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Biomaterials Advances Apr 2022Streptococcus mutans is one of the bacteria that initiates the colonization of the pellicle at the tooth surface. It forms a plaque, together with other bacteria, which...
Streptococcus mutans is one of the bacteria that initiates the colonization of the pellicle at the tooth surface. It forms a plaque, together with other bacteria, which gradually dissolves the pellicle and leaves the tooth surface unprotected against the acidic oral environment. Calcium phosphate ceramics are excellent synthetic materials for the study of biofilm formation in dentistry because they are comparable to teeth in chemical composition and structure. Calcium phosphates can be processed to achieve a variety of crystalline compounds with biologically relevant ionic substitutions and structures that allow study of the effect of the surface chemistry and the topography independently. In this article, we describe the preparation and characterization of three types of calcium phosphate-based materials as a suitable surface for the formation of the S. mutans biofilm: beta-tricalcium phosphate (β-TCP); sintered hydroxyapatite (SHA); and calcium-deficient hydroxyapatite (CDHA). The densest biofilms were formed on the surfaces of SHA and CDHA, with no significant differences due to the stoichiometry or microstructure. In contrast, β-TCP showed a lower susceptibility to S. mutans biofilm formation, suggesting that the crystalline structure is the controlling parameter. Subsequently, SHA was selected to develop a dental biofilm model that allowed study of S. mutans biofilm susceptibility to chlorhexidine and ethanol.
Topics: Biofilms; Calcium Phosphates; Ceramics; Durapatite; Streptococcus mutans
PubMed: 35929222
DOI: 10.1016/j.bioadv.2022.212750 -
Archives of Oral Biology Oct 2022The 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...
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.
Topics: Animals; Cattle; Cholesterol Esters; Dental Pellicle; Fatty Acids; Glycolipids; Humans; Phosphatidylcholines; Phospholipids; Saliva; Tandem Mass Spectrometry; Triglycerides
PubMed: 35901594
DOI: 10.1016/j.archoralbio.2022.105493 -
Journal of Dentistry Sep 2022The 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... (Randomized Controlled Trial)
Randomized Controlled Trial
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.
Topics: Animals; Calcium; Cattle; Dental Pellicle; Humans; Polyphenols; Tannins; Tea; Tooth Erosion
PubMed: 35850367
DOI: 10.1016/j.jdent.2022.104236 -
European Journal of Oral Sciences Oct 2022Salivary pellicle was modified with bioproducts and we assessed the change in tooth color and the protection of enamel to erosion. Human enamel specimens were assigned...
Salivary pellicle was modified with bioproducts and we assessed the change in tooth color and the protection of enamel to erosion. Human enamel specimens were assigned to one of three solutions: grape seed extract or black tea (bioproducts), or deionized water (negative control); after which one half the specimens underwent erosive challenges. The specimens underwent 15 cycles involving salivary pellicle formation (10 min, 37°C), incubation in solution (2 min, 25°C), subsequent pellicle formation (90 min, 37°C). Half of the specimens was kept in a humid chamber and the other half was submitted to erosion (2 min, 1% citric acid). After 15 such cycles, the pellicle was removed. Tooth color and the surface reflection intensity were assessed after every five cycles and after pellicle removal. For non-eroded specimens, the exposure to bioproducts promoted significantly greater color change than the deionized water, with increases in yellow appearance. After pellicle removal, the color was similar in all non-eroded specimens. The bioproducts increased the surface reflection intensity over cycles. For the erosion-exposed specimens, erosion itself resulted in color change. Black tea and deionized water resulted in increased yellow appearance. Exposure to the bioproducts resulted in higher relative surface reflection intensity values over time, but only grape seed extract resulted in higher relative surface reflection intensity value at the time of pellicle removal. The bioproducts caused transient staining effect, which was reduced after pellicle removal. For enamel submitted to erosion, grape seed extract resulted in less color change and better protection of enamel against erosion than black tea or water.
Topics: Citric Acid; Dental Pellicle; Grape Seed Extract; Humans; Tea; Tooth Erosion; Water
PubMed: 35839337
DOI: 10.1111/eos.12886 -
Clinical Oral Investigations Nov 2022This study evaluated the preventive effect of a chitosan gel containing CaneCPI-5 against enamel erosion and erosion + abrasion in situ. (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
This study evaluated the preventive effect of a chitosan gel containing CaneCPI-5 against enamel erosion and erosion + abrasion in situ.
METHODS
Sixteen volunteers participated in a crossover, double-blind protocol, comprising 4 phases: (1) no treatment (Nt); (2) chitosan gel (Cg); (3) chitosan gel + 12,300 ppm NaF (Cg + NaF); and (4) chitosan gel + 0.1 mg/mL CaneCPI-5 (Cg + Cane). Volunteers wore an appliance containing 4 specimens. Once/day, they applied the gel (except for Nt) (4 min/specimen). Erosive challenges were performed extra-orally (0.1% citric acid, 90 s, 4 × /day; ERO). Specimens were also abraded (toothbrush, 15 s/specimen, 2 × /day; ERO + ABR). Enamel wear was assessed by profilometry and relative surface reflection intensity (%SRI). Two-way RM-ANOVA/Sidak's tests and Spearman's correlation were used (p < 0.05).
RESULTS
For profilometry, ERO + ABR promoted significantly greater wear when compared with ERO. There was a significant difference among all treatments. The lowest enamel loss occurred for Cg + Cane, followed by Cg + NaF, Cg, and Nt (p < 0.05). The %SRI was significantly lower for ERO + ABR when compared to ERO, only for the Nt group. The greatest %SRI was found for the Cg + NaF and Cg + Cane groups, which did not differ significantly, regardless of the conditions. The lowest %SRI was found for the Nt and Cg groups, which did not differ from each other, regardless of the conditions. The Nt group did not differ significantly from the Cg + NaF (ERO). There was a significant correlation between both analyses.
CONCLUSION
The incorporation of CaneCPI-5 in the chitosan gel prevented erosive wear in situ.
CLINICAL RELEVANCE
These results open a new perspective for the use of CaneCPI-5 in other application vehicles, such as chitosan gel.
Topics: Humans; Chitosan; Dental Enamel; Sodium Fluoride; Tooth Abrasion; Tooth Erosion; Toothbrushing; Cross-Over Studies; Double-Blind Method
PubMed: 35829772
DOI: 10.1007/s00784-022-04600-z -
Journal of Microscopy Dec 2022Recent advances in atomic force microscopy (AFM) have allowed the characterisation of dental-associated biomaterials and biological surfaces with high resolution. In...
Recent advances in atomic force microscopy (AFM) have allowed the characterisation of dental-associated biomaterials and biological surfaces with high resolution. In this context, the topography of dental enamel - the hardest mineralised tissue in the body - has been explored with AFM-based approaches at the microscale. With age, teeth are known to suffer changes that can impact their structural stability and function; however, changes in enamel structure because of ageing have not yet been explored with nanoscale resolution. Therefore, the aim of this exploratory work was to optimise an approach to characterise the ultrastructure of dental enamel and determine potential differences in topography, hydroxyapatite (HA) crystal size, and surface roughness at the nanoscale associated to ageing. For this, a total of six teeth were collected from human donors from which enamel specimens were prepared. By employing intermittent contact (AC mode) imaging, HA crystals were characterised in both transversal and longitudinal orientation (respect to surface plane) with high resolution in environmental conditions. The external enamel surface displayed the presence of a pellicle-like coating on its surface that was not observable on cleaned specimens. Acid-etching exposed crystals that were imaged and morphologically characterised in high resolution at the nanoscale in both the external and internal regions of enamel in older and younger specimens. Our results demonstrated important individual variations in HA crystal width and roughness parameters across the analysed specimens; however, an increase in surface roughness and decrease in HA width was observed for the pooled older external enamel group compared to younger specimens. Overall, high-resolution AFM was an effective approach for the qualitative and quantitative characterisation of human dental enamel ultrastructure. Future work should focus on exploring the ageing of dental enamel with increased sample sizes to compensate for individual differences as well as other potential confounding factors such as behavioural habits and mechanical forces.
Topics: Humans; Aged; Microscopy, Atomic Force; Tooth; Durapatite; Dental Enamel; Surface Properties
PubMed: 35621144
DOI: 10.1111/jmi.13126 -
Journal of Veterinary Dentistry Sep 2022The 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...
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.
Topics: Animals; Chromatography, Liquid; Dental Pellicle; Durapatite; Humans; Proteins; Proteomics; Tandem Mass Spectrometry; Wolves
PubMed: 35549755
DOI: 10.1177/08987564221097188 -
Clinical Oral Investigations Aug 2022The aim of this study is to evaluate the adhesion of Streptococcus mutans on the surface of CAD/CAM materials with various surface treatments.
OBJECTIVES
The aim of this study is to evaluate the adhesion of Streptococcus mutans on the surface of CAD/CAM materials with various surface treatments.
METHODS
Vita Enamic, Lava Ultimate, and Cerasmart materials were used in this study. A total of 90 samples were prepared (n = 10). After various finishing and polishing procedures (non-polished, manual polished, and glazed), surface roughness (SR) measurements, surface free energy (SFE), and elemental and topographic analysis with FIB-SEM/EDX were used to evaluate the samples' surface properties. To improve bacterial adhesion, CAD/CAM materials were covered with sterile artificial saliva containing mucin for pellicle formation and incubated for 1 h at 37 °C. Bacteria were then inoculated into the pellicle-coated specimens, and incubation was performed at 37 °C for 24 h. Bacterial adhesion was determined as × 10 CFU/mL and monitored using FIB-SEM analysis. The Kolmogorov-Smirnov test was used to statistically analyze the normality of the distribution; the groups were then compared using one-way ANOVA and Tukey's test.
RESULTS
The SR of the control group was statistically higher in all materials (p < 0.05). There were no statistically significant differences in SR between all materials in the non-polished and manual polished groups (p > 0.05). The Vita Enamic control group exhibited the highest SFE value. The highest S. mutans adhesion was observed in non-polished (p < 0.05). Vita Enamic samples had higher CFU than other groups.
CONCLUSIONS
Non-polished surfaces showed higher SR and bacterial adhesion. Polishing processes affected the surface properties and bacterial adhesion.
CLINICAL RELEVANCE
Care must be taken in polishing restorations to minimize the risk of bacterial adhesion and recurrent caries.
TRIAL REGISTRATION
In this study, the materials used for dental treatments are in vitro evaluated. Due to that, this study is not registered to clinical trials.
Topics: Bacterial Adhesion; Ceramics; Computer-Aided Design; Dental Polishing; Dental Porcelain; Materials Testing; Surface Properties
PubMed: 35474553
DOI: 10.1007/s00784-022-04497-8 -
Microorganisms Mar 2022Engineering of the acquired enamel pellicle using salivary peptides has been shown to be a promising anticaries strategy. However, the mechanisms by which these peptides...
Engineering of the acquired enamel pellicle using salivary peptides has been shown to be a promising anticaries strategy. However, the mechanisms by which these peptides protect teeth against tooth decay are not fully understood. In this study, we evaluated the effect of the engineered salivary peptides DR9-DR9 and DR9-RR14 on enamel demineralization in two experimental conditions: (1) adsorbed onto the enamel surface forming the AEP, and (2) forming the AEP combined with their use to treat the biofilms 2×/day, using a validated cariogenic Streptococcus mutans in vitro biofilm model. Biofilms were grown for 144 h on enamel slabs and then collected to determine the bacterial viability (CFU/biofilm) and biofilm mass (mg protein/biofilm), and to extract cellular/extracellular proteins, which were characterized by mass spectrometry. The culture medium was changed 2×/day to fresh medium, and pH (indicator of biofilm acidogenicity) and calcium concentration (indicator of demineralization) was determined in used medium. DR9-RR14 peptide significantly reduced enamel demineralization (p < 0.0001) in both experimental conditions. However, this peptide did not have a significant effect on biofilm biomass (p > 0.05) nor did it modulate the expression of cellular and extracellular bacterial proteins involved in biofilm cariogenicity. These findings suggest that DR9-RR14 may control caries development mainly by a physicochemical mechanism.
PubMed: 35456793
DOI: 10.3390/microorganisms10040742 -
Biomedicines Mar 2022The oral microbiome, forming a biofilm that covers the oral structures, contains a high number of microorganisms. Biofilm formation starts from the salivary pellicle... (Review)
Review
The oral microbiome, forming a biofilm that covers the oral structures, contains a high number of microorganisms. Biofilm formation starts from the salivary pellicle that allows bacterial adhesion-colonization-proliferation, co-aggregation and biofilm maturation in a complex microbial community. There is a constant bidirectional crosstalk between human host and its oral microbiome. The paper presents the fundamentals regarding the oral microbiome and its relationship to modulator factors, oral and systemic health. The modern studies of oral microorganisms and relationships with the host benefits are based on genomics, transcriptomics, proteomics and metabolomics. Pharmaceuticals such as antimicrobials, prebiotics, probiotics, surface active or abrasive agents and plant-derived ingredients may influence the oral microbiome. Many studies found associations between oral dysbiosis and systemic disorders, including autoimmune diseases, cardiovascular, diabetes, cancers and neurodegenerative disorders. We outline the general and individual factors influencing the host-microbial balance and the possibility to use the analysis of the oral microbiome in prevention, diagnosis and treatment in personalized medicine. Future therapies should take in account the restoration of the normal symbiotic relation with the oral microbiome.
PubMed: 35327473
DOI: 10.3390/biomedicines10030671