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Archives of Oral Biology Nov 2020In the present study, we used an in vitro initial intrinsic erosion model to evaluate: (experiment 1) the influence of the degree of serine (Ser) phosphorylation of...
OBJECTIVES
In the present study, we used an in vitro initial intrinsic erosion model to evaluate: (experiment 1) the influence of the degree of serine (Ser) phosphorylation of peptides containing the 15 N-terminal residues of statherin and (experiment 2) the effect of different concentrations of the peptide with the best performance in experiment 1 on initial enamel erosion.
DESIGN
Bovine enamel specimens were divided into 6 groups (n = 15/group) for each experiment. In experiment 1, the peptides evaluated (at 1.88 × 10 M) were: not phosphorylated (StatSS), phosphorylated in Ser2 (StatpSS), phosphorylated in Ser3 (StatSpS) phosphorylated in Ser2 and Ser3 (StatpSpS). Phosphate buffer and human recombinant statherin were used as negative and positive controls, respectively. In experiment 2, StatpSpS was evaluated at different concentrations: 0.94, 1.88, 3.76 and 7.52 × 10 M. Phosphate buffer and 0.1 mg/mL CaneCPI-5 were employed as negative and positive controls, respectively. In each experiment, the specimens were incubated with the solutions for 2 h, then the AEP was allowed to form (under human pooled saliva) for 2 h. The specimens were then challenged with 0.01 M HCl for 10 s. Demineralization was evaluated by percentage of surface hardness change (%SHC). Data were analyzed by ANOVA and Tukey's test (p < 0.05).
RESULTS
In experiment 1, only StatpSpS significantly reduced the % SHC in comparison with control. In experiment 2, 1.88 × 10 M StatpSpS significantly reduced the %SHC in comparison with control.
CONCLUSIONS
This is the first study showing that statherin-derived peptide might protect against intrinsic erosion.
Topics: Animals; Cattle; Dental Enamel; Humans; In Vitro Techniques; Phosphorylation; Saliva; Salivary Proteins and Peptides; Serine; Tooth Erosion
PubMed: 32919104
DOI: 10.1016/j.archoralbio.2020.104890 -
Journal of Dental Research Apr 2017The acquired enamel pellicle is an oral, fluid-derived protein layer that forms on the tooth surface. It is a biologically and clinically important integument that...
The acquired enamel pellicle is an oral, fluid-derived protein layer that forms on the tooth surface. It is a biologically and clinically important integument that protects teeth against enamel demineralization, and abrasion. Tooth surfaces are exposed to different proteinaceous microenvironments depending on the enamel location. For instance, tooth surfaces close to the gingival sulcus contact serum proteins that emanate via this sulcus, which may impact pellicle composition locally. The aims of this study were to define the major salivary and serum components that adsorb to hydroxyapatite, to study competition among them, and to obtain preliminary evidence in an in vivo saliva/serum pellicle model. Hydroxyapatite powder was incubated with saliva and serum, and the proteins that adsorbed were identified by mass spectrometry. To study competition, saliva and serum proteins were labeled with CyDyes, mixed in various proportions, and incubated with hydroxyapatite. In vivo competition was assessed using a split-mouth design, with half the buccal tooth surfaces coated with serum and the other half with saliva. After exposure to the oral environment for 0 min, 30 min and 2 h, the pellicles were analyzed by SDS-PAGE. In pure saliva- or serum-derived pellicles, 82 and 84 proteins were identified, respectively. When present concomitantly, salivary protein adsorbers effectively competed with serum protein adsorbers for the hydroxyapatite surface. Specifically, acidic proline-rich protein, cystatin, statherin and protein S100-A9 proteins competed off apolipoproteins, complement C4-A, haptoglobin, transthyretin and serotransferrin. In vivo evidence further supported the replacement of serum proteins by salivary proteins. In conclusion, although significant numbers of serum proteins emanate from the gingival sulcus, their ability to participate in dental pellicle formation is likely reduced in the presence of strong salivary protein adsorbers. The functional properties of the acquired enamel pellicle will therefore be mostly dictated by the salivary component.
Topics: Adsorption; Biofilms; Blood Proteins; Chromatography, Liquid; Dental Enamel; Dental Pellicle; Durapatite; Humans; Male; Mass Spectrometry; Proteomics; Saliva; Salivary Proteins and Peptides; Surface Properties
PubMed: 27879420
DOI: 10.1177/0022034516680771 -
Scientific Reports Aug 2021Dental caries has been the most widespread chronic disease globally associated with significant health and financial burdens. Caries typically starts in the enamel,...
Dental caries has been the most widespread chronic disease globally associated with significant health and financial burdens. Caries typically starts in the enamel, which is a unique tissue that cannot be healed or regrown; nonetheless, new preventive approaches have limitations and no effective care has developed yet. Since enamel is a non-renewable tissue, we believe that the intimate overlaying layer, the acquired enamel pellicle (AEP), plays a crucial lifetime protective role and could be employed to control bacterial adhesion and dental plaque succession. Based on our identified AEP whole proteome/peptidome, we investigated the bioinhibitory capacities of the native abundant proteins/peptides adsorbed in pellicle-mimicking conditions. Further, we designed novel hybrid constructs comprising antifouling and antimicrobial functional domains derived from statherin and histatin families, respectively, to attain synergistic preventive effects. Three novel constructs demonstrated significant multifaceted bio-inhibition compared to either the whole saliva and/or its native proteins/peptides via reducing biomass fouling and inducing biofilm dispersion beside triggering bacterial cell death. These data are valuable to bioengineer precision-guided enamel pellicles as an efficient and versatile prevention remedy. In conclusion, integrating complementary acting functional domains of salivary proteins/peptides is a novel translational approach to design multifunctional customizable enamel pellicles for caries prevention.
Topics: Adult; Biofilms; Biomass; Biomimetics; Dental Caries; Dental Enamel; Durapatite; Fluorescence; Gentian Violet; Humans; Imaging, Three-Dimensional; Immobilized Proteins; Microbial Sensitivity Tests; Peptides; Proteins; Saliva; Streptococcus mutans
PubMed: 34417532
DOI: 10.1038/s41598-021-96622-y -
International Journal of Molecular... Jan 2022Extensive biofilm formation on materials used in restorative dentistry is a common reason for their failure and the development of oral diseases like peri-implantitis or...
Extensive biofilm formation on materials used in restorative dentistry is a common reason for their failure and the development of oral diseases like peri-implantitis or secondary caries. Therefore, novel materials and strategies that result in reduced biofouling capacities are urgently sought. Previous research suggests that surface structures in the range of bacterial cell sizes seem to be a promising approach to modulate bacterial adhesion and biofilm formation. Here we investigated bioadhesion within the oral cavity on a low surface energy material (perfluorpolyether) with different texture types (line-, hole-, pillar-like), feature sizes in a range from 0.7-4.5 µm and graded distances (0.7-130.5 µm). As a model system, the materials were fixed on splints and exposed to the oral cavity. We analyzed the enzymatic activity of amylase and lysozyme, pellicle formation, and bacterial colonization after 8 h intraoral exposure. In opposite to in vitro experiments, these in situ experiments revealed no clear signs of altered bacterial surface colonization regarding structure dimensions and texture types compared to unstructured substrates or natural enamel. In part, there seemed to be a decreasing trend of adherent cells with increasing periodicities and structure sizes, but this pattern was weak and irregular. Pellicle formation took place on all substrates in an unaltered manner. However, pellicle formation was most pronounced within recessed areas thereby partially masking the three-dimensional character of the surfaces. As the natural pellicle layer is obviously the most dominant prerequisite for bacterial adhesion, colonization in the oral environment cannot be easily controlled by structural means.
Topics: Bacteria; Bacterial Adhesion; Biofilms; Dental Pellicle; Humans; Models, Biological; Mouth; Surface Properties
PubMed: 35163081
DOI: 10.3390/ijms23031157 -
Biofouling 2014Sodium dodecyl sulphate (SDS) and sodium tripolyphosphate (STP) act to remove stained pellicle from dentition and loosen deposits on tooth surfaces that may become...
Sodium dodecyl sulphate (SDS) and sodium tripolyphosphate (STP) act to remove stained pellicle from dentition and loosen deposits on tooth surfaces that may become cariogenic over time. This study investigated how SDS and STP impact the salivary pellicle adsorbed onto hydroxyapatite and silica sensors using a dual polarisation interferometer and a quartz-crystal microbalance with dissipation. After the pellicle was exposed to SDS and STP the remaining pellicle, although weaker, due to the loss of material, became less dense but with a higher elastic component; suggesting that the viscous component of the pellicle was being removed. This would imply a structural transformation from a soft but dense structured pellicle, to a more diffuse pellicle. In addition, the majority of proteins displaced by both SDS and STP were identified as being acidic in nature; implying that the negatively charged groups of SDS and STP may be responsible for the displacement of the pellicle proteins observed.
Topics: Adult; Chromatography, Liquid; Dental Pellicle; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Polyphosphates; Quartz Crystal Microbalance Techniques; Saliva; Salivary Proteins and Peptides; Sodium Dodecyl Sulfate; Tandem Mass Spectrometry; Young Adult
PubMed: 25397690
DOI: 10.1080/08927014.2014.977268 -
Scientific Reports Jan 2020Pellicle is the initial proteinaceous layer that is formed almost instantaneously on all solid surfaces in the oral cavity. It is of essential relevance for any...
Pellicle is the initial proteinaceous layer that is formed almost instantaneously on all solid surfaces in the oral cavity. It is of essential relevance for any interactions and metabolism on the tooth surface. Up to now, there is no information on the metabolome of this structure. Accordingly, the present study aims to characterise the metabolomic profile of in-situ pellicle in children with different caries activity for the first time in comparison to saliva. Small molecules such as carbohydrates, amino acids, organic acids, and fatty acids, putatively involved in the formation of caries were quantified using mass spectrometry (MS)-based techniques, such as (stable isotope dilution analysis)-ultra-performance liquid chromatography-tandem MS and gas chromatography/electron ionisation-MS. Pellicle and corresponding saliva samples were collected from caries-active, caries-free and caries-rehabilitated 4- to 6-year-old children. The most abundant analytes in pellicle were acetic acid (1.2-10.5 nmol/cm), propionic acid (0.1-8.5 nmol/cm), glycine (0.7-3.5 nmol/cm), serine (0.08-2.3 nmol/cm), galactose (galactose + mannose; 0.035-0.078 nmol/cm), lactose (0.002-0.086 nmol/cm), glucose (0.018-0.953 nmol/cm), palmitic acid (0.26-2.03 nmol/cm), and stearic acid (0.34-1.81 nmol/cm). Significant differences depending on caries activity were detected neither in saliva nor in the corresponding pellicle samples.
Topics: Acetic Acid; Case-Control Studies; Child; Child, Preschool; Chromatography, Liquid; Dental Caries; Dental Pellicle; Glycine; Humans; Male; Metabolomics; Propionates; Saliva; Tandem Mass Spectrometry
PubMed: 31959821
DOI: 10.1038/s41598-020-57531-8 -
Clinical Oral Investigations Oct 2023The present study aimed to evaluate the potential of the salivary pellicle (SP) formed on titanium (Ti) surfaces to modulate the formation of a biofilm composed of...
OBJECTIVES
The present study aimed to evaluate the potential of the salivary pellicle (SP) formed on titanium (Ti) surfaces to modulate the formation of a biofilm composed of Streptococcus gordonii, Actinomyces naeslundii, Fusobacterium nucleatum, and Porphyromonas gingivalis.
MATERIALS AND METHODS
Ti substrates were incubated for 2 h with a pool of saliva samples obtained from 10 systemically and periodontally healthy subjects. Enamel substrates were included as a biological reference. Scanning electron microscopy (SEM) and Raman spectroscopy analysis were used to analyze the formation of the salivary pellicle. After the SP formation, the surfaces were incubated for 12 h with a mix of Streptococcus gordonii, Actinomyces naeslundii, Fusobacterium nucleatum, and Porphyromonas gingivalis. The number of bacterial cells attached to each surface was determined by the XTT assay while bacterial viability was analyzed by fluorescence microscopy using the LIVE/DEAD® BacLight kit.
RESULTS
The SEM and Raman spectroscopy analysis confirmed the presence of a salivary pellicle formed on the tested surfaces. Regarding the biofilm formation, the presence of the SP decreases the number of the bacterial cells detected in the test surfaces, compared with the uncover substrates. Even more, the SP-covered substrates showed similar bacterial counts in both Ti and enamel surfaces, meaning that the physicochemical differences of the substrates were less determinant than the presence of the SP. While on the SP-uncover substrates, differences in the bacterial adhesion patterns were directly related to the physicochemical nature of the substrates.
CONCLUSIONS
The salivary pellicle was the main modulator in the development of the biofilm consisting of representative oral bacteria on the Ti substrates.
CLINICAL RELEVANCE
The results of this study provide valuable information on the modulatory effect of the salivary pellicle on biofilm formation; such information allows us to understand better the events involved in the formation of oral biofilms on Ti dental implants.
Topics: Humans; Dental Pellicle; Titanium; Biofilms; Bacterial Adhesion; Streptococcus gordonii; Fusobacterium nucleatum; Surface Properties
PubMed: 37646908
DOI: 10.1007/s00784-023-05230-9 -
The Protein Journal Oct 2022Carbonic anhydrases (CA, EC 4.2.1.1) catalyze the hydration of carbon dioxide and take part in many essential physiological processes. In humans, 15 CAs are...
Carbonic anhydrases (CA, EC 4.2.1.1) catalyze the hydration of carbon dioxide and take part in many essential physiological processes. In humans, 15 CAs are characterized, including the only secreted isoenzyme CA VI. CA VI has been linked to specific processes in the mouth, namely bitter taste perception, dental caries, and maintenance of enamel pellicle, and implicated in several immunity-related phenomena. However, little is known of the mechanisms of the above. In this study, we characterized human CA VI purified from saliva and milk with biophysical methods and measured their enzyme activities and acetazolamide inhibition. Size-exclusion chromatography showed peaks of salivary and milk CA VI corresponding to hexameric state or larger at pH 7.5. At pH 5.0 the hexamer peaks dominated. SDS- PAGE of milk CA VI protein treated with a bifunctional crosslinker further confirmed that a majority of CA VI is oligomers of similar sizes in solution. Mass spectrometry experiments confirmed that both of the two putative N-glycosylation sites, Asn67 and Asn256, are heterogeneously glycosylated. The attached glycans in milk CA VI were di- and triantennary complex-type glycans, carrying both a core fucose and 1 to 2 additional fucose units, whereas the glycans in salivary CA VI were smaller, seemingly degraded forms of core fucosylated complex- or hybrid-type glycans. Mass spectrometry also verified the predicted signal peptide cleavage site and the terminal residue, Gln 18, being in pyroglutamate form. Thorough characterization of CA VI paves way to better understanding of the biological function of the protein.
Topics: Carbonic Anhydrases; Fucose; Humans; Milk, Human; Saliva
PubMed: 35947329
DOI: 10.1007/s10930-022-10070-9 -
PloS One 2022Polyphenol-rich solutions, such as plant extracts and teas, can modify the salivary pellicle and improve the protection against dental erosion. In this study, we further...
Polyphenol-rich solutions, such as plant extracts and teas, can modify the salivary pellicle and improve the protection against dental erosion. In this study, we further explored how these polyphenol-rich plant extracts solutions behave in the presence of fluoride. We distributed enamel specimens into 9 groups (n = 15): Control_No_F- (Deionized water); Control_F- (500 ppm F-), Grape_Seed_No_F- (Grape seed extract), Grape_Seed_F- (Grape seed extract + 500 ppm F-), Grapefruit_Seed_No_F- (Grapefruit seed extract), Grapefruit_Seed_F- (Grapefruit seed extract + 500 ppm F-), Blueberry_No_F- (Blueberry extract), Blueberry_F- (Blueberry extract + 500 ppm F-), and Sn2+/F-_Rinse (commercial solution containing 800 ppm Sn2+ and 500 ppm F-). The specimens were submitted to 5 cycles (1 cycle per day), and each cycle consisted of: salivary pellicle formation (human saliva, 30 min, 37°C), modification of the pellicle (2 min, 25°C), pellicle formation (60 min, 37°C), and an erosive challenge (1 min, citric acid). Between cycles, the specimens were kept in a humid chamber. Relative surface hardness (rSH), relative surface reflection intensity (rSRI) and calcium released to the acid were analysed, using general linear models, and Kruskal-Wallis with post-hoc Dunn's tests. We observed that the presence of fluoride in synergy with the extract solutions provided better protection than the groups containing extract or fluoride only. For rSH, we observed a significant main effect of extracts (F(4,117) = 9.20; p<0.001) and fluoride (F(1,117) = 511.55; p<0.001), with a significant interaction (F(3,117) = 6.71; p<0.001). Grape_Seed_F- showed the best protection, better than fluoride, and Sn2+/F-_Rinse. Calcium results also showed greater protection for the groups containing fluoride, whereas for rSRI, despite a significant interaction between extract and fluoride (F(3,117) = 226.05; p<0.001), the differences between the groups were not as clearly observed. We conclude that polyphenols from plant extracts, when combined with fluoride, improve the protective effect of salivary pellicles against enamel erosion.
Topics: Humans; Fluorides; Tooth Erosion; Sodium Fluoride; Calcium; Polyphenols; Grape Seed Extract; Dental Enamel
PubMed: 36383626
DOI: 10.1371/journal.pone.0277552 -
Biological Research Jan 2020The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of... (Comparative Study)
Comparative Study
BACKGROUND
The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins.
RESULTS
The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and α-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva.
CONCLUSIONS
Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.
Topics: Adult; Blotting, Western; Dental Pellicle; Electrophoresis, Polyacrylamide Gel; Female; Gingival Crevicular Fluid; Humans; Male; Mass Spectrometry; Proteins; Saliva
PubMed: 31941552
DOI: 10.1186/s40659-020-0271-2