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International Journal of Clinical Pediatric Dentistry logoLink to International Journal of Clinical Pediatric Dentistry
. 2013 Apr 26;6(1):22–25. doi: 10.5005/jp-journals-10005-1180

Effect of Propolis on Streptococcus mutans Counts: An in vivo Study

K Sundeep Hegde 1, Sham S Bhat 2,✉, Ajay Rao 3, Shaniya Sain 4
PMCID: PMC4034638  PMID: 25206182

ABSTRACT

Propolis, a natural antibiotic, is a resinous substance that honey bees (Apis mellifera) produce. The main chemical classes present in propolis are flavonoids, phenolics and other various aromatic compounds.

Aim: To evaluate the antibacterial action of propolis on the concentration of Streptococcus mutans colonizing the oral cavity of children.

Materials and methods: Thirty children performed the rinses, with no other changes in their oral hygiene and dietary habits. Saliva was collected at two time points: Before using the product, 1 hour after the rinse.

Results: Paired t-test was used for analysis of the results. A reduction in the concentration of Streptococcus mutans was observed in samples collected after use of the extract. There was a reduction in Streptococcus mutans count when compared to samples obtained in baseline. Significant reductions were seen at the end of 1 hour. The result was statistically significant. There were no side effects in soft and hard tissues of mouth.

Conclusion and clinical implication: The propolis possesses in vivo antimicrobial activity against Streptococcus mutans present in the oral cavity and might be used as a measure to prevent dental caries.

How to cite this article: Hegde KS, Bhat SS, Rao A, Sain S. Effect of Propolis on Streptococcus mutans Counts: An in vivo Study. Int J Clin Pediatr Dent 2013;6(1):22-25.

Keywords: Propolis, Streptococcus mutanscount, Saliva

INTRODUCTION

In the new era of globalization there is an evident paradigm shift in health care approaches. The critical aspect of focus and research is toward evolving innovative green perceptions in health care research and practices. The basis of these movements is anchored towards producing tangible process linked products that could be applicable on a global scale and carries with itself a label–Eco friendly. This line of thought has lead therapeutic research to experiment with medicinal properties of various plants and to evolve pharmaceutical products.

In similar context, propolis is a resinous substance. The word propolis (Russian Penicillin) is derived from the Greek word ‘pro’ before, polis ‘city’ or defender of the city. Honey bees (Apis mellifera) collect the resin from cracks in he bark of trees and leaf buds. It is masticated, salivary nzymes are added and the partially digested material is mixed with bee wax and used by bees to seal holes in their honeycombs, smooth out the internal walls and protect the entrance against external agents and contaminants.1 Propolis is composed of 50% resin and vegetable balsam, 30% wax, 10% essential and aromatic oils, 5% pollen and 5% various other substances, including organic debris depending on the place and time of collection.2,3 It is a natural antibiotic. The medicinal properties are due to the flavonoids, phenolics and various aromatic compounds. Flavonoids have antibacterial, antifungal, antiviral, antioxidant and anti-inflammatory proprieties. Galangin, pinocembrin and pinostrobin are known as the most effective flavonoids agents against bacteria. Ferulic acid and caffeic acid also contribute to the bactericidal action of propolis.4

Propolis has been widely used for clinical trials in dentistry for various purposes and seems to be promising. As an anti-inflammatory agent, propolis is shown to inhibit synthesis of prostaglandins, activate the thymus gland, aid the immune system by promoting phagocytic activity, stimulate cellular immunity and augment healing effects on epithelial tissues.5-7 Propolis also contains iron and zinc that are important for the synthesis of collagen.

Potential uses in dentistry are wound healing, storage media following avulsion,8 a pulp capping agent, intracanal rrigant, intracanal medicament, mouth rinse, cariostatic agent, n dentinal hypersensitivity, in treatment of periodontitis, has effect on Candida albicans, in treatment of denture stomatitis and has effect on recurrent aphthous stomatitis.9

Dental caries is the most prevalent disease affecting humans, and its susceptibility is much higher in childhood. During the initial phase of caries, Streptococcus mutans is the most frequently associated microorganism. In addition to its ability to adhere to teeth and survive in acid environment, Streptococcus mutans is transmissible, as first demonstrated by Keyes in his trials.10

AIM

To evaluate the antibacterial action of propolis on Streptococcus mutans colonizing the oral cavity of children.

OBJECTIVE

The objective of the study was to evaluate the in vivo antimicrobial activity of an extract prepared with propolis when used as mouth rinse on the colony-forming units of Streptococcus mutans present in the oral cavity of children.

MATERIAL AND METHOD

Determination of Antibacterial Efficacy

Streptococcus mutans MTCC 890 strains (Fig. 1) were used for the study. Serial dilutions of propolis (20, 10, 5, 3 and 2.5%) were used. Streptococcus mutans culture was swabbed evenly onto Trypticase soy agar (TSA) media (Fig. 2) and wells

Fig. 1.

Fig. 1

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MTCC-890 strain

Fig. 2.

Fig. 2

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Determination of antibacterial efficacy

Preparation of Propolis

Five percent propolis, commercially available as propolis platinum [K-Link Healthcare (India) Pvt Ltd Chennai] (Fig. 3) was diluted in sterile water (5 ml diluted in 90 ml of sterile water) and was used for the study (Fig. 4).

Fig. 3.

Fig. 3

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Commercially available propolis extract

Fig. 4.

Fig. 4

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Diluted propolis extract

Thirty children of both sexes, ranging in age from 5 to 10 years were enrolled for the study with the consent of parents. First samples of whole saliva were collected into sterile collection vials (3 ml on the average) and it was seeded in the laboratory (Fig. 5).

Fig. 5.

Fig. 5

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Collection of saliva

The children were then asked to rinse their mouth with 3 ml of the diluted propolis extract solution for 1 minute and a second saliva samples was collected 1 hour later in the same way as described for the first sample. The volunteers performed the mouth rinses with no other changes in their oral hygiene, dietary habits and day-to-day practices. The microbiological analysis was carried out using the selective Streptococcus mutans media. It permits semiquantitative analysis of this microorganism in salivary samples. Finally colony-forming units were counted (Figs 6 and Figs 7).

Fig. 6.

Fig. 6

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Colony counting grid

Fig. 7.

Fig. 7

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Pre and post samples with Streptococcus mutans

RESULT

Minimum inhibition concentration was seen to be 5%. Of the 30 saliva samples collected from the 30 volunteers, prebacterial counts ranged from 800 to 91,00,000 CFU/ml saliva (Table 1).

Table 1: Pre and post counts of Streptococcus mutans

    No.     Pre     Post (1 hour)     Log 1     Log 2     Log 1-Log 2
    1.         40000         2600         4.602         3.414         1.188    
    2.         6000         4000         3.778         3.602         0.1761    
    3.         20000         1500         4.301         3.176         1.125    
    4.         28000         14000         4.447         4.146         0.301    
    5.         30000         1000         4.477         3         1.447    
    6.         400000         30000         5.602         4.477         1.125    
    7.         500000         20000         5.698         4.301         1.397    
    8.         5000         0         3.698         0         3.698    
    9.         80000         4000         4.903         3.602         1.301    
    10.         32000         2800         4.505         4.447         0.103    
    11.         11000         5000         4.041         3.698         0.343    
    12.         270000         10000         5.431         4         1.431    
    13.         7000         2500         3.845         3.397         0.448    
    14.         20000         1300         4.301         3.113         1.188    
    15.         16000         7200         4.204         3.857         0.347    
    16.         300000         23000         5.477         4.361         1.116    
    17.         19000         8000         4.278         3.903         0.375    
    18.         800         800         2.903         2.903         0.000    
    19.         5000         400         3.698         2.602         1.096    
    20.         6000         200         3.778         2.301         1.477    
    21.         180000         90000         5.255         4.954         0.301    
    22.         86000         10000         4.934         4         0.934    
    23.         910000         43000         5.959         4.633         1.326    
    24.         6200         530         3.792         2.724         1.068    
    25.         98000         3000         4.991         3.477         1.514    
    26.         29000         1300         4.462         3.113         1.349    
    27.         60000         2300         4.778         3.361         1.417    
    28.         30000         1000         4.477         3         1.477    
    29.         3000         0         3.477         0         3.477    
    30.         400000         2200         5.602         3.342         2.260    
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Statistical analysis was done by the paired t-test. The results showed a significant difference in the number of S. mutans between collections 1 and 2 (mean ± SD: 1.1597 ± 0.8560; t = 2.045 and p < 0.05) i.e. an effect of propolis on bacterial growth both after the beginning (collection 1) and at the end of treatment (collection 2). These results indicate a reduction in the number of S. mutans (Graph 1). Analysis of variance to determine the relationship between the number of mouth rinses and bacterial counts indicated a significant difference.

Graph 1.

Graph 1

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Red line indicates pre and blue lines indicates post results

DISCUSSION

Majority of the samples showed a decrease in the colonies. A total of 90% showed reduction in bacterial load. In 6.6% there were no colony-forming units after mouth rinse. A total of 3.4% showed no reduction after rinsing with mouthwash. A significant reduction in the number of colonies in the samples is the result of the effect of the propolis extract on bacterial growth. Samples that showed no change in the number of bacteria between collections might have been influenced by overlapping factors, such as a delayed peak formation of colonies, i.e. more than 2 hours after the meal, together with the short period for an effective action and the transmissible nature of S. mutans.11,12 There were no side effects in soft and hard tissues of mouth.

Streptococcus mutans is not the only one organism which causes the disease. The etiology of dental caries clearly points out that dental caries is multifactorial. Many other microbial agents along with time, dietary factors and host factors results in caries.13

Studies had demonstrated that host binding characteristics are as important as the characteristics of bacterial adhesion in the process of colonization. It was suggested that salivary amylase may show the best binding to S. mutans. It was also seen that bacterial interactions have a key role in colonization.14

Propolis has activity against Gram-positive, Gram-negative organisms and even against Candida. Certain chemical components of propolis act on the cell wall of microorganisms causing functional and structural damages. It has mucoprotective effect so can be used efficiently in the oral cavity.15

Within the philosophy of health promotion, the extract of propolis may represent a new option showing long-term beneficial effects. Further clinical studies with large samples, long-term follow-up and comparison with conventional mouthwash is required.

CONCLUSION

The propolis possesses in vivo antimicrobial activity against S. mutans present in the oral cavity and might be used as an alternative measure to prevent dental caries.

Acknowledgments

The authors would like to thank Dr Arun Bhagwath and Dr Rekha Bhagwath, Yenepoya Research Centre, Yenepoya University for their technical assistance and analysis in the course of the study.

Footnotes

Source of support: Nil

Conflict of interest: None declared

Contributor Information

K Sundeep Hegde, Professor, Department of Pedodontics and Preventive Dentistry Yenepoya Dental College, Mangalore, Karnataka, India.

Sham S Bhat, Professor and Head, Department of Pedodontics and Preventive Dentistry, Yenepoya Dental College, Mangalore, Karnataka, India.

Ajay Rao, Reader, Department of Pedodontics and Preventive Dentistry, Yenepoya Dental College, Mangalore, Karnataka, India.

Shaniya Sain, Postgraduate Student, Department of Pedodontics and Preventive Dentistry, Yenepoya Dental College, Mangalore, Karnataka, India.

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