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Anaerobe
Vol. 40, 2016, Pages: 18–27

Copaifera reticulata oleoresin: Chemical characterization and antibacterial properties against oral pathogens

Danae Kala Rodríguez Bardají, Jonas Joaquim Mangabeira da Silva, Thamires Chiquini Bianchi, Daniele de Souza Eugênio, Pollyanna Francielli de Oliveira, Luís Fernando Leandro, Hervé Louis Ghislain Rogez, Rodrigo Cassio Sola Venezianni, Sergio Ricardo Ambrosio, Denise Crispim Tavares, Jairo Kenupp Bastos, Carlos Henrique G. Martins

Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.

Abstract

Oral infections such as periodontitis and tooth decay are the most common diseases of humankind. Oleoresins from different copaifera species display antimicrobial and anti-inflammatory activities. Copaifera reticulata is the commonest tree of this genus and grows abundantly in several Brazilian states, such as Pará, Amazonas, and Ceará. The present study has evaluated the chemical composition and antimicrobial potential of theCopaifera reticulata oleoresin (CRO) against the causative agents of tooth decay and periodontitis and has assessed the CRO cytotoxic potential. Cutting edge analytical techniques (GC-MS and LC-MS) aided the chemical characterization of CRO. Antimicrobial assays included determination of the Minimum Inhibitory Concentration (MIC), determination of the Minimum Bactericidal Concentration (MBC), determination of the Minimum Inhibitory Concentration of Biofilm (MICB50), Time Kill Assay, and Checkerboard Dilution. Conduction of XTT assays on human lung fibroblasts (GM07492-A cells) helped to examine the CRO cytotoxic potential. Chromatographic analyses revealed that the major constituents of CRO were β-bisabolene, trans-α-bergamotene, β-selinene, α-selinene, and the terpene acids ent-agathic-15-methyl ester, ent-copalic acid, and ent-polyalthic acid. MIC and MBC results ranged from 6.25 to 200 μg/mL against the tested bacteria. The time-kill assay conducted with CRO at concentrations between 50 and 100 μg/mL showed bactericidal activity against Fusobacterium nucleatum (ATCC 25586) and Streptococcus mitis (ATCC 49456) after 4 h, Prevotella nigrescens (ATCC 33563) after 6 h, Porphyromonas gingivalis (ATCC 33277) andLactobacillus casei (clinical isolate) after 12 h, and Streptococcus salivarius (ATCC 25975) and Streptococcus mutans (ATCC 25175) after 18 h. The fractional inhibitory concentration indexes (FICIs) revealed antagonistic interaction for Lactobacillus casei(clinical isolate), indifferent effect for Porphyromonas gingivalis (ATCC 33277),Fusobacterium nucleatum (ATCC 25586), Prevotella nigrescens (ATCC 33563), andStreptococcus salivarius (ATCC 25975), and additive effect for Streptococcus mutans(ATCC 25175) and Streptococcus mitis (ATCC 49456). Treatment of GM07492-A cells with CRO demonstrated that concentrations up to 39 μg/mL significantly reduced cell viability as compared to the negative control, being IC50 equal to 51.85 ± 5.4 μg/mL. These results indicated that CRO plays an important part in the search for novel sources of agents that can act against oral pathogens.

Keywords: Copaifera reticulata; Oleoresin; Oral pathogens; Antibacterial activity; Cytotoxicity; Chemical characterization.

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