Volume 22, Issue 4 (9-2019)                   J Arak Uni Med Sci 2019, 22(4): 16-27 | Back to browse issues page

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Iranpoor A, Bayani M, Arjomandzadegan M, Nakhostin A. Antibacterial Activity and Antibiofilm Properties of Satureja Essential Oil Against Periodontal Pathogens. J Arak Uni Med Sci 2019; 22 (4) :16-27
URL: http://jams.arakmu.ac.ir/article-1-6081-en.html
1- Dentist, Arak, Iran.
2- Department of Periodontics, School of Dentistry, Arak University of Medical Sciences, Arak, Iran.
3- Tuberculosis and Pediatrics Infectious Diseases Research Center, Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
4- Department of Restorative and Aesthetic Dentistry, School of Dentistry, Arak University of Medical Sciences, Arak, Iran. , afr_na_sa@yahoo.com
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1. Introduction
Tooth decay and periodontal disease are the most common infectious human diseases that are caused by oral bacteria such as Staphylococcus aureus, Micrococcus, Pseudomonas, Candida, etc. [1]. Various ways, such as chemical mouthwashes, toothpaste, and antibiotics, are used to prevent and treat periodontal disease and tooth decay [10, 11, 13].
Herbal and other natural antibacterial materials are now considered as useful and alternative antibacterial methods for oral and toothpaste washing [14]. Satureja has numerous compounds with antibacterial properties [15].
Since the oral environment contains many bacterial species and the use of chemical drugs in the oral cavity is associated with complications such as changes in the natural flora, in this study, we investigated the effect of Satureja oil on some common oral bacteria.
2. Materials and Methods
This experimental study was performed on microbial strains provided from Iran Microbial Bank (including Enterococcus faecalis, Streptococcus sanguis, Eikenella corrodens, and Actinomyces viscose). Some fresh Satureja was obtained from the Research Center for Medicinal Plants of Shahid Beheshti University, and its essential oil by water distillation or Clevenger apparatus.
The disk diffusion method was used to determine microbial susceptibility to essential oil after preparation of microbial suspension at half McFarland concentration and its culturing as aqueous media on Muller-Hinton Agar medium. The blank disk was immersed in 20 µL of essential oil with concentrations of 1 g/mL and 0.1 g/mL on the culture medium and placed at a suitable distance from the plate incubated at 37°C for 48 hours. After the incubation period, the bacterial growth inhibition zone diameter was measured in millimeters. The experiment was repeated 3 times, and the mean results was reported.
The microplate dilution method was also used to determine the antimicrobial effect of the essential oil. Median inhibitory and lethal concentrations were also determined in this study. The microplate method was also used to evaluate the effect of essential oil on biofilm formation inhibition.
3. Results
The results of microbial susceptibility to Satureja essential oil showed that the highest diameter of 30 mm growth inhibition zone was related to the effect of Satureja essential oil at a concentration of 1 g/mL on the growth of Streptococcus sanguis and Eikenella corrodens. The lowest diameter of 9 mm growth inhibition zone was due to the effect of essential oil of 0.1 g/mL on the growth of Eikenella corrodens. Table 1 presents the results of the evaluation of the median inhibitory concentration and median lethal concentration of the essential oil of the Satureja on the studied bacteria.

The median growth inhibitory concentration and the median growth inhibitory concentration were presented in μg/mL. The results of the microplate dilution test showed that the studied bacteria at higher concentrations of 1.562 mg/mL of Satureja essential oil lost their growth ability but still retain their growth potential at lower concentrations of 0.195 mg/mL of this essential oil.
The results of the biofilm inhibition test showed that the studied bacteria did not form biofilms at concentrations higher than or equal to 12.5 mg/mL, but the biofilms were formed at concentrations below 0.39 mg/mL.
4. Discussion
The present study was carried out to investigate the antibacterial activity of the essential oil of Satureja by disk diffusion and microplate dilution methods. The microplate technique was also used to examine the effect of essential oil on biofilm formation. The results of the disk diffusion method, considering the size of the bacterial growth inhibition zone, showed that pure Satureja essential oil (1 g/mL) had a strong antimicrobial activity compared to essential oil at 0.1 g/mL. 
By examining the results of the microplate dilution test, the median concentration of essential oil that could inhibit bacterial growth was 1.562 mg/mL for three bacteria of Enterococcus faecalis, Streptococcus sanguis, and Actinomyces viscose. However, this concentration was 0.39 mg/mL for Eikenella corrodens. These results show that Satureja essential oil has a better inhibitory effect on the growth of Eikenella corrodens than Enterococcus faecalis, Streptococcus sanguis, and Actinomyces viscose.
Overall, the results of this study showed that the essential oil of Satureja has antibacterial activity and can inhibit the growth of a group of periopathogens, including Enterococcus faecalis, Streptococcus sanguis, Eikenella corrodens, and Actinomyces viscose. It can also inhibit the growth of the mentioned bacteria at a concentration of 1.562 mg/mL and more Satureja essential oil. It also has an inhibitory effect on the biofilm formation of the mentioned bacteria so that at concentrations of 12.5 mg/mL and higher of Satureja essential oil, none of the studied bacteria was able to form a biofilm. Therefore, Satureja, as a natural and effective antibacterial, can have a potential role in reducing the probability of incidence and severity of periodontal diseases.
Ethical Considerations
Compliance with ethical guidelines

The Ethics Committee of the Arak University of Medical Sciences approved this study (Code: IR.ARAKMU.REC.1397.67).
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Authors' contributions
All authors met the standards of writing criteria based on guidelines of the  International Committee of Medical Journal Publishers (ICMJE).
Conflicts of interest
The authors declare no conflicts of interest.
The authors would like to thank the authorities of Arak University of Medical Sciences for their cooperation in conducting this research.
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Type of Study: Original Atricle | Subject: Basic Sciences
Received: 2019/05/20 | Accepted: 2019/09/7

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