In vitro evaluation of the antimicrobial activity of CTZ paste and its components on standard microorganisms

Farli Aparecida Carrilho Boer; Lucas Fernando Oliveira Tomáz Ferraresso; Karina Tomoe  Ajimura; Regina Lúcia  dos Santos; Wanda Terezinha Garbelini Frossard

doi:10.47990/vyj67q46

Authors

Farli Aparecida Carrilho Boer Clínica de Especialidades infantiles/ Bebê Clínica de la Universidad Estadual de Londrina https://orcid.org/0000-0003-1911-8247
Lucas Fernando Oliveira Tomáz Ferraresso Department of Preventive and Restorative Dentistry, School of Dentistry Araçatuba, São Paulo State University (Unesp), Araçatuba, Brazil. https://orcid.org/0000-0002-2225-9262
Karina Tomoe Ajimura Clínica de Especialidades infantiles/ Bebê Clínica de la Universidad Estadual de Londrina https://orcid.org/0009-0003-3123-8142
Regina Lúcia dos Santos Universidade Estadual de Londrina https://orcid.org/0009-0002-1527-6520
Wanda Terezinha Garbelini Frossard Clínica de Especialidades infantiles/ Bebê Clínica de la Universidad Estadual de Londrina https://orcid.org/0000-0002-8222-3771

DOI:

https://doi.org/10.47990/vyj67q46

Keywords:

Anti-Infective Agents, Dental Caries, Eugenol, Root Canal Filling Materials, Zinc Oxide

Abstract

CTZ paste (chloramphenicol, tetracycline, zinc oxide and eugenol) is used to treat deciduous molars with necrotic pulp. The antimicrobial activity (AA) of different concentrations of the components of the CTZ paste against different microorganisms was evaluated in vitro. Thus, the medications were divided into 6 groups: P1: 250 mg of tetracycline + 250 mg of chloramphenicol + 500 mg of zinc oxide (ZO) + 0.6 ml of eugenol (original proportion), P2: 500 mg of tetracycline + 500 mg of ZO + 0.6 ml eugenol, P3: 500 mg chloramphenicol + 500 mg ZO + 0.6 ml eugenol, P4: 1000 mg ZO + 0.6 ml eugenol, P5: 0.006 ml eugenol, P6: 1000 mg of ZO + saline solution (0.85%), and evaluated against individual standard microorganisms: Streptococcus mutans, Lactobacillus casei, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Bacillus subtilis. Agar diffusion tests (ADT) and direct exposure tests (DET) were applied. The data were subjected to ANOVA and Tukey test (P<5) were applied. P1 had greater AA compared to the other groups. For the evaluation of Escherichia coli, no statistically significant difference was observed between groups P1 and P3. Direct exposure test demonstrated antimicrobial effectiveness for groups P1, P2, P3, P4 and P5 in the time intervals evaluated and ineffectiveness for P6 due to the absence of eugenol. Therefore, it can be concluded that group P1 presented the highest antimicrobial activity among groups in the ADT. DET demonstrated antimicrobial effectiveness in groups with eugenol in its composition, which suggests a bactericidal effect of this component.

References

American Academy of Pediatric Dentistry. Pulp therapy for primary and immature permanent teeth. The Reference Manual of Pediatric Dentistry. Chicago, Ill.: American Academy of Pediatric Dentistry; 2023:457-65.

Ahmed HM. Anatomical challenges, electronic working length determination and current developments in root canal preparation of primary molar teeth. Int Endod J. 2013 Nov;46(11):1011-22. doi: 10.1111/iej.12134. Epub 2013 May 25. PMID: 23711096.

Moura LFAD, Lima MDM, Lima CCB, Bandeira AVL, Moura MS, Conde Júnior AM, Rizzo MDS. Cellular profile of primary molars with pulp necrosis after treatment with antibiotic paste. Int J Exp Pathol. 2018 Oct;99(5):264-268. doi: 10.1111/iep.12292. Epub 2018 Oct 15. PMID: 30324690; PMCID: PMC6302786.

Mohd Ariffin S, Dalzell O, Hardiman R, Manton DJ, Parashos P, Rajan S. Root canal morphology of primary maxillary second molars: a micro-computed tomography analysis. Eur Arch Paediatr Dent. 2020 Aug;21(4):519-525. doi: 10.1007/s40368-020-00515-z. Epub 2020 Feb 25. PMID: 32100200.

Fabiane P, Faraco-Junior IM, Estrela C. Antimicrobial activity of different root canal filling pastes used in deciduous teeth. Mater. Res. 2008, (11); 171-173.

Achanta A, Reche A, Dakhale R, Bharate RR. A Comprehensive Review of Lesion Sterilization and Tissue Repair: An Alternative for Pulpectomy in Deciduous Teeth. Cureus. 2023 Nov 3;15(11):e48218. doi: 10.7759/cureus.48218. PMID: 38054133; PMCID: PMC10694391.

Oliveira SCMD, Omena ALCSD, Lira GADL, Ferreira IA, Imparato JCP, Calvo AFB.. Do different proportions of antibiotics in the CTZ paste interfere with the antimicrobial action? In vitro study. Pesquisa Brasileira em Odontopediatria e Clínica Integrada. 2020, 19, e4801.

Cappiello J. Tratamentos pulpares em incisivos primários. Rev Assoc Odontol Argentina. 1964; 52:139-45

de Deus Moura L de F, de Lima M de D, Lima CC, Machado JI, de Moura MS, de Carvalho PV. Endodontic Treatment of Primary Molars with Antibiotic Paste: A Report of 38 Cases. J Clin Pediatr Dent. 2016;40(3):175-7. doi: 10.17796/1053-4628-40.3.175. PMID: 27472562.

Estrela C, Estrela CRA, Moura J, Bammann LL. Testing calcium hydroxide antimicrobial potential by different methods. J Dent Res. 2000;79, p.529, 2000.

Macià MD, Rojo-Molinero E, Oliver A. Antimicrobial susceptibility testing in biofilm-growing bacteria. Clin Microbiol Infect. 2014 Oct;20(10):981-90. doi: 10.1111/1469-0691.12651. Epub 2014 Jun 14. PMID: 24766583.

Ostrosky EA, Mizumoto MK, Lima ME, Kaneko TM, Nishikawa SO, Freitas BR. Métodos para avaliação da atividade antimicrobiana e determinação da concentração mínima inibitória (CMI) de plantas medicinais. Revista brasileira de Farmacognosia. 2008;18, 301-307.

Moore AM, Patel S, Forsberg KJ, Wang B, Bentley G, Razia Y, Qin X, Tarr PI, Dantas G. Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes. PLoS One. 2013 Nov 13;8(11):e78822. doi: 10.1371/journal.pone.0078822. PMID: 24236055; PMCID: PMC3827270.

Oliveira LM, Siqueira FS, Silva MT, Machado JVC, Cordeiro CF, Diniz LF, Campos MMA, Franco LL, Souza TB, Hawkes JA, Carvalho DT. Synthesis and antimicrobial activity of molecular hybrids based on eugenol and chloramphenicol pharmacophores. Folia Microbiol (Praha). 2023 Dec;68(6):823-833. doi: 10.1007/s12223-023-01057-9. Epub 2023 Apr 29. PMID: 37118368.

Marchese A, Barbieri R, Coppo E, Orhan IE, Daglia M, Nabavi SF, Izadi M, Abdollahi M, Nabavi SM, Ajami M. Antimicrobial activity of eugenol and essential oils containing eugenol: A mechanistic viewpoint. Crit Rev Microbiol. 2017 Nov;43(6):668-689. doi: 10.1080/1040841X.2017.1295225. Epub 2017 Mar 27. PMID: 28346030.

Ulanowska M, Olas B. Biological Properties and Prospects for the Application of Eugenol-A Review. Int J Mol Sci. 2021 Apr 1;22(7):3671. doi: 10.3390/ijms22073671. PMID: 33916044; PMCID: PMC8036490.