RESUMO
OBJECTIVES: The aim of the present study was to investigate three licorice-derived polyphenols (glabridin, licochalcone A, licoricidin) as well as cinnamon oil for their antimicrobial activities against major endodontic pathogens: Enterococcus faecalis, Streptococcus mutans, Actinomyces israelii, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas endodontalis, and Candida albicans. The synergistic interactions between the four compounds and chlorhexidine were assessed on E. faecalis. Lastly, the biocompatibility of the tested compounds was assessed using human gingival fibroblasts. DESIGN: Minimal inhibitory concentrations (MIC) and minimal microbicidal concentrations (MMC) were determined using a microplate dilution assay. A luminescence assay monitoring adenosine triphosphate was used to assess the antimicrobial activity of the tested compounds against E. faecalis biofilm. The synergistic effects of the tested compounds in association with chlorhexidine were evaluated using the checkerboard technique. Cytotoxicity toward human gingival fibroblasts was assessed by determination of cell metabolic activity using a colorimetric assay. RESULTS: Cinnamon oil showed the strongest microbicidal activity. Licochalcone A, licoricidin, and glabridin had MIC values ranging from 1.56 to 25 µg/mL against the six endodontic bacterial pathogens. The natural plant compounds were active to various extents against E. faecalis embedded in a biofilm. Synergistic antibacterial effects between chlorhexidine and the compounds, mainly glabridin, were observed against E. faecalis. Following a 2-h exposure, licochalcone A, licoricidin, and glabridin demonstrated no cytotoxicity toward gingival fibroblasts at concentrations up to 50 µg/mL, while cinnamon oil and, to a lesser extent, chlorhexidine displayed some cytotoxicity. CONCLUSIONS: The present study provides evidence that the natural plant compounds tested show promise as root canal disinfection agents.
Assuntos
Anti-Infecciosos , Clorexidina , Fibroblastos , Extratos Vegetais , Anti-Infecciosos/farmacologia , Biofilmes , Clorexidina/farmacologia , Enterococcus faecalis , Humanos , Testes de Sensibilidade Microbiana , Extratos Vegetais/farmacologia , Prevotella intermediaRESUMO
Tea, an aromatic beverage prepared with the leaves of the Camellia sinensis plant, is well known to contain bioactive polyphenols. Green tea contains mainly catechins such as epigallocatechin-3-gallate (EGCG), while black tea is characterized by the presence of theaflavins. TNF-α, which is a pro-inflammatory cytokine that activates the endogenous inflammatory cascade, plays a key role in periodontitis. In the present study, we investigated the ability of tea compounds to attenuate TNF-α-mediated activation of the host inflammatory response in monocytes/macrophages as well as the protective effect of green and black tea polyphenols on gingival keratinocyte barrier dysfunction induced by TNF-α. Tea compounds inhibited both the activation of NF-κB and caspase-1 as well as IL-1ß secretion by monocytes/macrophages. TNF-α time-dependently damaged keratinocyte tight junction barrier integrity, as determined by changes in transepithelial electrical resistance and FITC-dextran transport. Green tea extract, EGCG, theaflavins, and to a lesser extent, black tea extract protected keratinocytes against the TNF-α-mediated breakdown of barrier integrity. The treatment of keratinocytes with tea polyphenols markedly mitigated the morphological changes of tight junction proteins such as zonula occludens-1 and occludin compared to cells exposed only to TNF-α, as determined by immunofluorescence. Tea polyphenols also time-dependently decreased the paracellular flux of TNF-α-treated keratinocytes. In conclusion, the ability of tea polyphenols to exert an anti-inflammatory effect and to attenuate the gingival epithelial barrier dysfunction induced by TNF-α supports their potential for the prevention and treatment of periodontal disease.
Assuntos
Gengiva/efeitos dos fármacos , Inflamação/metabolismo , Queratinócitos/efeitos dos fármacos , Polifenóis/farmacologia , Chá/química , Junções Íntimas/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Biflavonoides/farmacologia , Catequina/análogos & derivados , Catequina/metabolismo , Catequina/farmacologia , Linhagem Celular Tumoral , Citocinas/metabolismo , Gengiva/metabolismo , Humanos , Queratinócitos/metabolismo , Macrófagos/metabolismo , Monócitos/metabolismo , NF-kappa B/metabolismo , Periodontite/metabolismo , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Células U937RESUMO
The gingival epithelium, a stratified squamous tissue that acts as an interface between the external environment and the underlying connective tissue, plays an active role in maintaining periodontal health. The aim of the present study was to investigate the ability of green tea catechins to enhance gingival epithelial barrier function and protect against the disruption of epithelial integrity induced by Porphyromonas gingivalis. Both the green tea extract and epigallocatechin-3-gallate (EGCG) dose- and time-dependently increased the transepithelial electrical resistance (TER) of a gingival keratinocyte model and decreased the permeability of the cell monolayer to fluorescein isothyocyanate-conjugated 4.4-kDa dextran. This was associated with the increased expression of zonula occludens-1 (ZO-1) and occludin, two tight junction proteins. Treating the gingival keratinocyte monolayer with P. gingivalis caused a reduction in TER and affected the distribution of ZO-1 and occludin, allowing P. gingivalis to translocate through the cell monolayer. These deleterious effects mediated by P. gingivalis were abolished by the green tea extract and EGCG. This protection may be in part related to the ability of tea catechins to inhibit the protease activities of P. gingivalis. Given the above properties, green tea catechins may represent promising preventive and therapeutic molecules against periodontal disease.
Assuntos
Antibacterianos/farmacologia , Catequina/análogos & derivados , Queratinócitos/efeitos dos fármacos , Porphyromonas gingivalis/efeitos dos fármacos , Chá/química , Adesinas Bacterianas/genética , Adesinas Bacterianas/metabolismo , Antibacterianos/isolamento & purificação , Translocação Bacteriana , Catequina/isolamento & purificação , Catequina/farmacologia , Linhagem Celular Transformada , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Dextranos/metabolismo , Impedância Elétrica , Ensaios Enzimáticos , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Cisteína Endopeptidases Gingipaínas , Gengiva/efeitos dos fármacos , Gengiva/metabolismo , Gengiva/microbiologia , Humanos , Queratinócitos/metabolismo , Queratinócitos/microbiologia , Modelos Biológicos , Ocludina/genética , Ocludina/metabolismo , Porphyromonas gingivalis/enzimologia , Porphyromonas gingivalis/crescimento & desenvolvimento , Porphyromonas gingivalis/patogenicidade , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
Fusobacterium nucleatum has been associated with both periodontal disease and inflammatory bowel disease. This Gram-negative bacterium possesses a high inflammatory potential that may contribute to the disease process. We hypothesized that green and black tea polyphenols attenuate the inflammatory response of monocytes/macrophages mediated by F. nucleatum. We first showed that the tea extracts, EGCG and theaflavins reduce the NF-κB activation induced by F. nucleatum in monocytes. Since NF-κB is a key regulator of genes coding for inflammatory mediators, we tested the effects of tea polyphenols on secretion of IL-1ß, IL-6, TNF-α, and CXCL8 by macrophages. A pre-treatment of macrophages with the tea extracts, EGCG, or theaflavins prior to a stimulation with F. nucleatum significantly inhibited the secretion of all four cytokines and reduced the secretion of MMP-3 and MMP-9, two tissue destructive enzymes. TREM-1 expressed by macrophages is a cell-surface receptor involved in the propagation of the inflammatory response to bacterial challenges. Interestingly, tea polyphenols inhibited the secretion/shedding of soluble TREM-1 induced by a stimulation of macrophages with F. nucleatum. The anti-inflammatory properties of tea polyphenols identified in the present study suggested that they may be promising agents for the prevention and/or treatment of periodontal disease and inflammatory bowel disease.