The role of saliva in the oral cavity is manifold; an important function is to serve as lubricant between hard (enamel) and soft (mucosal) tissues. Intraoral lubrication is of crucial importance in order to maintain functions such as deglutition, mastication and the faculty of speech. A large number of people suffer from impaired salivary functions, displaying symptoms such as 'dry mouth'. This results in a need for methods to assess the lubricating properties of both native saliva and potential artificial saliva formulations. Here, normal as well as lateral forces, acting between adsorbed salivary films, have been measured for the first time by means of colloidal probe atomic force microscopy (AFM). It was found that the presence of salivary pellicles between hard surfaces reduces the friction coefficient by a factor of 20. This reduction of friction is consistent with the long-range purely repulsive nature of the normal forces acting between the salivary films. The lubricating mechanism is presumably based on a full separation of the sliding surfaces by the salivary films. The friction between salivary films has been investigated at normal loads that cover the clinical jaw closing forces, and it can be concluded that the lubricating properties are maintained within this load interval. The present study indicates the usefulness of colloidal probe AFM, which offers a direct and quantitative measure of lubrication at a molecular level, in the study of biotribological phenomena. In particular, the results obtained here may have implications for the development of saliva substitutes.

Different physico-chemical properties (eg adsorption kinetics, thickness, viscoelasticity, and mechanical stability) of adsorbed salivary pellicles depend on different factors, including the properties (eg charge, roughness, wettability, and surface chemistry) of the substratum. Whether these differences in the physico-chemical properties are a result of differences in the composition or in the organization of the pellicles is not known. In this work, the influence of substratum wettability on the composition of the pellicle was studied. For this purpose, pellicles eluted from substrata of different but well-characterized wettabilities were examined by means of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that substratum hydrophobicity did not have a major impact on pellicle composition. In all substrata, the major pellicle components were found to be cystatins, amylases and large glycoproteins, presumably mucins. In turn, interpretation of previously reported data based on the present results suggests that variations in substratum wettability mostly affect the organization of the pellicle components.

The purpose of the present study was to characterise the structure dynamics of pure salivary secretions retained on controlled surfaces with different surface energies in the early stage of salivary film formation. Germanium prisms prepared to have either low surface energy of medium surface energy were incubated in fresh secretions of either human parotid saliva (HPS) or human submandibular/sublingual saliva (HSMSLS) for 15,90 and 180 min. After controlled rinsing with distilled water, the surfaces were air dried and thereafter imaged with atomic force microscopy (AFM). The amount of adsorbed material and the size of the structures detected increased with increased saliva exposure time. the film thicknesses varied from 10 to 160 nm, and both HPS and HSMSLS films contained structures with diameters varying from 40 nm to 2 microm. Some of these were clustered into special formations. The HPS films exhibited a more granular morphology than the HSMSLS films. Furthermore, branched lines were detected on the low surface energy germanium prisms incubated in saliva. the results indicate that exposure time, surface energy, and type of salivary secretion all are factors affecting the adsorption characteristics of salivary films.

A challenge for the next generation marine antifouling (AF) paints is to deliver minimum amounts of biocides to the environment. The candidate AF compound medetomidine is here shown to be released at very low concentrations, ie ng ml(-1) day(-1). Moreover, the release rate of medetomidine differs substantially depending on the formulation of the paint, while inhibition of barnacle settlement is independent of release to the ambient water, ie the paint with the lowest release rate was the most effective in impeding barnacle colonisation. This highlights the critical role of chemical interactions between biocide, paint carrier and the solid/aqueous interface for release rate and AF performance. The results are discussed in the light of differential affinity states of the biocide, predicting AF activity in terms of a high surface affinity and preserved bioavailability. This may offer a general framework for the design of low-release paint systems using biocides for protection against biofouling on marine surfaces.

BACKGROUND: Titanium implants in the oral cavity are covered with a saliva-derived pellicle to which early colonizing microorganisms such as Streptococcus oralis can bind. The protein profiles of salivary pellicles on titanium have not been well characterized and the proteins of importance for binding are thus unknown. Biofilm bacteria exhibit different phenotypes from their planktonic counterparts and contact with salivary proteins may be one factor contributing to the induction of changes in physiology. We have characterized salivary pellicles from titanium surfaces and investigated how contact with uncoated and saliva-coated titanium surfaces affects metabolic activity in adherent cells of S. oralis. METHODS: Salivary pellicles on smooth titanium surfaces were desorbed and these, as well as purified human saliva, were subjected to two-dimensional gel electrophoresis and mass spectroscopy. A parallel plate flow-cell model was used to study binding of a fresh isolate of S. oralis to uncoated and saliva-coated titanium surfaces. Metabolic activity was assessed using the BacLight CTC Vitality Kit and confocal scanning laser microscopy. Experiments were carried out in triplicate and the results analyzed using Student's t-test or ANOVA. RESULTS: Secretory IgA, α-amylase and cystatins were identified as dominant proteins in the salivary pellicles. Selective adsorption of proteins was demonstrated by the enrichment of prolactin-inducible protein and absence of zinc-α₂-glycoprotein relative to saliva. Adherence of S. oralis to titanium led to an up-regulation of metabolic activity in the population after 2 hours. In the presence of a salivary pellicle, this effect was enhanced and sustained over the following 22 hour period. CONCLUSIONS: We have shown that adherence to smooth titanium surfaces under flow causes an up-regulation of metabolic activity in the early oral colonizer S. oralis, most likely as part of an adaptation to the biofilm mode of life. The effect was enhanced by a salivary pellicle containing sIgA, α-amylase, cystatins and prolactin-inducible protein which was, for the first time, identified as an abundant component of salivary pellicles on titanium. Further studies are needed to clarify the mechanisms underlying the effect of surface contact on metabolic activity as well as to identify the salivary proteins responsible for enhancing the effect.

The aim of this paper was to elucidate the mechanisms behind salivary lubrication with special emphasis on the lubricity of three key components of the pellicle, viz human acidic proline-rich protein 1 (PRP-1), human statherin and bovine submaxillary mucin (BSM). The lubricating properties of the proteins have been assessed by means of colloidal probe atomic force microscopy, and are discussed in relation to their adsorption behaviour. To various extents, the proteins investigated all showed a lubricating effect when adsorbed to silica surfaces. For comparable concentrations, PRP-1 was found to have a more pronounced lubricating effect than BSM, which in turn showed a higher lubricity than statherin. The relative lubricity is in accordance with previously reported relative adsorbed amounts of the three proteins, within the investigated concentration interval. It is concluded that PRP-1 has the highest lubricating capacity as a pure fraction among the preparations investigated, and that the lubricating effect of PRP-1 as a pure fraction is notably large as compared to the lubricity of human whole saliva.

Ellipsometry was used to investigate the effect of polyaluminium chloride (PAC) formulations of different degrees of hydrolysation on an adsorbed mucin film. The results were compared to the effect of aluminium chloride (AlCl3) and ferric chloride. A compaction of the mucin film took place upon addition of the formulations and this occurred to different extents and at different concentrations for the different formulations. The compaction of PAC of a low degree of hydrolysis behaved similarly to AlCl3. PAC of a high degree of hydrolysis showed a greater compaction effect than the other aluminium formulations. The initial compaction concentration was found to be 0.001 mM which is less than previously found for aluminium–mucin complex formation in bulk. The reversibility of the compaction was also investigated. The compaction of the mucin film was found to be partly reversible for AlCl3 and PAC of low degree of hydrolysis. No reversibility was observed for the formulations of PAC of high hydrolysis grade or for ferric chloride. The results are consistent with previously observed effects of PAC of a low degree of hydrolysis on bacterial surfaces where a compaction of surface polymers was indicated by the reduced range of repulsive steric interactions.

The filamentous bacterium Microthrix parvicella causes severe separation and foaming problems at wastewater treatment plants (WWTPs). An effective control of the bacterium in activated sludge WWTPs can be accomplished by dosage with polyaluminium chloride (PAX-14). The purpose of this study was to investigate whether addition of PAX-14 affects surface properties such as the hydrophobicity of the bacterium and to study the exopolymers of M. parvicella that host surface-associated enzymes. To this end, force measurements by atomic force microscopy were carried out to measure the interactions between hydrophilic and hydrophobized tips and the bacterium surface. Addition of PAX-14 caused no changes in the hydrophobicity of the bacterium surface but the data indicate that it collapsed the polymeric layer likely due to electrostatic screening. It is concluded that the collapse of the polymeric layer may affect the transport of substrates (eg free fatty acids) to the bacterium and hence the competitiveness of M. parvicella compared to the other bacteria present in activated sludge.

The rates and plateau values of adsorption at the solid/ liquid interface from human whole resting saliva from three male donors were studied in vitro. The influence of saliva concentration, the total protein content and the wettability of the exposed solids was investigated. Ellipsometry was used to study the rate and plateau values of the adsorption process in situ. The solid surfaces used were hydrophilic, negatively charged, silica and hydrophobic methylated silica. Salivary films were formed by adsorption from solutions of 0.05%, 0.10%, 0.20%, 0.50%, 1.0% and 2.0% (v/v) saliva in phosphate buffered saline. The results revealed that adsorption onto both types of surfaces was a rapid and, as expected, a concentration dependent process. The adsorbed amounts were in the range of monomolecular layers and were found to be larger onto the hydrophobic than the hydrophilic surfaces (p≤0.0062). A strong concentration dependence was denoted in all cases (p< 0.0001). Inter-individual differences were found not to be significant (0.0636 ≤ p≤ 0.8508) at any of the saliva concentrations. However, an interaction was shown (p < 0.0001) between the protein concentrations and the donors, indicating qualitative differences in protein content and surface activity.

The influence of saliva concentration, saliva total protein content and the wetting characteristics of exposed solids on in vitro film formation was studied by the technique of in situ ellipsometry. The rates and plateau values of adsorption (45 min) at solid/liquid interfaces (hydrophilic silica and hydrophobic methylated silica surfaces) were determinated for human parotid (HPS) and submandibular/sublingual (HSMSLS) resting saliva solutions (0.1 and 1.0%, (v/v), saliva in phosphate buffered saline). Adsorption rates were related to a model assuming mass transport through an unstirred layer adjacent to the surface. The results showed that the adsorption was rapid, concentration dependent and higher on hydrophobic than on hydrophilic surfaces. Analysis of the influence of protein concentration on the adsorbed amounts demonstrated an interaction between protein concentration and the two surfaces for HPS and HSMSLS, respectively. This may indicate differences in binding mode. Inter-individual differences were found not to be significant at the 1% level of probability. Comparison of the observed adsorption and calculated diffusion rates suggest that on hydrophilic surfaces initial adsorption of proteins diffusing at rates corresponding to those of statherin and aPRPs takes place, whereas on hydrophobic surfaces lower molecular mass compounds appear to be involved.

The objective of the present study was to investigate the adsorption of PRP-1, PRP-3 and statherin to solid surfaces in terms of dependence on concentration, the presence of electrolyte and surface wettability. Time resolved in situ ellipsometry was used to determine the adsorbed amounts and adsorption rates of pure PRP-1, PRP-3 and statherin onto pure (hydrophilic) and methylated (hydrophobized) silica surfaces. The initial film build-up was fast and plateaus were reached within 10 min at all concentrations for both types of surfaces and all proteins. The observed adsorption and calculated diffusion rates of PRP-1, PRP-3 and statherin, respectively, indicated that the initial adsorption was mass transport controlled at low concentrations. At hydrophobic surfaces, isotherm shapes and adsorbed amounts were similar for PRP-1 and PRP-3, while statherin adsorbed to a higher extent. At hydrophilic surfaces only PRP-1 adsorbed substantially, while for PRP-3 and statherin adsorbed amounts were low. The presence of Ca 2+ ions in the phosphate buffer solution increased the adsorption of statherin and PRP-3 on hydrophobic surfaces, while PRP-1 was unaffected. On hydrophilic surfaces, all three proteins adsorbed in higher amounts in NaCl, compared to CaCl 2 at similar ionic strength. It is concluded that acidic PRPs (PRP-1 and PRP-3) and statherin readily form films on a variety of materials and solution conditions, showing that their functions may be fulfilled under a wide range of conditions.

The adsorption of human whole saliva (HWS) onto silica and hydroxyapatite surfaces (HA) was followed by quartz crystal microbalance with dissipation (QCM-D) and ellipsometry. The influence of different surface properties and adsorption media (water and PBS) on the HWS adsorption was studied. The viscoelastic properties of the saliva films formed onto the solid surfaces were estimated by the use of the Voigt-based viscoelastic film model. Furthermore, the efficiency of SDS and delmopinol to elute the adsorbed saliva film from the surfaces was investigated at different surfactant concentrations. It was observed a biphasic kinetic regime for the adsorption of saliva on the silica and HA surfaces, indicating the formation of a rigidly coupled first layer corresponding to an initial adsorption of small proteins and a more loosely bound second layer. The results further showed a higher adsorption of HWS onto the HA surfaces compared to the silica surfaces in both adsorption media (PBS and water). The adsorption in PBS led to higher adsorbed amounts on both surfaces as compared to water. SDS was found to be more efficient in removing the saliva film from both surfaces than delmopinol. The salivary film was found to be less tightly bound on to the silica surfaces since more saliva film could be removed with both SDS and delmopinol compared to from the HA surface. When saliva was adsorbed from PBS a higher energy dissipation was determined implying that a softer saliva layer is build up in PBS as opposed to in water. In the same media (water) a softer saliva layer was formed onto the HA surfaces, while in PBS the saliva layer formed at both surfaces seems to have similar structure.

This study investigated the effect of glow discharge treatment of titania surfaces on plasma protein adsorption, by means of ellipsometry and mechanically assisted SDS elution. The adsorption and film elution of three plasma proteins, viz. human serum albumin (HSA), human immunoglobulin G (IgG) and laminin-1, as well as competitive adsorption from a mixture of the three proteins, showed that the adsorbed amount of the individual proteins after 1 h increased in the order HSA

In situ ellipsometry was employed to study adsorption from human palatal saliva (HPalS) in terms of dependence on surface wettability and saliva concentration (<or=1%). Adsorbed amounts, kinetics, and elutability with buffer and sodium dodecyl sulphate (SDS) were determined. The low-molecular weight protein content of bulk HPalS was also investigated using two-dimensional gel electrophoresis, and this revealed the presence of a large group of proteins

The salivary film, denoted the pellicle, formed on oral surfaces is of great importance for oral health and comfort. The present study describes mechanically-assisted sodium dodecyl sulphate (SDS) elution of the in vivo pellicle formed on human enamel and visualisation of the desorbed pellicle proteins using two-dimensional gel electrophoresis (2-DE). To verify this removal of the pellicle, a combined mechanical and surfactant procedure was additionally performed on an in vitro pellicle formed on human enamel, and the effectiveness was validated by mechanical removal in combination with HCl. As indicated by protein quantitation and one dimensional gel electrophoresis, rubbing with polyamide fibre pellets soaked in a 0.5% SDS solution was optimal for completely removing the adsorbed proteins from the enamel surface, and yet provided separation of the proteins by 2-DE to enable identification in future studies.

This study utilized two-dimensional gel electrophoresis (2-DE) to illustrate compositional differences between in vitro salivary conditioning films (denoted pellicles) formed on human enamel as well as on the dental materials titanium and poly methyl methacrylate (PMMA). The salivary pellicles were formed by immersing each surface in individual tubes containing small volumes of freshly collected whole saliva. Saliva remaining in the tubes after 2 hours of pellicle formation was visualized by means of 2-DE and silver staining. The results showed that the protein patterns in 2-DE of the liquid phase of saliva left after exposure to the respective surfaces, regarding proteins below 100 kDa in size, were different depending on the surface used. Several protein groups and/or individual proteins were shown to be distinct for each surface used.

Ellipsometry and mechanically assisted sodium dodecyl sulphate elution was utilized to study the adsorption of human serum albumin (HSA), human immunoglobulin G (IgG), and laminin-1, as well as competitive adsorption from a mixture of these proteins on spin-coated and sintered hydroxyapatite (HA) surfaces, respectively. The HA surfaces were characterized with respect to wettability and roughness by means of water contact angles and atomic force microscopy, respectively. Both surface types were hydrophilic, and the average roughness (Sa) and surface enlargement (Sdr) were lower for the sintered compared to the spin-coated HA surfaces. The adsorbed amounts on the sintered HA increased as follows: HSA < laminin-1 < IgG < the protein mixture. For the competitive adsorption experiments, the adsorbed fractions increased accordingly: HSA < laminin-1 < IgG on both types of HA substratum. However, a higher relative amount of HSA and laminin-1 and a lower relative amount of IgG was found on the spin-coated surfaces compared to the sintered surfaces. The effects observed could be ascribed to differences in surface roughness and chemical composition between the two types of HA substratum, and could have an influence on selection of future implant surface coatings.