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Antimicrobial Activity of Essential Oil Against Oral Strain Original Article Int J Clin Prev Dent 2018;14(4):216-221ㆍhttps://doi.org/10.15236/ijcpd.2018.14.4.216 ISSN (Print) 1738-8546ㆍISSN (Online) 2287-6197 Antimicrobial Activity of Essential Oil against Oral Strain Chungmu Park1, Hyunseo Yoon2 Departments of 1Clinical Laboratory Science and 2Dental Hygiene, Dong-Eui University, Busan, Korea Objective: In this study, we tried to investigate the antimicrobial activity of natural herbaceous plant-derived essential oils against oral disease-causing bacterial strains and establish the scientific evidences to apply them to the dentifrice, mouth- wash, and other uses. Methods: We used the disk diffusion method with 23 types of 100% natural essential oils to verify the antimicrobial effect against Streptococcus mutans, Porphyromonas gingivalis, and Lactobacillus rhamnosus. After adhering to the surface of the plate culture medium, 20 l of the essential oil was prepared, and the diameter of the transparent ring was measured after in- cubation for 24 hours. All the experiments were repeated three times. Results: Among 23 types of oils, 13 were effective against all three strains: myrrh, ginger, basil, carrot seed, tea tree, patch- ouli, ylang ylang, cypress, lemongrass, cinnamon, peppermint, lavender, and eucalryptus. Seventeen oils were effective against S. mutans, and myrrh, basil, and carrot seed showed high antimicrobial activity. Eighteen oils were effective against P. gingivalis, and tea tree, carrot seed, and cinnamons showed high antimicrobial activity. Sixteen oils were effective against L. rhamnosus, and carrot seed and peppermint cinnamon showed high antimicrobial activity. Conclusion: Consequently, 13 essential oils showed the antimicrobial activity against three bacterial strains, which indicates these essential oils might be used as the effective materials to suppress the growth of oral-disease inducing microorganisms. Moreover, essential oils that have been analyzed activities in this study will be mixed up within the constant range and ana- lyzed their antimicrobial effects to examine the synergistic activity among them. Keywords: anti-microbial agents, essential oil, Lactobacillus rhamnosus, Porphyromonas gingivalis, Streptococcus mutans Introduction creased, people are paying more attention to natural extracts with fewer side effects than the synthetic additives or extracts As the interest in economic development and health has in- [1]. The natural extracts are obtained from medicinal plants and edible plants by various methods such as compression, Corresponding author Hyunseo Yoon freeze extraction, solvent extraction, and steam distillation, Department of Dental Hygiene, Dong-Eui University, 176 which were used for oral medication or skin treatment. Eongwang-ro, Busanjin-gu, Busan 47340, Korea. Tel: Among them, the essential oils that condense fragrances ex- +82-51-890-2688, Fax: +82-505-182-6878, E-mail: yoonhs@ tracted from herb and other plants are more useful because deu.ac.kr they contain many bioactive components [2,3]. These natural https://orcid.org/0000-0002-7455-5506 extracts or oils are used as the natural preservatives for food Received September 6, 2018, Revised November 2, 2018, and grains [4] or cosmetic functional additives [5]. Accepted December 4, 2018 The essential oil is the common product of extracts of flow, stem, and leave components of plants [6]. The essential oil has Copyright ⓒ 2018. Korean Academy of Preventive Dentistry. All rights reserved. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 216 Chungmu Park and Hyunseo Yoon：Antimicrobial Activitiy of Various Essential Oils been known to have pharmacological effects such as relieving is the leading strain causing periodontal disease [2]. Among pain and alleviating acne through antimicrobial and anti-in- essential oils, peppermint and lemongrass oils showed anti- flammatory activities against acne-causing bacteria [6-9]. microbial effects against P. gingivalis [20]. Moreover, it is proven to be an effective antioxidant in food First, diverse mint-flavored essential oils were selected spices and utilized in various fields such as seeking psycho- since they would be used for the additives for dentifrice or logical stability [10-12]. mouthwash. Then, their antimicrobial activities were ana- Thus far, chlorohexidine, cetylpyridinium chloride and flu- lyzed through the inhibited growth against three representa- orine have been commonly used for additive agents for tive microorganisms including S. mutans, P. gingivalis, and mouthwashing solution. By the way, they have various side ef- Lactobacillus rhamnosus which can evoke oral disease. fects such as teeth stain, unpleasant taste, and oral pain in spite Thourghout this study, essential oils that contain antimicrobial of their outstanding bacteriostatic activity [13]. Therefore, activities against oral-disease causing strains. many researchers have tried to search natural resources- de- rived materials which can inhibit the proliferation of micro- Materials and Methods organisms that cause oral diseases to overcome these prob- lems [1]. Specifically, the studies verified the antimicrobial 1. Materials activity of the plum extract [14], bamboo salt [15], green tea leaf, mulberry leaf, and matte leaf extract [16] against We used 100% natural essential oils sold by UK-based Streptococcus mutans which is the leading strain causing den- Aqua Oleum. There were 23 types of oils as shown in Table 1. tal caries. Other studies identified the antimicrobial activity They were mixed with Tween 20 (Anatrace Products, LLC, tea tree oil [17] and orange oil [18] among the essential oils. Maumee, OH, USA) and then diluted with triple distilled wa- Moreover, the studies also exhibited the antimicrobial activity ter into 50 % (v/v) to be the test sample. of Alnus firma extract [19], myrrh extract, rhatany extract, and chamomile extract against Porphyromonas gingivalis which Table 1. Characteristics of materials Extraction Case No. Oil name Botanical name Cultivated area methoda 1 Myrrh Commiphora myrrha Somalia 1 2Rosemary Rosemarinus officinalis France 1 3 Ginger Zingiber officinale Nigeria 1 4Pine Pinus Sylvestris Siberia 1 5Basil Ocimum basilicum Egypt 1 6 Carrot seed Daucus carota Belgium 1 7Tea tree Melaleuca alternifolia Australia 1 8Neroli Citrus aurantium subsp. amara or Bigaradia Tunisia 1 9 Patchouli Pogostemon patchouli Indonesia 1 10 Palmarosa Cymbopogon martini India 1 11 Ylang Ylang Cananga odorata Masagascar 1 12 Grape Fruit Citrus paradisi California 2 13 Cypress Cupressus sempervirens France 1 14 Lemongrass Cymbopogon citratus Guatemala 1 15 Majoram Origanum majorane Mediterranean Sea 1 16 Lemon Citratus limon Asia 3 17 Cederwood Cedrus atlantica Morocco 1 18 Cinnamon Cinnamomum zeylanicum Sri Lanka 1 19 Mandarin Citrus reticulata Italy 2 20 Peppermint Mentha piperita United Kingdom 1 21 Lavender Lavandula angustifolia France 1 22 Eucalryptus Eucalyptus globulus Spain 1 23 Spearmint Mentha spicata Europe 1 a1: water vapor distillation, 2: cooling extraction method, 3: cooling compression extraction method. www.ijcpd.org 217 International Journal of Clinical Preventive Dentistry patchouli, ylang ylang, grape fruit, cypress, lemongrass, lemon, 2. Tested strains and cultivation condition cederwood, cinnamon, peppermint, lavender, and spearmint. To measure the antimicrobial effect of the samples against Sixteen oils showed the antimicrobial effect against L. the strains causing oral diseases, we procured the S. mutans rhamnosus: myrrh, ginger, basil, carrot seed, tea tree, patch- (KCTC 3065) and P. gingivalis (5352) strains from Korean ouli, palmarosa, ylang ylang, cypress, lemongrass, marjoram, Collection for Type Cultures in Daejeon, Korea and the L. cinnamon, peppermint, lavender, eucalryptus, and spearmint. rhamnosus (KCOM 1213) strains from the Korean Collection 2. Comparison of antimicrobial activity of for Oral Microbiology S. mutans and L. rhamnosus strains essential oil against S. mutans were cultured on agar medium of brain heart infusion (Difco Laboratories Inc., Detroit, MI, USA) while P. gingivalis Table 3 shows the measurement results of the diameter of strains were cultured on tryptic soy agar (Difco) medium con- the transparent rings using the disk diffusion method to check taining 10% sheep blood, 0.1% hemin (Sigma, St. Louis, MO, the antimicrobial activity against S. mutans. Among 17 essen- USA), and containing vitamin K1 (Sigma). tial oils that had the antimicrobial activity against S. mutans, myrrh showed the highest antimicrobial activity at 18.34 mm 3. Measurement of antimicrobial activity using and followed by basil at 18.08 mm, and carrot seed at 17.91 disk diffusion method mm (p<0.001). We measured the antimicrobial activity of essential oil 3. Comparison of antimicrobial activity of against S. mutans, P. gingivalis, and L. rhamnosus using the essential oil against P. gingivalis disk diffusion method [21]. All strains were tested in accord- ance with NCCLS Guide Line M11-A6 (National Committee Table 4 shows the measurement results of the diameter of for Clinical Laboratory Standards, 2004). Each strain was in- the transparent rings using the disk diffusion method to check oculated on a plate medium, a sterilized paper disk (Advantec; the antimicrobial activity against P. gingivalis.
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