Efficacy of true cinnamon (Cinnamomum verum) leaf essential oil as a therapeutic alternative for Candida biofilm infections

Document Type : Original Article

Authors

Area of Microbiology and Immunology, Department of Oral Diagnosis, Piracicaba Dental School, State University of Campinas, SP, Brazil

Abstract

Objective(s): The essential oil (EO) extracted from Cinnamomum verum leaves has been used as an antimicrobial agent for centuries. But its antifungal and antibiofilm efficacy is still not clearly studied. The objective of this research was to evaluate the in vitro antifungal and antibiofilm efficacy of C. verum leaf EO against C. albicans, C. tropicalis, and C. dubliniensis and the toxicity of EO using an in vitro model.
Materials and Methods: The effect of EO vapor was evaluated using a microatmosphere technique. CLSI microdilution assay was employed in determining the Minimum Inhibitory (MIC) and Fungicidal Concentrations (MFC). Killing time was determined using a standard protocol. The effect of EO on established biofilms was quantified and visualized using XTT and Scanning Electron Microscopy (SEM), respectively. Post-exposure intracellular changes were visualized using Transmission Electron Microscopy (TEM). The toxicological assessment was carried out with the Human Keratinocyte cell line. The chemical composition of EO was evaluated using Gas Chromatography-Mass Spectrometry (GC-MS).
Results: All test strains were susceptible to cinnamon oil vapor. EO exhibited MIC value 1.0 mg/ml and MFC value 2.0 mg/ml against test strains. The killing time of cinnamon oil was 6 hr. Minimum Biofilm Inhibitory Concentration (MBIC50) for established biofilms was Conclusion: C. verum EO is a potential alternative anti-candida agent with minimal toxicity on the human host.

Keywords


1. Wijesinghe GK, Maia FC, de Oliveira TR, de Feiria SNB, Joia F, Barbosa JP, et al. Effect of Cinnamomum verum leaf essential oil on virulence factors of Candida species and determination of the in-vivo toxicity with Galleria mellonella model. Mem Inst Oswaldo Cruz 2020; 115:e200349.
2. Oliveira T, Teixeira A, Barbosa JP, de Feiria SNB, Boni GC, Maia F, et al. Melaleuca spp. essential oil and its medical applicability. A Brief Review. Braz J Nat Sci 2020; 3: 249.
3. Wijesinghe G, Jayarathna P, Gunasekara T, Fernando N, Kottegoda N, Weerasekera M. Antibacterial and anti-Candida activity of chlorhexidine gluconate, Triphala and Munamal pothu (bark of Mimusops elengi). Sri Lankan Journal of Infectious Diseases 2018; 8:25.
4. Gupta P, Daswani P, Birdi T. Approaches in fostering quality parameters for medicinal botanicals in the Indian context. Indian J Pharmacol 2014; 46:363-371.
5. Ranasinghe P, Pigera S, Premakumara GS, Galappaththy P, Constantine GR, Katulanda P. Medicinal properties of “true” cinnamon (Cinnamomum zeylanicum): a systematic review. BMC Complement Altern Med 2013; 13:275.
6. Wijesinghe GK, Dilhari A, Gayani B, Kottegoda N, Samaranayake L, Weerasekera M. Influence of Laboratory Culture Media on In-vitro Growth, Adhesion and Biofilm Formation of Pseudomonas aeruginosa and Staphylococcus aureus. Med Princ Pract 2019; 28:28-35.
7. Acker HA, Van DP, Coenye P. Molecular mechanisms of antimicrobial tolerance and resistance in bacterial and fungal biofilms. Trends Microbiol 2014; 22: 326-333.
8. Deveau A, Hogan DA. Linking Quorum Sensing Regulation and Biofilm Formation by Candida albicans. Methods Mol Biol 2011; 692:219-233.
9. Percival SL, Suleman L, Vuotto C, Donelli G. Healthcare-associated infections, medical devices and biofilms: risk, tolerance and control. J Med Microbiol 2015; 64: 323–334.
10. Weerasekera MM, Wijesinghe GK, Jayarathna TA, Gunasekara CP, Fernando N, Kottegoda N, et al. Culture media profoundly affect Candida albicans and Candida tropicalis growth, adhesion, and biofilm development. Mem Inst Oswaldo Cruz 2016; 111:697-702.
11. Nett JE. Future directions for anti-biofilm therapeutics targeting Candida. Expert Rev Anti Infect Ther 2014; 12: 375-382.
12. Lazar V, Ditu, LM, Curutiu C, Gheorghe I, Holban A, Popa M, et al. Impact of Dental Plaque Biofilms in Periodontal Disease: Management and Future Therapy. Periodontitis-A Useful Reference 2017.
13. Nomura R, Nakano K, Nemoto H, Fujita K, Inagaki S, Takahashi T, et al. Isolation and characterization of Streptococcus mutans in heart valve and dental plaque specimens from a patient with infective endocarditis. J Med Microbiol 2006; 55:1135-1140.
14. Maia FC, Wijesinghe GK, Oliveira TR, Barbosa JP, de Feiria SNB, Boni GC, et al. Phyllanthus niruri L. (stone breacher) as an alternative of anti-human diseases, antimicrobial agent, and its applicability to combat resistant microrganisms. A Brief Review. Braz J Nat Sci 2020; 3:342.
15. Spampinato C, Leonardi D. Candida Infections, Causes, Targets, and Resistance Mechanisms: Traditional and Alternative Antifungal Agents. Biomed Res Int 2013; 2013:204237.
16. de Oliveira Santos GC, Vasconcelos CC, Lopes AJO, de Sousa Cartágenes MdS, Filho AKDB, do Nascimento FRF, et al. Candida Infections and Therapeutic Strategies: Mechanisms of Action for Traditional and Alternative Agents. Front Microbiol 2018; 9:1351.
17. Wijesinghe GK (2019), Effect of True Cinnamon (Cinnamomum verum) Leaf Oil Against In Vitro Candida Biofilms and Its Cytotoxic Effect. Master’s Thesis. State university of Campinas, Brazil. (Available from: http://repositorio.unicamp.br/jspui/bitstream/REPOSIP/335816/1/Wijesinghe_GayanKanchana_M.pdf)
18. Serban ES, Ionescu M, Matinca D, Maier CS, Bojiţă MT. Screening of the antibacterial and antifungal activity of eight volatile essential oils. Farmacia 2011; 59:440–46.
19. CLSI. Performance Standards for Antifungal Susceptibility Testing of yeasts. 1st ed. CLSI Supplement M60. Wayne, PA: Clinical and Laboratory Standards Institute; 2017.
20. Khodavandi A, Alizadeh F, Vanda NA, Karimi G, Chong PP. Possible mechanisms of the antifungal activity of fluconazole in combination with terbinafine against Candida albicans. Pharm Biol 2014; 52: 1505-1509.
21. Souza CMC, Junior SAP, Moraes TDS, Damasceno JL, Mendes SA, Dias HJ, et al. Antifungal activity of plant-derived essential oils on Candida tropicalis planktonic and biofilms cells. Med Mycol 2016; 54 : 515-523.
22. Gayani B, Dilhari A, Wijesinghe GK, Kumarage S, Abayaweera G, Samarakoon SR, et al. Effect of natural curcuminoids-intercalated layered double hydroxide nanohybrid against Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis: A bactericidal, antibiofilm, and mechanistic study. Microbiologyopen 2019; 8:e00723.
23. Kapoor G, Saigal S, Elongavan A. Action and Resistance Mechanisms of Antibiotics: A Guide for Clinicians. J Anaesthesiol Clin Pharmacol 2017; 33: 300-305.
24. Zanette C, Pelin M, Crosera M, Adami G, Bovenzi M, Larese FF,  et al. Silver nanoparticles exert a long-lasting antiproliferative effect on human keratinocyte HaCaT cell line. Toxicology in Vitro 2011; 25: 1053-1060.
25. Tantengco OAG, Jacinto SD. Cytotoxic activity of crude extracts and fractions from Premna odorata (Blanco), Artocarpus camansi (Blanco) and Gliricidia sepium (Jacq.) against selected human cancer cell lines. Asian Pac J Trop Biomed 2015; 5: 1037-1041.
26. Siddiqui ZN, Farooq F, Musthafa TNM, Ahmad A, Khan AU. Synthesis, characterization and antimicrobial evaluation of novel halopyrazole derivatives. Journal of Saudi Chemical Society 2013; 17: 237-43.
27. Castro RD, Lima EO. Anti-Candida activity and chemical composition of Cinnamomum zeylanicum blume essential oil. Braz Arch Biol Technol 2013; 56:749–55.
28. Rangel MDL, Aquino SGD, Lima JMD Castellano LR, de Castro RD. In vitro effect of Cinnamomum zeylanicum blume essential oil on Candida spp. involved in oral infections. Evid based Complement Alternat Med 2018; 2018:4045013.
29. Duarte MC, Figueira GM, Sartoratto Um, Rehder VLG, Delarmelina C. Anti-Candida activity of Brazilian medicinal plants. J Ethnopharmacol 2005; 97: 305-311.
30. Bax HI, Bakker-Woudenberg IAJM, de Vogel CP, van der Meijden A, Verbon A, de Steenwinkel JEM. The role of the time-kill kinetics assay as part of a preclinical modeling framework for assessing the activity of anti-tuberculosis drugs. Tuberculosis 2017; 105: 80-85.
31. Da Silva PMB, Acosta EJTR, de Rezende Pinto L, Graeff M, Spolidorio DMP, Almeida RS, et al. Microscopical analysis of Candida albicans biofilms on heat-polymerised acrylic resin after chlorhexidine gluconate and sodium hypochlorite treatments. Mycoses 2011; 54: e712–e717.
32. Scheibler E, da Silva RM, Leite CE, Campos MM, Figueiredo MA, Salum FG, et al. Stability and efficacy of combined nystatin and chlorhexidine against suspensions and biofilms of Candida albicans. Archives of Oral Biology 2018; 89: 70-76.
33. Jantan IB, Karim Moharam BA, Santhanam J. Jamal JA.  Correlation between Chemical Composition and Antifungal Activity of the Essential Oils of Eight Cinnamomum Species. Pharm Biol 2008; 46: 406-412.
34. Choi O, Cho SK, Kim J, Park CG,  Kim J. In vitro antibacterial activity and major bioactive components of Cinnamomum verum essential oils against cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus. Asian Pacific Journal of Tropical Biomedicine 2016; 6:308-314.
35. Pavesi C, Banks LA, Hudaib T. Antifungal and antibacterial activities of eugenol and non-polar extract of Syzygium aromaticum L. J Pharma Sci Res 2018; 10: 337-339.