title: What interactions drive the salivary mucosal pellicle formation?
abstract: The bound salivary pellicle is essential for protection of both the enamel and mucosa in the oral cavity. The enamel pellicle formation is well characterised, however the mucosal pellicle proteins have only recently been clarified and what drives their formation is still unclear. The aim of this study was to examine the salivary pellicle on particles with different surface properties (hydrophobic or hydrophilic with a positive or negative charge), to determine a suitable model to mimic the mucosal pellicle. A secondary aim was to use the model to test how transglutaminase may alter pellicle formation. Particles were incubated with resting whole mouth saliva, parotid saliva and submandibular/sublingual saliva. Following incubation and two PBS and water washes bound salivary proteins were eluted with two concentrations of SDS, which were later analysed using SDS-PAGE and Western blotting. Experiments were repeated with purified transglutaminase to determine how this epithelial-derived enzyme may alter the bound pellicle. Protein pellicles varied according to the starting salivary composition and the particle chemistry. Amylase, the single most abundant protein in saliva, did not bind to any particle indicating specific protein binding. Most proteins bound through hydrophobic interactions and a few according to their charges. The hydrophobic surface most closely matched the known salivary mucosal pellicle by containing mucins, cystatin and statherin but an absence of amylase and proline-rich proteins. This surface was further used to examine the effect of added transglutaminase. At the concentrations used only statherin showed any evidence of crosslinking with itself or another saliva protein. In conclusion, the formation of the salivary mucosal pellicle is probably mediated, at least in part, by hydrophobic interactions to the epithelial cell surface.
authors.affiliation: Salivary Research Unit, King's College London Dental Institute, London SE1 9RT, UK. Electronic address: hannah.gibbins@kcl.ac.uk.
authors.forename: Hannah L
authors.initials: HL
authors.lastname: Gibbins
authors.affiliation: Australian Research Council Centre of Excellence in Plant Cell Walls, School of Chemical Engineering, The University of Queensland, Queensland 4072, Australia. Electronic address: g.yakubov@uq.edu.au.
authors.forename: Gleb E
authors.initials: GE
authors.lastname: Yakubov
authors.affiliation: Salivary Research Unit, King's College London Dental Institute, London SE1 9RT, UK. Electronic address: gordon.proctor@kcl.ac.uk.
authors.forename: Gordon B
authors.initials: GB
authors.lastname: Proctor
authors.affiliation: Unilever R&D Discover, Colworth Science Park, Sharnbrook MK44 1LQ, UK. Electronic address: steve.wilson@unilever.com.
authors.forename: Stephen
authors.initials: S
authors.lastname: Wilson
authors.affiliation: Salivary Research Unit, King's College London Dental Institute, London SE1 9RT, UK. Electronic address: guy.carpenter@kcl.ac.uk.
authors.forename: Guy H
authors.initials: GH
authors.lastname: Carpenter
mesh_terms: D015153:Blotting, Western; D044622:Dental Pellicle / Q000254:growth & development / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D009092:Mucous Membrane / Q000378:metabolism*; D011485:Protein Binding; D015203:Reproducibility of Results; D012471:Salivary Proteins and Peptides / Q000378:metabolism*; D013194:Staining and Labeling; D011503:Transglutaminases / Q000378:metabolism
chemical_list: D012471:Salivary Proteins and Peptides; D011503:Transglutaminases
keywords: Hydrophobic; Pellicle; Proteins; Saliva; Transglutaminase