Saussurea lappa inhibits the growth, acid production, adhesion, and water-insoluble glucan synthesis of Streptococcus mutans

In the present study, inhibitory effects of the ethanol extract of Saussurea lappa (S. lappa) on the growth, acid production, adhesion, and water-insoluble glucan synthesis of Streptococcus mutans (S. mutans) were examined. The growth and acid production of Streptococcus mutans were inhibited by the presence of ethanol extract of Saussurea lappa (0.5-4 mg/ml) significantly. The ethanol extract of Saussurea lappa (0.25-4 mg/ml) also significantly lowered the adherence of Streptococcus mutans in a dose dependent manner. In water-insoluble glucan synthesis assay, 2-4 mg/ml of the ethanol extract of Saussurea lappa significantly inhibited the formation of water-insoluble glucan. These results suggest that Saussurea lappa may inhibit the caries-inducing properties of Streptococcus mutans. Further studies are necessary to clarify the active constituents of Saussurea lappa responsible for such biomolecular activities.     

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 2 mg/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.     

Antimicrobial activity of Streblus asper leaf extract

Bactericidal activity was found in the 50% ethanol (v/v) extract of Streblus asper leaves. The extract possessed a selective bactericidal activity towards Streptococcus, especially to Streptococcus mutans which has been shown to be strongly associated with dental caries. The extract had no effect on cultures of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, Staphylococcus coagulase positive, Staphylococcus coagulase negative, Serratia marcescens, Klebsiella pneumoniae, Enterobacter, Pseudomonas aeruginosa, Burkholderia pseudomeallei and Candida albicans. The minimum growth inhibitory concentration and the minimum bactericidal concentration of S. asper extract against 10(8) CFU/mL of S. mutans was 2 mg/mL. The active compound is partially polar, partially heat labile, precipitated by 80% ammonium sulphate, and possesses a molecular weight larger than 10 000 Da. The potential for using S. asper extract as a natural product for controlling dental caries is discussed.     

Potential of the aqueous extract of Terminalia chebula as an anticaries agent

The aqueous extract from Terminalia chebula was tested for its ability to inhibit the growth and some physiological functions of Streptococcus mutans. The extract strongly inhibited the growth, sucrose induced adherence and glucan induced aggregation of S. mutans. Mouthrinsing with a 10% solution of the extract inhibited the salivary bacterial count and salivary glycolysis. Mouthrinsing with the extract significantly reduced total bacterial counts and the total streptococcal counts in the saliva samples obtained up to and including 3 h after rinsing, compared with the counts obtained prerinsing or after placebo rinsing. The extract successfully inhibited glycolysis of salivary bacteria for up to 90 min postrinsing.     

Effects of Mikania genus plants on growth and cell adherence of mutans streptococci

The present study evaluated the chemical composition and the antimicrobial activity of the extracts and fractions of Mikania laevigata and Mikania glomerata on growth and cell adherence of mutans streptococci. Ethanolic extract, hexane and ethyl acetate fractions of Mikania laevigata and Mikania glomerata were chemically identified by chromatographic methods and tested on mutans streptococci from culture collection and clinical isolates. Twenty-two compounds were identified in both Mikania extracts, including coumarin, 1-octadecene, and diterpenic, cupressenic and kaurenoic acids. Antimicrobial activity was assessed by determination of the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and inhibition of cell adherence to a glass surface. Hexane fraction from both plant extracts was the most effective in inhibiting the growth of the bacterial strains tested (MIC values between 12.5 microg/ml and 400 microg/ml, and MBC values between 25 microg/ml and 400 microg/ml). In addition, sub-MIC levels of the crude extracts and their hexane fractions significantly inhibited the adherence of the microorganisms to a glass surface. The data indicate that the biologically active compounds are present mostly in the hexane fraction of both Mikania species, which showed remarkable inhibitory activities against mutans streptococci. Mikania genus plant is a promising source for novel antimicrobial agents against oral pathogens.     

苏木不同提取物对两种口腔致龋菌的影响

目的观察苏木不同提取物对常见口腔致龋菌的影响。方法测定苏木不同提取物对粘性放线菌和变形链球菌的MIC、MBC值,观察苏木不同提取物对这两种菌的抑制产酸和抑制黏附的效果。结果苏木4种提取物对粘性放线菌和变形链球菌最低抑菌浓度(MIC)为1.25~2.50 g/L,最低杀菌浓度(MBC)均为5 g/L。1/2 MIC乙醇提取物对粘性放线菌的抑制产酸的效果最好,1/2 MIC水提取物对变形链球菌的抑制产酸效果最好,这两组效果与阳性对照组相比没有统计学差异。实验组中抑制黏附效果最好的是1/2 MIC乙醇提取物,1/2 MIC乙醇提取物对粘性放线菌和变形链球菌抑制黏附率分别为41.83%和46.40%。结论苏木4种提取物对粘性放线菌和变形链球菌都有很好的抑菌杀菌效果。其中1/2 MIC乙醇提物对粘性放线菌有好的抑制产酸效果,1/2 MIC水提取物对变形链球菌抑制产酸效果好,苏木不同提取物对粘性放线菌和变形链球菌都有抑制黏附的效果。     

Kuwanon G: an antibacterial agent from the root bark of Morus alba against oral pathogens

Kuwanon G was isolated from the ethyl acetate fraction of methanol extract of Morus alba and its structure was elucidated by 13C-NMR, 1H-NMR and FAB-MS. Antibacterial activity of kuwanon G was investigated by the minimum inhibitory concentration (MIC) test and the viable cell count method. MIC of kuwanon G against Streptococcus mutans causing dental caries was determined to be 8.0 microg/ml. The bactericidal test showed that kuwanon G completely inactivated S. mutans at the concentration 20 microg/ml in 1 min. Kuwanon G also significantly inhibited the growth of other cariogenic bacteria such as Streptococcus sobrinus and Streptococcus sanguis, and Porpyromonas gingivalis causing periodontitis. Transmission electron microscopy (TEM) of kuwanon G treated cells demonstrated remarkable morphological damage of the cell wall and condensation of the cytoplasm.     

In vitro inhibitory effect of Streblus asper leaf-extract on adhesion of Candida albicans to human buccal epithelial cells

This in vitro study aimed at determining the effects of a sublethal concentration of Streblus asper Lour (Moraceae) leaf ethanolic extract on adherence of Candida albicans to human buccal epithelial cells (HBEC). The minimum concentration of Streblus asper leaf ethanolic extract (SAE) that significantly reduced adherence (P<0.05) after a 1-h exposure was 15.6 mg/ml. However, there was a significant reduction (P<0.05) of candidal adhesion to HBEC after 1-min exposure to 125 mg/ml of SAE. Pre-treatment of either Candida or HBEC, or both, with 125 mg/ml of SAE for 1h resulted in reduced adherence. SAE at concentrations of 125 and 250 mg/ml also showed 41 and 61% inhibition of germ tube formation, respectively, which might affect adherence. These findings indicate that the sublethal concentration of SAE may modulate candidal colonization of the oral mucosa thereby suppressing the invasive potential of the pathogen.     

Inhibitory effect of some herbal extracts on adherence of Streptococcus mutans

The objective of this study was to investigate the inhibitory effect of the crude extracts from some herbs on adherence of Streptococcus mutans (S. mutans) ATCC 25175 and TPF-1 in vitro. Six herbs, Andrographis paniculata; Cassia alata; Chinese black tea (Camellia sinensis); guava (Psidium guajava); Harrisonia perforata and Streblus asper, were extracted with 50 or 95% ethanol and dried. Herbal extracted solution at 0.5% concentration (w/v) was initially tested for bacterial adherence on glass surfaces. In order to identify type and effective concentration of the extracts, the extracts that showed the inhibition on glass surfaces were then tested on saliva-coated hydroxyapatite by the use of radiolabeled bacteria. To study the mechanism of action, the effect of the extracts at such concentration on glucosyltransferase and glucan-binding lectin activities were examined. It was found that all extracts, but Streblus asper, showed significant inhibitory effect on bacterial adherence to glass surfaces. For the saliva-coated hydroxyapatite adherence assay, Andrographis paniculata, Cassia alata, Chinese black tea and Harrisonia perforata could inhibit adherence of S. mutans ATCC 25175. Chinese black tea was the strongest inhibitor followed by Andrographis paniculata, Cassia alata and Harrisonia perforata, respectively. For S. mutans TPF-1, adherence inhibition was observed from Andrographis paniculata and Cassia alata at similar levels. The lowest concentrations of the extracts that inhibited the adherence at least 50% were 0.5% of Andrographis paniculata, 0.5% of Cassia alata, 0.3% of Chinese black tea and 0.5% of Harrisonia perforata for S. mutans ATCC 25175. For S. mutans TPF-1, the effective concentrations were 0.5% of Andrographis paniculata and 0.4% of Cassia alata. All extracts at such concentrations decreased the activity of glucosyltransferase from both strains. Only Andrographis paniculata and Cassia alata eliminated or decreased the activity of glucan-binding lectin from both strains. These findings suggested that Andrographis paniculata, Cassia alata, Chinese black tea and Harrisonia perforata could inhibit adherence of S. mutans ATCC 25175, while Andrographis paniculata and Cassia alata had an effect on S. mutans TPF-1 in vitro at the concentrations employed in this study.     

Inhibitory effects of methanol extract of Cyperus rotundus rhizomes on nitric oxide and superoxide productions by murine macrophage cell line, RAW 264.7 cells

The rhizomes of Cyperus rotundus (C. rotundus) have been used in oriental traditional medicines for the treatment of stomach and bowel disorders, and inflammatory diseases. Nitric oxide (NO) and superoxide (O2-) are important mediators in the pathogenesis of inflammatory diseases. This study was undertaken to address whether the metanol (MeOH) extract of rhizomes of C. rotundus could modulate NO and O2- productions by murine macrophage cell line, RAW 264.7 cells. The MeOH extract of rhizomes of C. rotundus showed the inhibition of NO production in a dose-dependent manner by RAW 264.7 cells stimulated with interferon-gamma plus lipopolysaccharide. The inhibition of NO production by the extract was due to the suppression of iNOS protein, as well as iNOS mRNA expression, determined by Western and Northern blotting analyses, respectively. In addition, the MeOH extract suppressed the production of O2- by phorbol ester-stimulated RAW 264.7 cells in dose- and time-dependent manners. Collectively, these results suggest that the MeOH extract of rhizomes of C. rotundus could be developed as anti-inflammatory candidate for the treatment of inflammatory diseases mediated by overproduction of NO and O2-.     

In vitro effects of a fraction separated from Polygonum cuspidatum root on the viability, in suspension and biofilms, and biofilm formation of mutans streptococci

Polygonum cuspidatum (Polygonaceae) has traditionally been used in folk medicine to control oral diseases. Nevertheless, there are no reports related to its possible effect on the diseases, particularly on biofilm-related diseases such as dental caries. In this study, we evaluated in vitro effects of a fraction separated from Polygonum cuspidatum root on the viability, in both suspension and biofilms, and the biofilm formation of mutans streptococci. The separated fraction (F1) showed bacteriostatic and bactericidal activity against mutans streptococci in suspension, with minimum inhibitory concentration (MIC) range of 31.3-250 microg/ml and minimum bactericidal concentration (MBC) range of 0.5-1 mg/ml. At a concentration of 1.5 mg/ml, F1 killed approximately 2 log(10)CFU/ml of Streptococcus mutans and Streptococcus sobrinus after 2h of exposure. In biofilms, F1 also inhibited the viability of Streptococcus mutans and Streptococcus sobrinus, dependent on the biofilm age, the concentration of F1, and the treatment time. Four hours of exposure to 1.5 mg/ml F1 reduced the viable counts of Streptococcus mutans and Streptococcus sobrinus by greater than 2 log(10)CFU/disc. Furthermore, at sub-MIC levels, F1 inhibited biofilm formation by Streptococcus mutans and Streptococcus sobrinus in a dose-dependent fashion. Based on the preliminary phytochemical analysis, the activity of F1 may be related to the presence of anthraquinones, cardiac glycosides, terpenoids, and phenolics. These results indicate that F1 is probably useful in the control of oral biofilms and subsequent dental caries development.     

Administration of Cyperus rotundus tubers extract prevents weight gain in obese Zucker rats

Cyperus rotundus L. (Cyperaceae; C. rotundus) is an Indian medicinal plant demonstrated to exert multiple health benefits. The purpose of the present study was to test the biological efficacy of C. rotundus tubers extract on weight control in obese Zucker rats. It was demonstrated that administration of 45 or 220 mg/kg/day of C. rotundus tubers hexane extract for 60 days in Zucker rats induced a significant reduction in weight gain without affecting food consumption or inducing toxicity. In vitro, 250 microg/mL of this extract was able to stimulate lipolysis in 3T3-F442 adipocytes suggesting that this medicinal plant contains activators of beta-adrenoreceptors (AR). The binding assay performed on the rat beta3-AR isoform, known to induce thermogenesis, demonstrated that C. rotundus tubers extract can consistently and effectively bind to this receptor. These data suggest that the effect on weight gain exerted by C. rotundus tubers extract may be mediated, at least partially, through the activation of the beta3-AR. In conclusion, C. rotundus tubers extract prove to be a new herbal supplement for controlling body weight preferentially in beta3-AR sensitive species.     

Effect of Powerdental on caries-inducing properties of Streptococcus mutans and TNF-alpha secretion from HMC-1 cells

We studied the inhibitory effect of Powerdental on the growth and acid production of Streptococcus mutans as well as secretion of pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). The growth of Streptococcus mutans was reduced by the presence of the Powerdental (1 mg/ml) and NaCl (1 mg/ml) significantly, and the positive control group (1% NaF) also exhibited a significant antibacterial activity. The decrease of pH was significantly inhibited in the presence of Powerdental (1 mg/ml) compared to the control group. The decrease in pH was also inhibited in the presence of positive control (1% NaF), but the bamboo salt alone did not show inhibitory activity. We also found that Powerdental (0.01 mg/ml) inhibited significantly the secretion of TNF-alpha with 46.5+/-0.2% from human mast cells. Our results suggest that Powerdental contributes to the prevention or treatment of periodontitis and other oral diseases or inflammatory diseases.     

香附总黄酮的超声波提取工艺研究

目的 探讨香附总黄酮的提取及鉴别方法,以充分利用香附植物资源,避免资源的浪费.方法 采用超声波乙醇浸提法从香附中提取黄酮类物质,对所提取的黄酮类物质进行验证,并用分光光度法测定含量.结果 测得样品中总黄酮的含量C=0.186 4 mg/ml,回收率为102.7%,其纯度和产率均较高.结论 该方法采用全物理过程,无任何污染,是提取香附黄酮类物质的有效途径.     

Inhibition of Staphylococcus aureus by aqueous Goiaba extracts

The antimicrobial activity of an aqueous extract of three Brazilian medicinal plants, Alhodomato (Allium sativon), Traoeraba (Comnelina beghlensis) and Goiaba (Psidium guajava), was studied and compared with commercial antibiotics using three different methods: plate count, disk inhibition zone and turbidity techniques. Results indicated that Goiaba leaf extract at a concentration of 8 and 40 mg/ml showed promising results. As Goiaba leaf extract showed good antimicrobial activity against Staphylococcus aureus, nine different strains were tested using these methods. Results showed a complete inhibition of all strains tested at a concentration of 6.5 mg/ml. This indicates that Goiaba leaf extract could be an important source of food preservative and a new source of an antimicrobial agent against S. aureus.     

Screening of Taiwanese crude drugs for antibacterial activity against Streptococcus mutans

Preliminary antibacterial screening of local crude drugs was carried out using the cariogenic bacterium, Streptococcus mutans. Of 79 aqueous extracts tested, 6 crude drugs were shown to have significant antibacterial activity with minimal inhibitory concentration equal to or lower than 7.8 mg/ml (expressed in terms of dry starting material). Of these effective crude drugs, Morus australis, Ludwigia octovalvis and Thuja orientalis were very effective in inhibiting the growth of serotypes c and d of S. mutans (MIC less than or equal to 2.0-7.8 mg/ml). Elephantopus scaber, Artemisia vulgaris, Mosla chinensis and Orthosiphon aristatus also exhibited considerable antibacterial activity (MIC = 7.8-23.4 mg/ml) against both serotypes. In the presence of 5% sucrose, the antibacterial potency of the majority of the extracts did not change for type c, while the potency decreased about one-half for type d.     

Antidermatophyte activity of ether extract of Nigella sativa and its active principle, thymoquinone

The antifungal activity of ether extract of Nigella sativa seed and its active principle thymoquinone was tested against eight species of dermatophytes: four species of Trichophyton rubrum and one each of Trichophyton interdigitale, Trichophyton mentagrophytes, Epidermophyton floccosum and Microsporum canis. Agar diffusion method with serial dilutions of ether extract of Nigella sativa, thymoquinone and griseofulvin was employed. The incubation was carried out at 30 degrees C for 14 days. The diameter of fungal colonies and the percentage inhibition of the fungal growth at each dilution were determined, taking those of the controls as 100%. The minimum inhibitory concentration (MIC) was considered as the minimum concentration of the drug, which inhibited 80-100% of the fungal growth. The MICs of the ether extract of Nigella sativa and thymoquinone were between 10 and 40 and 0.125 and 0.25 mg/ml, respectively, while those of griseofulvin ranged from 0.00095 to 0.0155 mg/ml. These results denote the potentiality of Nigella sativa as a source for antidermatophyte drugs and support its use in folk medicine for the treatment of fungal skin infections.     

蛇莓对人食管癌细胞作用的研究

本文以体外长期培养的 Eca-109细胞系为实验材料,以分裂指数、集落形成、生成曲线、~3H-胸腺嘧啶核苷标记为观察指标,研究了中药蛇莓在体外对人食管癌细胞的生长、分裂、增殖、再繁殖能力和 DNA 合成的影响。结果表明,蛇莓水提取物相当于5mg/ml、10mg/ml、15mg/ml 时,对细胞生长有较强的抑制效应;蛇莓15mg/ml 作用48小时可使细胞完全丧失再繁殖能力。蛇莓10mg/ml、15mg/ml 作用48小时对细胞有丝分裂的抑制率分别为46%和49%,但对细胞 DNA 合成仅呈轻度抑制效应。     

In vitro study of the effects of khat (Catha edulis Forsk) extract on isolated mouse interstitial cells

Isolated mouse interstitial cells were incubated with different concentrations of khat (Catha edulis) extract (0.06 mg/ml, 0.6 mg/ml, 6 mg/ml, 30 mg/ml and 60 mg/ml) and cell viability as well as testosterone concentration measured at 30 min intervals over a 3 h incubation period. High concentrations of khat extract (30 mg/ml and 60 mg/ml) significantly inhibited testosterone production while low concentrations (0.06 mg/ml, 0.6 mg/ml and 6 mg/ml) significantly stimulated (P < 0.05) testosterone production by mouse interstitial cells. Similarly, at concentrations of 30 mg/ml and 60 mg/ml, there was a significant decrease in interstitial cell viability, whereas at 0.06 mg/ml, 0.6 mg/ml and 6 mg/ml there was no significant decrease. There was only a weak correlation (r = 0.39) between testosterone production and viable interstitial cells. We postulate that khat extract at high concentrations may cause reproductive function impairment in the user but at low concentrations, may enhance testosterone production with accompanying effects on reproductive functions in male mice.     

Inhibitory potential of neem (Azadirachta indica Juss) leaves on dengue virus type-2 replication.

In the present study we report in vitro and in vivo inhibitory potential of crude aqueous extract of neem leaves and pure neem compound (Azadirachtin) on the replication of Dengue virus type-2. In vitro antiviral activity of aqueous neem leaves extract assessed in C(6/36) (cloned cells of larvae of Aedes albopictus) cells employing virus inhibition assay showed inhibition in dose dependent manner. The aqueous extract of neem leaves at its maximum non-toxic concentration of 1.897 mg/ml completely inhibited 100-10,000 TCID(50) of virus as indicated by the absence of cytopathic effects. The in vivo protection studies with neem leaves extract at its maximum non-toxic concentrations 120-30 mg/ml resulted in inhibition of the virus replication as confirmed by the absence of Dengue related clinical symptoms in suckling mice and absence of virus specific 511 bp amplicon in RT-PCR. The pure neem i.e. Azadirachtin did not reveal any inhibition on Dengue virus type-2 replication in both in vitro and in vivo systems.