Copaifera reticulata oleoresin: Chemical characterization and antibacterial properties against oral pathogens.
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 the Copaifera 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) and Lactobacillus 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), and Streptococcus 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.
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Danae Kala Rodríguez
DK
Bardají
School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
Jonas Joaquim Mangabeira
JJ
da Silva
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Thamires Chiquini
TC
Bianchi
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Daniele
D
de Souza Eugênio
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Pollyanna Francielli
PF
de Oliveira
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Luís Fernando
LF
Leandro
School of Food Engineering, Institute of Technology, Federal University of Pará, Belém, Pará, Brazil.
Hervé Louis Ghislain
HL
Rogez
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Rodrigo Cassio Sola
RC
Venezianni
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Sergio Ricardo
SR
Ambrosio
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil.
Denise Crispim
DC
Tavares
School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
Jairo Kenupp
JK
Bastos
Nucleus of Research in Sciences and Technology, Research Laboratory Culture of Applied Microbiology, University of Franca, Franca, São Paulo, Brazil. Electronic address: carlos.martins@unifran.edu.br.
Carlos Henrique G
CH
Martins
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D001643:Bridged Bicyclo Compounds / Q000302:isolation & purification; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003731:Dental Caries / Q000382:microbiology; D007887:Fabaceae / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007780:Lacticaseibacillus casei / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification; D008826:Microbial Sensitivity Tests; D000081222:Monocyclic Sesquiterpenes; D010518:Periodontitis / Q000382:microbiology; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D016966:Porphyromonas gingivalis / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification; D045242:Prevotella nigrescens / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification; D012717:Sesquiterpenes / Q000302:isolation & purification; D013295:Streptococcus mutans / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification; D000070001:Streptococcus salivarius / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification; D013729:Terpenes / Q000302:isolation & purification
D000900:Anti-Bacterial Agents; D001643:Bridged Bicyclo Compounds; D000081222:Monocyclic Sesquiterpenes; D010936:Plant Extracts; D012717:Sesquiterpenes; D013729:Terpenes; C018620:oleoresins; C539285:selinene; C423904:alpha-bergamotene; C531191:beta-bisabolene
Antibacterial activity; Chemical characterization; Copaifera reticulata; Cytotoxicity; Oleoresin; Oral pathogens