Denture microbiome shift and changes of salivary inflammatory markers following insertion of 3D printed removable partial PMMA denture: a pilot study.

BACKGROUND: The aim of this study was to investigate the microbiome shift of denture biofilm formation, the incidence of Candida and changes of salivary inflammatory markers following insertion of 3-dimensional printed PMMA denture. METHODS: This was a pilot study of 6 patients provided with 3D printed (test group) or conventionally heat-cured (control group) removable partial acrylic dentures followed up for 6 weeks. 3 denture swabs were collected at week 1, 3 and 6; and saliva samples were collected at baseline, week 1, 3 and 6 following denture insertion. Microbial DNA was isolated, and the 16S rRNA gene was amplified and sequenced to assess the denture microbiota. The presence of Candida was determined using PCR assay. The levels of salivary biomarkers lactoferrin (LTF) and histatin (HTN) were determined using ELISA. RESULTS: Denture microbiome of both groups exhibited similar microbial compositions at weeks 1, 3 and 6. However, the percentage of Streptococcus in the test group was significantly lower at week 1 (p < 0.05). Greater bacterial diversity was detected in the test group (p < 0.05; pairwise Wilcoxon) after 6 weeks. The presence of Candida was only detected in one sample of the 3D printed denture (test group) after 6 weeks. The level of LTF biomarkers increased in both groups after 6 weeks, however, the level of HTN increased only in the control group. CONCLUSIONS: Denture microbiome of both groups demonstrated similar microbial compositions. After 6 weeks, 3D printed denture demonstrated higher diversity with delayed microbiome shift compared to conventional heat cure dentures. 3D printed denture may be a viable alternative to conventional denture and it has a potential to delay microbial dysbiosis. CLINICAL RELEVANCE: Among wearers, 3D printed denture can potentially reduce the risk of denture stomatitis. A greater diversity may delay or slow the transition of denture microbiome to dysbiosis.

Randomised-crossover clinical trial on the substantivity of a single application of a gel containing chlorhexidine and o-cymen-5-ol on the oral biofilm and saliva.

BACKGROUND: No clinical trials have evaluated the antimicrobial activity and substantivity of gel formulations containing chlorhexidine (CHX) and cymenol. OBJECTIVE: To compare the in situ antimicrobial effect and substantivity of a new 0.20% CHX + cymenol gel (test) with the current 0.20% CHX gel formulation (control) on salivary flora and dental plaque biofilm up to seven hours after a single application. METHODS: A randomised-crossover clinical trial was conducted with 29 orally healthy volunteers participating in the development of Experiments 1 (saliva) and 2 (dental plaque biofilm). All subjects participated in both experiments and were randomly assigned to receive either the test or control gels. Samples were collected at baseline and five minutes and one, three, five, and seven hours after a single application of the products. The specimens were processed using confocal laser scanning microscopy after staining with the LIVE/DEAD® BacLight™ solution. Bacterial viability (BV) was quantified in the saliva and biofilm samples. The BV was calculated using the DenTiUS Biofilm software. RESULTS: In Experiment 1, the mean baseline BV was significantly reduced five minutes after application in the test group (87.00% vs. 26.50%; p < 0.01). This effect was maintained throughout all sampling times and continued up to seven hours (40.40%, p < 0.01). The CHX control followed the same pattern. In Experiment 2, the mean baseline BV was also significantly lower five minutes after applying the test gel for: (1) the total thickness of biofilm (91.00% vs. 5.80%; p < 0.01); (2) the upper layer (91.29% vs. 3.94%; p < 0.01); and (3) the lower layer (86.29% vs. 3.83%; p < 0.01). The reduction of BV from baseline was observed for the full-thickness and by layers at all sampling moments and continued seven hours after application (21.30%, 24.13%, and 22.06%, respectively; p < 0.01). Again, the control group showed similar results. No significant differences between test and control gels were observed in either saliva or dental plaque biofilm at any sampling time. CONCLUSIONS: A 0.20% CHX + cymenol gel application demonstrates potent and immediate antimicrobial activity on salivary flora and de novo biofilm. This effect is maintained seven hours after application. Similar effects are obtained with a 0.20% CHX-only gel.

Current approaches to produce durable biomaterials: Trends in polymeric materials for restorative dentistry applications.

Dental caries continues to be a public health issue, especially more evident in underserved populations throughout the U.S. Unfortunately, especially with an aging population, hundreds of thousands of resin composite restorations are replaced each year due to recurring decay and fracture. According to several cohort studies, the average life span of this type of restoration is 10 years or less, depending on the caries risk level of the patient and the complexity of the restorative procedure. Any new material development must depart from the simple restoration of form paradigm, in which the filling is simply inert/biocompatible. This review will discuss novel antibiofilm structures, based on a targeted approach specifically against dysbiotic bacteria. Biofilm coalescence can be prevented by using glycosyl transferase - GTF inhibitors, in a non-bactericidal approach. On the tooth substrate side, MMP-inhibiting molecules can improve the stability of the collagen in the hybrid layer. This review will also discuss the importance of testing the materials in a physiologically relevant environment, mimicking the conditions in the mouth in terms of mechanical loading, bacterial challenge, and the presence of saliva. Ultimately, the goal of materials development is to achieve durable restorations, capable of adapting to the oral environment and resisting challenges that go beyond mechanical demands. That way, we can prevent the unnecessary loss of additional tooth structure that comes with every re-treatment. CLINICAL SIGNIFICANCE: While proper restorative technique and patient education in terms of diet and oral hygiene are crucial factors in increasing the longevity of esthetic direct restorations, materials better able to resist and interact with the conditions of the oral environment are still needed. Reproducing the success of dental amalgams with esthetic materials continues to be the Holy Grail of materials development.

Stability of the Dentin-Bonded Interface Using Self-Etching Adhesive Containing Diacrylamide after Bacterial Challenge.

Purpose/Aim: Acrylamides are hydrolytically stable at pH lower than 2, and were shown to preserve bonded interface integrity with two-step, total etch adhesives. The objective of this study was to leverage those two characteristics in self-etching primers containing the acidic monomer 10-MDP and test the microtensile bond strength before and after incubation with S. mutans incubation. Materials and Methods: Acidic primers (10 wt % 10-methacryloyloxydecyl dihydrogen phosphate─10-MDP; 45 wt % N,N-diethyl-1,3-bis(acrylamido)propane─DEBAAP, or 2-hydroxyethyl methacrylate─HEMA; 45 wt %, glycerol-dimethacrylate─GDMA) and adhesives (DEBAAP or HEMA/10-MDP/UDMA 45/10/45 wt %) were made polymerizable by the addition of 0.2 wt % camphorquinone, 0.8 wt % ethyl-4-dimethylaminobenzoate, 0.4 wt % diphenyliodonium hexafluorophosphate, and 0.1 wt % butylhydroxytoluene. Nonsolvated materials were characterized for flexural strength (FS), modulus (E), toughness, water sorption/solubility (WS/SL), contact angle, and vinyl conversion (DC). Viscosity was evaluated after adding 20 and 40 vol % ethanol to the primer and adhesive, respectively. The experimental materials or Clearfil SE Bond (CC─commercial control) were used to bond a commercial composite (Filtek Supreme) to the flat surface of human dentin. Microtensile bond strength (MTBS) was tested in 1 mm2 sticks for the 5 primer/bond combinations: CC (Clearfil Bond Primer and Bond), HH (HEMA/HEMA), DD (DEBAAP/DEBAAP), HD (HEMA/DEBAAP), and DH (DEBAAP/HEMA). Prior to testing, sticks were stored in water or biofilm-inducing culture medium with S. mutans for 1 week. Confocal images and FTIR-ATR evaluation evaluated the hybrid layer of the adhesives. Results were analyzed using Student's t-test (WS, SL, DC, contact angle, FS, E, toughness), one-way ANOVA/Tukey's test for viscosity, and two-way ANOVA/Tukey's test for MTBS (95%). Results: HEMA-based materials had lower contact angle (p = 0.004), higher WS (p < 0.001), and similar SL values compared to DEBAAP (p = 0.126). FS (p = 0.171) and E (p = 0.065) dry values were similar, but after one week of water storage, FS/E dropped more significantly for HEMA materials. Dry and wet toughness was greater for DEBAAP (p < 0.001), but it also had the greatest drop (46%). Clearfil bonds had the highest viscosity, followed by DEBAAP and HEMA, respectively (p = 0.002). For the primers, HEMA had the lowest viscosity (p = 0.003). As far as MTBS, all groups tested in water were statistically different when compared with HH (p < 0.001). After storage in biofilm, DH had the highest MTBS value, being statistically different from HH (p = 0.002), CC (p = 0.015), and DD (p = 0.027). Conclusions: The addition of a diacrylamide and its association with HEMA in self-etching adhesive systems provided greater bonding stability after bacterial challenge.

Production and characterization of the lipopeptide with anti-adhesion for oral biofilm on the surface of titanium for dental implants.

Titanium implants are subject to bacterial adhesion and peri-implantitis induction, and biosurfactants bring a new alternative to the fight against infections. This work aimed to produce and characterize the biosurfactant from Bacillus subtilis ATCC 19,659, its anti-adhesion and antimicrobial activity, and cell viability. Anti-adhesion studies were carried out against Streptococcus sanguinis, Staphylococcus aureus, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Proteus mirabilis as the minimum inhibitory concentration and the minimum bactericidal concentration. Cell viability was measured against osteoblast and fibroblast cells. The biosurfactant was classified as lipopeptide, with critical micelle concentration at 40 µg mL- 1, and made the titanium surface less hydrophobic. The anti-adhesion effect was observed for Staphylococcus aureus and Streptococcus sanguinis with 54% growth inhibition and presented a minimum inhibitory concentration of 15.7 µg mL- 1 for Streptococcus sanguinis and Aggregatibacter actinomycetemcomitans. The lipopeptide had no cytotoxic effect and demonstrated high potential application against bacterial biofilms.

Topical oxygen therapy as a novel strategy to promote wound healing and control the bacteria in implantology, oral surgery and periodontology: A review.

Globally, oral infections and inflammatory lesions persist as substantial public health concerns, necessitating the introduction of novel oral treatment protocols. Oral diseases are linked to various causative factors, with dental plaque/biofilm resulting from inadequate hygiene practices playing a predominant role. The strategic implementation of novel topical therapies holds promise for effectively controlling the biofilms, addressing oral infections and promoting enhanced oral wound healing. This review aims to providing a comprehensive overview of the available evidence pertaining to the potential efficacy of topical oxygen and lactoferrin-releasing biomaterials, exemplified by the blue®m formula, as novel oral care interventions within the scope of contemporary implantology, oral surgery and periodontology.

Characterization and morphological methods for oral biofilm visualization: where are we nowadays?

The oral microbiome represents an essential component of the oral ecosystem whose symbiotic relationship contributes to health maintenance. The biofilm represents a state of living of microorganisms surrounding themselves with a complex and tridimensional organized polymeric support and defense matrix. The substrates where the oral biofilm adhere can suffer from damages due to the microbial community metabolisms. Therefore, microbial biofilm represents the main etiological factor of the two pathologies of dental interest with the highest incidence, such as carious pathology and periodontal pathology. The study, analysis, and understanding of the characteristics of the biofilm, starting from the macroscopic structure up to the microscopic architecture, appear essential. This review examined the morphological methods used through the years to identify species, adhesion mechanisms that contribute to biofilm formation and stability, and how the action of microbicidal molecules is effective against pathological biofilm. Microscopy is the primary technique for the morphological characterization of biofilm. Light microscopy, which includes the stereomicroscope and confocal laser microscopy (CLSM), allows the visualization of microbial communities in their natural state, providing valuable information on the spatial arrangement of different microorganisms within the biofilm and revealing microbial diversity in the biofilm matrix. The stereomicroscope provides a three-dimensional view of the sample, allowing detailed observation of the structure, thickness, morphology, and distribution of the various species in the biofilm while CLSM provides information on its three-dimensional architecture, microbial composition, and dynamic development. Electron microscopy, scanning (SEM) or transmission (TEM), allows the high-resolution investigation of the architecture of the biofilm, analyzing the bacterial population, the extracellular polymeric matrix (EPS), and the mechanisms of the physical and chemical forces that contribute to the adhesion of the biofilm to the substrates, on a nanometric scale. More advanced microscopic methodologies, such as scanning transmission electron microscopy (STEM), high-resolution transmission electron microscopy (HR-TEM), and correlative microscopy, have enabled the evaluation of antibacterial treatments, due to the potential to reveal the efficacy of different molecules in breaking down the biofilm. In conclusion, evidence based on scientific literature shows that established microscopic methods represent the most common tools used to characterize biofilm and its morphology in oral microbiology. Further protocols and studies on the application of advanced microscopic techniques are needed to obtain precise details on the microbiological and pathological aspects of oral biofilm.

Label-Free Quantitative Proteomics of Oral Microbial Communities.

The oral cavity is a habitat for different microorganisms, of which bacteria are best described. Studying different bacterial taxa and their proteins is crucial to understanding their interactions with the host and other microbes. Also, for bacteria with virulence potential, identifying novel antigenic proteins is essential to finding candidates for the development of vaccines.Here, a workflow for gel-free and label-free protein analysis of oral bacterial species grown in vitro as a biofilm and a planktonic culture is described. Details on cultivation, protein extraction and digestion, peptide cleanup, LC-MS/MS run parameters, and subsequent bioinformatics analysis are included. Challenging steps in the workflow, such as growing different types of bacteria and selecting a suitable protein database, are also discussed. This protocol provides a valuable guide for metaproteomic experiments using multi-species models of oral bacteria.

Novel Resin-Based Antibacterial Root Surface Coating Material to Combat Dental Caries.

Root caries caused by cariogenic bacteria are a burden on a large number of individuals worldwide, especially the elderly. Applying a protective coating to exposed root surfaces has the potential to inhibit the development of caries, thus preserving natural teeth. This study aimed to develop a novel antibacterial coating to combat root caries and evaluate its effectiveness using the antibacterial monomer dimethylaminohexadecyl methacrylate (DMAHDM). DMAHDM was synthesized and incorporated into a resin consisting of 55.8% urethane dimethacrylate (UDMA) and 44.2% TEG-DVBE (UV) at a 10% mass fraction of glass filler. Multiple concentrations of DMAHDM were tested for their impact on the resin's mechanical and physical properties. S. mutans biofilms grown on resin disks were analyzed for antibacterial efficacy. Cytotoxicity was assessed against human gingival fibroblasts (HGFs). The results showed an 8-log reduction in colony-forming units (CFUs) against S. mutans biofilm (mean ± sd; n = 6) (p < 0.05) when 5% DMAHDM was incorporated into the UV resin. There was a 90% reduction in metabolic activity and lactic acid production. A low level of cytotoxicity against HGF was observed without compromising the physical and mechanical properties of the resin. This coating material demonstrated promising physical properties, potent antibacterial effects, and low toxicity, suggesting its potential to protect exposed roots from caries in various dental procedures and among elderly individuals with gingival recession.

Analysis of cariogenic biofilms by using a swept-source optical coherence tomography in vitro.

OBJECTIVE: The objective was to measure the thickness of Streptococcus mutans (S. mutans) biofilms forming in an oral biofilm reactor (OBR) by using a noninvasive swept-source optical coherence tomography (SS-OCT) system at every 4 h time interval until 20 h and analyze the correlations with the amounts of biofilms. METHODS: S. mutans biofilms were formed on square-shaped bovine enamel blocks inside an OBR. Biofilms were analyzed at every 4 h stage (4 h, 8 h, 12 h, 16 h and 20 h) using a SS-OCT system and a laser scanning confocal microscope (LSCM). The amounts of biofilms were measured at each stage by separating the water insoluble glucan (WIG) and bacterial cells. Co-relationships between the SS-OCT measured biofilm thickness and the amounts of adhered biofilms were analyzed. RESULTS: The thickness of biofilms detected on SS-OCT images at 4 h stage was 0.059 ± 0.029 (Av ± SD) mm which increased time-dependently in a linear fashion after 8 h stage and reached to 0.435 ± 0.159 mm at 20 h stage and the correlation coefficient was about 0.89. The amounts of biofilms; bacterial optical density (OD) and WIG concentration increased time-dependently were 0.035 ± 0.008 / mm2 and 10.328 ± 2.492 µg/ mm2 respectively at 20 h stage. Correlation coefficients of 0.66 between 'the amounts of bacteria' and 'biofilm thickness on OCT' and 0.67 between 'the amounts of WIG' and 'biofilm thickness on OCT' were obtained, suggesting that there was a relatively positive correlation between them. CONCLUSION: The SS-OCT can be a useful tool to measure time-dependent growth of biofilms. Further studies are needed in order to assess biofilms using SS-OCT more accurately.

Oxygen self-sufficient nanodroplet composed of fluorinated polymer for high-efficiently PDT eradicating oral biofilm.

Oral biofilm is the leading cause of dental caries, which is difficult to completely eradicate because of the complicated biofilm structure. What's more, the hypoxia environment of biofilm and low water-solubility of conventional photosensitizers severely restrict the therapeutic effect of photodynamic therapy (PDT) for biofilm. Although conventional photosensitizers could be loaded in nanocarriers, it has reduced PDT effect because of aggregation-caused quenching (ACQ) phenomenon. In this study, we fabricated an oxygen self-sufficient nanodroplet (PFC/TPA@FNDs), which was composed of fluorinated-polymer (FP), perfluorocarbons (PFC) and an aggregation-induced emission (AIE) photosensitizer (Triphenylamine, TPA), to eradicate oral bacterial biofilm and whiten tooth. Fluorinated-polymer was synthesized by polymerizing (Dimethylamino)ethyl methacrylate, fluorinated monomer and 1-nonanol monomer. The nanodroplets could be protonated and behave strong positive charge under bacterial biofilm acid environment promoting nanodroplets deeply penetrating biofilm. More importantly, the nanodroplets had extremely high PFC and oxygen loading efficacy because of the hydrophobic affinity between fluorinated-polymer and PFC to relieve the hypoxia environment and enhance PDT effect. Additionally, compared with conventional ACQ photosensitizers loaded system, PFC/TPA@FNDs could behave superior PDT effect to ablate oral bacterial biofilm under light irradiation due to the unique AIE effect. In vivo caries animal model proved the nanodroplets could reduce dental caries area without damaging tooth structure. Ex vivo tooth whitening assay also confirmed the nanodroplets had similar tooth whitening ability compared with commercial tooth whitener H2O2, while did not disrupt the surface microstructure of tooth. This oxygen self-sufficient nanodroplet provides an alternative visual angle for oral biofilm eradication in biomedicine.

Investigation of a novel biofilm model close to the original oral microbiome.

A more optimized culture medium used in vitro to mimic the bacterial composition of original oral flora as similar as possible remains difficult at present, and the goal of this study is to develop a novel oral biofilm medium to restore the original oral microbiome. Firstly, we conducted a systematic literature review by searching PubMed and summarized the current reported culture media in vitro. Seven culture media were found. We used mixed saliva as the origin of oral species to compare the effects of the above media in culturing oral multispecies biofilms. Results indicated that among the seven media brain heart infusion containing 1% sucrose (BHIs) medium, PG medium, artificial saliva (AS) medium, and SHI medium could obviously gain large oral biofilm in vitro. The nutrients contained in different culture media may be suitable for the growth of different oral bacteria; therefore, we optimized several novel media accordingly. Notably, results of crystal violet staining showed that the biofilm cultured in our modified artificial saliva (MAS) medium had the highest amount of biofilm biomass. 16S rRNA gene sequencing showed that the operational taxonomic units (OTUs) and Shannon index of biofilm cultured in MAS medium were also the highest among all the tested media. More importantly, the 16S rRNA gene sequencing analysis indicated that the biofilm cultured in MAS medium was closer to the original saliva species. Besides, biofilm cultured by MAS was denser and produced more exopolysaccharides. MAS supported stable biofilm formation on different substrata. In conclusion, this study demonstrated a novel MAS medium that could culture oral biofilm in vitro closer to the original oral microbiome, showing a good application prospect. KEY POINTS: • We compare the effects of different media in culturing oral biofilms • A novel modified artificial saliva (MAS) medium was obtained in our study • The MAS medium could culture biofilm that was closer to oral microbiome.

Nanomaterial-based drug-delivery system as an aid to antimicrobial photodynamic therapy in treating oral biofilm.

Diverse microorganisms live as biofilm in the mouth accounts for oral diseases and treatment failure. For decades, the prevention and treatment of oral biofilm is a global challenge. Antimicrobial photodynamic therapy (aPDT) holds promise for oral biofilm elimination due to its several traits, including broad-spectrum antimicrobial capacity, lower possibility of resistance and low cytotoxicity. However, the physicochemical properties of photosensitizers and the biological barrier of oral biofilm have limited the efficiency of aPDT. Nanomaterials has been used to fabricate nanocarriers to improve photosensitizer properties and thus enhance antimicrobial effect. In this review, we have discussed the challenges of aPDT used in dentistry, categorized the nanomaterial-delivery system and listed the possible mechanisms involved in nanomaterials when enhancing aPDT effect.

[Box: see text].

The Impact of Candida albicans in the Development, Kinetics, Structure, and Cell Viability of Biofilms on Implant Surfaces-An In Vitro Study with a Validated Multispecies Biofilm Model.

This study aimed to evaluate the impact of Candida albicans on subgingival biofilm formation on dental implant surfaces. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to compare biofilm structure and microbial biomass in the presence and absence of the fungus after periods of 24, 48, and 72 h. Quantitative polymerase chain reaction (qPCR) was used to quantify the number of viable and total micro-organisms for each of the biofilm-forming strains. A general linear model was applied to compare CLSM and qPCR results between the control and test conditions. The biofilm developed with C. albicans at 72 h had a higher bacterial biomass and a significantly higher cell viability (p < 0.05). After both 48 and 72 h of incubation, in the presence of C. albicans, there was a significant increase in counts of Fusobacterium nucleatum and Porphyromonas gingivalis and in the cell viability of Streptococcus oralis, Aggregatibacter actinomycetemcomitans, F. nucleatum, and P. gingivalis. Using a dynamic in vitro multispecies biofilm model, C. albicans exacerbated the development of the biofilm grown on dental implant surfaces, significantly increasing the number and cell viability of periodontal bacteria.

Characterization of MdpS: an in-depth analysis of a MUC5B-degrading protease from Streptococcus oralis.

Oral biofilms, comprising hundreds of bacteria and other microorganisms on oral mucosal and dental surfaces, play a central role in oral health and disease dynamics. Streptococcus oralis, a key constituent of these biofilms, contributes significantly to the formation of which, serving as an early colonizer and microcolony scaffold. The interaction between S. oralis and the orally predominant mucin, MUC5B, is pivotal in biofilm development, yet the mechanism underlying MUC5B degradation remains poorly understood. This study introduces MdpS (Mucin Degrading Protease from Streptococcus oralis), a protease that extensively hydrolyses MUC5B and offers an insight into its evolutionary conservation, physicochemical properties, and substrate- and amino acid specificity. MdpS exhibits high sequence conservation within the species and also explicitly among early biofilm colonizing streptococci. It is a calcium or magnesium dependent serine protease with strict physicochemical preferences, including narrow pH and temperature tolerance, and high sensitivity to increasing concentrations of sodium chloride and reducing agents. Furthermore, MdpS primarily hydrolyzes proteins with O-glycans, but also shows activity toward immunoglobulins IgA1/2 and IgM, suggesting potential immunomodulatory effects. Significantly, MdpS extensively degrades MUC5B in the N- and C-terminal domains, emphasizing its role in mucin degradation, with implications for carbon and nitrogen sequestration for S. oralis or oral biofilm cross-feeding. Moreover, depending on substrate glycosylation, the amino acids serine, threonine or cysteine triggers the enzymatic action. Understanding the interplay between S. oralis and MUC5B, facilitated by MdpS, has significant implications for the management of a healthy eubiotic oral microenvironment, offering potential targets for interventions aimed at modulating oral biofilm composition and succession. Additionally, since MdpS does not rely on O-glycan removal prior to extensive peptide backbone hydrolysis, the MdpS data challenges the current model of MUC5B degradation. These findings emphasize the necessity for further research in this field.

Pine-Oil-Derived Sodium Resinate Inhibits Growth and Acid Production of Streptococcus mutans In Vitro.

S. mutans is a key pathogen in dental caries initiation and progression. It promotes oral biofilm dysbiosis and biofilm acidification. Sodium resinate is a salt of pine-oil-derived resin which has antimicrobial properties. Pine-oil-derived resin consists of terpenes, diterpenes, and abietic acids. The aim of this study was to determine the effects of pine (Pinus sylvestris) oil resinate (RS) on growth and acid production of cariogenic S. mutans strains in planktonic form and biofilm. The S. mutans type strain NCTC10449 and clinical isolate CI2366 were grown on 96-well plates for testing of RS effects on growth and biofilm formation, and on plates with integrated pH-sensitive optical ensors for real-time measurements of the effects of RS on bacterial acid production. We found that even short-time exposure to RS inhibits the growth and acid production of S. mutans in the planktonic phase and biofilms. In addition, RS was able to penetrate the biofilm matrix and reduce acid production inside S. mutans biofilm. RS thus shows potential as a novel antibacterial agent against cariogenic bacteria in biofilm.

The impact of arthritogenic viruses in oral tissues.

Arthritis and periodontitis are inflammatory diseases that share several immunopathogenic features. The expansion in the study of virus-induced arthritis has shed light on how this condition could impact other parts of the human body, including the mouth. Viral arthritis is an inflammatory joint disease caused by several viruses, most notably the alphaviruses Chikungunya virus (CHIKV), Sindbis virus (SINV), Ross River virus (RRV), Mayaro virus (MAYV), and O'nyong'nyong virus (ONNV). These viruses can induce an upsurge of matrix metalloproteinases and immune-inflammatory mediators such as Interleukin-6 (IL6), IL-1ß, tumor necrosis factor, chemokine ligand 2, and receptor activator of nuclear factor kappa-B ligand in the joint and serum of infected individuals. This can lead to the influx of inflammatory cells to the joints and associated muscles as well as osteoclast activation and differentiation, culminating in clinical signs of swelling, pain, and bone resorption. Moreover, several data indicate that these viral infections can affect other sites of the body, including the mouth. The human oral cavity is a rich and diverse microbial ecosystem, and viral infection can disrupt the balance of microbial species, causing local dysbiosis. Such events can result in oral mucosal damage and gingival bleeding, which are indicative of periodontitis. Additionally, infection by RRV, CHIKV, SINV, MAYV, or ONNV can trigger the formation of osteoclasts and upregulate pro-osteoclastogenic inflammatory mediators, interfering with osteoclast activation. As a result, these viruses may be linked to systemic conditions, including oral manifestations. Therefore, this review focuses on the involvement of alphavirus infections in joint and oral health, acting as potential agents associated with oral mucosal inflammation and alveolar bone loss. The findings of this review demonstrate how alphavirus infections could be linked to the comorbidity between arthritis and periodontitis and may provide a better understanding of potential therapeutic management for both conditions.

Identification of a glutathione transporter in A. actinomycetemcomitans.

IMPORTANCE: Microbes produce a large array of extracellular molecules, which serve as signals and cues to promote polymicrobial interactions and alter the function of microbial communities. This has been particularly well studied in the human oral microbiome, where key metabolites have been shown to impact both health and disease. Here, we used an untargeted mass spectrometry approach to comprehensively assess the extracellular metabolome of the pathogen Aggregatibacter actinomycetemcomitans and the commensal Streptococcus gordonii during mono- and co-culture. We generated and made publicly available a metabolomic data set that includes hundreds of potential metabolites and leveraged this data set to identify an operon important for glutathione secretion in A. actinomycetemcomitans.

pH-responsive polymeric nanomaterials for the treatment of oral biofilm infections.

Bacterial and fungal pathogens forming oral biofilms present significant public health challenges due to the failure of antimicrobial drugs. The ability of biofilms to lower pH levels results in dental plaque, leading to gingivitis and cavities. Nanoparticles (NPs) have attracted considerable interest for drug delivery and, thus, as a solution to biofilm-related microbial infections. A novel strategy in this regard involves using pH-responsive polymeric NPs within the acidic microenvironment of oral biofilms. The acidity of the oral biofilm microenvironment is governed by carbohydrate metabolism, accumulation of lactic acid, and extracellular DNA of extracellular polymeric substances by oral biofilm-forming microbial pathogens. This acidity also provides an opportunity to enhance antibacterial activity against biofilm cells using pH-responsive drug delivery approaches. Thus, various polymeric NPs loaded with poorly soluble drugs and responsive to the acidic pH of oral biofilms have been developed. This review focuses on various forms of such polymeric NPs loaded with drugs. The fundamental mechanisms of action of pH-responsive polymeric NPs, their cytological toxicity, and in vivo efficacy testing are thoroughly discussed.

Oral microbiota in human health and disease: A perspective.

The evolution of medical knowledge about oral microbiota has increased awareness of its important role for the entire human body health. A wide range of microbial species colonizing the oral cavity interact both with each other and with their host through complex pathways. Usually, these interactions lead to a harmonious coexistence (i.e. eubiosis). However, several factors - including diet, poor oral hygiene, tobacco smoking, and certain medications, among others - can disrupt this weak homeostatic balance (i.e. dysbiosis) with potential implications on both oral (i.e. development of caries and periodontal disease) and systemic health. This article is thus aimed at providing an overview on the importance of oral microbiota in mediating several physiological and pathological conditions affecting human health. In this context, strategies based on oral hygiene and diet as well as the role of probiotics supplementation are discussed.