Antibiofilm efficacy of tea tree oil and of its main component terpinen-4-ol against Candida albicans

Abstract Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.

Interactions between Terpinen-4-ol and Nystatin on biofilm of Candida albicans and Candida tropicalis

Abstract The aim of this study was to evaluate the antifungal activity of Terpinen-4-ol associated with nystatin, on single and mixed species biofilms formed by Candida albicans and Candida tropicalis, as well as the effect of terpinen-4-ol on adhesion in oral cells and the enzymatic activity. The minimum inhibitory concentrations and minimum fungicide concentrations of terpinen-4-ol and nystatin on Candida albicans and Candida tropicalis were determined using the microdilution broth method, along with their synergistic activity ("checkerboard" method). Single and mixed species biofilms were prepared using the static microtiter plate model and quantified by colony forming units (CFU/mL). The effect of Terpinen-4-ol in adhesion of Candida albicans and Candida tropicalis in coculture with oral keratinocytes (NOK Si) was evaluated, as well as the enzymatic activity by measuring the size of the precipitation zone, after the growth agar to phospholipase, protease and hemolysin. Terpinen-4-ol (4.53 mg mL-1) and nystatin (0.008 mg mL-1) were able to inhibit biofilms growth, and a synergistic antifungal effect was showed with the drug association, reducing the inhibitory concentration of nystatin up to 8 times in single biofilm of Candida albicans, and 2 times in mixed species biofilm. A small decrease in the adhesion of Candida tropicalis in NOK Si cells was showed after treatment with terpinen-4-ol, and nystatin had a greater effect for both species. For enzymatic activity, the drugs showed no action. The effect potentiated by the combination of terpinen-4-ol and nystatin and the reduction of adhesion provide evidence of its potential as an anti-fungal agent.

Resumo O objetivo desse estudo foi avaliar a atividade antifúngica do Terpinen4-ol associado à nistatina em biofilmes simples e misto, formados por Candida albicans e Candida tropicalis, bem como o efeito do terpinen-4-ol na adesão em células orais e atividade enzimática. As concentrações inibitórias mínimas e as concentrações fungicidas mínimas do terpinen-4-ol e da nistatina em Candida albicans e Candida tropicalis foram determinadas pelo método de microdiluição em caldo, juntamente com a atividade sinérgica (método do tabuleiro de "xadrez"). Biofilmes simples e misto foram preparados usando o modelo de placa de microtitulação estática e quantificados por unidades formadoras de colônias (CFU/mL). O efeito do Terpinen-4-ol na adesão de Candida albicans e Candida tropicalis em co-cultura com queratinócitos orais (NOK Si) foi avaliado, bem como a atividade enzimática, medindo o tamanho da zona de precipitação, após o crescimento em ágar fosfolipase, protease e hemolisina. O terpinen-4-ol (4.53 mg mL-1) e a nistatina (0,008 mg mL-1) conseguiram inibir o crescimento de biofilmes e um efeito antifúngico sinérgico foi demonstrado com a associação de fármaco, reduzindo a concentração inibidora de nistatina até 8 vezes em biofilme simpes de Candida albicans e 2 vezes em biofilme misto. Uma pequena diminuição na adesão de Candida tropicalis em células NOK Si foi mostrada após o tratamento com terpinen-4-ol e a nistatina teve um efeito maior para ambas as espécies. Para a atividade enzimática, as drogas não apresentaram ação. O efeito potencializado pela combinação de terpinen-4-ol e nistatina e a redução de adesão evidenciam seu potencial como agente anti-fúngico.

Analyses of Biofilm on Implant Abutment Surfaces Coating with Diamond-Like Carbon and Biocompatibility

Abstract The aim of this study was to evaluate the surface free energy (SFE), wetting and surface properties as well as antimicrobial, adhesion and biocompatibility properties of diamond-like carbon (DLC)-coated surfaces. In addition, the leakage of Escherichia coli through the abutment-dental implant interface was also calculated. SFE was calculated from contact angle values; R a was measured before and after DLC coating. Antimicrobial and adhesion properties against E. coli and cytotoxicity of DLC with human keratinocytes (HaCaT) were evaluated. Further, the ability of DLC-coated surfaces to prevent the migration of E. coli into the external hexagonal implant interface was also evaluated. A sterile technique was used for the semi-quantitative polymerase chain reaction (semi-quantitative PCR). The surfaces showed slight decreases in cell viability (p<0.05), while the SFE, R a, bacterial adhesion, antimicrobial, and bacterial infiltration tests showed no statistically significant differences (p>0.05). It was concluded that DLC was shown to be a biocompatible material with mild cytotoxicity that did not show changes in R a, SFE, bacterial adhesion or antimicrobial properties and did not inhibit the infiltration of E. coli into the abutment-dental implant interface.

Resumo O objetivo deste trabalho foi avaliar a energia livre de superfície (ELS), molhabilidade e propriedades de superfície assim como propriedades antimicrobianas, de adesão e biocompatibilidade de superfícies recobertas com Diamond-Like Carbon (DLC). Além disso, investigou-se a infiltração de Escherichia coli por meio da interface abutment-implante dentário. ELS foi calculada a partir dos valores de ângulo de contato; Ra foi medida antes e depois do revestimento com DLC. Foram avaliadas propriedades antimicrobianas e de adesão contra E. coli e citotoxicidade do DLC utilizando queratinócitos humanos (HaCaT). Além disso, também avaliamos a capacidade para impedir a migração de E. coli na interface do implante hexágono externo. Uma técnica estéril foi utilizada para a reação em cadeia da polimerase semi-quantitativa (PCR semi-quantitativo). As superfícies mostraram uma ligeira diminuição da viabilidade celular (p<0,05), enquanto a ELS, R a , adesão bacteriana, testes antimicrobianos e de infiltração não apresentaram diferenças estatisticamente significativas (p>0,05). Concluiu-se que o DLC demonstrou ser um material biocompatível levemente citotóxico que não mostra alterações na Ra , ELS, adesão bacteriana ou propriedades antimicrobianas e não inibiu a infiltração de E. coli na interface abutment-implante dentário.