Antibiofilm effect and cytotoxicity of calcium hydroxide associated with the Melaleuca alternifolia essential oil

Authors

  • Ana Clara Kuerten Gil Universidade do Sul de Santa Catarina
  • Taynara Santos Goulart Universidade do Sul de Santa Catarina
  • Michelli Buratto Universidade Federal de Santa Catarina
  • Emily Marques dos Reis Universidade Federal de Santa Catarina
  • Luismar Marques Porto Universidade Federal de Santa Catarina
  • Josiane de Almeida Cava da Silveira Universidade do Sul de Santa Catarina

DOI:

https://doi.org/10.7308/aodontol/2020.56.e29

Keywords:

Endodontics , Enterococcus faecalis, Fibroblasts , Tea tree oil, Oils, volatile

Abstract

Aim: To evaluate and compare the antimicrobial and antibiofilm effect, as well as the cytotoxicity of calcium hydroxide (CH) associated with the Melaleuca alternifolia (MA) essential oil, in different concentrations, and with propylene glycol.

Methods: The following medications composed the experimental groups: G1) CH/MA 1%; G2) CH/MA 5%; G3) CH/MA 10%; G4) CH/MA 20%; and G5) CH/PG. Saline solution and culture medium DMEM were used as a control in antimicrobial and cytotoxicity tests in human periodontal ligament fibroblasts (PDLF), respectively. The antimicrobial activity (n = 12) was evaluated by the disk-diffusion agar method. The immediate antibiofilm effect (n = 12) of the medications was evaluated for bacterial viability in 72 hours-biofilms of E. faecalis, formed on the dentin disc surface and treated for seven days with medications. After microbiological sampling of the remaining biofilm, the dentin discs were immersed in sterile culture medium and stored for another seven days, for analysis of the residual antibiofilm effect of the medications, when a new microbiological sampling was performed. PDLF viability was evaluated by MTS colorimetric assay (n = 9). The mean values of the inhibition halos, in mm, the colony forming units, and the metabolic cell activity percentage were analyzed by means of Kruskal-Wallis and post hoc Dunn (α = 5%) tests.

Results: All of the experimental medications presented higher antimicrobial and antibiofilm effects, when compared to the saline solution control (p < 0.05), and maintained the PDLF feasible, similar to the DMEM control (p > 0.05).

Conclusions: The association of the Melaleuca alternifolia essential oil, at concentrations of 1%, 5%, 10%, and 20%, with calcium hydroxide promoted an excellent antimicrobial and antibiofilm activity, and biocompatibility with fibroblasts, similarly to the association with propylene glycol.

Downloads

Download data is not yet available.

References

Diogenes A, Ruparel NB. Regenerative endodontic procedures: clinical outcomes. Dent Clin North Am. 2017;61(1):111-25.

Associação Americana de Endodontia. Scope of endodontics: regenerative endodontics [homepage na Internet]. AAE Position Statement; 2018. Disponível em: https://f3f142zs0k2w1kg84k5p9i1owpengine.netdna-ssl.com/specialty/wp-content/ uploads/sites/2/2018/07/2018AAE_Scope_of_ Endo_Regenerative_Endodontics.pdf.

Nagata JY, Gomes BPFA, Lima TFR, Murakami LS, Faria DE, Campos GR, et al. Traumatized immature teeth treated with 2 protocols of pulp revascularization. J Endod. 2014;40:606-12.

Porter MLA, Munchow EA, Albuquerque MTP, Spolnik KJ, Hara AT, Bottino MC. Effects of novel 3-dimensional antibiotic-containing electrospun scaffolds on dentin discoloration. J Endod. 2016;42:106–12.

Chen MYH, Chen KL, Chen CA, Tayebaty F, Rosenberg PA, Lin LM. Responses of immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscess to revascularization procedures. Int Endod J. 2012;45:294-305.

Asnaashari M, Ashraf H, Rahmati A, Amini N. A comparison between effect of photodynamic therapy by LED and calcium hydroxide therapy for root canal disinfection against Enterococcus faecalis: a randomized controlled trial. Photodiagn Photodyn. 2017;17:226-32.

Sathorn C, Parashos P, Messer H. Antibacterial efficacy of calcium hydroxide intracanal dressing: a systematic review and meta-analysis. Int Endod J. 2007;40:2-10.

Prada I, Micó-Muñoz P, Giner-Lluesma T, Micó-Martínez P, Muwaquet-Rodríguez S, Albero-Monteagudo A. Update of the therapeutic planning of irrigation and intracanal medication in root canal treatment. A literature review. J Clin Exp Dent. 2019;11:e185-93.

Jacobs JC, Troxel A, Ehrlich Y, Spolnik K, Bringas JS, Gregory RL, et al. Antibacterial effects of antimicrobials used in regenerative endodontics against biofilm bacteria obtained in mature and immature teeth with necrotic pulps. J Endod. 2017;43:575-9.

Distel JW, Hatton JF, Gillespie MJ. Biofilm formation in medicated root canals. J Endod. 2002;28:689–93.

Portenier I, Haapasalo H, Rye A, Waltimo T, Ørstavik D, Haapasalo M. Inactivation of root canal medicaments by dentine, hydroxylapatite and bovine serum albumin. Int Endod J. 2001;34:184–8.

Zand V, Mokhtari H, Hasani A, Jabbari G. Comparison of the penetration depth of conventional and nano-particle calcium hydroxide into dentinal tubules. Iran Endod J. 2017;3:366-70.

Peters LB, Van Winkelhoff AJ, Buijs JF, Wesselink PR. Effects of instrumentation, irrigation and dressing with calcium hydroxide on infection in pulpless teeth with periapical bone lesions. Int Endod J. 2002;35:13–21.

Vianna ME, Horz HP, Conrads G, Feres M, Gomes BPFA. Comparative analysis of endodontic pathogens using checkerboard hybridization in relation to culture. Oral Microbiol Immunol. 2008;23:282–90.

Del Carpio-Perochena A, Kishen A, Felitti R, Bhagirath AY, Medapati MR, Lai C, et al. Antibacterial properties of chitosan nanoparticles and propolis associated with calcium hydroxide against single- and multispecies biofilms: an in vitro and in situ study. J Endod. 2017;43:1332-6.

Giongo M, Santos RAM, Maciel SM, Fracasso MLC, Victorino FR. Analysis of pH and release of calcium of association between Melaleuca alternifolia oil and calcium hydroxide. Rev Odont Unesp. 2017;46:104-8

Lahijani MSS, Kateb HRR, Heady R, Yazdani D. The effect of German chamomile (Marticaria recutita L.) extract and tea tree (Melaleuca alternifolia L.) oil used as irrigants on removal of smear layer: a scanning electron microscopy study. Int Endod J. 2006;39:190-5.

Carvalho NC, Guedes SAG, AlbuquerqueJúnior RLC, Albuquerque DS, Souza Araújo AA, Paranhos LR, et al. Analysis of Aleo vera citotoxicity and genotoxicity associated with endodontic medication and laser photobiomodulation. J Photochem Photobiol B. 2018;178:348-54.

Yoshpe M, Einy S, Ruparel N, Lin S, Kaufman AY. Regenerative endodontics: a potential solution for external root resorption (Case Series). J Endod. 2020;46:192-9.

Galler KM, Krastl G, Simon S, Gorp GV, Meschi N, Vahedi B, et al. European Society of Endodontology position statement: revitalization procedures. Int Endod J. 2016;49:717-23.

Ruparel NB, Teixeira FB, Ferraz CCR, Diogenes A. Direct effect of intracanal medicaments on survival of stem cells of the apical papilla. J Endod. 2012;38:1372-5.

Alasqah M, Khan SIR, Alfouzan K, Jamleh A. Regenerative endodontic management of an immature molar using calcium hydroxide and triple antibiotic paste: a two-year follow-up. Case Rep Dent. 2020;1–5.

Hammer KA, Carson CF, Riley TV, Nielsen JB. A review of the toxicity of Melaleuca alternifolia (tea tree) oil. Food Chem Toxicol. 2006;44:616–25.

Homeyer DC, Sanchez CJ, Mende K, Beckius ML, Murray CK, Wenke JC, et al. In vitro activity of Melaleuca alternifolia (tea tree) oil on filamentous fungi and toxicity to human cells. Med Mycol. 2015;53:285-94.

Nicolić MM, Jovanović KK, Marković TL, Marković DL, Gligorijević NN, Radulović SS, et al. Antimicrobial synergism and cytotoxic properties of Citrus limon L., Piper nigrum L. and Melaleuca alternifolia (Maiden and Betche) Cheel essential oils. J Pharm Phamacol. 2017;69:1606.

Casarin M, Pazinatto J, Santos RCV, Zanatta FB. Melaleuca alternifolia and its application against dental plaque and periodontal diseases: a systematic review. Phytother Res. 2018;32:230-42.

Deng DM, Hoogenkamp MA, Exterkate RA, Jiang LM, Van Der Sluis LW, Ten Cate JM, et al. Influence of Streptococcus mutans on Enterococcus faecalis biofilm formation. J Endod. 2009;35:1249–52.

Morotomi T, Washio A, Kitamura C. Current and future options for dental pulp therapy. Jpn Dent Sci Rev. 2019;55:5-11.

Estrela C, Holland R. Calcium hydroxide: study based on scientific evidences. J Appl Oral Sci. 2003;11:269-82.

Silva S, Alves N, Silva P, Vieira T, Maciel P, Castellano LR, et al. Antibacterial activity of Rosmarinus Officinalis, Zingiber Officinale, Citrus Aurantium bergamia, and Copaifera officinalis alone and in combination with Calcium Hydroxide against Enterococcus faecalis. Biomed Res Int. 2019;1-7.

Asnaashari M, Eghbal MJ, Yaghmayi AS, Shokri M, AzariMarhabi S. Comparison of antibacterial effects of photodynamic therapy, modified triple antibiotic paste and calcium hydroxide on root canals infected with Enterococcus faecalis: an in vitro study. J Lasers Med Sci. 2019;10:S23.

Manzur A, González AM, Pozos A, Silva-Herzog D, Friedman S. Bacterial quantification in teeth with apical periodontitis related to instrumentation and different intracanal medications: a randomized clinical trial. J Endod. 2007;33:114-8.

McGurkin-Smith R, Trope M, Caplan D, Sigurdsson A. Reduction of intracanal bacteria using GT rotary instrumentation, 5.25% NaOCl, EDTA, and Ca (OH) 2. J Endod. 2005;31:359-63.

Raoof M, Khaleghi M, Siasar N, Mohannadalizadeh S, Haghani J, Amanpour S. Antimicrobial activity of methanolic extracts of Myrtus Communis L. and Eucalyptus Galbie and their combination with calcium hydroxide powder against Enterococcus Faecalis. J Dent (Shiraz). 2019;20:195-202.

Lima RKP, Guerreiro-Tanomaru JM, FariaJúnior NB, Tanomaru-Filho M. Effectiveness of calcium hydroxide-based intracanal medicaments against Enterococcus faecalis. Int Endod J. 2012;45:311-6.

Menezes MM, Valera MC, Jorge AOC, Koga-Ito CY, Camargo HR, Mancini MNG. In vitro evaluation of the effectiveness of irrigants and intracanal medicaments on microorganisms within root canals. Int End J. 2004;37:311-9.

Love RM. Enterococcus faecalis – a mechanism for its role in endodontic failure. Int Endod J. 2001;34:399-405.

Shupping GB, Orstavik D, Sigurdsson A. Trope M. Reduction of intracanal bacteria using nickel-titanium rotary instrumentation and various medications. J Endod. 2000;26:751-5.

Siqueira Jr JF, Magalhães KM, Roças IN. Bacterial reduction in infected root canals treated with 2.5% NaOCl as an irrigant and calcium hydroxide/camphorated paramonochlorophenol paste as an intracanal dressing. J Endod. 2007;33:667-72.

Abbaszadegan A, Dadolahi S, Gholami A, Moein MR, Hamedani S, Ghasemi Y, et al. Antimicrobial and cytotoxic activity of Cinnamomum zeylanicum, Calcium Hydroxide, and triple antibiotic paste as root canal dressing materials. J Contemp Dent Pract. 2016;17:105–13.

Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev. 1999;12:564-82.

Gupta A, Duhan J, Tewari S, Sangwan P, Yadav A, Singh G, et al. Comparative evaluation of antimicrobial efficacy of Syzygium aromaticum, Ocimum sanctum and Cinnamomum zeylanicum plant extracts against Enterococcus faecalis: a preliminary study. Int Endod J. 2013;46:775-83.

Cavalcanti YW, Pérez ALAL, Xavier GDR, Almeida LFD. Efeito inibitório de óleos essenciais sobre microrganismos do canal radicular. Rev Odontol UNESP. 2011;40:208-14.

Monteiro DU, Azevedo MI, Weiblen C, Botton SA, Funk NL, Da Silva CB, et al. In vitro and ex vivo activity of Melaleuca alternifolia against protoscoleces of Echinococcus ortleppi. Cambridge University Press. 2017;144:214-9.

Soderberg TA, Johansson A, Gref R. Toxic effects of some conifer resin acids and tea tree oil on human epithelial and fibroblast cells. J Toxicol. 1996;107:99-109.

Graziano TS, Calil CM, Sartoratto A, Franco GC, Groppo FC, Cogo-Müller K. In vitro effects of Melaleuca alternifolia essential oil on growth and production of volatile sulphur compounds by oral bacteria. J Appl Oral Sci. 2016;24:582-9.

Oliveira ACM, Fontana A, Negrini TC, Nogueira MNM, Bedran TBL, Andrade CR, et al. Emprego do óleo de Melaleuca alternifolia Cheel (Myrtaceae) na odontologia: perspectivas quanto à utilização como antimicrobiano alternativo às doenças infecciosas de origem bucal. Rev Bras Plantas Med. 2011;13:492-9.

Vishwanat L, Duong R, Takimoto K, Phillips L, Espitia CO, Diogenes A, et al. Effect of bacterial biofilm on the osteogenic differentiation of stem cells of apical papilla. J Endod.2017;43:916–22.

Widbiller M, Althumairy RI, Diogenes A. Direct and indirect effect of chlorhexidine on survival of stem cells from the apical papilla and its neutralization. J Endod. 2019;45:156-60.

Published

2020-12-16

How to Cite

Gil, A. C. K., Goulart, T. S., Buratto, M., Reis, E. M. dos, Porto, L. M., & Silveira, J. de A. C. da. (2020). Antibiofilm effect and cytotoxicity of calcium hydroxide associated with the Melaleuca alternifolia essential oil. Arquivos Em Odontologia, 56. https://doi.org/10.7308/aodontol/2020.56.e29

Issue

Section

Artigos