Engineering mucins for biomedical applications Thomas Crouzier biopolymersforlife

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

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

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

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

Mucins Thomas Crouzier, 2015 Page 5

Mucins Author unknown Thomas Crouzier, 2015 Page 6

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

Mucins cover our wet epithelium Thomas Crouzier, 2015 Page 8

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

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

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

Assembling mucins into materials nm Mucin coatings Crouzier et al. Biomacromolecules 14, 3010– 3016 (2013). Co et al. Adv. Mater. Interfaces 2, (2015). µm Mucin thin films 1 2 Polak et al. Biomacromolecules 15, 3093– 3098 (2014). Crouzier et al. Biomacromolecules 13, 3401– 3408 (2012). mm Mucin hydrogel Duffy et al. Acta Biomater. 20, 51– 59 (2015). Thomas Crouzier, 2015 Page 12

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

Controlled drug release Drug delivery in soft tissue engineering. 2011, Vol. 8(9) p 1175 -1188 Thomas Crouzier, 2015 Page 14

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

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

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

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

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

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

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

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

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

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

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

Sustained release from BSM-MA gels Dextran (5 k. Da) PGA (25 k. Da) Hydrophobic drug (0. 8 k. Da) Hydrophilic drug (1. 3 k. Da) Thomas Crouzier, 2015 Page 26

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

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

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

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

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

Thank you for listening! Thomas Crouzier, 2015 Page 32

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

Thomas Crouzier, 2015