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Investigating mucin interactions with diverse surfaces for biomedical applications
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience. (Crouzier)ORCID iD: 0000-0001-6359-2216
2019 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

Mucous membranes are covered with mucus, a viscoelastic hydrogel that plays an essential role in their protection from shear and pathogens. The viscoelasticity of mucus is owing to mucins, a group of densely glycosylated proteins. Mucins can interact with a wide range of surfaces; thus, there is big interest in exploring and manipulating such interactions for biomedical applications. This thesis presents investigations of mucin interactions with hydrophobic surfaces in order to identify the key features of mucin lubricity, as well as describes the development of materials that are optimized to interact with mucins.

 

In Paper I we investigated the domains which make mucins outstanding boundary lubricants. The results showed that the hydrophobic terminal domains of mucins play a crucial role in the adsorption and lubrication on hydrophobic surfaces. Specifically, protease digestion of porcine gastric mucins and salivary mucins resulted in the cleavage of these domains and the loss of lubricity and surface adsorption. However, a “rescue” strategy was successfully carried out by grafting hydrophobic phenyl groups to the digested mucins and enhancing their lubricity. This strategy also enhanced the lubricity of polymers which are otherwise bad lubricants.

 

In Paper II we developed mucoadhesive materials based on genetically engineered partial spider silk proteins. The partial spider silk protein 4RepCT was successfully functionalized with six lysines (pLys-4RepCT), or the Human Galectin-3 Carbohydrate Recognition Domain (hGal3-4RepCT). These strategies were aiming to either non-specific electrostatic interactions between the positive lysines and the negative mucins, or specific binding between the hGal3 and the mucin glycans. Coatings, fibers, meshes and foams were prepared from the new silk proteins, and the adsorption of porcine gastric mucins and bovine submaxillary mucins was measured, demonstrating enhanced adsorption.

 

The work presented demonstrates how mucin-material interactions can provide us with valuable information for the development of new biomaterials. Specifically, mucin-based and mucin-inspired lubricants could provide desired lubrication to a wide range of surfaces, while our new silk based materials could be valuable tools for the development of mucosal dressings.

Abstract [sv]

Slemhinnor täckts av slem, en viskoelastisk hydrogel som spelar en viktig roll för att skydda mot mekanisk nötning och patogener. Muciner, en grupp av tätt glykosylerade proteiner, spelar en viktig roll i viskoelasticiteten av slem. Eftersom muciner kan interagera med diverse ytor är det av stort intresse att utforska och manipulera sådana interaktioner för biomedicinska tillämpningar. Denna avhandling presenterar undersökningar av mucininteraktioner med hydrofoba ytor för att identifiera de viktigaste egenskaperna hos mucinsmörjning, samt beskriver utveckling av material som optimerades för att interagera med muciner.

 

I Artikel I undersökte vi de domäner som bidrar till  mucinernas enastående kapacitet som smörjmedel. Resultaten visade att mucinernas hydrofoba terminaldomäner spelar en avgörande roll vid adsorption och smörjning på hydrofoba ytor. Mer specifikt, proteasklyvning av svinmagemuciner och salivmuciner resulterade i klyvningen av dessa domäner och förlust av smörjning och ytadsorption. Genom att länka hydrofobiska fenylgrupper till de uppbrutna mucinerna, lyckades deras smörjningsegenskaper förbättras. Denna strategi förbättrade också smörjningsegenskaper hos andra polymerer som annars har  dåliga smörjningsegenskaper.

 

I Artikel II utvecklade vi mukoadhesiva material baserade på genetiskt modifierade partiella spindelsilkeproteiner. Spindelsilkeproteinet 4RepCT funktionaliserades framgångsrikt med tillsats av sex lysiner (pLys-4RepCT), eller den mänskliga Galectin-3 karbohydrat igenkänningsdomänen (hGal3-4RepCT). Syftet med dessa strategier var antingen att öka ospecifika elektrostatiska interaktioner mellan de positiva lysinerna och de negativa mucinerna, eller den specifika bindningen mellan hGal3 och mucin-glykanerna. Beläggningar, fibrer, nät och skum framställdes från de nya silkeproteinerna. Efter att adsorption av svinmagsmuciner och bovina submaxillära muciner uppmätts, visade de nya silkeproteinerna förbättrad mucin adsorption.

 

Detta arbete visar hur interaktioner mellan mucin-material kan bidra med värdefull information för utvecklingen av nya biomaterial. Mucinbaserade och mucininspirerade smörjmedel kan ge önskad smörjning till ett brett spektrum av ytor, medan vår nya silkesbaserad material kan vara ett värdefullt verktyg för utvecklingen av slemhinneförband.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2019. , p. 41
Series
TRITA-CBH-FOU ; 2019:24
Keywords [en]
mucus, mucin, protein adsorption, biomaterials, lubrication, mucoadhesion, 4RepCT
National Category
Biomaterials Science
Research subject
Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-249469ISBN: 978-91-7873-172-5 (print)OAI: oai:DiVA.org:kth-249469DiVA, id: diva2:1304413
Presentation
2019-05-06, FA31, Roslagstullsbacken 21, 114 21 (plan 3), Stockholm, 13:00 (English)
Opponent
Supervisors
Note

QC 20190412

Available from: 2019-04-12 Created: 2019-04-12 Last updated: 2019-04-12Bibliographically approved
List of papers
1. Mucin-Inspired Lubrication on Hydrophobic Surfaces
Open this publication in new window or tab >>Mucin-Inspired Lubrication on Hydrophobic Surfaces
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2017 (English)In: Biomacromolecules, ISSN 1525-7797, E-ISSN 1526-4602, Vol. 18, no 8, p. 2454-2462Article in journal (Refereed) Published
Abstract [en]

In the human body, high-molecular-weight glycoproteins called mucins play a key role in protecting epithelial surfaces against pathogenic attack, controlling the passage of molecules toward the tissue and enabling boundary lubrication with very low friction coefficients. However, neither the molecular mechanisms nor the chemical motifs of those biomacromolecules involved in these fundamental processes are fully understood. Thus, identifying the key features that render biomacromolecules such as mucins outstanding boundary lubricants could set the stage for creating versatile artificial superlubricants. We here demonstrate the importance of the hydrophobic terminal peptide domains of porcine gastric mucin (MUCSAC) and human salivary mucin (MUCSB) in the processes of adsorbing to and lubricating a hydrophobic PDMS surface. Tryptic digestion of those mucins results in removal of those terminal domains, which is accompanied by a loss of lubricity as well as surface adsorption. We show that this loss can in part be compensated by attaching hydrophobic phenyl groups to the glycosylated central part of the mucin macromolecule. Furthermore, we demonstrate that the simple biopolysaccharide dextran can be functionalized with hydrophobic groups which confers efficient surface adsorption and good lubricity on PDMS to the polysaccharide.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2017
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-214344 (URN)10.1021/acs.biomac.7b00605 (DOI)000407869400024 ()28635258 (PubMedID)2-s2.0-85027351851 (Scopus ID)
Note

QC 20170912

Available from: 2017-09-12 Created: 2017-09-12 Last updated: 2019-04-13Bibliographically approved
2. Genetically Engineered Mucoadhesive Spider Silk
Open this publication in new window or tab >>Genetically Engineered Mucoadhesive Spider Silk
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2018 (English)In: Biomacromolecules, ISSN 1525-7797, E-ISSN 1526-4602, Vol. 19, no 8, p. 3268-3279Article in journal (Refereed) Published
Abstract [en]

Mucoadhesion is defined as the adhesion of a material to the mucus gel covering the mucous membranes. The mechanisms controlling mucoadhesion include nonspecific electrostatic interactions and specific interactions between the materials and the mucins, the heavily glycosylated proteins that form the mucus gel. Mucoadhesive materials can be used to develop mucosal wound dressings and noninvasive transmucosal drug delivery systems. Spider silk, which is strong, biocompatible, biodegradable, nontoxic, and lightweight would serve as an excellent base for the development of such materials. Here, we investigated two variants of the partial spider silk protein 4RepCT genetically engineered in order to functionalize them with mucoadhesive properties. The pLys-4RepCT variant was functionalized with six cationically charged lysines, aiming to provide nonspecific adhesion from electrostatic interactions with the anionically charged mucins, while the hGal3-4RepCT variant was genetically fused with the Human Galectin-3 Carbohydrate Recognition Domain which specifically binds the mucin glycans Gal beta 1-3GlcNAc and Gal beta 1-4GlcNAc. First, we demonstrated that coatings, fibers, meshes, and foams can be readily made from both silk variants. Measured by the adsorption of both bovine submaxillary mucin and pig gastric mucin, the newly produced silk materials showed enhanced mucin binding properties compared with materials of wild-type (4RepCT) silk. Moreover, we showed that pLys-4RepCT silk coatings bind mucins through electrostatic interactions, while hGal3-4RepCT silk coatings bind mucins through specific glycan-protein interactions. We envision that the two new mucoadhesive silk variants pLys-4RepCT and hGal3-4RepCT, alone or combined with other biofunctional silk proteins, constitute useful new building blocks for a range of silk protein-based materials for mucosal treatments.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2018
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:kth:diva-234195 (URN)10.1021/acs.biomac.8b00578 (DOI)000441852400011 ()29932649 (PubMedID)2-s2.0-85049259614 (Scopus ID)
Note

QC 20180920

Available from: 2018-09-20 Created: 2018-09-20 Last updated: 2019-04-13Bibliographically approved

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