Anti-quorum sensing and cytotoxic activity of elemi essential oil

Essential oils have several biological activities such as antimicrobial, antioxidant, proliferative, and anti-inflammatory. This study aimed identification of bioactive compounds found in Elemi essential oil (EO) and to determine the anti-quorum sensing and cytotoxic activities of EO. In this study, bioactive compounds of EO were analyzed using GC-MS, and the antibacterial activity of elemi was screened against Staphylococcus aureus ATCC 25923, Methicillin-Resistant Staphylococcus aureus ATCC 43300, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Pseudomonas aeruginosa PAO1. Anti-biofilm activity and pyocyanin production on P. aeruginosa PAO1 were also investigated. The effect of EO on cell viability was also analyzed by thiazolyl blue tetrazolium bromide (MTT) and neutral red uptake (NR) assay in fibroblast cells. According to GC results, the major component of EO was determined as limonene (55%). A sub-MIC of elemi essential oil inhibited biofilm formation and pyocyanin production by 43% and 56%, respectively. On the other hand, EO also had an acute effect on the mitochondrial and lysosomal activities of fibroblast cell lines. Mitochondrial and lysosomal activities were significantly decreased when EO concentrations were applied for 24 and 48 hours (p<0.05). In conclusion, EO has inhibitory activity on biofilm formation and pyocyanin production, and also the lower doses of oil have no toxic effects on fibroblast cells. However, higher doses of EO have more cytotoxic effects on mitochondrial activity rather than the lysosomal activity of fibroblast cell lines. It is thought that EO exhibits these activities due to the amount of limonene in its content.

Anahtar Kelimeler:

Essential oil, Elemi oil, Anti-bacterial, Anti-quorum, Cellular activity

Anti-quorum sensing and cytotoxic activity of elemi essential oil

Keywords:

Essential oil, Elemi oil, Anti-bacterial, Anti-quorum, Cellular activity,

PDF

___

Abisado, R.G., Benomar, S., Klaus, J.R., Dandekar, A.A., & Chandler, J.R. (2018). Bacterial quorum sensing and microbial community interactions. MBio, 9(3), e02331-17. https://doi.org/10.1128/mBio.02331-17
Algburi, A., Comito, N., Kashtanov, D., Dicks, L.M.T., & Chikindas, M.L. (2017). Control of biofilm formation: antibiotics and beyond. Applied and Environmental Microbiology, 83(3), e02508-16. https://doi.org/10.1128/AEM.02508-16
Alva, P.P., Suresh, S., Gururaj, M.P., & Premanath, R. (2019). Evaluation of anti-quorum sensing activity of indigenous dietary plants against Pseudomonas aeruginosa. European Journal of Integrative Medicine, 30, 100931. https://doi.org/10.1016/j.eujim.2019.100931
Banu, N., & Mary, R.N.I. (2016). Review on Anti Quorum Sensing Activity in Pseudomonas Aeruginosa Using Plants Extracts. 18(3), 663–671.
Bhuiyan, F.R., Howlader, S., Raihan, T., and Hasan, M. (2020). Plants metabolites: Possibility of natural therapeutics against the COVID-19 pandemic. Frontiers in Medicine, 7, 444. https://doi.org/10.3389/fmed.2020.00444
Bilenler, T., & Gökbulut, İ. (2013). Determination of Sensitivity of Hospital Pathogens Against Commercial Essential Oil. Gümüşhane University Journal of Science and Technology, 9(4), 716–723. https://doi.org/0.17714/gumusfenbil.537244
Borenfreund, E., Babich, H., & Martin-Alguacil, N. (1988). Comparisons of two in vitro cytotoxicity assays—the neutral red (NR) and tetrazolium MTT tests. Toxicology in Vitro, 2(1), 1–6. https://doi.org/10.1016/0887-2333(88)90030-6
Ceylan, O., Uğur, A., Saraç, N., Ozcan, F., & Baygar, T. (2014). The in vitro antibiofilm activity of Rosmarinus officinalis L. essential oil against multiple antibiotic resistant Pseudomonas sp. and Staphylococcus sp. J. Food. Agric. Environ, 12, 82–86.
Dănilă, E., Moldovan, Z., Popa, M., Chifiriuc, M.C., Kaya, A.D., & Kaya, M.A. (2018). Chemical composition, antimicrobial and antibiofilm efficacy of C. limon and L. angustifolia EOs and of their mixtures against Staphylococcus epidermidis clinical strains. Industrial Crops and Products, 122, 483 492. https://doi.org/10.1016/j.indcrop.2018.06.019
Essar, D.W., Eberly, L.E.E., Hadero, A., & Crawford, I.P. (1990). Identification and characterization of genes for a second anthranilate synthase in Pseudomonas aeruginosa: interchangeability of the two anthranilate synthases and evolutionary implications. Journal of Bacteriology, 172(2), 884–900. https://doi.org/10.1128/jb.172.2.884-900.1990
Fotakis, G., & Timbrell, J.A. (2006). In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicology Letters, 160(2), 171 177. https://doi.org/10.1016/j.toxlet.2005.07.001
Galovičová, L., Valková, V., Štefániková, J., & Kačániová, M. (2020). Essential oils and their application in a food model. Slovak Journal of Food Sciences, 14, 1088–1096. https://doi.org/10.5219/1490
Gürağaç Dereli, F.T., Önem, E., Özaydın, A.G., Arın, E., & Muhammed, M.T. (2022). Persea americana Mill.: As a potent quorum sensing inhibitor of Pseudomonas aeruginosa PAO1 virulence. International Journal of Secondary Metabolite, 9(1), 14 26. https://doi.org/10.21448/ijsm.1029610
Holder, I.A., & Boyce, S.T. (1994). Agar well diffusion assay testing of bacterial susceptibility to various antimicrobials in concentrations non-toxic for human cells in culture. Burns, 20(5), 426–429. https://doi.org/10.1016/0305-4179(94)90035-3
Hyldgaard, M., Mygind, T., & Meyer, R.L. (2012). Essential oils in food preservation: mode of action, synergies, and interactions with food matrix components. Frontiers in Microbiology, 3, 12. https://doi.org/10.3389/fmicb.2012.00012
John, K.M.M., Bhagwat, A.A., & Luthria, D.L. (2017). Swarm motility inhibitory and antioxidant activities of pomegranate peel processed under three drying conditions. Food Chemistry, 235, 145–153. https://doi.org/10.1016/j.foodchem.2017.04.143
Kačániová, M., Terentjeva, M., Štefániková, J., Žiarovská, J., Savitskaya, T., Grinshpan, D., Kowalczewski, P.Ł., Vukovic, N., & Tvrdá, E. (2020). Chemical composition and antimicrobial activity of selected essential oils against Staphylococcus spp. isolated from human semen. Antibiotics, 9(11), 765. https://doi.org/10.3390/antibiotics9110765
Koutsoudaki, C., Krsek, M., & Rodger, A. (2005). Chemical composition and antibacterial activity of the essential oil and the gum of Pistacia lentiscus Var. chia. Journal of Agricultural and Food Chemistry, 53(20), 7681–7685. https://doi.org/10.1021/jf050639s
Langeveld, W.T., Veldhuizen, E.J.A., & Burt, S.A. (2014). Synergy between essential oil components and antibiotics: a review. Critical Reviews in Microbiology, 40(1), 76–94. https://doi.org/ 10.3109/1040841X.2013.763219
Lewis, K. (2001). Riddle of biofilm resistance. Antimicrobial Agents and Chemotherapy, 45(4), 999–1007. https://doi.org/10.1128/AAC.45.4.999-1007.2001
Millezi, A.F., Piccoli, R.H., Oliveira, J.M., & Pereira, M.O. (2016). Anti-biofim and antibacterial effect of essential oils and their major compounds. Journal of Essential Oil Bearing Plants, 19(3), 624–631. https://doi.org/10.1080/0972060X.2014.960262
Mogana, R., & Wiart, C. (2011). Canarium L.: a phytochemical and pharmacological review. J Pharm Res, 4(8), 2482–2489. https://doi.org/10.4061/2011/768673
Morohoshi, T., Shiono, T., Takidouchi, K., Kato, M., Kato, N., Kato, J., & Ikeda, T. (2007). Inhibition of quorum sensing in Serratia marcescens AS-1 by synthetic analogs of N-acylhomoserine lactone. Applied and Environmental Microbiology, 73(20), 6339–6344. https://doi.org/10.1128/AEM.00593-07
Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65(1–2), 55–63. https://doi.org/ 10.1016/0022-1759(83)90303-4
Mukhtar, Y.M., Adu-Frimpong, M., Xu, X., & Yu, J. (2018). Biochemical significance of limonene and its metabolites: future prospects for designing and developing highly potent anticancer drugs. Bioscience Reports, 38(6), BSR20181253. https://doi.org/10.1042/BSR20181253
Murthy, K.N.C., Jayaprakasha, G.K., & Patil, B.S. (2012). D-limonene rich volatile oil from blood oranges inhibits angiogenesis, metastasis and cell death in human colon cancer cells. Life Sciences, 91(11–12), 429–439. https://doi.org/10.1016/j.lfs.2012.08.016
Nannapaneni, R., Chalova, V.I., Crandall, P.G., Ricke, S.C., Johnson, M.G., & O’Bryan, C.A. (2009). Campylobacter and Arcobacter species sensitivity to commercial orange oil fractions. International Journal of Food Microbiology, 129(1), 43 49. https://doi.org/10.1016/j.ijfoodmicro.2008.11.008
Nikolic, M., Smiljkovic, M., Markovic, T., Cirica, A., Glamoclija, J., Markovic, D., & Sokovic, M. (2016). Sensitivity of clinical isolates of Candida to essential oils from Burseraceae family. Excli Journal, 15, 280. https://doi.org/10.17179/excli2014-621
Onem, E., Soyocak, A., Muhammed, T.M., Ak, Ayşe.(2021). In Vitro and in Silico Assessment of the Potential of Niaouli Essential Oil as a Quorum Sensing Inhibitor of Biofilm Formation and its Effects on Fibroblast Cell Viability. Brazilian Archives of Biology and Technology, 64. https://doi.org/10.1590/1678-4324-2021200782
O’Toole, G.A. (2011). Microtiter dish biofilm formation assay. Journal of Visualized Experiments: JoVE, 47, 2437. https://doi.org/10.3791/2437
Paibon, W., Yimnoi, C., Tembab, N., Boonlue, W., Jampachaisri, K., Nuengchamnong, N., Waranuch, N., & Ingkaninan, K. (2011). Comparison and evaluation of volatile oils from three different extraction methods for some Thai fragrant flowers. International Journal of Cosmetic Science, 33(2), 150–156. https://doi.org/10.1111/j.1468-2494.2010.00603.x
Perez, M.G., Fourcade, L., Mateescu, M.A., & Paquin, J. (2017). Neutral Red versus MTT assay of cell viability in the presence of copper compounds. Analytical Biochemistry, 535, 43–46. https://doi.org/ 10.1016/j.ab.2017.07.027
Prabuseenivasan, S., Jayakumar, M., & Ignacimuthu, S. (2006). In vitro antibacterial activity of some plant essential oils. BMC Complementary and Alternative Medicine, 6(1), 1–8. https://doi.org/ 10.1186/1472-6882-6-39
Repetto, G., Del Peso, A., & Zurita, J.L. (2008). Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature Protocols, 3(7), 1125. https://doi.org/10.1038/nprot.2008.75
Riss, T.L., & Moravec, R.A. (2006). Cell proliferation assays: improved homogeneous methods used to measure the number of cells in culture. In Cell biology (pp. 25–31). Elsevier. https://doi.org/ 10.1016/B978-012164730-8/50005-8
Roy, R., Tiwari, M., Donelli, G., & Tiwari, V. (2018). Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action. Virulence, 9(1), 522–554. https://doi.org/10.1080/21505594.2017.1313372
Russo, R., Ciociaro, A., Berliocchi, L., Cassiano, M.G., Rombolà, L., Ragusa, S., Bagetta, G., Blandini, F., Corasaniti, M.T. (2013). Implication of limonene and linalyl acetate in cytotoxicity induced by bergamot essential oil in human neuroblastoma cells. Fitoterapia, 89, 48-57. https://doi.org/10.1016/j.fitote.2013.05.014
Russo, R., Cassiano, M.G.V., Ciociaro, A., Adornetto, A., Varano, G.P., Chiappini, C., Berliocchi, L., Tassorelli, C., Bagetta, G., & Corasaniti, M.T. (2014). Role of D-Limonene in autophagy induced by bergamot essential oil in SH-SY5Y neuroblastoma cells. PloS One, 9(11), e113682. https://doi.org/10.1371/journal.pone.0113682
Sobrinho, A.C.N., Morais, S.M., Souza, E.B., Albuquerque, M.R.J.R., Santos, H.S., Cavalcante, C.S.P., et al. (2020). Antifungal and antioxidant activities of Vernonia Chalybaea Mart. ex DC. essential oil and their major constituent ß-caryophyllene. Brazilian Archives of Biology and Technology, 63. https://doi.org/10.1590/1678-4324-2021200782
Senthil Kumar, K.J., Gokila Vani, M., Wang, C.-S., Chen, C.-C., Chen, Y.-C., Lu, L.-P., Huang, C.-H., Lai, C.-S., & Wang, S.-Y. (2020). Geranium and lemon essential oils and their active compounds downregulate angiotensin-converting enzyme 2 (ACE2), a SARS-CoV-2 spike receptor-binding domain, in epithelial cells. Plants, 9(6), 770. https://doi.org/ 10.3390/plants9060770
Tahir, H., Muhammad, N., Intisar, A., Din, M.I., Qaisar, U., Qadir, M.A., Ain, N.U., Ahmad, Z., Aziz, P., & Shahzad, M.K. (2020). Essential oil composition and antibacterial activity of Canarium strictum Roxb. resin. Plant Biosystems-An International Journal Dealing with All Aspects of Plant Biology, 155(6), 1198 1202. https://doi.org/10.1080/11263504.2020.1869114
Tokur, O., & Aksoy, A. (2017). In vitro sitotoksisite testleri. Harran Üniversitesi Veteriner Fakültesi Dergisi, 6(1), 112–118. https://doi.org/ 10.31196/huvfd.325794
Vieira, A.J., Beserra, F.P., Souza, M.C., Totti, B.M., & Rozza, A.L. (2018). Limonene: Aroma of innovation in health and disease. Chem Biol Interact, 283, 97–106. https://doi.org/ 10.1016/j.cbi.2018.02.007
Yang, L.P., Gu, X.L., Chen, J.X., Yang, J., Tan, S.Y., Duan, W.J. (2018). Chemical constituents from Canarium album Raeusch and their anti-influenza A virus activities. J Nat Med,72(3), 808-815. https://doi.org/ 0.1007/s11418-018-1208-8
Yu, X., Lin, H., Wang, Y., Lv, W., Zhang, S., Qian, Y., Deng, X., Feng, N., Yu, H., & Qian, B. (2018). D-limonene exhibits antitumor activity by inducing autophagy and apoptosis in lung cancer. Onco Targets Ther, 11, 1833. https://doi.org/10.2147/OTT.S155716