Mutans Biofilm Research Articles

Epigallocatechin gallate reduces the virulence of cariogenic Streptococcus mutans biofilm by affecting the synthesis of biofilm matrix components

IntroductionThere have been reports on the effects of epigallocatechin gallate (EGCG) against Streptococcus mutans viability and acidogenesis. However, the effects of EGCG on the virulence of S. mutans biofilm development have yet to be fully investigated using validated cariogenic biofilm models. ObjectiveThus, this study aimed to evaluate the effects of EGCG on S. mutans biofilm virulence using a validated cariogenic model and clinically relevant treatment regimens, twice a day for 1.5 min. MethodsEffects of EGCG on bacterial viability, polyssacharide synthesis and biofilm acidogenesis were evaluated. The morphology and 3D structure of the biofilms were evaluated by scanning electron (SEM) and confocal laser scanning microscopy, respectively. ResultsNo significant change in S. mutans viability or culture medium pH were observed when comparing EGCG-treated and NaCl-treated biofilms. EGCG significantly reduced the accumulation of soluble and insoluble polysaccharides, resulting in the formation of a biofilm with interspaced exopolysaccharide-microcolony complexes unevenly distributed on enamel. The SEM images of the biofilm treated with EGCG depict multilayers of cells arranged in short chains of microorganisms adhered to an unstructured matrix, which is not continuous and does not enmesh or protect the microorganisms entirely. Importantly, confocal images demonstrated that treatment with EGCG affected the 3D structure and organization of S. mutans biofilm, which presented a biofilm matrix more confined to the location of the microcolonies. ConclusionIn conclusion, EGCG lowered the virulence of S. mutans matrix-rich biofilm by reducing the synthesis of biofilm matrix components, altering the biofilm matrix structure, organization, and distribution.

Anti-oral streptococci and anti-biofilm properties of Etlingera pavieana essential oil and its bioactive compounds proposed for an alternative herbal mouthwash

Oral streptococci are the major group of bacteria in the oral cavity. Some of their species cause oral diseases that may lead to tooth loss and quality-of-life reduction, such as dental caries. One of prevention techniques to promote oral health is rinsing mouthwash after toothbrushing. This study aimed to determine the potential uses of local food, also remedy, plant in Thailand called Reaw-Horm or Etlingera pavieana for alternative herbal mouthwash. The essential oil from E. pavieana rhizome (Eo) is used for anti-streptococci including Streptococcus mutans and Streptococcus sobrinus and anti-biofilm activities. The main components of Eo are methyl chavicol (MC) and trans-anethole (TA). The disk diffusion method showed the inhibition zone of Eo in a dose-dependent manner. The minimum inhibitory concentration (MIC) of Eo and TA was >1.6 % v/v, and 0.4 % v/v of MC. Regarding anti-biofilm activities, MC showed nearly equal anti-biofilm formation of S. mutans and S. sobrinus, whereas Eo and TA acted toward S. sobrinus more than S. mutans biofilm. Sub-MIC killing effects on cells under biofilm were observed in Eo and MC. Therefore, MC was recommended as an active compound for anti-streptococci activities. Biocompatibility of Eo and MC were shown to be safe for epidermal cell lines. Herbal mouthwashes containing Eo were developed and had antioxidant and antimicrobial actions with established for 3 months. This study provides in vitro support on the use of herbal mouthwash with antioxidant and antimicrobial activities for dental caries prevention and well-being of individuals.

Chemical Characterization and Effect of a Lactobacilli-Postbiotic on Streptococcus mutans Biofilm In Vitro.

Postbiotic is the term used to define the soluble factors, metabolic products, or byproducts released by live probiotic bacteria or after its lysis. The objective of this study was to carry out the chemical characterization of the postbiotic of Lacticaseibacillus rhamnosus LR-32 and to evaluate its in vitro effect on the development of the Streptococcus mutans biofilm. After the cultivation of the probiotic strain, the postbiotic was extracted by centrifuging the culture and filtering the supernatant. This postbiotic was characterized by using gas chromatography coupled with mass spectrometry (GC-MS), and then it was used to determine the growth inhibition of S. mutans in its planktonic form; additionally, its effects on the following parameters in 48 h biofilm were evaluated: viable bacteria, dry weight, and gene expression of glucosyltransferases and VicR gene. The control group consisted of the biofilm without any treatment. A paired t-test was performed for statistical analysis, with the p-value set at 5%. Seventeen compounds of various chemical classes were identified in the postbiotic, including sugars, amino acids, vitamins, and acids. The treatment with the postbiotic led to an inhibition of the growth of S. mutans in its planktonic form, as well as a decrease in the number of viable bacteria, reduction in dry weight, and a negative regulation of the gene expression of gtfB, gtfC, gtfD, and vicR in its biofilm state, compared with the nontreated group (p < 0.05). The postbiotic of L. rhamnosus impaired the development of S. mutans biofilm.

Antimicrobial and Antibiofilm Effects of Silver-Copper Nanoparticles Obtained by Green Synthesis Against Streptococcus mutans.

Ag-Cu NPs were synthesized using a green chemical method and characterized using a scanning electron microscope (SEM), ultraviolet-visible (U-vis) spectrophotometer, and Fourier Transform Infrared Spectroscopy (FT-IR). The minimum inhibitory concentration (MIC) against S. mutans was determined using the broth microdilution method, while the antibiofilm effect was assessed by the crystal violet method. The synthesized Ag-Cu NPs demonstrated antimicrobial activity with an MIC of 128 µg/ml, significantly inhibiting S. mutans biofilm formation by up to 94% at a concentration of 256 µg/ml. Characterization studies confirmed the successful synthesis of Ag-Cu NPs with spherical morphology. These findings highlight the potential of Ag-Cu NPs as a novel approach for combating dental caries by targeting S. mutans biofilms. Further research is warranted to explore their safety and efficacy in clinical settings.

Tetracyclic homoisoflavanoid (+)-brazilin: a natural product inhibits c-di-AMP-producing enzyme and Streptococcus mutans biofilms.

The tenacious biofilms formed by Streptococcus mutans are resistant to conventional antibiotics and current treatments. There is a growing need for novel therapeutics that selectively inhibit S. mutans biofilms while preserving the normal oral microenvironment. Previous studies have shown that increased levels of cyclic di-AMP, an important secondary messenger synthesized by diadenylate cyclase (DAC), favored biofilm formation in S. mutans. Thus, targeting S. mutans DAC is a novel strategy to inhibit S. mutans biofilms. We screened a small NCI library of natural products using a fluorescence detection assay. (+)-Brazilin, a tetracyclic homoisoflavanoid found in the heartwood of Caesalpinia sappan, was identified as one of the 11 "hits," with the greatest reduction (>99%) in fluorescence at 100 µM. The smDAC inhibitory profiles of the 11 "hits" established by a quantitative high-performance liquid chromatography assay revealed that (+)-brazilin had the most enzymatic inhibitory activity (87% at 100 µM) and was further studied to determine its half maximal inhibitory concentration (IC50 = 25.1 ± 0.98 µM). (+)-Brazilin non-competitively inhibits smDAC's enzymatic activity (Ki = 140.0 ± 27.13 µM), as determined by a steady-state Michaelis-Menten kinetics assay. In addition, (+)-brazilin's binding profile with smDAC (Kd = 11.87 µM) was illustrated by a tyrosine intrinsic fluorescence quenching assay. Furthermore, at low micromolar concentrations, (+)-brazilin selectively inhibited the biofilm of S. mutans (IC50 = 21.0 ± 0.60 µM) and other oral bacteria. S. mutans biofilms were inhibited by a factor of 105 in colony-forming units when treated with 50 µM (+)-brazilin. In addition, a significant dose-dependent reduction in extracellular DNA and glucan levels was evident by fluorescence microscopy imaging of S. mutans biofilms exposed to different concentrations of (+)-brazilin. Furthermore, colonization of S. mutans on a representative model of enamel using suspended hydroxyapatite discs showed a >90% reduction with 50 µM (+)-brazilin. In summary, we have identified a drug-like natural product inhibitor of S. mutans biofilm that not only binds to smDAC but can also inhibit the function of smDAC. (+)-Brazilin could be a good candidate for further development as a potent therapeutic for the prevention and treatment of dental caries.IMPORTANCEThis study represents a significant advancement in our understanding of potential therapeutic options for combating cariogenic biofilms produced by Streptococcus mutans. The research delves into the use of (+)-brazilin, a natural product, as a potent inhibitor of Streptococcus mutans' diadenylate cyclase (smDAC), an enzyme crucial in the formation of biofilms. The study establishes (+)-brazilin as a non-competitive inhibitor of smDAC while providing initial insights into its binding mechanism. What makes this finding even more promising is that (+)-brazilin does not limit its inhibitory effects to S. mutans alone. Instead, it demonstrates efficacy in hindering biofilms in other oral bacteria as well. The broader spectrum of anti-biofilm activity suggests that (+)-brazilin could potentially serve as a versatile tool in a natural product-based treatment for combating a range of conditions caused by resilient biofilms.

Formulation of arginine-loaded mesoporous silica nanoparticles (Arg@MSNs) modified orthodontic adhesive

ObjectivesThe objective of this study was to synthesize arginine loaded mesoporous silica nanoparticles (Arg@MSNs), develop a novel orthodontic adhesive using Arg@MSNs as modifiers, and investigate the adhesive performance, antibacterial activity, and biocompatibility. MethodsArg@MSNs were synthesized by immobilizing arginine into MSNs and characterized using transmission electron microscope (TEM), dynamic light scattering (DLS), and Fourier Transform Infrared Spectrometer (FT-IR). Arg@MSNs were incorporated into Transbond XT adhesive with different mass fraction to form functional adhesives. The degree of conversion (DC), arginine release behavior, adhesive performance, antibacterial activity against Streptococcus mutans biofilm, and cytotoxicity were comprehensively evaluated. ResultsTEM, DLS, and FT-IR characterizations confirmed the successful preparation of Arg@MSNs. The incorporation of Arg@MSNs did not significantly affect DC and exhibited clinically acceptable bonding strength. Compared to the commercial control, the Arg@MSNs modified adhesives greatly suppressed the metabolic activity and polysaccharide production while increased the biofilm pH values. The cell counting kit (CCK)-8 test indicated no cytotoxicity. ConclusionsThe novel orthodontic adhesive containing Arg@MSNs exhibited significantly enhanced antibacterial activities and inhibitory effects on acid production compared to the commercial adhesive without compromising their bonding strength or biocompatibility. Clinical significanceThe novel orthodontic adhesive containing Arg@MSNs exhibits potential clinical benefits in preventing demineralization of enamel surfaces around or beneath orthodontic brackets due to its enhanced antibacterial activities and acid-producing inhibitory effects.

Lactobacillus plantarum NCU001774 microcapsules target and inhibit cariogenic Streptococcus mutans

This study aims to develop a cariogenic bacteria-targeting delivery system to release lactobacilli at infectious sites. Firstly, L. plantarum NCU001774 was selected as anti-S. mutans strain from 385 lactic acid bacteria by evaluating its inhibitory effects on S. mutans, tolerance to lysozyme, and protective effect on hydroxyapatite. Subsequently, NCU001774 microcapsules composed of gelatin/chitosan in a ratio of 1:5 (2%, W/V), trehalose as protectant (2%, W/V), were successfully prepared. The NCU001774 microcapsules were mainly 1–5 μm, with the t1/2 of 12.01 min at pH4.5 and 21.41 min at pH7.2. When the environmental pHwasbelow6.5, NCU001774 microcapsules were positively charged, which enabled the targeting effect on S. mutans biofilm through electrostatic interaction. Fluorescence staining and scanning electronic microscopy assay further confirmed the targeting effect of NCU001774 on the surface of S. mutans biofilm. The formation of S. mutans biofilm was reduced by 62% with the treatment of NCU001774 microcapsules. Meanwhile, the number of S. mutans decreased by NCU001774 microcapsules from 8.50 to 2.20 (lgCFU/mL) at 24 h. The microcapsules developed in this study can release lactobacilli at targeted infectious sites and inhibit growth and biofilm formation of S. mutans, demonstrating great potential in prevention of S. mutans associated infection including dental caries.

Examination of the Structure and Formation Streptococcus mutans Biofilm Induced by Glucose, Lactose, Soy Protein, and Iron.

Streptococcus mutans, the main causative agent of caries, have the ability to form biofilms on the surface of teeth. The availability of nutrients such as glucose, lactose, soy protein, and iron can influence S. mutans in biofilm formation. All four sources of nutrients have been shown to increase the formation of S. mutans biofilms. The purpose of this study was to determine the structure and thickness of S. mutans biofilms induced by glucose, lactose, soy protein, and iron. This experimental laboratory study aimed to examine the formation of biofilm structures (chemical elements) and determine the thickness of S. mutans biofilms induced by glucose, lactose, soy protein, and iron. The structures (chemical elements) were examined using scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) analysis. Confocal laser scanning microscopy (CLSM) was used to determine the thickness of S. mutans biofilms with an Olympus FV1000 microscope, and the findings were analyzed using Olympus Fluoview Ver. 4.2a software. It was established that the results of SEM-EDX examination of the structure of S. mutans biofilms induced by glucose had oxygen (O) as the dominant chemical element (30.24 w%); lactose reported oxygen (O) as the dominant element (29.65 w%); soy protein had carbon (C) as the dominant element (34.31 w%); and iron showed oxygen (O) as the dominant element (32.51 w%). The thickness (measured by the CLSM examination) of biofilms induced by glucose, lactose, soy protein, and iron were 17,666, 12,666, 18,000, and 15,666 nm, respectively. The structure of S. mutans biofilms induced by glucose, lactose, and iron contain the following elements in amounts from the highest to lowest: O, C, N, P, and S; the biofilm produced by S. mutans induced by soy protein in amounts from the highest to lowest comprised the elements: C, O, N, S, and P. The S. mutans biofilms induced by soy protein had the maximum thickness, followed by those induced by glucose, iron, and lactose.

Versatility of copper-iron bimetallic nanoparticles fabricated using Hibiscus rosa-sinensis flower phytochemicals: various enzymes inhibition, antibiofilm effect, chromium reduction and dyes removal.

Bimetallic nanoparticles (NPs) are considered superior in terms of stability and function with respect to its monometallic counterparts. Hence, in the present study Hibiscus rosa-sinensis flower extract was used to synthesis copper-iron bimetallic nanoparticles (HF-FCNPs). HF-FCNPs was characterized and its applications (biological and environmental) were determined. HF-FCNPs were spherical in shape with high percentage of copper inducted into the NPs. HF-FCNPs inhibited mammalian glucosidases [maltase (IC50: 548.71 ± 61.01µg/mL), sucrase (IC50: 441.34 ± 36.03µg/mL), isomaltase (IC50: 466.37 ± 27.09µg/mL) and glucoamylase (IC50: 403.12 ± 14.03µg/mL)], alpha-amylase (IC50: 16.27 ± 1.73µg/mL) and acetylcholinesterase [AChE (IC50: 0.032 ± 0.004µg/mL)] activities. HF-FCNPs showed competitive inhibition against AChE, maltase and sucrase activities; mixed inhibition against isomaltase and glucoamylase activities; whereas non-competitive inhibition against α-amylase activity. HF-FCNPs showed zone of inhibition of 16 ± 2mm against S. mutans at 100µg/mL concentration. HF-FCNPs inhibited biofilm formation of dental pathogen, S. mutans. SEM and confocal microscopy analysis revealed the disruption of network formation and bacterial cell death induced by HF-FCNPs treatment on tooth model of S. mutans biofilm. HF-FCNPs efficiently removed hexavalent chromium in pH-independent manner and followed first order kinetics. Through Langmuir isotherm fit the qmax (maximum adsorption capacity) was determined to be 62.5mg/g. Further, HF-FCNPs removed both anionic and cationic dyes. Altogether, facile synthesis of HF-FCNPs was accomplished and its biological (enzyme inhibition and antibiofilm activity) and environmental (catalyst to remove pollutants) applications have been understood.

Antibacterial effect of novel dental resin composites containing rod-like zinc oxide

Abstract Dental resin composite materials are widely used as dental fillings; however, the accumulation of microbes and the resulting secondary caries often leads to filling failure. ZnO, an inorganic antibacterial material, exhibits effective antibacterial properties and is considered safe for use. In this study, rod-like ZnO was prepared by using the atmospheric-pressure hydrothermal method, and its microstructure and antibacterial effects on Streptococcus mutans were studied. Subsequently, we created modified resins by incorporating rod-like ZnO at varying mass fractions and analyzed their morphological characteristics and elemental distributions. The antibacterial effectiveness, biocompatibility, and mechanical properties of these materials were examined using in vitro experiments. The results indicated that the rod-like ZnO exhibited a complete hexagonal wurtzite structure, with columnar dimensions of approximately 2.5 μm in length, 0.8 μm in diameter, and a lattice spacing of 0.2544 nm. The growth, biofilm formation, and biofilm destruction of S. mutans were significantly inhibited at 1/4, 1/2, 3/4, and 1 times the minimum inhibitory concentration. The rod-like modified resin, with mass fractions of 2.5, 5, and 7.5 wt%, exhibited evident inhibitory effects on S. mutans biofilm formation. These modified resins demonstrated no cytotoxicity toward HGF-1 cells and exhibited enhanced compressive strength. Therefore, rod-like ZnO modified resin has promising potential for the treatment of dental caries.

Periodontal ligament stem cell-derived exosome-loaded Emodin mediated antimicrobial photodynamic therapy against cariogenic bacteria

BackgroundThis study was conducted to investigate the efficiency of periodontal ligament (PDL) stem cell-derived exosome-loaded Emodin (Emo@PDL-Exo) in antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans and Lactobacillus acidophilus as the cariogenic bacteria.Materials and methodsAfter isolating and characterizing PDL-Exo, the study proceeded to prepare and verify the presence of Emo@PDL-Exo. The antimicrobial effect, anti-biofilm activity, and anti-metabolic potency of Emo, PDL-Exo, and Emo@PDL-Exo were then evaluated with and without irradiation of blue laser at a wavelength of 405 ± 10 nm with an output intensity of 150 mW/cm2 for a duration of 60 s. In addition, the study assessed the binding affinity of Emodin with GtfB and SlpA proteins using in silico molecular docking. Eventually, the study examined the generation of endogenous reactive oxygen species (ROS) and changes in the gene expression levels of gelE and sprE.ResultsThe study found that using Emo@PDL-Exo-mediated aPDT resulted in a significant decrease in L. acidophilus and S. mutans by 4.90 ± 0.36 and 5.07 log10 CFU/mL, respectively (P < 0.05). The study found that using Emo@PDL-Exo for aPDT significantly reduced L. acidophilus and S. mutans biofilms by 44.7% and 50.4%, respectively, compared to untreated biofilms in the control group (P < 0.05). Additionally, the metabolic activity of L. acidophilus and S. mutans decreased by 58.3% and 71.2%, respectively (P < 0.05). The molecular docking analysis showed strong binding affinities of Emodin with SlpA and GtfB proteins, with docking scores of -7.4 and -8.2 kcal/mol, respectively. The study also found that the aPDT using Emo@PDL-Exo group resulted in the most significant reduction in gene expression of slpA and gtfB, with a decrease of 4.2- and 5.6-folds, respectively, compared to the control group (P < 0.05), likely due to the increased generation of endogenous ROS.DiscussionThe study showed that aPDT using Emo@PDL-Exo can effectively reduce the cell viability, biofilm activity, and metabolic potency of S. mutans and L. acidophilus. aPDT also significantly reduced the expression levels of gtfB and slpA mRNA due to the increased endogenous ROS generation. The findings suggest that Emo@PDL-Exo-mediated aPDT could be a promising antimicrobial approach against cariogenic microorganisms.

Inhibitory and preventive effects of Arnebia euchroma (Royle) Johnst. root extract on Streptococcus mutans and dental caries in rats

BackgroundDental caries is one of the prevalent conditions that threaten oral health. Arnebia euchroma (Royle) Johnst. root (AR) extracts exhibit anti-inflammatory, anti-cancer, and antibacterial properties. This study was designed to investigate the antibacterial impact of AR extract on Streptococcus mutans (S. mutans) UA159 and the anti-caries effect on rats.MethodsThe antibacterial activity of AR extract against S. mutans and its biofilm was determined using the bacterial sensitivity test, the biofilm sensitivity test, and the live-dead staining technique. By fluorescently tagging bacteria, the influence of bacterial adhesion rate was determined. Using a rat caries model, the anti-caries efficacy and safety of AR extract were exhaustively investigated in vivo.ResultsAR extract inhibit not only the growth of S. mutans, but also the generation of S. mutans biofilm, hence destroying and eliminating the biofilm. Moreover, AR extract were able to inhibit S. mutans’ adherence to saliva-encapsulated hydroxyapatite (HAP). Further, in a rat model of caries, the AR extract is able to greatly reduce the incidence and severity of caries lesions on the smooth surface and pit and fissure of rat molars, while exhibiting excellent biosafety.ConclusionsAR extract exhibit strong antibacterial activity against S. mutans and can lower the incidence and severity of dental cavities in rats. These findings suggest that Arnebia euchroma (Royle) Johnst. could be utilized for the prevention and treatment of dental caries.

Enhancing the Antibacterial Effect of Erythrosine-Mediated Photodynamic Therapy with Ethylenediamine Tetraacetic Acid

This study evaluated the additive impact of ethylenediamine tetraacetic acid (EDTA) on erythrosine-mediated photodynamic therapy (PDT) against &lt;i&gt;Streptococcus mutans&lt;/i&gt; (&lt;i&gt;S. mutans&lt;/i&gt;) biofilm by measuring colony-forming units and applying confocal laser scanning microscopy. Fifty-six bovine incisors, free from dental caries or structural defects, were utilized in this study. Dentin specimens were created by cutting with a low-speed diamond disk under a continuous flow of water, resulting in dimensions of 6.0 mm × 3.0 mm × 2.0 mm. The specimens were categorized into 4 groups: Control, EDTA, PDT, and EDTA + PDT. &lt;i&gt;S. mutans&lt;/i&gt; ATCC 25175 was employed to establish biofilm on the dentin specimens. A 17% EDTA solution was applied for 1 min. For PDT, erythrosine served as the photosensitizer. Finally, a light-emitting diode source (385 - 515 nm) was employed in this study. The PDT group exhibited a significantly lower bacterial count than both the control and EDTA groups (&lt;i&gt;p&lt;/i&gt; &lt; 0.001). The EDTA + PDT group demonstrated a significantly reduced bacterial count compared to the other 3 groups (&lt;i&gt;p&lt;/i&gt; &lt; 0.001). This study demonstrated that EDTA enhances the antimicrobial efficacy of PDT on S. mutans biofilm. Even at a low concentration of photosensitizer, the combination of EDTA and PDT yields a significant antibacterial effect.

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.

Suppression of Streptococcus mutans Biofilm Formation and Gene Expression by PRG Barrier Coat: A Molecular and Microscopic Study for Preventing Dental Caries.

This study aimed to investigate the inhibitory effect of a PRG Barrier Coat on biofilm formation and structure by Streptococcus mutans and propose an effective method for preventing dental caries. Streptococcus mutans MT8148 biofilms were obtained from hydroxyapatite disks with and with- out a PRG Barrier Coat. Scanning electron microscopy (SEM) was used to observe the 12- and 24-h-cultured biofilms, while reverse-transcription polymerase chain reaction (qRT-PCR) was used to quantify caries-related genes. Biofilm adhe- sion assessments were performed on glass. Statistical analysis was performed using a two-sample t-test. A statistically significant difference in Streptococcus mutans biofilm adhesion rate was observed between the con- trol and PRG Barrier Coat-coated samples (p < 0.01). However, there was no statistically significant difference in total bacter- ial count or biofilm volume (p > 0.05). SEM revealed that the PRG Barrier Coat inhibited biofilm formation by Streptococcus mutans. Real-time RT-PCR revealed that the material restricted the expression of genes associated with caries-related bio- film formation. However, the suppression of gtfD and dexB differed from that of other genes. PRG Barrier Coat suppressed biofilm formation by Streptococcus mutans by inhibiting the expression of in- soluble glucan synthase, which is associated with primary biofilm formation. The material also affected gene expression and altered the biofilm structure. Tooth surface-coating materials, such as PRG Barrier Coat, may improve caries preven- tion in dental practice.

Comparative evaluation of anti-biofilm and anti- adherence potential of plant extracts against Streptococcus mutans: A therapeutic approach for oral health

Dental caries predominantly attributed to the cariogenic nature of Streptococcus mutans, continue to pose a substantial global challenge to oral health. In response to this challenge, this study aimed to evaluate the effectiveness of leaf extracts (LEs) and essential oils (EOs) derived from different medicinal plants in inhibiting the growth of Streptococcus mutans biofilm. In vitro and in silico approaches were employed to identify active compounds and assess their inhibitory effects on S. mutans. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were measured to determine the anti-biofilm and anti-adherence activity against S. mutans. Biofilm viability (CFU/mL) and extracellular polymeric substance (EPS) concentration were quantified. GC-MS analysis was utilized to identify active compounds in the most effective plant extracts exhibiting anti-S. mutans activity. A high-throughput screening focused on the interaction between these compounds and the target enzyme SortaseA (SrtA) using molecular docking was performed. Results indicated that Cymbopogon citratus displayed the highest efficacy in reducing S. mutans biofilm formation and adhesion activity, achieving 90 % inhibition at an MIC value of 12 μg/mL. Among the 12 bioactive compounds identified, trans-Carvyl acetate exhibited the lowest binding energy with SrtA (-6.0 Kcal/mole). Trans-Carvyl acetate also displayed favorable pharmacokinetic properties. This study provides novel insights into the anti-S. mutans properties of C. citratus and suggests its potential as a therapeutic approach for oral health. Further research is needed to explore the combined effect of plant extracts for enhanced protection against dental caries.

Antimicrobial effect of probiotic bacteriocins on Streptococcus mutans biofilm in a dynamic oral flow chamber model – an in vitro study

ABSTRACT Aim To determine the antimicrobial activity of the bacteriocin-producing probiotic strains Streptococcus salivarius K12 and Streptococcus salivarius M18 alone or in combination against caries-associated Streptococcus mutans. Methods Antimicrobial activity of S. salivarius K12 and/or S. salivarius M18 against S. mutans ATCC 25175 growth and biofilm formation on hydroxyapatite (HA) discs was determined in a flow chamber model by recording the colony forming units (CFU/ml) after 48 h of co-cultivation. The biofilm was analyzed by scanning electron microscopy (SEM) and by confocal laser scanning microscopy (CLSM). Additionally, the simultaneous antagonism assay was used to assess the inhibitory effect of S. salivarius K12 and/or S. salivarius M18 against S. mutans ATCC 25175 and 21 clinical isolates of S. mutans. Results Co-cultivation of S. mutans and S. salivarius K12 and/or S. salivarius M18 led to the inhibition of S. mutans viability, thereby, preventing its biofilm formation on HA discs. Furthermore, S. salivarius K12 and S. salivarius M18 exhibited antimicrobial activity against most clinical isolates of S. mutans. Conclusion The in vitro flow chamber system used in this study allows the simulation of time-dependent administration of S. salivarius probiotic strains, either alone or in combination, to investigate the prevention of S. mutans biofilm formation in a standardized model.

Antibiofilm Properties and Demineralization Suppression in Early Enamel Lesions Using Dental Coating Materials.

This study aimed to investigate the effects of dental coating materials on Streptococcus mutans biofilm formation. The test materials were PRG Barrier Coat (PRG), BioCoat Ca (BioC), and FluorDental Jelly (FluorJ). Bovine enamel specimens were demineralized to mimic early enamel lesions. The biofilm was developed on a specimen treated with one of the materials by using a modified Robbins device flow-cell system. Scanning electron and fluorescence confocal laser scanning microscopy, viable and total cell counts, and gene expression assessments of the antibiofilm were performed. Ion incorporation was analyzed using a wavelength-dispersive X-ray spectroscopy electron probe microanalyzer. All materials allowed biofilm formation but reduced its volume. FluorJ was the only material that inhibited biofilm accumulation and had a bactericidal effect, revealing 0.66 log CFU in viable cells and 1.23 log copy reduction in total cells compared with the untreated group after 24 h of incubation. The ions released from PRG varied depending on the element. BioC contributed to enamel remineralization by supplying calcium ions while blocking the acid produced from the biofilm. In summary, the dental coating materials physically prevented acid attacks from the biofilm while providing ions to the enamel to improve its mechanical properties.

Inhibition of Streptococcus mutans growth and biofilm formation through protein acetylation.

Numerous cellular processes are regulated in response to the metabolic state of the cell, and one such regulatory mechanism involves lysine acetylation. Lysine acetylation has been proven to play an important role in the virulence of Streptococcus mutans, a major cariogenic bacterial species. S. mutans' glucosyltransferases (Gtfs) are responsible for synthesizing extracellular polysaccharides (EPS) and contributing to biofilm formation. One of the most common nonsteroidal anti-inflammatory drugs is acetylsalicylic acid (ASA), which can acetylate proteins through a nonenzymatic transacetylation reaction. Herein, we investigated the inhibitory effects of ASA on S. mutans. ASA treatment was observed to impede the growth of S. mutans, leading to a reduction in the production of water-insoluble EPS and the formation of biofilm. Moreover, ASA decreased the enzyme activity of Gtfs while increasing the protein acetylation level. The in vivo anticaries efficacy of ASA has further been proved using the rat caries model. In conclusion, ASA as an acetylation agent attenuated the cariogenic virulence of S. mutans, suggesting the potential value of protein acetylation on antimicrobial and anti-biofilm applications to S. mutans.

Chitosan nanoparticles loaded with epigallocatechin-3-gallate: synthesis, characterisation, and effects against Streptococcus mutans biofilm

This study evaluated the effects of chitosan nanoparticles loaded with epigallocatechin-3-gallate (EGCG) against Streptococcus mutans biofilm. EGCG-loaded chitosan (Nchi + EGCG) nanoparticles and Chitosan (Nchi) nanoparticles were prepared by ion gelation process and characterised regarding particle size, polydispersion index, zeta potential, and accelerated stability. S mutans biofilms were treated twice daily with NaCl 0.9% (negative control), Nchi, Nchi + EGCG, and chlorhexidine (CHX) 0.12% (positive control). After 67 h, the biofilms were evaluated for acidogenesis, bacterial viability and dry weight. Biofilm morphology and structure were analysed by scanning electron microscopy. The nanoformulations presented medium to short-term stability, size of 500 nm, and polydispersion index around 0.400. Treatments affected cell morphology and biofilm structure. However, no effects on microbial viability, biofilm dry weight, and acidogenesis were observed. Thus, the nanoformulations disassembled the biofilm matrix without affecting microbial viability, which makes them promising candidates for the development of dental caries preventive and therapeutic agents.