ORIGINAL ARTICLE


https://doi.org/10.5005/jp-journals-10063-0048
CODS Journal of Dentistry
Volume 11 | Issue 2 | Year 2019

Evaluation of Antibacterial Efficacy of Sesame Seed on Periodontal Pathogens: An In Vitro Study


Vangara Aditya1, Gunjiganur Ajjappa Babitha2, Shobha Prakash3, Juturu Timmasetty4

1–3Department of Periodontics, College of Dental Sciences, Rajiv Gandhi University of Health Sciences, Davangere, Karnataka, India
4Department of Pharmaceutics, Bapuji Pharmacy College, Davangere, Karnataka, India

Corresponding Author: Vangara Aditya, Department of Periodontics, College of Dental Sciences, Rajiv Gandhi University of Health Sciences Davangere, Karnataka, India, Phone: +91 9148188580, e-mail: dr.vangaraaditya@gmail.com

How to cite this article Aditya V, Babitha GA, Prakash S, et al. Evaluation of Antibacterial Efficacy of Sesame Seed on Periodontal Pathogens: An In Vitro Study. CODS J Dent 2019;11(2):44–47.

Source of support: Nil

Conflict of interest: None

ABSTRACT

Aim and objective: The aim and objective of this study was to identify the minimum inhibitory concentration (MIC) of pure concentrated sesame seed powder require to inhibit growth of periodontal pathogens like Porphyromonas (P.) gingivalis (Pg), Prevotella (P.) intermedia (Pi), Fusobacterium (F) nucleatum (Fn), Tannerella (T.) forsythia (Tf).

Materials and methods: Sesame seeds powder was obtained and minimum inhibitory concentration required to inhibit the growth of periodontal pathogens like P. gingivalis, P. intermedia, F. nucleatum, T. forsythia was determined by using the tube dilution method.

Results: MIC of sesame seed for Pg showed sensitive at 3.12 mg/mL, Pi at 6.25 mg/mL, Fn at 3.12 mg/mL, Tf at 50 mg/mL. Fn was sensitive until 3.12 mg/mL showed resistance to further dilution by illuminating its MIC. Hence, MIC values were considered as Tf < Pi < Pg = Fn which sensitive at these concentration.

Conclusion: Many studies in dentistry have shown its antibacterial efficacy on aerobic bacteria however, the present study findings conclude that the sesame seed powder exerts antimicrobial actions against major periodontal pathogens like Pg, Pi, Fn and Tf. So, further extensive in vivo studies are required to confirm the findings of the present study and explore therapeutic implications of sesame seed powder in the management of periodontal diseases.

Clinical significance: The antibacterial efficacy of sesame seed powder used as local drug delivery as chip, gel at particular concentration which will help to prevent further progression of periodontal pocket after scaling and root planning such as reduction of periodontal pocket.

Keywords: Bacteria, Indicator dilution techniques, Pedaliaceae, Periodontal pocket, Periodontitis, Seeds, Sesamum, Virulence factor.

INTRODUCTION

A periodontal disease is a group of localized microbial-induced infections that involve gingival and supporting tissues of teeth. This is not just a mere pathological deepening of gingival sulcus but may represent a sequel of microbial- and inflammatory-mediated degradation of collagenous connective tissues and alveolar bone.1 Periodontitis is caused by an increase in the virulence factors of the infecting organisms and the decreased resistance of the host.2 For centuries, various parts of the plants and plant products had been used for treating various systemic and oral ailments. The major vigor of these natural herbs is that they are time-proven to be safe without any adverse reactions.

Sesamum indicum is also known as Sesamum or benniseed or sesame, a member of the Pedaliaceae family, and it is an ancient oilseed crop. It plays a vital role in human health and nutrition.3 The qualitative and quantitative determination of sesamin bioactive compound in defatted ethanol extract was performed by high-performance liquid chromatography (HPLC) analysis, which was carried out and compared with standard sesamin. The results shown were that the concentration of sesamin was 79.9% of ethanolic extract according to the total peak area.4 Most sesame seeds grown were primarily used for oil extraction and remaining seeds were in edible form.4 These seeds were added for flavor and to garnish various food items, though they were primarily used as oil and wine.4 Once the extraction of oil from the seeds came out, some of it can be used as cake and also as manure. The seeds color may vary from cream-white to charcoal-black but it is usually white or black form. The extraction of oil from sesame seed was used to heal wounds from many years since the dawn of civilization.3

Two breeds of sesame seeds are usually cultivated in regions of Tiv and Idoma (Nigeria’s Benue state) they are Sesamum radiatum and S. indicum mainly for their seeds and leaves.5 It is a staple food consumed locally in regions (south-west, middle belt areas) of Nigeria where they are cultivated by local farmers5 and it accounts for high fecundity of people among the adult male population.5 These seeds can be consumed either as oil or in roast form or as animal feeds.5

Since many studies have been performed on sesame seeds but then there is a lack of evidence that gave an idea to find out its antimicrobial action of sesame seeds on anaerobic bacteria in this study.

MATERIALS AND METHODS

Commercially available sesame seeds were sent to the pharmaceutical laboratory (Fig. 1). The sesame seeds were converted into fine powder using motor and pestle. Before packing, ziplock bags were placed in a green pouch, autoclaved. This fine powder was packed (Fig. 2) in ziplock bags and sent to the microbiology laboratory for analysis. Quantity of sesame seed powder was measured using a weighing machine and 10 mg was taken for the study.

Fig. 1: Black sesame seeds

Preparation of Bacterial Suspension

From the maintained frozen stock cultures of Porphyromonas gingivalis (Pg) (ATCC no—33277), Prevotella intermedia (Pi) (ATCC no—25568), Tannerella forsythia (Tf) (ATCC no—700191), Fusobacterium nucleatum (Fn) (ATCC no—25586), a small number of cells were recovered and subcultured. The culture medium in this study was brain heart infusion (BHI) medium as a preferred medium for the growth of anaerobic bacteria.

The culture medium was then transferred into tubes containing 2 mL of the BHI medium to get culture suspension of Pg, Pi, Fn, and Tf, respectively. Now the test bacterial strains were selected and adjusted to 0.5 McFarland turbidity standards (108 colony-forming units/mL) then check for MIC of sesame seed powder against these bacteria.6 ATCC means American Type Culture Collection, Manassas, VA, USA.

Himedia M 210-500G

Brain heart infusion broth—500 g

Ingredientsg/L
Calf brain, infusion from200.00
Beef heart, infusion from250.00
Proteose peptone  10.00
Dextrose    2.00
Sodium chloride    5.00
Disodium phosphate    2.50

Final pH (at 25°C) 7.4 ± 0.2

MICROTITER BROTH DILUTION METHOD

Transfer of this sesame seed powder into an initial Eppendorf tube. A total of nine dilutions are required for the identification of each anaerobic bacteria in this study. Each dilution in an Eppendorf tube contains sesame seed powder along with BHI broth required for MIC. From this initial tube, 20 μL of sesame seed powder was added into 380 μL of BHI broth. For this, 200 μL of BHI broth was transferred into the next nine tubes separately. Now, the initial tube contains 400 μL that was prepared in total only 200 μL was transferred to the first tube containing 200 μL of BHI broth to make a dilution of 10−1 dilution required for this study. From this 10−1 diluted tube, 200 μL was then transferred to the second tube to make 10−2 dilution. Then, serial dilution was repeated until 10−9 dilution for each drug. Here from maintained stock cultures of required organisms, 5 μL was taken and added to 2 mL of BHI broth. From a serially diluted tube, 200 μL of above culture suspension was added. These tubes were incubated for 24 hours and observed for turbidity. Similarly, the procedure should be repeated separately for each bacterial strain. For facultative anaerobes, the temperature required to incubate the tube was 37°C for 48–72 hours in CO2 jar. For strict anaerobes, tubes were incubated in anaerobic jars for 48–72 hours (Figs 3 to 6). The turbidity for each tube was compared with the positive control, which contains only pure bacterial culture. The least concentration of a drug in the tube does not show any turbidity as it was considered as MIC to test for particular organism.7

Fig. 2: Sesame seeds powder

RESULTS

In Vitro Antibacterial Activity

In the present study, Pg, Pi, Fn, and Tf were sensitive to pure sesame seed powder. Porphyromonas gingivalis was sensitive until 3.12 mg/mL dilution, Pi was sensitive until 6.25 mg/mL, Tf was sensitive until 50 mg/mL, and Fn was sensitive until 3.12 mg/mL shows resistance to further dilution by illuminating its MIC. So, the MIC values were considered as Tf < Pi < Pg = Fn which were sensitive at these concentrations. Then, the mean value for MIC considered will be 50 mg/mL (Table 1).

DISCUSSION

The use of antibiotics as an adjunct to anti-infective therapies had been a gold standard for the treatment of periodontitis.8 A present dilemma was the resistance of periodontal pathogens to antibiotics which are currently in use. Persistent attempts were made to bring about natural, biocompatible, and efficient antibacterial agents against periodontal pathogens. Sesame oil is known as “gingelly oil or til oil” had antifungal, antioxidant, and health-promoting activities.9 A study by Ahmed et al. had revealed that methanolic extract of sesame shows a mild antimicrobial activity at a concentration of 60.7 μg/mL for Staphylococcus aureus 47.6 μg/mL for Candida albicans and Streptococcus pneumoniae.10 An experimental study by Reza et al. shows that olive oil and sesame oil were extracted by cold press assay and then both oils were mixed at an equal proportions. They concluded that the MIC of 128 and 32 mg/mL was effective against S. aureus.11 A randomized controlled trial conducted by Singla et al. on the efficacy of sesame, coconut, and olive oil on controlling the gingival inflammation and agents were compared with chlorhexidine (CHX).12 The study concluded that these herbs had potent antibacterial and efficacy to CHX against pathogens.12 A study by Rao et al. stated that an aqueous extract of sesame oil had an antimicrobial activity for Pseudomonas aeruginosa, Escherichia coli, S. aureus, Klebsiella pneumoniae, and C. albicans.13 The results obtained from this study showed that at full concentration extracts of S. indicum seed had inhibitory effects on Pg, Pi, Fn, and Tf, at various concentrations.

This study is also in tandem with other studies proved its antibacterial effects on natural products.14 To summarize, sesame seed powder has been evaluated to know its degree of antimicrobial activity against major anaerobic periodontal pathogens (Pg, Pi, Fn, and Tf).

Fig. 3: Serial dilution of Porphyromonas gingivalis

Fig. 4: Serial dilution of Prevotella intermedia

Fig. 5: Serial dilution of Fusobacterium nucleatum

Fig. 6: Serial dilution of Tannerella forsythia

Table 1: Minimum inhibitory concentration
Sesame seed powder100 mg/mL502512.56.253.121.60.80.4
PgSSSSSSRRR
PiSSSSSRRRR
FnSSSSSSRRR
TfSSRRRRRRR

Fn, Fusobacterium nucleatum; Pg, Porphyromonas gingivalis; Pi, Prevotella intermedia; R, resistance; S, sensitive; Tf, Tannerella forsythia

CONCLUSION

Many studies in dentistry have shown its antibacterial efficacy on aerobic bacteria; however, the present study findings conclude that the sesame seed powder exerts antimicrobial actions against major periodontal pathogens, such as Pg, Pi, Fn, and Tf. So, further extensive in vivo studies are required to confirm the findings of the present study and explore the therapeutic implications of sesame seed powder in the management of periodontal diseases.

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