 28

29. Thomas Crouzier, 2015 BSM-MA (-) + polymyxin Antimicrobial effect of BSM-MA gels Page 29

30. Thomas Crouzier, 2015 No gelBSM-MA BSM-MA + Paclitaxel Antimicrobial effect of BSM-MA gels Page 30

31. Thomas Crouzier, 2015 Katharina Ribbeck Connor Duffy Funding: Thanks to: Page 31 Xueying Zhong Victoria Linderberg Sujit Kootala

32. Thomas Crouzier, 2015 Thank you for listening! Page 32

33. Thomas Crouzier, 2015 Measuring drug release Prepare BSM-MA gel with fluorescently labeled drug mixed in Release by incubating with PBS at 37ºC Measure fluorescence of supernatant Thomas Crouzier, 2014 Page 33

34. Thomas Crouzier, 2015