RESUMEN
OBJECTIVES: To evaluate in vivo 1) the bioavailability of trans-resveratrol when administered through sublingual capsules; 2) the effect of resveratrol on the protein composition of the acquired enamel pellicle (AEP). DESIGN: Ten volunteers received a sublingual capsule containing 50 mg of trans-resveratrol. Unstimulated saliva was then collected after 0, 30, 60, and 120 min and AEP was collected after 120 min following administration of the capsule. In the next week, the volunteers received a placebo sublingual capsule, and saliva and AEP were collected again. Saliva samples were analyzed for free trans-resveratrol using high-performance liquid chromatopgraphy (HPLC), and AEP samples were subjected to proteomic analysis (nLC-ESI-MS/MS). RESULTS: Trans-resveratrol was detected in saliva at all the time points evaluated, with the peak at 30 min. A total of 242 proteins were identified in both groups. Ninety-six proteins were increased and 23 proteins were decreased in the Resveratrol group. Among the up-regulated proteins, isoforms of cystatins, PRPs, Mucin-7, Histatin-1, Lactotrasnferrin and Lysozyme-C were increased and the isoforms of Protein S100, Neutrophil defensins, Albumin, PRPs, and, Statherin were decreased in Resveratrol group. CONCLUSION: The sublingual capsule is effective at increasing the bioavailability of trans-resveratrol in saliva. Several proteins involved in important processes to maintain systemic and oral health homeostasis were identified. These proteins differently expressed due to the presence of trans-resveratrol deserve attention for future studies, since they have important functions, mainly related to antimicrobial action.
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Cápsulas , Película Dental , Resveratrol , Saliva , Humanos , Resveratrol/farmacología , Resveratrol/farmacocinética , Resveratrol/administración & dosificación , Saliva/metabolismo , Saliva/química , Masculino , Adulto , Película Dental/metabolismo , Película Dental/química , Cromatografía Líquida de Alta Presión , Femenino , Disponibilidad Biológica , Estilbenos/farmacocinética , Estilbenos/farmacología , Estilbenos/administración & dosificación , Proteómica , Espectrometría de Masas en Tándem , Proteínas y Péptidos Salivales/metabolismoRESUMEN
Using the surface characterization techniques of quartz crystal microbalance with dissipation, atomic force microscopy, and scanning electron microscopy, the structure of the salivary pellicle was investigated before and after it was exposed to dairy proteins, including micellar casein, skim milk, whey protein isolate (WPI), and a mixture of skim milk and WPI. We have shown that the hydration, viscoelasticity, and adsorbed proteinaceous mass of a preadsorbed salivary pellicle on a PDMS surface are greatly affected by the type of dairy protein. After interaction with whey protein, the preadsorbed saliva pellicle becomes softer. However, exposure of the saliva pellicle to micellar casein causes the pellicle to partially collapse, which results in a thinner and more rigid surface layer. This structure change correlates with the measured lubrication behavior when the saliva pellicle is exposed to dairy proteins. While previous studies suggest that whey protein is the main component in milk to interact with salivary proteins, our study indicates interactions with casein are more important. The knowledge gained here provides insights into the mechanisms by which different components of dairy foods and beverages contribute to mouthfeel and texture perception, as well as influence oral hygiene.
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Película Dental , Proteínas y Péptidos Salivales , Película Dental/química , Película Dental/metabolismo , Proteínas y Péptidos Salivales/química , Proteínas y Péptidos Salivales/metabolismo , Adsorción , Caseínas/química , Caseínas/metabolismo , Propiedades de Superficie , Proteína de Suero de Leche/química , Humanos , Animales , Microscopía de Fuerza Atómica , Saliva/química , Saliva/metabolismo , Tecnicas de Microbalanza del Cristal de CuarzoRESUMEN
OBJECTIVE: This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10- 5M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization. MATERIALS AND METHODS: In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (H2O-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed. RESULTS: The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS. CONCLUSIONS: The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization. CLINICAL RELEVANCE: Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.
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Caries Dental , Desmineralización Dental , Humanos , Película Dental/química , Película Dental/microbiología , Caries Dental/prevención & control , Proteómica , Biopelículas , Hemoglobinas/análisis , Desmineralización Dental/prevención & controlRESUMEN
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® BacLightTM 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.
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Biopelículas , Titanio , Humanos , Película Dental/química , Película Dental/microbiología , Titanio/química , Adhesión Bacteriana , Streptococcus gordonii , Fusobacterium nucleatum , Propiedades de SuperficieRESUMEN
OBJECTIVES: To evaluate and compare the proteomic profile of acquired pellicle on smooth bovine tooth and tooth-coloured restorative materials, including resin composite (RC), glass ionomer cement (GIC), and casein phosphopeptide-amorphous calcium phosphate modified GIC (CPP-ACP GIC). METHODS: Two-hour in situ pellicles on tooth/materials specimens mounted in oral appliances worn by ten healthy adults were investigated. Pellicle proteins and corresponding unstimulated whole saliva were quantitatively analysed through liquid chromatography-tandem mass spectrometry. RESULTS: Significantly higher amounts of protein were adsorbed onto tooth surface than restorative materials tested (4.11 ± 0.69 vs. 2.54 ± 0.38/2.98 ± 0.80/3.01 ± 0.37 µg, RC/GIC/CPP-ACP GIC). From the ten participants, 1,444 (487-1,086/person), 1,454 (645-1,051/person), 1,731 (454-1,475/person), or 1,597 (423-1,261/person) pellicle proteins were detected at least once on bovine tooth, RC, GIC, or CPP-ACP GIC, respectively, and with 1,072 (304-793/person) salivary proteins identified. Comparative quantification revealed minor differences between tooth and restorative materials pellicle profiles. High inter-individual variations in pellicle protein composition were demonstrated. Compared to the salivary protein profile, 214/57 proteins showed significantly increased/decreased abundance in pellicle formed on at least one substrate (fold change > 3.325/fold change < 0.301). Gene Ontology enrichment analysis showed some pellicle proteins detected with increased affinity to tooth/material surface were identified as being related to "calcium-dependent protein binding" or "cell-cell adhesion mediator activity". CONCLUSION: Similar protein quantity and composition was observed in 2 h in situ pellicles formed on different smooth restorative material surfaces. The proteomic profile of pellicles appeared distinct from that of the corresponding unstimulated whole saliva. CLINICAL SIGNIFICANCE: Host backgrounds appeared more influential on the proteomic profile of the in situ acquired pellicle than the underlying substrate characteristics among systemically and orally healthy adults. Pellicle proteins preferentially adsorbed on tooth/materials were putatively associated with calcium ion homeostasis or host-microbiota interaction.
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Calcio , Proteómica , Adulto , Humanos , Animales , Bovinos , Calcio/análisis , Materiales Dentales/química , Película Dental/química , Resinas Compuestas , Proteínas y Péptidos Salivales/análisis , Cementos de Ionómero Vítreo/químicaRESUMEN
While the ultrastructure of the enamel pellicle and its erosion protective properties are well studied, the dentin pellicle is still neglected in dental research. Therefore, the ultrastructure and erosion protective properties of a pellicle formed on bovine dentin specimens were investigated in the present study. The dentin pellicle was formed in situ for 3, 30, 120, and 360 min at buccal or palatal oral sites of 3 subjects and analyzed by transmission electron microscopy. In order to clarify the impact of an erosive challenge to the ultrastructure of the pellicle and the underlying dentin, specimens were exposed to the oral cavity and eroded in vivo with 0.1% or 1% citric acid either immediately or after 30 min of pellicle formation. Specimens that were eroded without exposure to the oral cavity served as control. In another trial, specimens with a 30-min pellicle were exposed to the oral cavity for a further 60 min after the erosive challenge to investigate the effect of saliva on the impaired pellicle and dentin. Transmission electron micrographs reveal a globular and granular structured pellicle layer, which was thicker when the pellicle was formed buccally or with longer formation times. Erosion with citric acid reduced the thickness of the pellicle and interrupted its continuity. The dentin was also affected by erosion, which was represented by a lower electron density and formation of demineralized lacunae. These were infiltrated by a granular structured material when specimens were exposed to the oral cavity. After further intraoral exposure, the infiltration was more pronounced, indicating a significant impact of saliva on the demineralized dentin. A reformation of the dentin pellicle on the other hand did not occur. In conclusion, the dentin pellicle is neither acid-resistant nor able to effectively protect dentin from erosion.
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Esmalte Dental , Erosión de los Dientes , Humanos , Bovinos , Animales , Película Dental/química , Erosión de los Dientes/prevención & control , Ácido Cítrico/efectos adversos , DentinaRESUMEN
OBJECTIVE: The present study aimed to systematically analyse the complete lipid profile of the in situ pellicle in comparison to saliva. For the first time, the modern sensitive methods GC-EI/MS and HPLC MS/MS were to be used for this purpose. DESIGN: Bovine enamel slabs were exposed to the oral cavity of 12 subjects by customized splints (3 min, 30 min or 120 min). Afterwards, the pellicle samples were obtained and further investigated in vitro. Additionally, corresponding unstimulated saliva samples were collected. GC-EI/MS was performed to qualitatively and quantitatively determine all fatty acids contained in the investigated samples. The individual lipid classes of phospholipids, triacylglycerols, glycolipids, cholesterol and cholesterol esters were analysed qualitatively by HPLC MS/MS. RESULTS: A characteristic fatty acid profile of the in situ pellicle was proven. Furthermore, triacylglycerols with the major fatty acids 16:0, 18:0, 18:1, 18:2, and phospholipids were detected as integral components in the pellicle. There were four groups of phospholipids: Lyso-phosphatidylcholines, phosphatidylcholines, phosphatidylethanol-amines, and phosphatidylinositols. Differences between saliva and pellicle were evident in the composition of the fatty acid- and the phospholipid profile. Glycolipids, cholesterol and cholesterol esters could neither be detected in pellicle- nor in saliva samples. CONCLUSION: The lipid profiles of the in situ pellicle and saliva were successfully characterised. Differences in the phospholipid and fatty acid composition between pellicle and saliva indicate a selective pellicle formation process. The results provide an important reference and core data for further investigation of the complex surface interactions in the oral cavity, especially concerning hydrophobic substances.
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Ésteres del Colesterol , Espectrometría de Masas en Tándem , Animales , Bovinos , Ésteres del Colesterol/análisis , Película Dental/química , Ácidos Grasos , Glucolípidos/análisis , Humanos , Fosfatidilcolinas/análisis , Fosfolípidos/análisis , Saliva/química , TriglicéridosRESUMEN
OBJECTIVE: The aim of the present study was to investigate the impact of periodic polyphenol treatment on the ultrastructure and anti-erosive potential of an in-situ formed pellicle. METHODS: Subjects wore intraoral appliances with buccally and palatally fixed bovine enamel specimens. During 6 h of intraoral pellicle formation, 100 ml black tea or tannic acid was applied ex-vivo every 25 min for 5 min. Untreated pellicles served as control. After the trial, specimens were immersed in 0.1% or 1% citric acid for 60 s and analysed for calcium release with atomic adsorption spectrometry and ultrastructure with transmission electron microscopy. RESULTS: Specimens covered by pellicles treated with black tea or tannic acid released less calcium than untreated pellicles. Ultrastructural analyses reveal an increase in pellicle's thickness and density after treatment with polyphenols. CONCLUSIONS: Periodic polyphenol treatment of the pellicle modify its ultrastructure and increase its anti-erosive potential. CLINICAL SIGNIFICANCE: Consumption of polyphenolic beverages can enhance the anti-erosive potential of the enamel pellicle.
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Polifenoles , Erosión de los Dientes , Animales , Calcio/análisis , Bovinos , Película Dental/química , Humanos , Polifenoles/análisis , Polifenoles/farmacología , Taninos/farmacología , Té , Erosión de los Dientes/tratamiento farmacológico , Erosión de los Dientes/prevención & controlRESUMEN
The acquired enamel pellicle (AEP) is a multi-protein film attached to the surface of teeth, which functions to lubricate the dental surface, form an anti-erosive barrier and exhibits antimicrobial properties. The initiation of AEP formation occurs within seconds of exposure to saliva, a biofluid rich in protein species. While there have been many publications on the formation of human AEP there is little research on the composition of canine AEP during its acquisition. The aim of these studies was to explore the composition of canine AEP formation, utilising hydroxyapatite (HA) discs as a tooth substitute matrix, over time. Qualitative and quantitative proteomics techniques using tandem mass tag labelled peptides and LC-MS/MS were used to follow the formation of canine AEP on hydroxyapatite discs over the course of an hour. Proteins adsorbed to the HA surface included highly abundant proteins in canine saliva, antimicrobial proteins, protease inhibitors and the buffering agent carbonic anhydrase. Greater understanding of the canine AEP deepens fundamental knowledge of the early processes driving bacterial colonisation of the tooth surface and subsequent plaque accumulation.
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Proteómica , Lobos , Animales , Cromatografía Liquida/veterinaria , Película Dental/química , Película Dental/metabolismo , Durapatita/análisis , Durapatita/química , Durapatita/metabolismo , Humanos , Proteínas/análisis , Proteínas/metabolismo , Proteómica/métodos , Espectrometría de Masas en Tándem/veterinariaRESUMEN
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.
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Bacterias/metabolismo , Adhesión Bacteriana , Biopelículas/crecimiento & desarrollo , Película Dental/fisiología , Modelos Biológicos , Boca/fisiología , Bacterias/crecimiento & desarrollo , Película Dental/química , Película Dental/microbiología , Humanos , Boca/química , Boca/microbiología , Propiedades de SuperficieRESUMEN
OBJECTIVE: To study the proteomic alterations in the initial AEP after rinsing with CaneCPI-5, StN15 or Hb or their combination. MATERIALS AND METHODS: In five crossover phases, after prophylaxis, 10 volunteers in 5 consecutive days, rinsed (10 mL, 1 min) with the following solutions: deionized water (H2O- negative control- 1), 0.1 mg/mL CaneCPI-5 (2), 1.88×10-5 M StN15 (3), 1.0 mg/mL Hb (4) or their combination (5). The AEP formed after 3 min was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. RESULTS: Rinsing with the proteins/peptide increased the amounts of proteins in the AEP. The total numbers of proteins identified after rinsing with CaneCPI-5, StN15, Hb or their combination versus water, were 131, 167, 148 and 142, respectively. The treatment with the proteins/peptide or their combination increased proteins that bind calcium, phosphate and interact with distinct proteins, as well as proteins with antimicrobial and acid-resistant properties, such as, Cornifin-B (7.7, 12.6, and 4.3-fold for CaneCPI-5, StN15 and Hb, respectively), isoforms of Cystatin (2.2-2.4-fold for CaneCPI-5 and StN15), Proline-rich-protein 4 (4.3-fold; StN15), Histatin-1 (2.8-fold; StN15) and Hemoglobin (7.7-25-fold for Hb and Combination). Immunoglobulin, Keratin and Histone were exclusively identified upon treatment with the proteins/peptide, alone or combined. CONCLUSION: Rinsing with proteins/peptide, alone or combined, increased protective proteins in the initial AEP. CLINICAL RELEVANCE: Our results suggest that rinsing with the proteins/peptide or their combination increases the proteins capable of enhancing the protective function of the basal layer of AEP.
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Proteínas , Proteómica , Película Dental/química , Humanos , Péptidos , AguaRESUMEN
In the oral environment, the acquired salivary pellicle (ASP) on the tooth surface comprises proteins, glycoproteins, carbohydrates, and lipids. The ASP can specifically and rapidly adsorb on the enamel surface to provide effective lubrication, protection, hydration, and remineralisation, as well as be recognised by various bacteria to form a microbial biofilm (plaque). The involved proteins, particularly various phosphoproteins such as statherins, histatins, and proline-rich proteins, are vital to their specific functions. This review first describes the relationship between the biological functions of these proteins and their structures. Subsequently, recent advances in functional biomedical materials derived from these proteins are reviewed in terms of dental/bone therapeutic materials, antibacterial materials, tissue engineering materials, and coatings for medical devices. Finally, perspectives and challenges regarding the rational design and biomedical applications of ASP-derived materials are discussed.
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Materiales Biocompatibles/química , Película Dental/química , Proteínas/química , Humanos , Ensayo de MaterialesRESUMEN
This study reports the differences in the protein composition of salivary pellicles formed under in situ conditions on two Titanium (Ti) surfaces, with different roughness and wettability. Smooth pretreatment Ti surfaces (Ti-PT) with an average roughness (Ra) of 0.45 µm and a water contact angle (WCA) of 92.4°, as well as a more rough sandblasted, large grit, acid-etched treatment Ti surfaces (Ti-SLA) with a Ra of 3.3 µm and WCA of 131.8°, were tested. The salivary pellicles were quantitatively analyzed by bicinchoninic acid assays, and the protein identification was performed by Nano-LC-MS/MS (nano mass spectrometry). Protein levels of 2.5, and 9.1 µg/ml were quantified from the detached salivary pellicle formed on the Ti-PT and Ti-SLA surfaces, respectively. Using Nano-LC-MS/MS, a total of 597 proteins were identified on all the substrates tested; 43 proteins were identified only on the Ti-PT, and 226 proteins were adsorbed solely on the Ti-SLA substrates. The physicochemical characteristics of the Ti implant surfaces modified the amount and the identity of the salivary proteome of the pellicles formed, confirming the high selectivity of the protein pellicle formed on a surface once is exposed in the oral cavity.
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Película Dental/química , Prótesis e Implantes , Titanio/química , HumectabilidadRESUMEN
OBJECTIVE: Aim of this in vitro study was to investigate erosive tooth loss in dependence of the enamel surface structure and presence of an acquired pellicle. METHODS: Enamel specimens from 19 bovine incisors (4 specimens/incisor) were allocated to four experimental groups (n = 19). The surfaces of half of the specimens were polished (two groups), while the other half was left native (two groups). Specimens of one polished and one native group were placed in pooled human saliva (30 min) for the formation of an acquired pellicle. Thereafter, all specimens were demineralized by superfusion with hydrochloric acid (17 min, pH 2.3) with collection of the superfluent. Erosive substance loss was determined by measuring the dissolved calcium content using a colorimetric assay with Arsenazo III reagent. Differences in erosive substance loss were statistically analyzed with respect to enamel surface and pellicle. A linear mixed effects model was fitted to the data and pairwise differences between groups were evaluated (significance level α= 0.05). RESULTS: Enamel surface structure (p < 0.001) and presence of pellicle (p = 0.01) had a significant effect on erosive substance loss. Polished surfaces with pellicle showed the lowest cumulative calcium release [nmol Ca/mm2] (means ± standard deviation: 48+/-5), followed by polished specimens without (51+/-9) and native specimens with pellicle (54+/-10). No significant differences were found between these groups. Highest cumulative calcium release was found for native specimens without pellicle (61+/-9; p < 0.05). CONCLUSIONS: Both enamel surface structure and the acquired pellicle are important determinants of the susceptibility to erosive tooth loss.
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Esmalte Dental/química , Película Dental/química , Erosión de los Dientes , Animales , Bovinos , Ácido Cítrico , Solubilidad del Esmalte Dental , Humanos , Ácido Clorhídrico , SalivaRESUMEN
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/cm2), propionic acid (0.1-8.5 nmol/cm2), glycine (0.7-3.5 nmol/cm2), serine (0.08-2.3 nmol/cm2), galactose (galactose + mannose; 0.035-0.078 nmol/cm2), lactose (0.002-0.086 nmol/cm2), glucose (0.018-0.953 nmol/cm2), palmitic acid (0.26-2.03 nmol/cm2), and stearic acid (0.34-1.81 nmol/cm2). Significant differences depending on caries activity were detected neither in saliva nor in the corresponding pellicle samples.
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Caries Dental/metabolismo , Película Dental/química , Metabolómica/métodos , Saliva/química , Ácido Acético/análisis , Estudios de Casos y Controles , Niño , Preescolar , Cromatografía Liquida , Glicina/análisis , Humanos , Masculino , Propionatos/análisis , Espectrometría de Masas en TándemRESUMEN
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.
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Película Dental/química , Líquido del Surco Gingival/química , Proteínas/análisis , Saliva/química , Adulto , Western Blotting , Electroforesis en Gel de Poliacrilamida , Femenino , Humanos , Masculino , Espectrometría de MasasRESUMEN
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.
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Humanos , Masculino , Femenino , Adulto , Saliva/química , Proteínas/análisis , Líquido del Surco Gingival/química , Película Dental/química , Espectrometría de Masas , Western Blotting , Electroforesis en Gel de PoliacrilamidaRESUMEN
This study aimed to analyse if the erosion-protective potential of the salivary pellicle is different between female and male subjects. Bovine enamel and dentin specimens (each n = 3) were exposed to the oral cavity of healthy female or male volunteers (each n = 25, females: 25.8 ± 3.5 years, males: 26.7 ± 4.0 years) for 120 min to form a salivary pellicle. Subsequently, each 2 enamel and 2 dentin specimens were eroded with hydrochloric acid (pH 2.6, 60 s). Specimens of the control group (each n = 30) were eroded without presenting a salivary pellicle. Calcium release into the acid was determined photometrically. Additionally, total protein content in the pellicle (each n = 1 enamel and dentin specimen/volunteer) and different salivary parameters (flow rate, pH, buffer capacity, protein, albumin, calcium, phosphate, fluoride) were assessed. Statistical analyses were performed by one-way ANOVA, t tests, multiple linear regressions and Pearson correlations (p < 0.05). The erosion-protective capacity was not significantly different among female (calcium release [% of control]: enamel: 82.6 ± 28.1, dentin: 80.7 ± 24.0) and male (enamel: 76.0 ± 27.5, dentin: 87.1 ± 34.9) subjects. The protein content of the pellicle was not different between female and male subjects. The protein content and pH of unstimulated saliva were significantly reduced in female compared to male volunteers. Calcium release was neither correlated with the protein content of the salivary pellicle nor with salivary parameters. Under the conditions of the present study, the erosion-protective capacity of the salivary pellicle of female and male subjects is not different.
Asunto(s)
Película Dental/química , Saliva/química , Erosión de los Dientes/prevención & control , Animales , Bovinos , Esmalte Dental/química , Solubilidad del Esmalte Dental , Dentina/química , Femenino , Humanos , Masculino , Factores Sexuales , Adulto JovenRESUMEN
The chemical composition of biomaterials can drive their biological responses; therefore, this in vitro study aimed to evaluate the proteomic profile of the salivary pellicle formed on titanium (Ti) alloys containing niobium (Nb) and zirconium (Zr). The experimental groups consisted of Ti35NbxZr (x = 5 and 10 wt%) alloys, and commercially pure titanium (cpTi); titanium aluminium vanadium (Ti6Al4V) alloys were used as controls. The physical and chemical characteristics of the Ti materials were analysed. The proteomic profile was evaluated by liquid chromatography coupled with tandem mass spectrometry. Bacterial adhesion (2 h) of mixed species (Streptococcus sanguinis and Actinomyces naeslundii) was investigated as colony-forming units (n = 6). This paper reports the finding that salivary pellicle composition can be modulated by the composition of the Ti material. The Ti35NbxZr group showed a significant ability to adsorb proteins from saliva, which can favour interactions with cells and compatibility with the body.
Asunto(s)
Aleaciones/química , Película Dental/química , Niobio/química , Proteoma/análisis , Proteínas y Péptidos Salivales/análisis , Titanio/química , Circonio/química , Adsorción , Adhesión Bacteriana , Materiales Biocompatibles/química , ProteómicaRESUMEN
The proteinaceous pellicle layer, which develops upon contact with saliva on the surface of teeth, is important for the formation of oral biofilms and for the protection of teeth from abrasion and chemically induced erosion. Astringent food ingredients comprising polyphenols, cationic macromolecules, and multivalent metal salts are known to interact with the pellicle. However, astringent-induced changes in the physicochemical properties of the tooth-saliva interphase are not yet completely understood. Here we provide comprehensive insights into interfacial charging, ultrastructure, thickness, and surface roughness of the pellicles formed on the model substrates silicon oxide (SiO2), Teflon® AF, and hydroxyapatite, as well as on bovine enamel before and after incubation with the astringents epigallocatechin gallate, tannic acid, iron(III) salt, lysozyme, and chitosan. Quartz crystal microbalance with dissipation monitoring demonstrated viscous behavior of untreated pellicles formed in vitro on the different materials. Electrokinetic (streaming current) measurements revealed that cationic astringents reverse the charge of native pellicles, whereas polyphenols did not change the charge under physiological pH condition. In addition, transmission electron microscopy and atomic force microscopy showed a concentration-dependent increase in average film thickness and pellicle surface roughness as induced by astringents. These multifaceted alterations of the salivary pellicle may come along with an increase in roughness perceived on the teeth, which is part of the complex sensations of oral astringency.