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Proteomics. Clinical Applications May 2022The use of dental restorative materials is a routine task in clinical dentistry. Upon exposure to the oral cavity, continuous adsorption of salivary proteins and other...
PURPOSE
The use of dental restorative materials is a routine task in clinical dentistry. Upon exposure to the oral cavity, continuous adsorption of salivary proteins and other macromolecules to all surfaces occurs, representing the first step in dental biofilm formation. Different physico-chemical properties of substrate materials potentially influence the composition of the initial biofilm, termed pellicle. This study aimed at characterizing and comparing the individual proteomic composition of the 3-min pellicle formed on bovine enamel and six restorative materials.
EXPERIMENTAL DESIGN
After chemical elution, pellicle proteins were identified by nano-LC-HR-MS/MS. Proteomic profiles were analyzed in terms of molecular weights, isoelectric points, molecular functions and compared to saliva to reveal substrate material-specific adsorption patterns.
RESULTS
A total of 1348 different pellicle proteins were identified, with 187-686 proteins in individual 3-min pellicles. Unexpectedly, this yielded quite similar distribution patterns independent of the substrate materials. Furthermore, overall similar fold changes were obtained for the major part of commonly enriched or depleted proteins in the pellicles.
CONCLUSIONS AND CLINICAL RELEVANCE
The current results point to a minor role of the substrate material on the proteomic composition of the 3-min pellicle and represent core data for understanding the complex surface interactions in the oral cavity.
Topics: Animals; Cattle; Dental Pellicle; Proteomics; Saliva; Salivary Proteins and Peptides; Tandem Mass Spectrometry
PubMed: 35195368
DOI: 10.1002/prca.202100109 -
The International Journal of Oral &... 2021The aim of this study was to compare the Streptococcus oralis biofilm formation on titanium machined turned surfaces and sandblasted surfaces that were previously...
PURPOSE
The aim of this study was to compare the Streptococcus oralis biofilm formation on titanium machined turned surfaces and sandblasted surfaces that were previously characterized for their superficial topographies.
MATERIALS AND METHODS
Two commercially pure titanium surfaces were analyzed and compared: machined (turned surfaces subjected to a process of decontamination that also included a double acid attack) and sandblasted (sandblasted surfaces, cleaned with purified water, enzymatic detergent, acetone, and alcohol). The characterization of the samples at the nanolevel was performed using atomic force microscopy, which permitted calculation of the superficial nanoroughness (Ra). The sessile drop method was used to measure the water contact angle in both groups and allowed information to be gained about their wetting properties. Scanning electron microscope and energy-dispersive x-ray spectroscopy analysis allowed comparison of the microtopographic geometry and the chemical composition of the samples. Then, the disks were pre-incubated with saliva in order to form an acquired pellicle. Streptococcus oralis was put on the disks, and both groups were tested at 24 and 48 hours for biofilm biomass evaluation, colony-forming units (CFUs), and live/dead staining for cell viability.
RESULTS
The sandblasted samples were characterized by a significantly higher level of superficial oxides, superficial roughness, and hydrophilicity, compared with the machined turned samples. Although there were topographic differences, the Streptococcus oralis biofilm formation, quantified in CFUs, and biomass formation at 24 and 48 hours were similar in both groups. With the live/dead staining, the sandblasted disks were characterized by an increased percentage of dead cells compared with the machined disks.
CONCLUSION
Although significant topographic differences were present between machined and sandblasted disks, the Streptococcus oralis biofilm formation seems to not be significantly affected.
Topics: Biofilms; Dental Implants; Microscopy, Electron, Scanning; Streptococcus oralis; Surface Properties; Titanium
PubMed: 34698719
DOI: 10.11607/jomi.8739 -
Journal of Dental Research Jun 2020The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse... (Review)
Review
The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse which can be ascribed to the variety of niches available in the mouth. Previous studies have focused on the microflora in disease-either caries or periodontitis-and only recently have they considered factors that maintain the normal microflora. This has led to the perception that the microflora proliferate in nutrient-rich periods during oral processing of foods and drinks and starves in between times. In this review, evidence is presented which shows that the normal flora are maintained on a diet of salivary factors including urea, lactate, and salivary protein degradation. These factors are actively secreted by salivary glands which suggests these factors are important in maintaining normal commensals in the mouth. In addition, the immobilization of SIgA in the mucosal pellicle indicates a mechanism to retain certain bacteria that does not rely on the bacterial-centric mechanisms such as adhesins. By examining the salivary metabolome, it is clear that protein degradation is a key nutrient and the availability of free amino acids increases resistance to environmental stresses.
Topics: Dental Pellicle; Immunoglobulin A, Secretory; Microbiota; Mouth; Saliva; Salivary Proteins and Peptides
PubMed: 32283990
DOI: 10.1177/0022034520915486 -
Journal of Clinical Medicine Jun 2021Studies on small quantity, highly complex protein samples, such as salivary pellicle, have been enabled by recent major technological and analytical breakthroughs.... (Review)
Review
Studies on small quantity, highly complex protein samples, such as salivary pellicle, have been enabled by recent major technological and analytical breakthroughs. Advances in mass spectrometry-based computational proteomics such as Multidimensional Protein Identification Technology have allowed precise identification and quantification of complex protein samples on a proteome-wide scale, which has enabled the determination of corresponding genes and cellular functions at the protein level. The latter was achieved via protein-protein interaction mapping with Gene Ontology annotation. In recent years, the application of these technologies has broken various barriers in small-quantity-complex-protein research such as salivary pellicle. This review provides a concise summary of contemporary proteomic techniques contributing to (1) increased complex protein (up to hundreds) identification using minute sample sizes (µg level), (2) precise protein quantification by advanced stable isotope labelling or label-free approaches and (3) the emerging concepts and techniques regarding computational integration, such as the Gene Ontology Consortium and protein-protein interaction mapping. The latter integrates the structural, genomic, and biological context of proteins and genes to predict protein interactions and functional connections in a given biological context. The same technological breakthroughs and computational integration concepts can also be applied to other low-volume oral protein complexes such as gingival crevicular or peri-implant sulcular fluids.
PubMed: 34202147
DOI: 10.3390/jcm10132801 -
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 Sep 2020Dental composites are routinely placed as part of tooth restoration procedures. The integrity of the restoration is constantly challenged by the metabolic activities of...
Dental composites are routinely placed as part of tooth restoration procedures. The integrity of the restoration is constantly challenged by the metabolic activities of the oral microbiome. This activity directly contributes to a less-than-desirable half-life for the dental composite formulations currently in use. Therefore, many new antimicrobial dental composites are being developed to counteract the microbial challenge. To ensure that these materials will resist microbiome-derived degradation, the model systems used for testing antimicrobial activities should be relevant to the in vivo environment. Here, we summarize the key steps in oral microbial colonization that should be considered in clinically relevant model systems. Oral microbial colonization is a clearly defined developmental process that starts with the formation of the acquired salivary pellicle on the tooth surface, a conditioned film that provides the critical attachment sites for the initial colonizers. Further development includes the integration of additional species and the formation of a diverse, polymicrobial mature biofilm. Biofilm development is discussed in the context of dental composites, and recent research is highlighted regarding the effect of antimicrobial composites on the composition of the oral microbiome. Future challenges are addressed, including the potential of antimicrobial resistance development and how this could be counteracted by detailed studies of microbiome composition and gene expression on dental composites. Ultimately, progress in this area will require interdisciplinary approaches to effectively mitigate the inevitable challenges that arise as new experimental bioactive composites are evaluated for potential clinical efficacy. Success in this area could have the added benefit of inspiring other fields in medically relevant materials research, since microbial colonization of medical implants and devices is a ubiquitous problem in the field.
Topics: Anti-Infective Agents; Biofilms; Composite Resins; Dental Pellicle; Humans; Microbiota; Mouth; Streptococcus mutans
PubMed: 32479134
DOI: 10.1177/0022034520927690 -
Caries Research 2023The aim of this study was to evaluate the effect of film-forming polymer solutions of different concentrations and pH values, either associated or not with sodium...
The aim of this study was to evaluate the effect of film-forming polymer solutions of different concentrations and pH values, either associated or not with sodium fluoride (F; 225 ppm F-), when applied during the initial stage of salivary pellicle formation, to prevent the dissolution of hydroxyapatite (HA), which was determined by the pH-stat method. Polyacrylic acid (PA), chitosan, sodium linear polyphosphate (LPP), polyvinyl methyl ether/maleic anhydride (PVM/MA), and propylene glycol alginate (PGA) were tested in three concentrations (lower, medium, and higher), two pH values (native or adjusted), and either associated or not with F. Distilled water, F, and stannous ion+fluoride (Sn/F; 225 ppm F- and 800 ppm Sn2+, as SnCl2) solutions were the controls, totalizing 63 groups. HA crystals were pretreated with human saliva for 1 min to allow pellicle formation, then immersed in the experimental solutions (1 min), and exposed to saliva for another 28 min. Subsequently, they were added to a 0.3% citric acid solution (pH = 3.8), connected to a pH-stat system that added aliquots of 28 μL 0.1 N HCl for a total reaction time of 5 min. Data were analyzed with one-way ANOVA and Tukey's tests (α = 0.05). For PA alone, the concentrations of 0.1% (native pH), 0.06%, and 0.08% (both pH adjusted) showed significantly lower HA dissolution than the negative control. PA concentrations of 0.1% and 0.08%, of both pH values, improved the effect of F against HA dissolution to a near-identical value as Sn/F. All solutions containing chitosan and LPP significantly reduced HA dissolution in comparison with the control. For chitosan, the concentration of 0.5% (in both pH values) improved the effect of F. LPP at 0.5% (native pH) and all associations of LPP with F outperformed the effect of F. Some PVM/MA solutions significantly reduced HA dissolution but PVM/MA could not improve the protection of F. PGA was incapable of reducing HA dissolution or improving F effect. It was concluded that chitosan, LPP, and some PA and PVM/MA solutions used alone were capable of reducing HA dissolution. Only PA, chitosan, and LPP were able to enhance fluoride protection, but for PA and chitosan, this was influenced by the polymer concentration.
Topics: Humans; Fluorides; Durapatite; Polymers; Chitosan; Tooth Erosion; Sodium Fluoride; Tin Fluorides; Polyphosphates; Polyvinyls
PubMed: 37848001
DOI: 10.1159/000533546 -
Journal of Dentistry May 2024Previous studies on short- and long-term pellicles showed that the enamel pellicle provides partial protection against erosion. The aim of the present study was to...
OBJECTIVES
Previous studies on short- and long-term pellicles showed that the enamel pellicle provides partial protection against erosion. The aim of the present study was to investigate the protective properties of clinically relevant pellicles formed within 2 to 24 h. The hypothesis was that factors such as pellicle formation time, intraoral location, and acidic challenge severity would not influence the erosion-protective properties of the pellicle.
METHODS
Six subjects participated in the study. Bovine enamel specimens were prepared and intraorally exposed at buccal or palatal sites for 2, 6, 12, and 24 h to allow pellicle formation, followed by erosion using 0.1 % or 1 % citric acid. Calcium release and surface microhardness were measured, and specimens were analysed using scanning and transmission electron microscopy. Quantitative data were statistically analysed with three-way ANOVA and Tuckey's multiple comparison test (p = 0.05).
RESULTS
Pellicle formation time and intraoral location did not significantly influence the erosion-protective properties of the pellicle, while citric acid concentration significantly affected enamel erosion. The pellicle thickness increased with longer formation times and on buccal sites, but decreased or was entirely removed following treatment with 0.1 % or 1 % citric acid, respectively. The enamel surface exhibited a characteristic erosion pattern.
CONCLUSIONS
This study underscores the importance of investigating pellicle properties within the critical 2- to 24-h timeframe and highlights the significance of pellicle thickness in acid resistance.
CLINICAL SIGNIFICANCE
These findings provide valuable insights into the factors influencing the protective properties of enamel pellicles and could guide preventive measures in dental practice.
PubMed: 38815730
DOI: 10.1016/j.jdent.2024.105103 -
Frontiers in Public Health 2019The objective of this systematic review was to assess the scope and breadth of publicly available prospective cohort and randomized controlled trial (RCT) literature on...
The objective of this systematic review was to assess the scope and breadth of publicly available prospective cohort and randomized controlled trial (RCT) literature on 100% fruit juice and dental caries or tooth erosion in humans. We performed a systematic search in MEDLINE/PubMed, EMBASE, and Web of Science for studies published from inception through May 2018, and the Cochrane Library databases for reports published through January 2018. Prospective cohort studies or RCTs conducted on dental health and 100% fruit juice, and published in English were selected. No restrictions were set for age, sex, geographic location, or socioeconomic status. Eight publications representing five independent prospective cohort studies and nine publications on nine RCTs were included. All prospective cohort studies were in children or adolescents, and all RCTs were in adults. Prospective cohort studies on tooth erosion found no association between juice intake and tooth erosion, while those on dental caries incidence reported either no association or an inverse association between 100% fruit juice intakes and dental caries incidence. RCTs on tooth erosion showed decreased microhardness, increased surface enamel loss, increased erosion depth, greater enamel softening, and/or increased pellicle layer with 100% fruit juice, and those on dental caries showed increased demineralization of enamel slabs with 100% fruit juice. The existing evidence on 100% fruit juice intake and caries and tooth erosion are not conclusive. Overall, prospective cohort studies in children and adolescents found no association between 100% fruit juice intake and tooth erosion or dental caries, but, RCT data in adults suggests that 100% fruit juice could contribute to tooth erosion and dental caries. The RCT data, however, were from small, short-term studies that utilized intra-oral devices generally devoid of normal plaque or saliva action, and generally employed conditions that are not reflective of normal juice consumption.
PubMed: 31355175
DOI: 10.3389/fpubh.2019.00190 -
Experimental and Therapeutic Medicine Sep 2020Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that...
Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that horseradish peroxidase (HRP) adsorbs onto the membrane of the major oral streptococci, and (). However, the mechanism of interaction between HRP and the bacterial cell wall component is unclear. Dental plaques containing salivary glycoproteins and extracellular microbial products are visualized with 'dental plaque disclosing agent', and are controlled within dental therapy. However, current 'dental plaque disclosing agents' are difficult to evaluate with just dental plaques, since they stain and disclose not only dental plaques but also pellicle formed with salivary glycoproteins on a tooth surface. In this present study, we have demonstrated that HRP interacted with the cell wall component of the major gram-positive bacterial peptidoglycan, but not the major cell wall component of gram-negative bacteria lipopolysaccharide. Furthermore, we observed that the adsorbed HRP labeled with fluorescence was detected on the major oral gram-positive strains and (), but not on a gram-negative strain, (). Furthermore, we have demonstrated that the combination of HRP and chromogenic substrate clearly disclosed the dental plaques and the biofilm developed by , and the major gram-postive bacteria on tooth surfaces, and slightly disclosed the biofilm by . The combination of HRP and chromogenic substrate did not stain either the dental pellicle with the salivary glycoprotein mucin, or naked tooth surfaces. These results have suggested the possibility that the adsorption activity of HRP not only contributes to the evaluation of dental plaque, but that enzymatic activity of HRP may also contribute to improve dental hygiene.
PubMed: 32765778
DOI: 10.3892/etm.2020.9016