Skip to main content
Download PDF

Design and study of anticaries effect of different medicinal plants against S.mutans glucosyltransferase

BMC Complementary and Alternative Medicine volume 19, Article number: 197 (2019) Cite this article

Abstract

Background

The present study was aimed to evaluate the molecular level anticaries effect of different medicinal plants against Streptococcus mutans (S.mutans) glucosyltransferases (gtf).

Methods

A total of six natural sources named as Terminalia chebula (T.chebula), Psidium guajava (P.guajava), Azadirachta indica (A.indica) and Pongamia pinnata (P.pinnata); two essential oils, clove (Syzygium aromaticum) and peppermint oil (Mentha piperita) were selected as test samples. Hydroalcoholic plant extracts and essential oils were examined for their inhibitory potential on gtf isolated from S.mutans. Polyherbal mouth wash was prepared and its effect on gtf activity was compared with commercial chlorhexidine mouth wash (5%w/v). Enzyme kinetic study was carried out in order to explore the molecular mechanism of enzyme action.

Results

Out of six natural sources tested, A.indica has shown maximum inhibitory effect of 91.647% on gtf and T.chebula has shown IC50 of 1.091 mg/ml which is significant when compared to standard chlorhexidine. From the final result of kinetic analysis it was found that T.chebula, P.guajava and P.pinnata have show uncompetitive inhibition where as A.indica has shown non-competitive inhibition. Surprisingly, both essential oils have shown allosteric inhibition (sigmoidal response). The polyherbal moutwash has shown significant inhibitory potential on gtf (95.936%) when compared to commercial chlorhexidine mouthwash (p < 0.05).

Conclusion

All the tested samples have shown considerable gtf inhibitory action. Moreover polyherbal mouth wash has shown promising noncompetitive inhibitory activity against gtf and it could be the future formulation to combat dental caries.

Peer Review reports

Introduction

Fermentation of carbohydrates presents in a food by oral bacteria results in a decrease in the pH of plaque and demineralization of enamel and finally formation of dental caries [1,2,3,4]. Streptococcus mutans (S.mutans) is considered to be the principal cariogenic bacterium in humans [5, 6]. It possesses several virulence factors that are associated with its cariogenicity [7]. An essential factor of S.mutans is its sucrose-dependent and glucan-mediated colonization of tooth surfaces. Glucans are glucose polymers synthesized by extracellular glucosyltransferases (gtfs) [1]. There are three S.mutans gtf’s: gtf B and gtf C synthesize primarily water-insoluble glucans (WIG) [8, 9] and gtf D, synthesize water-soluble glucans (WSG) [10]. Gtfs are involved in many biological processes such as cell-cell communications, signal transduction, immune response, microbial adhesion and infection [11]. Biofilm or dental plaque is formed through two different sequential steps: initial and reversible cell-to-surface attachment and subsequent sucrose-dependent adhesion of the microorganisms, which is firm and irreversible, gtfs are strongly involved in the latter step [12]. There are multiple ways that compounds can exert anticaries action in addition to gtf inhibition. In view of essential roles played by gtfs, intervention of gtfs has attracted remarkable interest among other ways for drug development since inhibitors of gtfs can potentially interfere with the pathological processes [13, 14]. Targeting gtf enzymes prevents the synthesis of extracellular polysaccharides (glucans) and is an attractive strategy for the development of anti-biofilm compounds, as the glucans are extremely important in the processes of biofilm formation and stabilization [15].

The use of medicinal plants or natural products has been one of the most successful strategies for the discovery of new medicines [16]. It already been reported that plant extracts and their components have significant antibacterial activity on oral bacteria, especially on S.mutans [17,18,19,20,21]. Phytochemical rich extracts and their associated compounds have repeatedly shown inhibitory effects against adhesion, plaque and biofilm formation of S.mutans [22,23,24,25,26,27]. As inhibition of essential virulence factor (gtf) is the primary goal for the prevention of dental carries and possibly other plaque related diseases, present study is aimed to report the anti-caries effects of four plant extracts (Gallnut, Guava, Neem, Indian beech) and two essential oils (Clove oil, Peppermint oil) against S.mutans gtfs.

Material and methods

Brain Heart Infusion broth (BHI), sucrose, ethanol, sodium dodecyl sulfate, acetic acid, sodium acetate, tween 80, dimethyl formamide (DMF) and ammonium sulfate (all are of analytical grade) were used in the present study and purchased from Merck, Mumbai.

Design of the study

Four medicinal flora, Terminalia chebula (T.chebula), Psidium guajava (P.guajava), Azadirachta indica (A.indica) and Pongamia pinnata (P.pinnata); two essential oils, clove (Syzygium aromaticum) and peppermint oil (Mentha piperita) were selected as test samples for the present study. By keeping in survey of the folklore use of these plants in dentistry and their reported antibacterial activity [28,29,30,31], specific parts of the above mentioned plants like fruits of T.chebula, leaves of P.guajava, twigs of A.indica and P.pinnata were selected. In a similar way essential oils of clove and peppermint were selected as essential oils have stood out as a promising source of bioactive molecules with potential application in the management of dental caries [32, 33].

Collection and authentication of plant material

Fruits of T. chebula (068) were purchased from local commercial market. The leaves of P. guajava (081), twigs of A. indica (0125) and P. pinnata (001) were collected from the institutional medicinal garden (cultivated), Bharat Institute of Technology, Hyderabad. All the collected plant materials were authenticated by Department of Botany, Osmania University, Hyderabad and voucher specimens were submitted in the same place. Clove oil and peppermint oil were procured from essential oil manufacturer (PSC aromatics), Ooty, Tamilnadu.

Preparation of hydroalcoholic extracts of plants

Plant materials were shade dried and pulverized separately. Each plant material was extracted separately using an equimolar ratio of ethanol and water (50:50) by using a soxhlet extraction method. The hydroalcoholic extracts were concentrated using a rotary evaporator and the dried extracts were preserved at 4 °C for further use.

Culture collection

S.mutans MTCC 49 strain used for the present study was facultatively anaerobic, Gram-positive coccus-shaped bacterium, procured from the centre for cellular and molecular biology (CCMB), Tarnaka, Hyderabad. A stock of S.mutans MTCC 49 was prepared in glycerol and preserved at 0 °C until further use.

Production and partial purification of gtf

A partially purified gtf required for enzyme inhibition studies was prepared according to the modified method of Tomita et.al [34]. BHI broth containing 5% sucrose was prepared and S.mutans MTCC 49 were inoculated into the media. Furthermore tween 80 (1 mg/ml) was incorporated into the media to increase the enzyme production [35]. The inoculated broth was then incubated for 24 h at 37 °C. Bacteria grown in 600 ml BHI broth were centrifuged (1800Xg, 30 min) and supernatant fluid obtained was precipitated using 2/3 volume of ammonium sulfate (60%). The precipitate was centrifuged and dissolved in 30 ml phosphate buffer (pH 7.4) and dialyzed for 24 h against the same solution. The insoluble material was removed by centrifugation. The dialysate was preserved at 0 °C until further use.

Determination of cell free gtf activity

Gtf activity was determined by modified methods of Fukushima et.al [36].

Assay for WIG forming activity (method 1)

The gtf activity was determined based on the amount of glucans formed, expressed as the absorbance at 340 nm (ΔA340/min). The reaction mixture composed of 100 μM citrate buffer (pH 6.2), 5% sucrose, phosphate buffer of pH 7.4 and 0.5 ml of partially purified gtf extract was incubated at 25 °C for 10 min. The increase in absorbance at 340 nm was determined using UV-VIS spectrophotometer (Shimadzu 1800). The activity (ΔA340/min) of gtfs was determined from the slope of the linear part of the time course curve.

Calorimetric assay for WIG and WSG forming activity (method 2)

In this method gtf activity was determined based on the amount of glucans formed, expressed as the glucose content per minute. The reaction mixture composed of 100 μM citrate buffer (pH 6.2), 5% sucrose, phosphate buffer of pH 7.4 (control), 0.5 ml of partially purified gtf extract was incubated at 37 °C for 2 h. The reaction was terminated by the addition of 0.6 M sodium dodecyl sulfate. WIG pellet was precipitated by centrifugation at 10,000×g for 5 min. The supernatant was then transferred to another tube and WSG was precipitated with 3 volumes of ethanol at 4 °C over night. The washed polysaccharide (WIG & WSG) was quantified based on the glucose content using phenol-sulfuric acid method [37]. For the quantification of WIG and WSG, 50 μl of the sample was used in the assay.

Determination of the effect of plant extracts on gtf activity

The gtf inhibition studies using different concentrations of plant extracts were carried out by method 1. Plant extracts were dissolved in phosphate buffer with gentle heating where necessary and diluted to give final concentrations of 0.312, 0.625, 1.25, 2.5, 5 and 10 mg/ml in the reaction mixture. The reaction mixture used in the method 1 was incubated replacing phosphate buffer with various concentrations of the plant extracts for 10 min at 25 °C. The amount of glucan formed was determined according to the procedure described in method 1. To adjust for the quantification errors due to carbohydrates of plant extracts which will precipitate with the glucans, a parallel series of mixtures without the enzyme was prepared and absorbance value was subtracted from test assay readings.

Determination of the effect of essential oils on gtf activity

The effect of essential oils on gtf was determined by method 2 to overcome the solubility problem of essential oils with enzyme solution in method 1. The two essential oils used in the present study were diluted with DMF separately to make concentrations of 3.125, 6.25, 12.5, 25, 50 and 100 mg/ml. The reaction mixture used in the method 2 was incubated with various concentrations of the test oils for 2 h at 37 °C. The amount of WIG and WSG was estimated by the method 2 described above.

Formulation of poly herbal mouth rinse [38]

The poly herbal mouth rinse formulation was designed based on IC50 values obtained for each plant extract and essential oils (Table 1). It was evaluated for pH and clarity test. Finally the effect of mouth rinse against gtf activity was determined and compared with commercial chlorhexidine mouthwash.

Table 1 Formulation of polyherbal mouth rinse
Full size table

Enzyme kinetics

For kinetic analysis KM, Vmax and hill coefficient values were measured with non-linear regression analysis using Graphpad Prism 5.0 version.

Statistical analysis

All the experiments were carried out in triplicates and data was presented as mean ± SEM. Statistical analysis and enzyme kinetics were performed using Graph pad Prism 5.0 software.

Results and discussion

The percentage yield of hydro alcoholic extract of T.chebula, P.guajava, A.indica and P.pinnata was found to be 0.480, 0.27, 0.87 and 1.177%w/w respectively.

Dental caries is a biofilm-mediated, sugar driven, multifactorial, dynamic disease that results in the phases of demineralization and remineralization of dental hard tissues. The synthesis of extracellular polysaccharide, including WIG & WSG is one of the most important virulence factors of S.mutans [39]. The WIG promotes the adhesive interactions of bacteria with the tooth surfaces and contributes the formation of dental biofilm [40]. Accordingly, in the present study, we examined whether selected plant extracts inhibits the synthesis of WIG and whether clove and peppermint oils inhibits the synthesis of WSG & WIG by crude gtfs as there is no previous molecular level mechanistic report on these samples. Our results were in correlation with the literature reports of the anti S.mutans activity of the test samples.

The hydroalcoholic extracts of T.chebula, P.guajava, A.indica and P.pinnata inhibit WIG synthesis by inhibiting S.mutans gtf activity in a dose dependent manner (Table 2). A.indica extract has shown highest percentage inhibition of 91.647 ± 0.445% at 10 mg/ml concentration. Order of gtfs inhibitory activity of the four plant extracts at their highest concentration of 10 mg/ml was A.indica > T.chebula > P.pinnata > P.guajava. T.chebula extract has shown predominant inhibitory action among tested extracts from 0.312 to 5 mg/ml (Fig. 1). It means gtf synthesizing WIG from sucrose was almost suppressed upon addition of 10 mg/ml concentration of A.indica extract. Highest correlation coefficient (R2) value is observed with A.indica when compared to other extracts. However, according to the 50 % inhibitory doses (IC50) T.chebula would be considered as an effective gtf inhibitor as it has shown effect at lower concentration (1.091 mg/ml) when compared to other three plant extracts. It has been reported that gtf B and C are important factor for synthesis of WIG by S.mutans, gtf B synthesize primary WIG where as gtf C synthesizes WIG & WSG [41]. Although we identified that T.chebula, P.guajava, A.indica and P.pinnata extracts may inhibits gtf B by inhibiting WIG synthesis where as clove and peppermint oil inhibits both gtf B and gtf C by inhibiting WIG & WSG synthesis.

Table 2 Gtf inhibitory effect of different plant extracts at various concentrations
Full size table
Fig. 1
figure 1

Glucosultransferase inhibitory effects of T.chebula, P.guajava, A.indica and P.pinnata

Full size image

To study the molecular level mechanism responsible for the anticaries activity of clove oil and peppermint oil, the effect of these on the WIG & WSG synthesis by cell free extracellular gtf of S.mutans were examined. Various concentrations ranging from 3.125-100 mg/ml were used for both the samples and their concentrations are different from that of concentrations used for plant extracts (based on previously reported MIC against S.mutans). Dose dependent inhibition of gtf was observed in both the cases (Table 3) and especially from 25 to 100 mg/ml there was a drastic increase in % inhibition. IC50 values of peppermint oil (44.693%) was significant when compared to clove oil (63.697%) (Fig. 2). Out of six test samples used in the present study, T.chebula extract has shown the low IC50 value (Fig. 3).

Table 3 Gtf inhibitory effect of clove and peppermint oils at various concentrations
Full size table
Fig. 2
figure 2

Gtf inhibitory effects of clove and peppermint oils at various concentrations

Full size image
Fig. 3
figure 3

IC50 values of plant extracts and essential oils

Full size image

R2 value represents square root of the correlation coeffecient and A.indica has R2 value of 0.9700 indicates 0.97002 × 100 (97%), the higher the percentage better the correlation between results. F-value is a numerical indicator of whether or not the result expressed by the method is coincidental, the higher the value, the less likely the result is due to chance (A.indica has highest F-value of 248 among tested extracts).

From kinetic analysis it was observed that T.chebula, P.guajava, A.indica and P.pinnata have shown the exponential curves (Fig. 4) indicative of deviation from normal response (hyperbolic) of the enzyme. From their Michael Menton constant (KM) and Vmax values, it was identified that T.chebula, P.guajava and P.pinnata have shown uncompetitive inhibition as their KM and Vmax values have been decreased when compared to control (without inhibitor). In case of A.indica it was found that KM unchanged and Vmax decreased indicated that the pattern was of non-competitive inhibition (Table 4).

Fig. 4
figure 4

Exponential response curves of T.chebula, P.guajava, A.indica and P.pinnata extracts

Full size image
Table 4 Kinetic parameters of the inhibitory effect of test samples on the activity of gtf
Full size table

Similar analyses of clove and peppermint oils revealed the fact that response was sigmoidal and the pattern was of allosteric inhibition (Fig. 5). Sigmoid response curve suggests the cooperativity between the inhibitor and the enzyme and hill coefficient provides a way to quantify the degree of binding between inhibitor and enzyme. Both clove and peppermint oil have shown hill coefficient > 1 represents positive, cooperative binding (Table 4). Hill coefficient value is more (4.60 ± 0.59) for clove oil when compared to peppermint oil (1.92 ± 0.46) indicates that clove oil has more affinity for gtf. The present study mainly focused on the effect of plant extracts and essential oils on gtf inhibition and comparison of the same with commercial anti-carries agent. Here the criteria is to compare molecular level gtf inhibitory mechanism of tested samples so that whether they possess significant anti carries property or not.

Fig. 5
figure 5

Sigmoidal response curves of clove oil and peppermint oil

Full size image

The bilologically active phytoconstituents or secondary metabolites present in all four plant extracts have been responsible for their gtf inhibitory activity. Moreover gtf activity of bioactive compounds like tannins [42], flavonoids [43], terpenoids [44] were reported previously. The tannins in T.chebula fruits, terpenoids in A.indica twigs, isoflavones in twigs of P.pinnata and flavonoids and tannins present in leaves of P.guajava may be the attributing factors for S.mutans gtf inhibition.

The antibacterial properties of plant essential oils against bacteria found in the oral cavity are also well documented [45]. While the commercial mouthwashes containing essential oils are useful in the long-term control of plaque and mild to moderate gingivitis and are preferred to use in chlorhexidine containing mouth washes for long-term daily use [46]. Therefore, essential oils may be suitable additives in pharmaceutical products.

There has been reports about the menthol present in Mentha piperata (peppermint oil) and eugenol in Syzygium aromaticum (clove oil) and are considered as outstanding compounds exhibiting an antibacterial potential [47] which could be responsible for S.mutans gtf inhibition.

The present study also focused on the formulation of the polyherbal mouth rinse and its IC50 value was found to be 7.938%w/v. The % inhibition at its maximum concentration of 15%w/v was found to be 95.936 ± 1.518 and it was very high when compared to 53.80 ± 1.83% inhibition of chlorohexidine commercial mouthwash. Dose dependent gtf inhibitory effect was observed in case of polyherbal mouth wash and chlorhexidine did not follow this pattern. From the results it was observed that gtf inhibitory potential of the polyherbal mouth rinse was significant (p < 0.05) compared to chlorhexidine mouthwash at tested concentrations (Fig. 6). From the shape of the exponential curve (concentration of inhibitor vs enzyme velocity) it was evident that polyherbal mouth rinse competitively inhibits gtf where as chlorhexidine shows non-competitive inhibition (Fig. 7).

Fig. 6
figure 6

Gtf inhibitory effects of Polyherbal mouth rinse and Chlorhexidine mouth rinse at various concentrations

Full size image
Fig. 7
figure 7

Exponential response curves of Polyherbal mouth rinse and Chlorhexidine mouth

Full size image

Conclusions

From the studies it was concluded that among the four plant extracts and two essential oils tested against gtf activity A.indica extract has shown maximum percentage inhibition and T.chebula extract has shown the significant IC50 value when compared to others. The results of kinetic analysis proposed that T.chebula, P.guajava, P.pinnata have shown uncompetitive gtf inhibition where as A.indica has shown non-competitive inhibition. Clove oil and Peppermint oil have proposed allosteric inhibition and the standard chlorhexidine has shown non-competitive inhibition. Moreover the polyherbal mouth rinse prepared from all four plant extracts and two essential oils has shown predominant gtf inhibition (95.936%) when compared to 54% shown by chlorohexidine mouthwash. Further clinical investigations are recommended for this polyherbal mouth rinse to utilize as anticaries agent.

Availability of data and materials

All data and materials used in this research are available from the corresponding author on reasonable request.

Abbreviations

A.indica :

Azadirachta indica

BHI broth:

Brain heart infusion broth

DMF:

Dimethyl formamide

Gtfs:

Glucosyltransferases

MIC:

Minimum inhibitory concentration

P.guajava :

Psidium guajava

P.pinnata :

Pongamia pinnata

S.mutans :

streptococcus mutans

T. chebula :

Terminalia chebula

WIG:

Water insoluble glucans

WSG:

Water soluble glucans

References

  1. Mundorff SA. Bibby BG cariogenic potential of food. Caries Res. 1990;24:344–9.

    Article  CAS  Google Scholar 

  2. Clarkson BH. Introduction to cariology: the discipline of cariology; art or science dent. Clin North Am. 1999;43(4):569–77.

    CAS  Google Scholar 

  3. Baneriee A, Watson TF, Kidd EAM. Dentin caries: take it or leave it? Dent Update. 2000;27:272–6.

    Article  Google Scholar 

  4. Ramya R, Srinivasan R. Clinical operational dentistry, principles and practice. 1st ed. EMM ESS Medical Publishers; 2007.

  5. Loesche WJ. Role of Streptococcus mutans in human dental decay. Microbial Rev. 1986;50:353–80.

    CAS  Google Scholar 

  6. Tanzer JM, Livingston J, Thompson AM. The microbiology of primary dental carries in humans. J Dent Educ. 2001;65:1028–37.

    CAS  PubMed  Google Scholar 

  7. Banas JA. Virulence properties of Streptococcus mutans. Front Biosci. 2004;9:1267–77.

    Article  CAS  Google Scholar 

  8. Aoki H, Shiroza T, Hayakawa M, Sato S, Kuramitsu HK. Cloning of a Streptococcus mutans glucosyltransferases gene, coding for insoluble glucan synthesis. Infect Immun. 1986;53:587–94.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Hanada N, Kuamitsu HK. Isolation and characterization of the Streptococcus mutans gtf C gene, coding for synthesis of both soluble and insoluble glucans. Infect Immun. 1988;56:1999–2005.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Hanada N, Kuamitsu HK. Isolation and characterization of the Streptococcus mutans gtf D gene, coding for primer dependent soluble glucan synthesis. Infect Immun. 1989;57:2079–85.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Varki A, Cummings RD, Esko JD, Freeze HH, Stanley P. Essentials of glycobiology. 2nd ed. Cold spring harbor (NY): Cold spring harbor laboratory press; 2009.

  12. Kawabata S, Hamada S. Studying biofilm formation of mutans streptococci: Mthods in enzymology 1999;30:513–523.

  13. Gloster TM, Vocadlo DJ. Developing inhibitors of glycan processing enzymes as tools for enabling glycobiology. Nat Chem Biol. 2012;8:683–94.

    Article  CAS  Google Scholar 

  14. Wagner GK, Pesnol T. Glucosyltransferases and their assays. Chembiochem. 2010;11:1939–49.

    Article  CAS  Google Scholar 

  15. Branda SS, Vik S, Friedman L, Kolter R. Biofilms: the matrix revisited. Trends Microbiol. 2005;13:20–6.

    Article  CAS  Google Scholar 

  16. Harvey A. Strategies for discovering drugs from previously unexplored natural products. Drug Discov Today. 2000;5:294–300.

    Article  CAS  Google Scholar 

  17. Namba T, Tsunezuka M, Dissanayake DMRB, Pilapitiya U. Saikok, Kakiuchi N, Hattori M. studies on dental caries prevention by traditional medicines part VII. Screening of ayurvedic medicines for anti-plaque action. Shoyakugoku Zasshi. 1985;39(2):146–53.

    CAS  Google Scholar 

  18. Chen CP, Lin CC, Namba T. Screening of Taiwanese crude drugs for antibacterial activity against Streptococcus mutans. Ethanopharm. 1989;27:285–95.

    Article  CAS  Google Scholar 

  19. Saeki Y, Ito Y, Shibata M, Sato Y, Okuda K, Takazoe I. Antimicrobial action of natural substances on oral bacteria. Bull Tokoyo Dent Coll. 1989;30(3):129–35.

    CAS  Google Scholar 

  20. Osawa K, Matsumoto T, Maruyama J, Takiguchi T, Okuda K, Takazoe T. Studies of the antibacterial activity of plant extracts and their constituents against periodonpatho bacteria. Bull Tokyo Dent Coll. 1990;31(1):1721.

    Google Scholar 

  21. Yamaki M, Kashihara M, Takagi S. Activity of Ku Shen compounds against Streptococcus aureus and Streptococcus mutans. Phytother Res. 1991;4(6):235–6.

    Article  Google Scholar 

  22. Nostro A, Cannatelli MA, Crisafi G, Musolino AD, Procopio Alonzo V. Modifications of hydrophobicity, in vitro adherence and cellular aggregation of Streptococcus mutans by Helichrysum italicum extract. LAM Lett Appl Microbiol. 2004;38:423–7.

    Article  CAS  Google Scholar 

  23. Yamanaka A, Kimizuka R, Kato T, Okuda K. Inhibitory effects of cranberry juice on attachment of oral streptococci and biofilm formation. Oral Microbiol Immunol. 2004;19:150–4.

    Article  CAS  Google Scholar 

  24. Percival RS, Devine DA, Duggal MS, Chartron S, Marsh PD. The effect of cocoa polyphenols on the growth, metabolism, and biofilm formation by Streptococcus mutans and Streptococcus sanguinis. EOS Eur J Oral Sci. 2006;114:343–8.

    Article  CAS  Google Scholar 

  25. Furiga A, Lonvaud Funel A, Dorignac G, Badet C. In vitro anti-bacterial and anti-adherence effects of natural polyphenolic compounds on oral bacteria. JAM J Appl Microbiol. 2008;105:1470–6.

    Article  CAS  Google Scholar 

  26. Rahim ZHA, Khan HBSG. Comparative studies on the effect of crude aqueous (ca) and solvent (cm) extracts of clove on the cariogenic properties of Streptococcus mutans. J Oral Sci. 2006;48:117–23.

    Article  Google Scholar 

  27. Matsumoto M, Tsuji M, Okuda J, Sasaki H, Nakano K, Osawa K, Shimura S, Ooshima T. Inhibitory effects of cacao bean husk extract on plaque formation in vitro and in vivo. Eur J Oral Sci. 2004;112:249–52.

    Article  Google Scholar 

  28. Nayak SS, Ankola AV, Metgud SC, Bolmal UK. An in vitro study to determine the effect of Terminalia chebula extract and its formulation on Streptococcus mutans. J Contemp Dent Prac. 2014;15(3):278–82.

    Article  Google Scholar 

  29. Garode AM, Waghode SM. Antibacterial activity of guava leaves extracts against S.mutans. Inter J Bioassays. 2014;3(10):3370–2.

    Google Scholar 

  30. Lakshmi T, Krishnan V, Rajendran R, Madhusudhanan N. Azadirachta indica: a herbal panacea in dentistry-an update. Pharmacogn Rev. 2015;9(17):41–4.

    Article  CAS  Google Scholar 

  31. Imran M, Bashir S, Sher M, Shah HS, Iqbal S, Asad M. Anticariogenic activity, possible mechanism and preliminary characterization of twigs of Pongamia pinnata. J Pharm Sci Pharmacol. 2015;2(1):57–63.

    Article  Google Scholar 

  32. Lang G, Buchbauer GA. A review on recent research results (2008-2010) on essential oils as antimicrobials and antifungals. A review. Flavour Fragr J. 2012;27:13–39.

    Article  CAS  Google Scholar 

  33. Lobo PL, Fonteles CS, Marques LA, Jamacaru FV, Fonseca SG, De Carvalho CB, De Movaes ME. The efficacy of three formulations of lippie sicloides Cham essential oil in the reduction of salivary Streptococcus mutans in children with caries: a randomized double blind, controlled study. Phytomed. 2014;21:1043–7.

    Article  CAS  Google Scholar 

  34. Tomita Y, Zhu X, Ochiai K, Namiki Y, Okada T, Ikemi T, Fukushima K. Evaluation of three individual glucosyl transferases produced by streptococcus mutans using monoclonal antibodies. FEMS Microbial Lett. 1996;145:427–32.

    Article  CAS  Google Scholar 

  35. Umesaki Y,* Kawai Y, Mutai M. Effect of tween 80 on glucosyltransferase production in Streptococcus mutans App Env Microbiol. 1977;3(2):115–119.

    Google Scholar 

  36. Fukushima K, Motoda R, Takada K, Ikeda T. Resolution of Streptococcus mutans glycosyltransferases into two components essential to water-insoluble glucan synthesis. FEBS Lett. 1981;128:213–6.

    Article  CAS  Google Scholar 

  37. Dubo's M, Gilles A, Hamilton JK, Rebers PA, Smith F. Colorimetric method for determination of sugars and related substances. Ann N Y Acad Sci. 1956;121:404–27.

    Google Scholar 

  38. Chiedozie EI, Ahamefule OF, Ukamaka AA. Anti-inflammatory, antimicrobial and stability studies of poly-herbal mouthwashes against Streptococcus mutans. J Pharmacog Phytochem. 2016;5(5):354–61.

    Google Scholar 

  39. Schilling KM, Bowen WH. Glucans synthesized in situ in experimental salivary pellicle formation as specific binding sites for Streptococcus mutans. Infect Immun. 1992;60:284–95.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Madisan KM, Bowen WH, Pearson SK, Falany JL. Enhancing the virulence of S.sorbinus in rats. J Dent Res. 1991;70:38–43.

    Article  Google Scholar 

  41. Yamachita Y, Bowen WH, Burne RA, Kuramitsu HK. Role of the S.mutans gtf genes in caries induction in the specific- pathogen-free rat model. Infect Immun. 1993;61:3811–7.

    Google Scholar 

  42. kakiuchi N, Hattori m, Nizhizawa M, Yamagishi T, kuda T, Namba IAT. Studies on dental caries prevention by traditional medicines VIII inhibitory effect of various tannins on glucan synthesis by glucosyltransferase from Streptococcus mutans. Chem Pharm Bull. 1986;34(2):720–5.

    Article  CAS  Google Scholar 

  43. Hyunk O, Pedro L. Rosalen, William H, Bowen. Effects of compounds found in propolis on S.mutans growth and on glucosyl transferase activity. Antimicrob Agents Chemother. 2002;46(5):1302–9.

    Article  Google Scholar 

  44. William H. Bowen, Hyun Koo, Yong kun park, Jaime a parecido Cury, Pedroluiz Rosalen. University of Rochester, Universidade Estadual De compinas WO 2002047615 A2; 2002.

  45. Kalemba D, Kunicka A. Antibacterial and antifungal properties of essentialoils. Cur Med Chem. 2003;10(10):813–29.

    Article  CAS  Google Scholar 

  46. Ciancio IS. Improving oral health: Current considerations. J Clin Periodont. 2003;30(3):4–6.

    Article  Google Scholar 

  47. Freires IA, Denny C, Benso B, de Alencar SI, Rosalen P. Antibacterial activity of essential oils and their isolated constituents against cariogenic bacteria: a systematic review. Molecules. 2015;20:7329–73.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Authors are thankful to Sri C. Venugopal Reddy, Secretary, Bharat Institutions and Dr. Sudheer B, IICT, Hyderabad for their kind help and constant encouragement towards this research.

Declaration

The experimental work described in this article was conducted in the Bharat institute of technology, Department of Pharmaceutical Chemistry and Pharmaceutical Biotechnology, Hyderabad, India, from December 2016 to May 2017. These studies are the result of our own investigations, except where the work of others is acknowledged and have not been submitted in any other form to another journal.

Funding

These analyses were done with self-funding.

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, Bharat Institute of Technology, Mangalpally, JNTUH, R.R. District, Hyderabad, Telangana, 501510, India

    Kiranmai Mandava, Shravya Kakulavaram, Shulamithi Repally, Ishwarya Chennuri, Srinivas Bedarakota & Namratha Sunkara

  2. Department of Pharmaceutical Biotechnology, Bharat Institute of Technology, Mangalpally, JNTUH, R.R. District, Hyderabad, 501510, India

    Uma Rajeswari Batchu

Authors
  1. Kiranmai Mandava
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Uma Rajeswari Batchu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shravya Kakulavaram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shulamithi Repally
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ishwarya Chennuri
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Srinivas Bedarakota
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Namratha Sunkara
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

KM and BUR designed and reviewed the research and wrote the paper, SK, SR, ICH and BS conducted the research, NS edited the analytical work. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Kiranmai Mandava.

Ethics declarations

Ethics approval and consent to participate

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Mandava, K., Batchu, U.R., Kakulavaram, S. et al. Design and study of anticaries effect of different medicinal plants against S.mutans glucosyltransferase. BMC Complement Altern Med 19, 197 (2019). https://doi.org/10.1186/s12906-019-2608-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12906-019-2608-3

Keywords

  • S.mutans
  • Glucosyltransferase
  • Azadirachta indica
  • Terminalia chebula
  • Polyherbal mouth wash
  • Anticaries agent

BMC Complementary Medicine and Therapies

ISSN: 2662-7671

Contact us