Antimicrobial activity of rhodomyrtone isolated from Rhodomyrtus tomentosa (Aiton) Hassk.

Rhodomyrtone was isolated from the leaves of Rhodomyrtus tomentosa (Aiton) Hassk grown in Vietnam using chromatographic methods. Its chemical structure was confirmed by means of spectroscopic data analysis. The pH drop measurement, enzyme activity assays and fluorescence stain were used to examine rhodomyrtone anticaries activity. It was found that rhodomyrtone suppressed acid production by Streptococcus mutans, a major cariogenic agent in human by inhibiting enzyme activities responsible for acid production and tolerance, including membrane bound enzymes F-ATPase and phosphotransferase system (PTS), as well as glycolysis enzymes glyceraldehyphosphate dehydrogenase (GAPDH) and pyruvate kinase (PK) in cytoplasm with the IC50 values of 24 µM, 19 µM, 23 µM and 28 µM, respectively. Moreover, 50 µM rhodomyrtone reduced biofilm biomass formed by S. mutans up to 59% (p < 0.05). Fluorescent images indicated that cells on the biofilms were significantly killed. Thus, rhodomyrtone is a new and potential anticaries agent against S. mutans.

Inhibitory effect of Aralia continentalis on the cariogenic properties of Streptococcus mutans.

ETHNOPHARMACOLOGICAL RELEVANCE: Aralia continentalis has been used in traditional Korean medicine for dental diseases such as toothache, dental caries, periodontal disease and gingivitis, and also has been used for neuralgia, analgesia, sweating, and as an antirheumatic. AIM OF THE STUDY: The present study was designed to investigate the inhibitory effect of Aralia continentalis extract on cariogenic properties of Streptococcus mutans, which is one of the most important bacteria in the formation of dental caries and dental plaque. MATERIALS AND METHODS: The inhibitory effects of Aralia continentalis extract on the growth, acid production, water-insoluble glucan synthesis, and adhesion were investigated in Streptococcus mutans. The biofilm formation of Streptococcus mutans was determined by scanning electron microscopy (SEM) and safranin staining. RESULTS: The ethanol extract of Aralia continentalis showed concentration dependent inhibitory activity on the growth of Streptococcus mutans and significant inhibition of acid production at the concentrations of 0.25, 0.5, 1, 2 and 4 mg/ml compared to the control group. The synthesis of water-insoluble glucan by glucosyltransferase (GTFase) was decreased in the presence of 0.5-4 mg/ml of the extract of Aralia continentalis. The extract markedly inhibited Streptococcus mutans adherence to saliva-coated hydroxyapatite beads (S-HAs). The extract of Aralia continentalis has an inhibitory effect on the formation of Streptococcus mutans biofilms at the concentrations higher than 2mg/ml. CONCLUSIONS: These results suggest that Aralia continentalis may inhibit cariogenic properties of Streptococcus mutans, and also may support the scientific rationale that native inhabitants used the extract for the treatment of dental diseases.

Novel compound from Trachyspermum ammi (Ajowan caraway) seeds with antibiofilm and antiadherence activities against Streptococcus mutans: a potential chemotherapeutic agent against dental caries.

AIM: The objective of this study was to isolate and characterize the active compound from Trachyspermum ammi seeds, exhibiting antibiofilm activity against Streptococcus mutans, a major causal organism of dental caries. METHODS AND RESULTS: Purification of the active compound from the seeds was performed by silica gel chromatography, and spectroscopic methods (FTIR, NMR and MS) were employed for its identification and structure determination. Antibiofilm and antiadherence activities of the active compound against S. mutans were analysed. Confocal microscopy was performed to visualize the effect of the compound on biofilm structure of S. mutans. Around 50% reduction was observed in adherence at 39·06 µg ml(-1) and in biofilm at 78·13 µg ml(-1) . It was found effective against adherent cells of S. mutans, reduced water-insoluble glucan synthesis and inhibited the reduction in pH. Confocal microscopy revealed scattered cells at sub-MIC concentration of the compound, resulting in distorted biofilm architecture in contrast to clustered cells seen in control. CONCLUSION: This study revealed a novel compound, a naphthalene derivative, isolated first time from T. ammi seeds with antibiofilm activity against S. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: Trachyspermum ammi represents an interesting source of a novel compound, (4aS, 5R, 8aS) 5, 8a-di-1-propyl-octahydronaphthalen-1-(2H)-one, with a great potential to be used as a therapeutic agent against dental caries.

Combined electrocoagulation and electroflotation for removal of fluoride from drinking water.

A combined electrocoagulation (EC) and electroflotation (EF) process was proposed to remove fluoride from drinking water. Its efficacy was investigated under different conditions. Experimental results showed that the combined process could remove fluoride effectively. The total hydraulic retention time required was only 30 min. After treatment, the fluoride concentration was reduced from initial 4.0-6.0mg/L to lower than 1.0mg/L. The influent pH value was found to be a very important variable that affected fluoride removal significantly. The optimal influent pH range is 6.0-7.0 at which not only can effective defluoridation be achieved, but also no pH readjustment is needed after treatment. In addition, it was found that SO(4)(2-) had negative effect; Ca(2+) had positive effect; while Cl(-) had little effect on the fluoride removal. The EC charge loading, EF charge loading and energy consumption were 3.0 Faradays/m(3), 1.5 Faradays/m(3), and 1.2 kWh/m(3), respectively, under typical conditions where fluoride was reduced from initial 4.0 to 0.87 mg/L.

In vitro anti-biofilm activity of macelignan isolated from Myristica fragrans Houtt. against oral primary colonizer bacteria.

In early dental plaque formation, oral primary colonizers such as Streptococcus mutans, Streptococcus sanguis and Actinomyces viscosus are initially attached to the pellicle-coated tooth surface to form a biofilm. The study aimed to determine the efficacy of macelignan, isolated from nutmeg (Myristica fragrans Houtt.), in removing each single oral primary biofilm in vitro on a polystyrene 96-well microtiter plate. Four biofilm growth phases (4, 12, 20 and 24 h) were evaluated in this study after treatment with macelignan at various concentrations (0.2, 2 and 10 microg/mL) and exposure times (5, 10 and 30 min). Anti-biofilm activity of macelignan was measured as the percentage of the remaining biofilm absorbance after macelignan treatment in comparison with the untreated control. At 24 h of biofilm growth, S. mutans, A. viscosus and S. sanguis biofilms were reduced by up to 30%, 30% and 38%, respectively, after treatment with 10 microg/mL macelignan for 5 min. Increasing the treatment time to 30 min resulted in a reduction of more than 50% of each of the single primary biofilms. The results indicate that macelignan is a potent natural anti-biofilm agent against oral primary colonizers.

Anticariogenic activity of macelignan isolated from Myristica fragrans (nutmeg) against Streptococcus mutans.

The occurrence of dental caries is mainly associated with oral pathogens, especially cariogenic Streptococcus mutans. Preliminary antibacterial screening revealed that the extract of Myristica fragrans, widely cultivated for the spice and flavor of foods, possessed strong inhibitory activity against S. mutans. The anticariogenic compound was successfully isolated from the methanol extract of M. fragrans by repeated silica gel chromatography, and its structure was identified as macelignan by instrumental analysis using 1D-NMR, 2D-NMR and EI-MS. The minimum inhibitory concentration (MIC) of macelignan against S. mutans was 3.9 microg/ml, which was much lower than those of other natural anticariogenic agents such as 15.6 microg/ml of sanguinarine, 250 microg/ml of eucalyptol, 500 microg/ml of menthol and thymol, and 1000 microg/ml of methyl salicylate. Macelignan also possessed preferential activity against other oral microorganisms such as Streptococcus sobrinus, Streptococcus salivarius, Streptococcus sanguis, Lactobacillus acidophilus and Lactobacillus casei in the MIC range of 2-31.3 microg/ml. In particular, the bactericidal test showed that macelignan, at a concentration of 20 microg/ml, completely inactivated S. mutans in 1 min. The specific activity and fast-effectiveness of macelignan against oral bacteria strongly suggest that it could be employed as a natural antibacterial agent in functional foods or oral care products.

Dental caries-protective agents in milk and milk products: investigations in vitro.

OBJECTIVES: To characterise and identify the anti-dental caries components that exist in milk and milk products. METHODS: Standard enamel or hydroxyapatite demineralisation tests were devised to simulate the action of acid on tooth mineral, and they were used to show which constituents of milk possessed a potential protective action against acid attack. RESULTS: Milk and milk products were fractionated and tested, revealing that minerals, including calcium and phosphorus, played a part in this protective process. The findings also drew attention to the effectiveness of minor milk protein or protein-associated components, the structures of which are mostly known, which were separated and characterised by gel electrophoresis as proteose-peptone fractions 3 and 5. The strength of adsorption of the protein or polypeptides in these fractions to dental enamel was measured and found to be sufficient to reduce the extent of demineralisation of enamel by acid buffer solutions. CONCLUSIONS: The removal of lactose, fat, casein and other proteins had little influence on the protective effect of the milk fractions. Besides calcium and phosphorus, milk contains other more powerful protective factors, which were identified as proteose-peptone fractions 3 and 5. Details of their composition and the strength of their adherence to the surface of dental mineral are given.

Identification of cariostatic substances in the cacao bean husk: their anti-glucosyltransferase and antibacterial activities.

The cacao bean husk has been shown to possess two types of cariostatic substances, one showing anti-glucosyltransferase (GTF) activity and the other antibacterial activity, and to inhibit experimental dental caries in rats infected with mutans streptococci. In the present study, chromatographic purification revealed high-molecular-weight polyphenolic compounds and unsaturated fatty acids as active components. The former, which showed strong anti-GTF activity, were polymeric epicatechins with C-4beta and C-8 intermolecular bonds estimated to be 4636 in molecular weight in an acetylated form. The latter, which showed bactericidal activity against Streptococcus mutans, were determined to be oleic and linoleic acids, and demonstrated a high level of activity at a concentration of 30 microgram/mL. The cariostatic activity of the cacao bean husk is likely caused by these biologically active constituents.

Anticaries effect of a component from shiitake (an edible mushroom).

The caries-inhibiting effect of the extract from shiitake (Lentinus edodes), the most popular edible mushroom in Japan, was studied both in vitro and in vivo. Shiitake extract showed an inhibitory effect on water-insoluble glucan formation from sucrose by crude glucosyltransferases of Streptococcus mutans JC-2 and Streptococcus sobrinus OMZ-176. The firmly adherent plaque in the artificial plaque formation test was strongly inhibited by shiitake extract. The reduction of firmly adherent plaque caused an increase in the incidence of non- and loosely adherent plaque and a decrease in total plaque formation. A significantly lower caries score was observed in specific pathogen-free rats infected with S. mutans JC-2 and fed with a cariogenic diet containing 0.25% shiitake extract as compared with controls fed the cariogenic diet without shiitake extract.