Anticariogenic Activity Research Articles

Macelignan suppresses Porphyromonas gingivalis supernatant-stimulated urokinase-type plasminogen activator expression via signal transduction in human KB oral cells

Macelignan, a bioactive compound isolated from Myristica fragrans Houtt. or nutmeg, has been reported for its anti-cariogenic and anti-biofilm activities for dental plaque control [1,2]. However, its efficacy to block the expression of urokinase-type plasminogen activator (uPA), a serine protease that is expressed in various inflamed and normal healing cell types in response to cytokines and bacterial products, for periodontal inflammation treatment has not been investigated. This study was aimed to examine whether macelignan suppressed Porphyromonas gingivalis supernatant-stimulated uPA expression through regulation of mitogen-activated protein kinase (MAPK) and activating protein (AP)-1 signalings in human KB oral cells by performing casein zymography, reverse transcription-PCR, Western blotting, and reporter gene assays. The main caseinolytic band secreted from the cells was found to be migrated at 54 kDa and represented uPA. Macelignan dose-dependently inhibited the expression of uPA activity, protein, and gene in KB cells in response to P. gingivalis supernatant. In accordance with these findings, macelignan effectively decreased phosphorylation of p38 and c-jun N terminal kinase (JNK) in P. gingivalis supernatant-stimulated KB cells. The levels of c-jun phosphorylation and c-fos expression, which composed of AP-1 transcription factor for uPA gene expression, were also reduced by macelignan in KB cells exposed to P. gingivalis supernatant. In linear with these results, macelignan was found to block P. gingivalis supernatant-stimulated AP-1 activity in KB cells. These results suggest that macelignan decreased P. gingivalis supernatant-stimulated uPA expression by blocking AP-1 activity which may be facilitated by inhibiting phosphorylation of p38 and JNK in KB cells.

Protective effect of yogurt extract on dental enamel demineralization in vitro

Casein phosphopeptides (CPPs) are phosphorylated casein-derived peptides produced synthetically by proteolytic digestion of alpha(s1)-, alpha(s2)- and beta-casein. The anticariogenic activity of CPPs is due to their ability to stabilize high levels of amorphous calcium phosphate (ACP) on tooth surface, preventing demineralization and enhancing remineralization of enamel caries. The aim of this study was to test the in vitro ability of natural CPPs (contained in yogurt) to prevent demineralization and promote remineralization of dental enamel. Eighty human molars were used. After standardizing an in vitro demineralization procedure for producing artificial caries (Group 1: pH 4.8; Group 2: pH 3.97), this procedure was used on teeth, but with the addition of natural CPPs (Group 3: pH 4.8; Group 4: pH 3.97). The effects of these procedures were evaluated by quantitative analysis (change in weight and calcium titration) and qualitative analysis (SEM). Statistical analysis of the results was performed using ANOVA. Statistical analysis showed significant differences in weight changes between the groups with and without natural CPPs. The SEM observation showed the protective effects of natural CPPs. The results demonstrated that CPPs contained in yogurt have an inhibitory effect on demineralization and promote the remineralization of dental enamel.

Preparation of calcium fluoroaluminosilicate glasses containing sodium and phosphorus by the nonhydrolytic sol–gel method

Aluminum and silicon oxide-based inorganic matrices have been extensively studied because of their countless applications. Dental cements consist of aluminum and silicon-based amorphous polymeric materials containing fluoride, sodium, phosphorus and calcium, which are also good candidates for bone replacement. The nonhydrolytic sol–gel method has emerged as an alternative route for the preparation of these materials under milder conditions than those employed in traditional methods, such as oxide fusion. The main advantages of the nonhydrolytic sol–gel method include the use of low temperatures and the ready availability and easy purification of the precursors. Together, these factors have contributed to the production of highly pure materials with controlled porosity and nanometric particles. Dental restorations based on aluminosilicate matrices are known as glass ionomer cements. These materials have interesting physical and dental properties, mainly because they display anticariogenic activity and exhibit prolonged adhesiveness to the dental structure. The base of the ionomer is an aluminosilicate that is industrially synthesized by the fusion of SiO 2, Al 2O 3, AlF 3, CaF 2, NaF and AlPO 4, in various concentrations. The characterizations conducted in this study reveal that this ionomer displays interesting properties, so its use as a precursor of dental cement and a biomaterial for bone replacement is highly recommended.

구강병인균에 대한 마와 꿀풀추출물의 항균.항우식효과

Yam, Prunella was stepwise extracted with hexane, chloroform, ethyl acetate, butanol, and water. Anti-microbial activity of each extract was investigated. Hexane extract was tested for anti-microbial effect on Streptocaccus mutans, one of causative factor of dental caries. Methanol extracts of 7 plants were investigated to anti-microbial effects on S. mutans KCTC 5316, P. gingivalis KCTC 5352, S. aureus KCTC 1927 by means of agar diffusion method. Methanol extract of Yam and Prunella revealed anti-microbial activity against S. mutans, P. gingivalis, and S. aureus. Also, hexane fraction of Yam revealed anti-microbial activity against S. mutans. In sequence of hexane, chloroform, ethylacetate, butanol fraction by Prunelia acted as potent anti-microbial agent on P. gingivalis. The measured MIC of hexane fraction of Yam and Prunella on S. mutans KCTC 5316 strain was 0.25 mg/ml and 0.5 mg/ml and the MIC of hexane fraction of Prunella on S. aureus was 0.5 mg/ml. The hexane fraction of Yam and Prunella suppressed viable ceil counts(VCC) of S. mutans, especially after 24 hrs. The Prunella hexane fraction suppressed VCC of S. aureus, after 12 and 24 hrs. Tested concentrations were 0.1, 0.25 and 0.5 mg/ml. the results were compared with control (0 mg/ml). The pH of S. mutans media and GTase activity were determined to evaluate the anticariogenic activity of Yam, Prunella hexane fraction. The pH were increased from 5.6 to 7.0-7.2 in concentration of 2.0 mg/ml. Yam hexane extraction revealed 35% inhibition to GTase activity and Punella inhibited 25% of GTase. These results suggest that the hexane extracts of Yam and prunella have Antibacterial activities against S. mutans, P. gingivalis, S. aureus and have preventive effect on dental caries.

Development of commercial mouthwash combined with herbal extracts for anticariogenic activity against Streptococcus mutans

The aim of the present study was to develope the commercial mouthwash combined with herbal extracts against Streptococcus mutans ATCC 25175. The 50% ethanol herbal extracts were from Cassia alata L., Quercus infectoria Oliver., Harrisonia perforata Merr., Camellia sinensis L. (oolong and green tea) and Psidium guajava L.,

The Crude Aqueous Extract of Piper betle L. and its Antibacterial Effect Towards Streptococcus mutans

In this study, the antimicrobial influence of crude aqueous extract of Piper betle L. on Streptococcus mutans (S. mutans) was investigated. The focus of the antimicrobial effects includes the ultrastructure and acid producing properties of S. mutans. Transmission electron microscopy (TEM) was used to determine the effect of the extract on the ultrastructure of S. mutans. Analysis of the effect on the acid producing properties was analyzed by pH drop assay. The investigation was further carried out to determine the possible chemical components of the extract using thin layer chromatography (TLC), bioautography and gas chromatography mass spectrometry (GCMS). From the micrographs of the transmission electron, it was found that the crude extract of Piper betle L. leaves causes plasma cell membrane damage and coagulation of the nucleoid. The extract was found to significantly reduce acid producing properties of the bacteria. Chemical analysis of the extract showed that hydroxychavicol, fatty acids (stearic and palmitic) and hydroxy fatty acid esters (stearic, palmitic and myristic) as the main components. It was suggested from the results obtained that the crude extract of Piper betle L. leaves may exert anticariogenic activities that are related to decrease in acid production and changes to the ultrastructure of S. mutans. Further study will be carried out to determine if the effect observed is attributed to the presence of hydroxychavicol, fatty acids and hydroxy fatty acid esters in the extract.

In vitro anti-cariogenic activity of dichloromethane fraction fromRheum undulatum L. root

This study aimed to evaluate in vitro effects of Rheum undulatum L. root on the development of dental caries, especially its effects on viability, dental plaque formation, and glycolytic acid production of Streptococcus mutans and Streptococcus sobrinus. Methanol extract of Rheum undulatum L. root and its fractions were prepared and tested. Among the test extract and fractions, dichloromethane fraction (DF) showed the most active antibacterial activity (inhibition zone: 13-17 mm) against S. mutans and S. sobrinus in a disc diffusion method. Minimal inhibitory concentrations (MICs) of DF against these bacteria ranged from 0.25 to 0.5 mg/mL. Furthermore, DF significantly inhibited the caries-inducing factors of these bacteria. At sub-MIC levels, DF inhibited in vitro dental plaque formation by S. mutans and S. sobrinus (IC50= 0.079 and 0.142 mg/mL, respectively), which was caused, in part, by the inhibitory effect on the activity of glucosyltransferases. A significant reduction of glycolytic acid production was found at the concentration as low as 0.032 mg/mL for S. mutans and 0.063 mg/mL for S. sobrinus. The possible bioactive compounds that are inducing in vitro anti-cariogenic activity of DF are unknown. Based on the preliminary phytochemical analysis, the activity of DF may be related to the presence of anthraquinones, cardiac glycosides, coumarines, sterols/terpenes, and phenolics. These results indicate that DF is probably useful for the control of dental plaque formation and subsequent dental caries development.

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.

Anticariogenic activity of some tropical medicinal plants against Streptococcus mutans

The methanol extracts of five tropical plants, Baeckea frutescens, Glycyrrhiza glabra, Kaempferia pandurata, Physalis angulata and Quercus infectoria, exhibited potent antibacterial activity against the cariogenic bacterium Streptococcus mutans. In particular, G. glabra, K. pandurata and P. angulata conferred fast killing bactericidal effect against S. mutans in 2 min at 50 μg/ml of extract concentration.

충치균 (Streptococcus mutans)에 대한 다시마 추출물의 항균활성

우리 나라 연안에서 채취한 해조류 27종의 <TEX>$80\%$</TEX> ethanol 추출물이 Streptococcus mutans KCTC 3300의 증식에 미치는 영향을 검색하고, 다시마의 추출용매별 항균성과, <TEX>$80\%$</TEX> ethanol 추출물을 용매분획하였을 때 획분별 MIC를 측정하였다. S. mutans KCTC 3300에 대한 해조류 추출물의 항균활성은 다시마가 가장 강하였고, 갈파래, 청각, 감태, 곰피 및 미역도 비교적 높은 활성을 나타내었으며 홍조류보다 갈조류와 녹조류에 속하는 종이 대체적으로 강하였다 다시마의 항균활성은 추출용매에 따라 차이가 있었으나 용매별 가용성 획분의 량과 항균성간에는 뚜렷한 관련이 나타나지 않았다. 5. mutans KCTC 3300과 S. sobrinus KCTC 3307에 대한 해조류의 <TEX>$80\%$</TEX> ethanol 추출물의 MIC는 해조류의 종뿐만 아니라 동일한 해조류 추출물의 경우에도 균종에 따라 차이가 있었다. 다시마의 <TEX>$80\%$</TEX> ethanol 추출물을 유기용매를 사용하여 순차 분획하였을 때 항균활성 물질의 대부분은 ether 획 분에 이행하여 이 획분의 MIC는 105<TEX>$\mu$</TEX>g/mL이었고, 수용성 획분은 약 5,500<TEX>$\mu$</TEX>g/mL의 농도에서도 균의 증식을 저지하지 못하였다. In previous reports the authors have screened the inhibition effects of marine algae extracts on halitosis, and demonstrated that a brown algae, Eisenia bicyclis (' Daehwang') possess not only strong deodorant effect bug also considerable anticariogenic activities. In this study, we screened antibacterial effects of various marine algae, and measured minimum inhibitory concentration (MIC) value of them against mutans streptococci in vitro. Among the 27 species of marine algae, <TEX>$80\%$</TEX> ethanol extract of dried sea-mustard, Laminaria japonica ('Dasima') showed the strongest inhibition activity against Streptococcus mutans KCTC 3300. The extracts of Ulva lactuca ('Galparae'), Codium fragile ('Cheonggak'), Ecklonia cava ('Gamtae'), E. stolonifera ('Gompi') and Undalia Pinnatifida ('Miyeok') showed slightly weaker inhibitory potency than L. japonica. Differences of MIC values in <TEX>$80\%$</TEX> ethanol extract of some species of marine algae were observed depending on test bacterial species, i.e., S. mutans KCTC 3300 or S. sobrinus KCTC 3307. Eighty percent ethanol extract of dried L japonica was fractionated with diethyl ether, chloroform, ethyl acetate, n-buthanol and water successively, The ether-soluble fraction had inhibitory effect on S. mutans KCTC 3300, however the inhibitory effects were not found in the other fractions. The MIC values of <TEX>$80\%$</TEX> ethanol extract and ether fraction were 180 and 105 <TEX>$\mu$</TEX>g/mL respectively, while no significant inhibition activity of water-soluble fraction was found even when the fraction was added up to 5,500 <TEX>$\mu$</TEX>g/mL.

Anti-cariogenic properties of tea (Camellia sinensis).

Various components in green and black tea, the beverages made by infusing appropriately processed dried leaves of Camellia sinensis, notably simple catechins, have properties in vitro that suggest an anti-cariogenic activity. These include: a direct bactericidal effect against Streptococcus mutans and S. sobrinus; prevention of bacterial adherence to teeth; inhibition of glucosyl transferase, thus limiting the biosynthesis of sticky glucan; inhibition of human and bacterial amylases. Studies in animal models show that these in-vitro effects can translate into caries prevention. A limited number of clinical trials in man suggest that regular tea drinking may reduce the incidence and severity of caries. If substantiated, this could offer a very economical public health intervention.

Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: A review

Using laboratory, animal, and human in situ caries models, investigators have shown that casein phosphopeptide amorphous calcium phosphate complexes (CPP-ACP) exhibit an anticariogenic activity. The casein phosphopeptides (CPP) are produced from a tryptic digest of the milk protein casein by aggregation with calcium phosphate and purification by ultrafiltration. The CPP have a remarkable ability to stabilize calcium phosphate in solution and substantially increase the level of calcium phosphate in dental plaque. Through their multiple phosphoseryl residues, the CPP bind to forming clusters of amorphous calcium phosphate (ACP) in metastable solution, preventing their growth to the critical size required for nucleation and precipitation. The proposed mechanism of anticariogenicity for the CPP-ACP is that they localize ACP in dental plaque, which buffers the free calcium and phosphate lon activities, thereby helping to maintain a state of supersaturation with respect to tooth enamel depressing demineralization and enhancing remineralization. The CPP-ACP, unlike fluoride, can be added to sugar-containing foods and therefore have commercial potential as an additive to foods as well as to toothpastes and mouthwashes for the control of dental caries.

Phosphopeptides from Grana Padano cheese: nature, origin and changes during ripening.

Casein phosphopeptides (CPP) which develop in Grana Padano cheese at different ages were isolated by precipitation with Ba2+ and analysed by HPLC. Profiles were complex throughout the period between 4 and 38 months. CPP in a cheese sample 14 months old were identified by a combination of fast atom bombardment-mass spectrometry and Edman degradation. They were found to consist of a mixture of components derived from three parent peptides, beta-CNf(7-28)4P, alpha s1-CNf(61-79)4P and alpha s2-CNf(7-21)4P. In total, 45 phosphopeptides were identified: 24 from beta-CN, 16 from alpha s1-CN and 5 from alpha s2-CN. The presence of aminopeptidase activity during cheese ripening was deduced from the presence of a number of CPP of different lengths with the loss of one or more residues from the N-terminus. The longest had C-terminal lysine and seemed to be progressively hydrolysed by carboxypeptidases A and B to shorter peptides. CPP in cheese appeared to be shortened plasmin-mediated products. Moreover, those most resistant to further hydrolysis contained at least three closely located phosphoserine residues. The anticariogenic activity of CPP is also discussed.

Hard-tissue substrates for evaluation of cariogenic and anti-cariogenic activity in situ.

Hard-tissue substrates include primarily human and bovine enamel and human dentin. They have been used for in situ studies in a natural or sound condition, as well as flattened or containing an in vitro-formed caries-like lesion. Human enamel and dentin are generally the substrates of choice for studies of coronal and root-surface caries, respectively, but bovine enamel appears to offer a suitable alternative for many studies of enamel caries. Substrates with caries-like lesions will respond more rapidly to changes in the intra-oral mineral equilibrium and will allow both demineralization and remineralization to be determined. Findings from some studies suggest that caries-like lesions may respond somewhat differently, depending upon the degree of mineralization of the surface layer. Because in vitro findings with dentin show it to be significantly more soluble in acid than enamel, results from a study that used dentin may not be directly applicable to enamel. Both enamel and dentin substrates can be used in thin-section models. Hard-tissue substrates can also differ, depending upon their intra-oral location. Locations that result in the accumulation of plaque will behave differently from those that are plaque-free. So that plaque would accumulate, substrates have been placed approximally, beneath a fabric or steel mesh, in a protected trough, beneath a metal band or within a depression on the buccal surface. For studies requiring a determination of both demineralization and remineralization, human enamel or dentin containing a surface-softened caries-like lesion and covered with a uniform natural plaque are the substrates of choice.

Findings from an in situ thin-section sandwich model for evaluating cariogenic and anti-cariogenic activity.

This report summarizes the findings from eight individual remineralization studies that used the thin-section sandwich model. This model uses thin sections of human enamel (or dentin) containing caries-like lesions sandwiched between sheets of thin plastic which are then implanted into the buccal surfaces of molars in removable partial mandibular dentures. Toothbrushing with a sodium fluoride-positive control dentifrice for two-week periods always produced positive remineralization of the lesions, and similar use of a placebo always produced mean mineral loss. In most studies, the difference was significant at p less than 0.01. However, variation among specimens placed within an individual mouth were often large and even included cases where one lesion gained mineral while a closely adjacent lesion lost mineral. Similarly, mean mineral changes produced by an individual subject were not consistent from study to study and ranged from net demineralization in one study to net remineralization in another study. Nevertheless, mean mineral changes from all eight studies showed that all subjects produced more remineralization than demineralization when using a sodium fluoride-positive control dentifrice. Data are presented showing remineralization of thin-section lesions from a number of other fluoride and non-fluoride dentifrices and mouthrinses and showing that a product with reduced anti-caries activity (as measured by several tests) also resulted in reduced remineralization by the thin-section model.

Anticariogenic Activity of Licorice and Glycyrrhizine I: Inhibition of In Vitro Plaque Formation by Streptococcus mutans

The effect of licorice and its active sweet component glycyrrhizin was tested on the growth and adherence to glass of the cariogenic Streptococcus mutans. Neither licorice nor glycyrrhizin promoted growth or induced plaque formation. In the presence of sucrose, glycyrrhizin did not affect bacterial growth, but the adherence (plaque formation) was markedly inhibited. At 0.5–1% glycyrrhizin, inhibition was almost complete. These results support our previous suggestions that glycyrrhizin might serve as an efficient vehicle for topical oral medications.

Anticariogenic effects of tea in rats.

Teas varying in fluoride and tannin concentration were evaluated in rats for anticariogenic activity. There was a direct correlation between fluoride in tea and the inhibition of sulcal caries in rats, whereas no relationship was observed between tannin and this type of lesion. Teas also had a significant effect on caries progression and imparted a black stain to the teeth.

Studies on the anticariogenic activity of oat hulls.

Studies on the anticariogenic activity of oat hulls.