Mutans Biofilm Research Articles

On-Demand Free Radical Release by Laser Irradiation for Photothermal-Thermodynamic Biofilm Inactivation and Tooth Whitening.

Dental diseases associated with biofilm infections and tooth staining affect billions of people worldwide. In this study, we combine photothermal agents (MoS2@BSA nanosheets, MB NSs), a thermolysis free-radical initiator (AIPH), and carbomer gel to develop laser-responsive hydrogel (MBA-CB Gel) for biofilm inactivating and tooth whitening. Under a physiological temperature without laser irradiation, MB NSs can eliminate free radicals generated from the slow decomposition of AIPH due to their antioxidative activity, thereby avoiding potential side effects. A cytotoxicity study indicates that MB NSs can protect mammalian cells from the free radicals released from AIPH without laser irradiation. Upon exposure to laser irradiation, MB NSs promote the rapid decomposition of AIPH to release free radicals by photothermal effect, suggesting their on-demand release ability of free radicals. In vitro experimental results show that the bacteria inactivation efficiency is 99.91% (3.01 log units) for planktonic Streptococcus mutans (S. mutans) and 99.98% (3.83 log units) for planktonic methicillin-resistant Staphylococcus aureus (MRSA) by the mixed solution of MB NSs and AIPH (MBA solution) under 808 nm laser irradiation (1.0 W/cm2, 5 min). For S. mutans biofilms, an MBA solution can inactivate 99.97% (3.63 log units) of the bacteria under similar laser irradiation conditions. Moreover, MBA-CB Gel can whiten an indigo carmine-stained tooth under laser irradiation after 60 min of laser treatment, and the color difference (ΔE) in the teeth of the MBA-CB Gel treatment group was 10.9 times that of the control group. This study demonstrates the potential of MBA-CB Gel as a promising platform for biofilm inactivation and tooth whitening. It is worth noting that, since this study only used stained models of extracted teeth, the research results may not fully reflect the actual clinic situation. Future clinical research needs to further validate these findings.

Antimicrobial activity of probiotic Streptococcus salivarius LAB813 on in vitro cariogenic biofilms

ObjectiveTo investigate the antimicrobial activity of a novel commensal strain of Streptococcus salivarius, LAB813, against Streptococcus mutans biofilms. MethodsThe inhibitory activity of LAB813 towards S. mutans was tested using mono-, dual-, and multi-species cariogenic biofilms formed on three types of orthodontic appliances (metal, ceramic, aligner). The activity of the commercially available probiotic, BLIS M18™ was used as control. ResultsLAB813 significantly inhibited S. mutans biofilms with cell killing approximating 99% for all materials. LAB813 showed effectiveness at inhibiting S. mutans in more complex multi-species biofilms with cell killing approximating 90% for all three materials. When comparing the killing kinetics of the probiotics, LAB813 had a faster rate of killing biofilms than M18. Experiments conducted with cell-free culture supernatant confirmed the presence of an inhibitory substance of proteinaceous nature. The addition of xylitol, a common sugar substitute used for human consumption, potentiated the inhibitory effects of LAB813 against S. mutans embedded in a more complex fungal-bacterial biofilm. ConclusionsLAB813 possesses strong antimicrobial activity, potent anti-biofilm properties, and enhanced antimicrobial activity in the presence of xylitol. The identification and characterization of strain LAB813 exhibiting antimicrobial activity towards S. mutans hold exciting promise for this novel strain to be developed as an oral probiotic for use in the prevention of dental caries.

The study on the effect of amino acid porphyrin conjugate-mediated antimicrobial photodynamic therapy on Streptococcus mutans biofilm

Antimicrobial Photodynamic Therapy (aPDT) based on the action of visible light and photosensitizers has emerged as a promising microbial reduction and alternative to antibiotics resistance to cariogenic pathogens. The present research aims to evaluate the antimicrobial effect of aPDT mediated by a new photosensitizer (amino acid porphyrin conjugate 4i) on Streptococcus mutans (S. mutans) biofilm. Qualitative morphologic characteristics of S. mutans biofilms are shown by scanning electron microscopy (SEM). The colony plate counting method is used to measure the dark toxicity and the phototoxicity of different concentrations of 4i-aPDT to S. mutans biofilms. MTT assay is conducted to investigate the effect of 4i mediated aPDT on the metabolic activity of S. mutans biofilm. Changes in structure morphology, bacterial density and extracellular matrix of S. mutans biofilm are observed by SEM. The distribution of living and dead bacteria in biofilm is detected using Confocal laser microscopy (CLSM). The results indicate that single laser irradiation has no antibacterial effect on S. mutans biofilms. With the increase of 4i concentration or the prolongation of laser irradiation time, the antibacterial effect of 4i-mediated aPDT on S. mutans biofilm is more statistically significant compared to the control. When the concentration of 62.5 µmol/L 4i is continuously illuminated for 10 min, the logarithm of the colonies in the biofilm shows a reduction of 3.4 log10. MTT assay detected absorbance values of biofilm by 4i-mediated aPDT are the lowest, indicating a significant decrease in biofilm metabolic activity. SEM analysis shows that 4i mediated aPDT reduced the quantity and density of S. mutans. A dense red fluorescence image of the 4i-aPDT treated biofilm is observed under CLSM, indicating that the dead bacteria are widely distributed.

Investigation of Components in Roasted Green Tea That Inhibit Streptococcus mutans Biofilm Formation.

Streptococcus mutans form oral biofilms (BFs) and cause dental caries. Roasted green tea (RGT) is prepared by roasting the tea plant, and RGT-specific polyphenols are produced during the roasting process. Catechins, polyphenols in green tea, have BF inhibitory activity against S. mutans; therefore, RGT-specific polyphenols are also expected to have this activity. However, there are few reports on the structural and functional properties of RGT. This study aimed to investigate the inhibitory activity of RGT against S. mutans BF formation and to investigate the active compounds. RGT extract fractionation and BF inhibitory assay were performed. Strong activity was confirmed in the RGT fractions that had medium-high hydrophobicity, were rich in phenolic hydroxyl groups, and lacked catechins. A peak comprising compounds with molecular weights of 918 (mw918) and 1050 (mw1050) was purified from the fraction. Since BF inhibitory activity was confirmed for this peak, these compounds were considered to be part of the active ingredients. The mw918 polyphenol was detected only in RGT and it was thought to be produced during the roasting process. The results of this research will serve as a basis for the future application of RGT as a safe and effective anti-caries agent.

Novel Dental Low-Shrinkage-Stress Composite with Antibacterial Dimethylaminododecyl Methacrylate Monomer.

Current dental resins exhibit polymerization shrinkage causing microleakage, which has the potential to cause recurrent caries. Our objectives were to create and characterize low-shrinkage-stress (LSS) composites with dimethylaminododecyl methacrylate (DMADDM) as an antibacterial agent to combat recurrent caries. Triethylene glycol divinylbenzyl ether and urethane dimethacrylate were used to reduce shrinkage stress. DMADDM was incorporated at different mass fractions (0%, 1.5%, 3%, and 5%). Flexural strength, elastic modulus, degree of conversion, polymerization stress, and antimicrobial activity were assessed. The composite with 5% DMADDM demonstrated higher flexural strength than the commercial group (p < 0.05). The addition of DMADDM in BisGMA-TEGDMA resin and LSS resin achieved clinically acceptable degrees of conversion. However, LSS composites exhibited much lower polymerization shrinkage stress than BisGMA-TEGDMA composite groups (p < 0.05). The addition of 3% and 5% DMADDM showed a 6-log reduction in Streptococcus mutans (S. mutans) biofilm CFUs compared to commercial control (p < 0.001). Biofilm biomass and lactic acid were also substantially decreased via DMADDM (p < 0.05). The novel LSS dental composite containing 3% DMADDM demonstrated potent antibacterial action against S. mutans biofilms and much lower polymerization shrinkage-stress, while maintaining excellent mechanical characteristics. The new composite is promising for dental applications to prevent secondary caries and increase restoration longevity.

Amyloid hexapeptide prevent dental caries by antibiofilm formation

ObjectivesBiofilm formed by cariogenic microbes is the direct cause of dental caries, therefore, prevention of dental caries should be anti-biofilm-based. Previously, we found the amyloid hexapeptides efficiently inhibited biofilm formation by aggregating into amyloid fibrils agglutinating microbes. This study aimed to select the most stable amyloid hexapeptide GIDLKI (GI6) and study its anti-caries effect. MethodsBiofilms of multi-species bacteria, derived from mixed saliva, were cultured to evaluate the anti-biofilm formation effect of GI6. And then, the primary cariogenic bacterium Streptococcus mutans (S.mutans) was cultured in BHI with various pH, gradient concentrations of sucrose, glucose, and calcium ions to evaluate the anti-biofilm formation effects of GI6. Then models of human enamel block caries and twenty male SPF-SD rat caries induced by S. mutans biofilm were constructed, and confocal laser scanning microscopy, scanning electron microscopy, and micro-computed tomography were applied to investigate the anti-biofilm formation, anti-caries effects and use safety of GI6. ResultsGI6 could inhibit the multi-species bacteria biofilm formation and remained effective in anti-biofilm activity against S. mutans in environments closely related to caries. GI6 suppressed S. mutans biofilm formation and thus prevented or alleviated the development of caries in human tooth blocks and rat teeth. GI6 did not affect the intestinal flora, serum biochemical parameters, and the pathological changes of various organs. ConclusionsAmyloid hexapeptides, including but not limited to GI6, are novel effective anti-caries agents that can be used to prevent dental caries safely. Clinical significanceThis study explored the anti-biofilm formation and anti-caries effect of GI6 in vitro, highlighting the anti-biofilm formation therapy for dental caries and setting a foundation for the practical application of GI6 for the treatment of dental caries.

Effectiveness of Xylitol Toothpaste on the Growth and Biofilm Formation of Streptococcus mutans ATCC 25175

Background: Several toothpaste ingredients have also been proven to prevent dental caries. An alternative material like xylitol can reduce biofilms and caries’ formation. Objective: This study aims to test the antibacterial xylitol toothpaste against Streptococcus mutans biofilm. Materials and Methods: The type of this research was true experimental by using two kinds of toothpaste with xylitol active ingredient (samples 1 and 2) and toothpaste with fluoride (sample 3). The toothpaste was diluted using the serial dilution method, and then the minimum inhibitory concentration (MIC), minimum bactericide concentration (MBC), minimum biofilm eradication concentration (MBEC), and minimum biofilm inhibition concentration (MBIC) were examined. The data were analyzed with ANOVA and Pearson correlation with a significance value of p0.005. Results: The MIC values of samples 1, 2, and 3 were 0.39%, 0.78%, and 0.39%, respectively. The MBC of samples 1, 2, and 3 were 12.5%, 25%, and 6.25%, respectively. The results of MBIC samples 1, 2, and 3 were 1.56%, 1.56%, and 0.78%, respectively. The results of MBEC samples 1, 2, and 3 were 6.25%, 6.25%, and 1.56%, respectively. There was a difference in the decrease of the mass of Streptococcus mutans ATCC 25175 biofilm after administration using xylitol and fluoride toothpaste, with a p-value of 0.0295. Additionally, there is a correlation between the time of administration of xylitol toothpaste with the changes in the mass of the biofilm Streptococcus mutans ATCC 25175. Conclusion: Xylitol toothpaste can inhibit the growth and eradication of Streptococcus mutans biofilm ATCC 25175.

3,3′-Diindolylmethane (DIM): A Potential Therapeutic Agent against Cariogenic Streptococcus mutans Biofilm

Indole, a metabolite of the amino acid tryptophan, has been proven to act as a signal molecule in bacteria, acting in different aspects of biofilm formation. The oral biofilm is a type of biofilm that has consequences for human health. It is a complex, three-dimensional structure that develops on the surface of teeth via the attachment of primary microbial colonizers. Many oral infections are caused by an imbalance occurring in the microorganisms naturally found in oral biofilms and are considered major public health concerns. In this study, we test the effect of a natural bis-indole, 3,3′-Diindolylmethane (DIM), in mitigating the pathogenicity of the oral biofilm inhabiting bacterium Streptococcus mutans, a bacterium that is considered to be a principal etiological agent in dental caries. Our study found that DIM was able to attenuate S. mutans biofilm formation by 92%. Additionally, treatment with DIM lowered extracellular polymeric substance (EPS) production and decreased its durability significantly under acidic conditions. Therefore, the anti-biofilm and anti-virulence properties of DIM against S. mutans bacteria in an “oral setting” provides evidence for its usefulness in reducing biofilm formation and potentially for caries attenuation.

Saliva Decreases Sucrose-Induced Cariogenicity in an Experimental Biological Caries Model.

Whether a minimum quantity of saliva inhibit the caries process remains uncertain. This study aimed to investigate the impact of saliva dilutions on an in vitro caries model using Streptococcus mutans (S. mutans) biofilms. S. mutans biofilms were cultivated on enamel and root dentin slabs, in culture media containing different proportions of saliva (v/v): 0%, 5%, 10%, 25%, 50%, 75%, and 100% saliva, and exposed to a 10% sucrose solution (5 min, 3x/day), with appropriate controls. After 5 (enamel) and 4 (dentin) days, demineralization, biomass, viable bacteria, and polysaccharide formation were analyzed. The acidogenicity of the spent media was monitored overtime. Each assay was performed in triplicate across two independent experiments (n = 6). In both enamel and dentin, an inverse relationship was observed between acidogenicity, demineralization, and the proportion of saliva. Even small quantities of saliva incorporated into the media led to a noticeable reduction in enamel and dentin demineralization. Saliva presence resulted in significant reductions in biomass, viable S. mutans cells, and polysaccharides, with the effects being concentration-dependent for both tissues. High quantities of saliva can almost completely inhibit sucrose-induced cariogenicity, while even small amounts exhibit a dose-dependent caries-protective effect.

Surface properties and Streptococcus mutans biofilm adhesion of ion-releasing resin-based composite materials

ObjectiveTo evaluate the adhesion of Streptococcus mutans (S. mutans) and related surface properties of ion-releasing resin-based composite (RBC) restorative materials. MethodsTwo ion-releasing RBCs, Activa (ACT) and Cention-N (CN), were compared to a conventional RBC (Z350) and a resin-modified glass ionomer cement (Fuji-II-LC). Ten disk-shaped specimens were fabricated for each material (n = 40). After standardized surface polishing procedure, the surface properties of the specimens were evaluated using surface roughness measurements by a profilometer and hydrophobicity using water contact angle measurements. To assess bacterial adhesion, the number of S. mutans bacteria was calculated from colony-forming units (CFU). Confocal laser scanning microscope analysis was done for qualitative & quantitative assessment. The data were analyzed using One-way ANOVA followed by Tukey's post-hoc test to compare the mean values of surface roughness, water contact angle and CFU values. To compare the mean dead cell percentage Kruskal-Wallis rank test and Conover test were used. A p-value of ≤ 0.05 was used to report the statistical significance. ResultsZ350 and ACT had the smoothest surfaces, followed by CN, and the roughest surface was seen in FUJI-II-LC. The lowest water contact angles were seen in CN, and Z350, and the highest were in ACT. S. mutans counts were the highest in ACT and the lowest in Z350 and CN. CN and Fuji-II-LC registered the highest percentage of dead bacterial cells, while the lowest were in ACT. SignificanceSurface properties did not significantly influence bacterial adhesion. More S. mutans bacteria accumulated on ACT than on the nanofilled composite and on CN. CN had antibacterial effects against Streptococcus mutans biofilms.

Effects of blue-light LED toothbrush on reducing dental plaque and gingival inflammation in orthodontic patients with fixed appliances: a crossover randomized controlled trial

BackgroundPatients with fixed orthodontic appliances have higher plaque accumulation and gingival inflammation. Our aim was to compare the effectiveness of a light emitting diode (LED) toothbrush with a manual toothbrush in reducing dental plaque and gingival inflammation in orthodontic patients with fixed appliances, and to investigate the effect of the LED toothbrush on Streptococcus mutans (S. mutans) biofilm in vitro.MethodsTwenty-four orthodontic patients were recruited and randomly assigned into 2 groups: (1) started with manual and (2) started with LED toothbrushes. After a 28-day usage and 28-day wash-out period, the patients switched to the other intervention. The plaque and gingival indices were determined at baseline and 28 days after each intervention. The patients’ compliance and satisfaction scores were collected using questionnaires. For the in vitro experiments, S. mutans biofilm was divided into 5 groups (n = 6) with 15-, 30-, 60-, or 120-sec LED exposure, and without LED exposure as a control group.ResultsThere was no significant difference in the gingival index between the manual and LED toothbrush groups. The manual toothbrush was significantly more effective in reducing the plaque index in the proximal area on the bracket side (P = 0.031). However, no significant difference was found between the two groups in other areas around the brackets or on the non-bracket side. After LED exposure in vitro, the percentages of bacterial viability after LED exposure for 15–120 s were significantly lower compared with the control (P = 0.006).ConclusionClinically, the LED toothbrush was not more effective in reducing dental plaque or gingival inflammation than the manual toothbrush in orthodontic patients with fixed appliances. However, the blue light from the LED toothbrush significantly reduced the number of S. mutans in biofilm when it was exposed to the light for at least 15 s in vitro.Clinical Trial RegistrationThai Clinical Trials Registry (TCTR20210510004). Registered 10/05/2021.

Relationships between composite roughness and Streptococcus mutans biofilm depth under shear in vitro

ObjectiveTo investigate the effect of substrate, surface roughness, and hydraulic residence time (HRT) on Streptococcus mutans biofilms growing on dental composites under conditions relevant to the oral cavity. MethodsDental composites were prepared with varying amounts of polishing and incubated in a CDC bioreactor with an approximate shear of 0.4 Pa. S. mutans biofilms developed in the bioreactors fed sucrose or glucose and at 10-h or 40-h HRT for one week. Biofilms were characterized by confocal laser microscopy (CLM). Composite surface roughness was characterized by optical profilometry, and pre- and post-incubation composite surface fine structure and elemental composition were determined using scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS). ResultsPolishing had a significant impact on surface roughness, varying by a factor of 15 between the polished samples and the unpolished control. S. mutans biofilms grew statistically significantly thicker on the unpolished composites. Biofilm thickness was greater at shorter 10-h HRT compared to 40-h HRT. In most cases, biofilm thickness was not statistically significantly greater in sucrose-fed bioreactors than in glucose-fed bioreactors. SEM-EDS analysis did not identify any significant change in elemental composition after aging. ConclusionsAccurate characterization of oral cavity biofilms must consider shear forces and the use of techniques that minimize alteration of the biofilm structure. Under shear, surface smoothness is the most important factor determining S. mutans biofilm thickness followed by HRT, while sucrose presence did not result in significantly greater biofilm thickness. Clinical SignificanceThe obvious patterns of S. mutans growth along sub-micron scale grooving created by the polishing process suggested that initial biofilm attachment occurred in the shear-protected grooves. These results suggest that fine polishing may help prevent the initial formation of S. mutans biofilms compared to unpolished/coarse polished composites.

Inhibitory Activity of Flower Extracts from Salvia deserta Schang on Streptococcus mutans

To examine the in vitro inhibitory effect of flower extracts from Salvia deserta Schang (SFE) on Streptococcu smutans ( S. mutans). The inhibitory effect of SFE on planktonic S. mutans and the effect of SFE on the growth process of planktonic S. mutans were determined by the agar drilling method and the microdilution method. Crystal violet staining and MTT reduction assay were conducted to determine the effect of SFE on S. mutans biofilm formation. The effect of SFE on the production of exopolysaccharides (EPS) in S. mutans biofilm was determined by anthrone-sulfuric acid method. The intracellular lactate dehydrogenase (LDH) activity in S. mutans was determined by LDH colorimetric assay. The effects of SFE on the acid-producing capacity of S. mutans was determined by pH meter. The minimum inhibitory concentration (MIC) of SFE against S. mutans was 14 μg/μL. SFE of the the concentration between 1/8 MIC and MIC could inhibit the growth rate of S. mutans within 30 h and it could significantly inhibit the LDH activity compared with the control group ( P<0.0001). SFE of the concentration between 4 MIC and 1/4 MIC had an inhibitory effect on the acid production of S. mutans ( P<0.001). Moreover, it could effectively restrain the formation of S. mutans biofilm and significantly reduce the amount of EPS produced by biofilm ( P<0.01). SFE can effectively inhibit the activity of S. mutans and its biofilm. The mechanism of inhibiting S. mutans by SFE was preliminarily discussed as follows, it interferes with microbial adhesion and aggregation by reducing the production of bacterial EPS, thus inhibiting the formation of bacterial biofilms. In addition, it interferes with glycolysis of S. mutans by reducing the LDH activity of bacteria, thus inhibiting the acid production of S. mutans.

Development, characterization and antimicrobial activity of multilayer silica nanoparticles with chlorhexidine incorporated into dental composites.

In this study a dentistry nanocomposite with prolonged antibacterial activity using silica nanoparticles (SNPs) loaded with chlorhexidine (CHX) was developed. SNPs were coated with the Layer-by-Layer technique. Dental composites were prepared with organic matrix of BisGMA/TEGDMA and SNPs with or without CHX (0, 10, 20 or 30% w/w). The physicochemical properties of the developed material were evaluated and agar diffusion method was used to test the antibacterial. In addition, the biofilm inhibitory activity of the composites was evaluated against S. mutans. SNPs were rounded with diameters about 50nm, the organic load increased with increasing deposited layers. Material samples with SNPs loaded with CHX (CHX-SNPs) showed the highest values of post-gel volumetric shrinkage, that ranged from 0.3% to 0.81%. Samples containing CHX-SNPs 30% w/w showed the highest values of flexural strength and modulus of elasticity. Only samples containing SNPs-CHX showed growth inhibition against S. mutans, S. mitis and S. gordonii in a concentration-dependent manner. The composites with CHX-SNPs reduced the biofilm formation of S. mutans biofilm at 24h and 72h. The nanoparticle studied acted as fillers and did not interfere with the evaluated physicochemical properties while providing antimicrobial activity against streptococci. Therefore, this initial study is a step forward to the synthesis of experimental composites with improved performance using CHX-SNPs.

Study of Streptococcus mutans in Early Biofilms at the Surfaces of Various Dental Composite Resins.

Biofilm deposit on the composite restoration is a common phenomenon and bacterial growth follows the deposition. The study aims to evaluate Streptococcus mutans (S. mutans) early biofilm formation on the surfaces of various dental composite resins by using the real-time quantitative polymerase chain reaction (qPCR) technique. Thirty-two discs, where eight discs were in each group of Filtek Supreme Ultra (FSU; 3M, St. Paul, MN), Clearfil AP-X (APX; Kuraray Noritake Dental Inc., Tokyo, Japan), Beautifil II (BE2; Shofu, Inc., Kyoto, Japan), and Estelite Sigma Quick (ESQ; Tokuyama Dental, Tokyo, Japan), were fabricated and subjected to S. mutans biofilm formation in an oral biofilm reactor for 12 hours. Contact angles (CA) were measured on the freshly fabricated specimen. The attached biofilms underwent fluorescent microscopy (FM). S. mutans from biofilms were analyzed using a qPCR technique. Surface roughness (Sa) measurements were taken before and after biofilm formation. Scanning electron microscopy (SEM), including energy dispersive X-ray spectrometer (EDS) analysis, was also performed for detecting relative elements on biofilms. The study showed that FSUdemonstrated the lowest CA while APXpresented the highest values. FMrevealed that condensed biofilm clusters were most on FSU. The qPCR results indicated the highest S. mutans DNA copies in the biofilm were on FSU while BE2was the lowest (p < 0.05). Sa test signified that APX was significantly the lowest among all materials while FSU was the highest (p < 0.05). SEM displayed areas with apparently glucan-free S. mutans more on BE2 compared to APX and ESQ, while FSU had the least. Small white particles detected predominantly on the biofilms of BE2 appeared to be Si, Al, and F extruded from the resin. Differences in early biofilm formation onto various composite resins are dependent on the differences in materialcompositions and their surface properties. BE2 showed the lowest quantity of biofilm accumulation compared to other resin composites (APX, ESQ, and FSU). This could be attributed to BE2 proprieties as a giomer and fluoride content.

Evaluation of photodynamic therapy on nanoparticles and films loaded-nanoparticles based on chitosan/alginate for curcumin delivery in oral biofilms

Nanoparticles and nanoparticle-loaded films based on chitosan/sodium alginate with curcumin (CUR) are promising strategies to improve the efficacy of antimicrobial photodynamic therapy (aPDT) for the treatment of oral biofilms. This work aimed to develop and evaluate the nanoparticles based on chitosan and sodium alginate encapsulated with CUR dispersed in polymeric films associated with aPDT in oral biofilms. The NPs were obtained by polyelectrolytic complexation, and the films were prepared by solvent evaporation. The photodynamic effect was evaluated by counting Colony Forming Units (CFU/mL). Both systems showed adequate characterization parameters for CUR release. Nanoparticles controlled the release of CUR for a longer period than the nanoparticle-loaded films in simulated saliva media. Control and CUR-loaded nanoparticles showed a significant reduction of 3 log10 CFU/mL against S. mutans biofilms, compared to treatment without light. However, biofilms of S. mutans showed no photoinactivation effect using films loaded with nanoparticles even in the presence of light. These results demonstrate the potential of chitosan/sodium alginate nanoparticles associated with aPDT as carriers for the oral delivery of CUR, offering new possibilities to improve the treatment of dental caries and infections. This work will contribute to advances in the search for innovative delivery systems in dentistry.

Effect of different finishing and polishing procedures on surface roughness and microbial adhesion on highly-filled composites for injectable mold technique.

To evaluate the effect of different finishing and polishing procedures on surface roughness and microbial adhesion and viability of Streptococcus mutans on novel highly-filled composites for injectable mold technique. One hundred sixty specimens were divided into four material groups: FSF (Filtek Supreme Flowable Restorative), TE (Tetric EvoFlow), GUI (G-aenial Universal Injectable) and GUF (G-aenial Universal Flo). Within each group, specimens were split regarding finishing and polishing (F/P) procedures into: SLD (Sof-Lex Discs), SLS (Sof-Lex Spirals), OG (OneGloss) and PG (PoGo). Surface roughness was analyzed using profilometer (Ra and Rz) and scanning electron microscopy (SEM) and Strep. mutans biofilm formation was analyzed using colony forming unit (CFU) and cell viability assay. Two-way analysis of variance (ANOVA) followed by Tukey's post hoc test were used for comparison among groups, Pearson's coefficient was applied for the correlation between Ra and CFU/ml and all data were presented as mean ± SD. Both materials and F/P procedures affect Ra, Rz and Strep. mutans CFU/ml values (p ≤ 0.05). Considering the Ra and Rz, GUI and GUF revealed lower values, compared to FSF and TE and SLD and SLS revealed lower values, compared to OG and PG. Considering the Strep. mutans CFU/ml and viability, GUI and TE presented lower values, compared to GUF and FSF and SLD and SLS presented lower values, compared to OG and PG. Moderate positive correlation was found between Ra and CFU/ml (r = 0.552). The smoothest surfaces possess GUI and GUF, among materials and SLD and SLS, among F/P procedures. GUI adhered the lowest amount of Strep. mutans, due to the smoothest surfaces. FSF and GUF revealed the highest amount of Strep. mutans, due to bis-GMA, bis-MEPP and TEGDMA in their composition. The findings of the present study may be beneficial for the proper selection of highly-filled composites and an adequate finishing and polishing procedure when performing the injectable mold composite resin veneer technique.

Comparison Effect of Leaves and Bark Extract of Eucalyptus (Melaleuca leucadendra), Sappan (Caesalpinia sappan), and Cinnamon (Cinnamomum zeylanicum) to Reduce Streptococcus mutans Biofilm Formation

To evaluate the effect of eucalyptus, sappan, and cinnamon leaf and bark extract on the percentage of S. mutans biofilm formation. The test group was divided into a negative control (1% Dimethyl sulfoxide), a treatment group (eucalyptus, sappan, and cinnamon leaves and bark at concentrations of 50%, 25%, and 12.5%, respectively), and a positive control (0.2% chlorhexidine). Each concentration of eucalyptus, sappan, and cinnamon leaves and barks extract, Brain Heart Infusion Broth (BHI-B), bacteria according to the McFarland 0.5 standard, a positive control, and a negative control were added to a 96-well microplate. They were incubated at 37 °C for twenty-four hours before being rinsed with phosphate buffer saline (PBS) and stained with 0.1% crystal violet. The optical density was then measured using a microplate reader with a 540 nm wavelength. The absorbance value is then factored into the percentage of bacterial adhesion inhibition formula. A statistical test revealed a significant difference (p &lt; 0.05) in the percentage of inhibition between the three extracts of leaves and bark and the negative control group (1% DMSO). There is a significant difference between all leaves and barks concentration and the positive control group except the 50% concentration of sappan leaf group and 50% concentration of cinnamon leaf group. The sappan leaf and cinnamon leaf at 50% concentration exhibit no significant difference (p &gt; 0.05) with the positive control. Eucalyptus, sappan, cinnamon leaves and barks at a concentration of 50% demonstrated effectiveness of the extractant in inhibiting the formation of biofilm masses by S. mutans in comparison with the other group.

Anti-Bacterial and Anti-Biofilm Activities of Anandamide against the Cariogenic Streptococcus mutans

Streptococcus mutans is a cariogenic bacterium in the oral cavity involved in plaque formation and dental caries. The endocannabinoid anandamide (AEA), a naturally occurring bioactive lipid, has been shown to have anti-bacterial and anti-biofilm activities against Staphylococcus aureus. We aimed here to study its effects on S. mutans viability, biofilm formation and extracellular polysaccharide substance (EPS) production. S. mutans were cultivated in the absence or presence of various concentrations of AEA, and the planktonic growth was followed by changes in optical density (OD) and colony-forming units (CFU). The resulting biofilms were examined by MTT metabolic assay, Crystal Violet (CV) staining, spinning disk confocal microscopy (SDCM) and high-resolution scanning electron microscopy (HR-SEM). The EPS production was determined by Congo Red and fluorescent dextran staining. Membrane potential and membrane permeability were determined by diethyloxacarbocyanine iodide (DiOC2(3)) and SYTO 9/propidium iodide (PI) staining, respectively, using flow cytometry. We observed that AEA was bactericidal to S. mutans at 12.5 µg/mL and prevented biofilm formation at the same concentration. AEA reduced the biofilm thickness and biomass with concomitant reduction in total EPS production, although there was a net increase in EPS per bacterium. Preformed biofilms were significantly affected at 50 µg/mL AEA. We further show that AEA increased the membrane permeability and induced membrane hyperpolarization of these bacteria. AEA caused S. mutans to become elongated at the minimum inhibitory concentration (MIC). Gene expression studies showed a significant increase in the cell division gene ftsZ. The concentrations of AEA needed for the anti-bacterial effects were below the cytotoxic concentration for normal Vero epithelial cells. Altogether, our data show that AEA has anti-bacterial and anti-biofilm activities against S. mutans and may have a potential role in preventing biofilms as a therapeutic measure.

&lt;i&gt;In vitro&lt;/i&gt; Evaluation of Antibiofilm Activity of Methanolic Leaf Extract of &lt;i&gt;Azadirachta indica&lt;/i&gt; on Cariogenic &lt;i&gt;Streptococcus mutans&lt;/i&gt;

Dental caries is a biofilm-dependent disease, and Streptococcus mutans is the primary etiological agent involved in the initiation of the disease. The extensive use of a limited range of antimicrobial drugs in dentistry has led to the development of drug-resistant bacteria. There is an increasing need to find new alternatives against drug-resistant bacteria. Globally, there is a continuous effort towards identifying natural anti-caries agents for the prevention and better management of caries. The objective of the present study was to evaluate the antibiofilm potential of Azadirachta indica leaf methanolic (ALM) extract against S. mutans biofilm. The study employed a standard reference strain of S. mutans MTCC 497, for in vitro standardisation of biofilm by microtiter plate assay. The antibiofilm activity of the ALM extract was evaluated against the S. mutans strain, and the same was confirmed by light and scanning electron microscopy (SEM). The in vitro biofilm standardisation results demonstrated that 50 µl/ml of S. mutans inoculum concentration exhibited a much superior biofilm formation than the other concentrations employed. Light microscopy and SEM images revealed that ALM extract at 100 mg/ml concentration significantly inhibited the S. mutans biofilm. To conclude, the study reports that the A. indica leaf extract is a potential source to inhibit the S. mutans biofilm. Further studies are warranted to identify the phytochemicals responsible for the antibiofilm activity of ALM extract against S. mutans biofilm that aid in the design of natural anti-caries products.