-
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 -
Microorganisms Jan 2022Since the modification of the proteinaceous components of the Acquired Enamel Pellicle (AEP) could influence the adhesion of the most cariogenic bacteria, to dental...
Since the modification of the proteinaceous components of the Acquired Enamel Pellicle (AEP) could influence the adhesion of the most cariogenic bacteria, to dental surfaces, we assessed if engineered salivary peptides would affect the adherence and modulate the bacterial proteome upon adherence. Single-component AEPs were formed onto hydroxyapatite (HAp) discs by incubating them with statherin, histatin-3, DR9, DR9-DR9, DR9-RR14, RR14, and parotid saliva. Then, the discs were inoculated with UA159 and the bacteria were allowed to adhere for 2 h, 4 h, and 8 h ( = 12/treatment/time point). The number of bacteria adhered to the HAp discs was determined at each time point and analyzed by two-way ANOVA and Bonferroni tests. Cell-wall proteins were extracted from adhered, planktonic, and inoculum (baseline) bacteria and proteome profiles were obtained after a bottom-up proteomics approach. The number of adhered bacteria significantly increased over time, being the mean values obtained at 8 h, from highest to lowest, as follows: DR9-RR14 > statherin > RR14 = DR9-DR9 > DR9 = histatin3 > saliva ( < 0.05). Treatments modulated the bacterial proteome upon adherence. The findings suggested a potential use of our engineered peptide DR9-DR9 to control biofilm development by reducing bacterial colonization.
PubMed: 35208678
DOI: 10.3390/microorganisms10020223 -
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 -
Caries Research 2022This study evaluated the combination of a sugarcane cystatin (CaneCPI-5) and sodium fluoride (NaF) in acquired pellicle engineering for the prevention of dental erosion...
This study evaluated the combination of a sugarcane cystatin (CaneCPI-5) and sodium fluoride (NaF) in acquired pellicle engineering for the prevention of dental erosion in vitro. Seventy-five human enamel specimens were prepared and divided into 5 treatment groups (n = 15/group): Deionized water (Control); Elmex™ (SnCl2/NaF/AmF); 0.1 mg/mL CaneCPI-5; 500 ppm NaF; and CaneCPI-5+NaF (Combination). The specimens were individually treated (200 μL; 2 min; 37°C), then incubated in human saliva (200 μL; 1 h, at 37°C) for acquired pellicle formation. Afterward, the specimens were submitted to an erosive challenge (1% citric acid [CR], pH 3.6, 10 mL, 2 min, 25 °C). This sequence was conducted 5 times. Percentage of surface microhardness change (%SMC), relative surface reflection intensity (rSRI), and calcium released to the CR were measured and analyzed by one-way ANOVA followed by Tukey's test (p < 0.05). In general, all the treatments (SnCl2/NaF/AmF, CaneCPI-5, NaF, and Combination) significantly protected the enamel when compared the control group. Regarding %SMC and rSRI, the Combination was the most effective treatment, reducing the %SMC significantly (p < 0.01) when compared to all the other treatments, although this difference was not significant in the CR analysis. All treatments demonstrated a protective effect on enamel against dental erosion; however, the combination of CaneCPI-5 with NaF showed a greater protection.
Topics: Cystatins; Dental Pellicle; Fluorides; Humans; Saccharum; Sodium Fluoride; Tooth Erosion
PubMed: 35168236
DOI: 10.1159/000522490 -
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 -
International Journal of Dental Hygiene Feb 2023The aim of this study was to investigate the direct and indirect influence of fluoridated toothpastes and fluoride-free toothpaste with hydroxyapatite (HAP) as active...
OBJECTIVE
The aim of this study was to investigate the direct and indirect influence of fluoridated toothpastes and fluoride-free toothpaste with hydroxyapatite (HAP) as active ingredient on initial bacterial colonization on enamel in situ.
METHODS
For this clinical-experimental pilot study, eight subjects were instructed to brush their teeth with three different toothpastes (Elmex : 1400 ppm AmF, Meridol : 1400 ppm AmF +SnF2, Karex : HAP), using each for two consecutive days. As a control, brushing without toothpaste was performed. To evaluate bacterial colonization, subject wore splints with buccally placed bovine enamel platelets overnight. Two modes were tested. In a first pass (regimen A), the splints were inserted after toothbrushing to examine the indirect effects of the dentifrices. In order to investigate the direct effects, the specimens were brushed in situ in a second pass (regimen B). Biofilm formation was visualized and quantified using fluorescence microscopy (DAPI and BacLight) and transmission electron microscopy (TEM).
RESULTS
For brushing regimen A (indirect effect of dentifrices), no statistical differences were detected between any of the tested dentifrices or the control. Likewise, no statistically significant differences were recorded for brushing regimen B (direct effect of dentifrices). Furthermore, no differences between the different brushing techniques were determined with regard to the ultrastructure of the overnight biofilm.
CONCLUSION
Within the limitations of the present pilot study, it can be concluded that in patients with good oral hygiene, dentifrices and their chemical composition have no statistically significant effect on the initial bacterial colonization of enamel platelets in situ, irrespectively of the mode of application.
Topics: Humans; Animals; Cattle; Toothpastes; Dentifrices; Pilot Projects; Fluorides; Dental Enamel; Toothbrushing; Sodium Fluoride
PubMed: 35075780
DOI: 10.1111/idh.12581 -
BMC Oral Health Dec 2021Early childhood caries is the most common infectious disease in childhood, with a high prevalence in developing countries. The assessment of the variables that influence...
BACKGROUND
Early childhood caries is the most common infectious disease in childhood, with a high prevalence in developing countries. The assessment of the variables that influence early childhood caries as well as its pathophysiology leads to improved control of this disease. Cystatin S, as one of the salivary proteins, has an essential role in pellicle formation, tooth re-mineralization, and protection. The present study aims to assess salivary cystatin S levels and demographic data in early childhood caries in comparison with caries-free ones using statistical analysis and machine learning methods.
METHODS
A cross-sectional, case-control study was undertaken on 20 cases of early childhood caries and 20 caries-free children as a control. Unstimulated whole saliva samples were collected by suction. Cystatin S concentrations in samples were determined using human cystatin S ELISA kit. The checklist was collected from participants about demographic characteristics, oral health status, and dietary habits by interviewing parents. Regression and receiver operating characteristic (ROC) curve analysis were done to evaluate the potential role of cystatin S salivary level and demographic using statistical analysis and machine learning.
RESULTS
The mean value of salivary cystatin S concentration in the early childhood caries group was 191.55 ± 81.90 (ng/ml) and in the caries-free group was 370.06 ± 128.87 (ng/ml). T-test analysis showed a statistically significant difference between early childhood caries and caries-free groups in salivary cystatin S levels (p = 0.032). Investigation of the area under the curve (AUC) and accuracy of the ROC curve revealed that the logistic regression model based on salivary cystatin S levels and birth weight had the most and acceptable potential for discriminating of early childhood caries from caries-free controls. Furthermore, using salivary cystatin S levels enhanced the capability of machine learning methods to differentiate early childhood caries from caries-free controls.
CONCLUSION
Salivary cystatin S levels in caries-free children were higher than the children with early childhood caries. Results of the present study suggest that considering clinical examination, demographic and socioeconomic factors, along with the salivary cystatin S levels, could be usefull for early diagnosis ofearly childhood caries in high-risk children; furthermore, cystatin S is a protective factor against dental caries.
Topics: Case-Control Studies; Child; Child, Preschool; Cross-Sectional Studies; Dental Caries; Dental Caries Susceptibility; Humans; Machine Learning; Saliva; Salivary Cystatins
PubMed: 34922509
DOI: 10.1186/s12903-021-02016-x -
Archives of Oral Biology Feb 2022The objective of this study was to determine whether differences in the abundance of mucins 5b and 7 as well as secretory IgA exist in the oral acquired pellicle between...
OBJECTIVE
The objective of this study was to determine whether differences in the abundance of mucins 5b and 7 as well as secretory IgA exist in the oral acquired pellicle between children with active caries and caries-free children.
DESIGN
Pellicle formation was performed for 10 min in-situ on ceramic slabs in the oral cavity of children (5-7 years of age) with caries (n = 15) and without signs of caries (n = 13). Furthermore, unstimulated saliva was collected. Concentrations of Muc5b, Muc7 and sIgA were measured in desorbed pellicle eluates and in saliva.
RESULTS
Significantly larger concentrations of Muc5b, Muc7 and sIgA were detected in the pellicle obtained from children with caries compared to caries-free children. However, in the salivary samples concentrations of mucins Muc5b and Muc7 as well as sIgA did not differ significantly between the two groups.
CONCLUSIONS
All three pellicle components Muc5b, Muc7 as well as sIgA could be identified as potential biomarkers for early childhood caries with high sensitivity and specificity. This could contribute to a better understanding of the different caries susceptibility in children.
Topics: Biomarkers; Child; Child, Preschool; Dental Caries Susceptibility; Dental Pellicle; Humans; Immunoglobulin A, Secretory; Mucin-5B; Mucins; Saliva; Salivary Proteins and Peptides
PubMed: 34861462
DOI: 10.1016/j.archoralbio.2021.105314 -
Microorganisms Oct 2021Caries is one of the most prevalent diseases worldwide, which is caused by the degradation of the tooth enamel surface. In earlier research the opportunistic pathogen...
Caries is one of the most prevalent diseases worldwide, which is caused by the degradation of the tooth enamel surface. In earlier research the opportunistic pathogen has been associated with the formation of caries in children. Colonization of teeth by starts with the initial adhesion of individual yeast cells to the tooth enamel surface. In this study, we visualized the initial colonization of yeast cells on pellicle-covered enamel by scanning electron microscopy. To quantitatively unravel the initial adhesion strength, we applied fluidic force microscopy-based single-cell force spectroscopy to examine the key adhesion parameters adhesion force, rupture length and de-adhesion work. We analyzed single saliva-treated or untreated yeast cells on tooth enamel specimens with or without salivary pellicle. Under all tested conditions, adhesion forces in the lower nanonewton range were determined. Furthermore, we have found that all adhesion parameters were enhanced on the pellicle-covered compared to the uncovered enamel. Our data suggest that initial adhesion occurs through a strong interaction between yeast cell wall-associated adhesins and the salivary pellicle. Future SCFS studies may show whether specific management of the salivary pellicle reduces the adhesion of on teeth and thus contributes to caries prophylaxis.
PubMed: 34835339
DOI: 10.3390/microorganisms9112213 -
Journal of Dentistry Jan 2022To study the proteomic alterations in the initial AEP after rinsing with CaneCPI-5, StN15 or Hb or their combination.
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 (HO- negative control- 1), 0.1 mg/mL CaneCPI-5 (2), 1.88×105 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.
Topics: Dental Pellicle; Humans; Peptides; Proteins; Proteomics; Water
PubMed: 34798481
DOI: 10.1016/j.jdent.2021.103874