Terpinen-4-ol and nystatin co-loaded precursor of liquid crystalline system for topical treatment of oral candidiasis.

This study was performed to develop a liquid crystalline system (LCS) incorporated with terpinen-4-ol and nystatin to evaluate its antifungal, antibiofilm, and synergistic/modulatory activity against Candida albicans. The LCS was composed of a dispersion containing 40% propoxylated and ethoxylated cetyl alcohol, 40% oleic acid, and 0.5% chitosan dispersion. According to analysis by polarized light microscopy, rheology, and mucoadhesion studies, the incorporation of 100% artificial saliva increased the pseudoplasticity, consistency index, viscosity, and mucoadhesion of the formulation. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity; the LCS containing terpinen-4-ol and nystatin effectively inhibited C. albicans growth at a lower concentration, displaying a synergistic action. Therefore, LCS incorporated with terpinen-4-ol and nystatin is a promising alternative for preventing and treating infections and shows potential for the development of therapeutic strategies against candidiasis.

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

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.

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.

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.

Antimicrobial effects of terpinen-4-ol against oral pathogens and its capacity for the modulation of gene expression.

This study evaluated the antibacterial activity of terpinen-4-ol against Streptococcus mutans and Lactobacillus acidophilus and its influence on gbpA (S. mutans) and slpA (L. acidophilus) gene expression. As measured by XTT assay, the concentrations of terpinen-4-ol that effectively inhibited the biofilm were 0.24% and 0.95% for S. mutans and L. acidophilus, respectively. Confocal microscopy revealed the presence of a biofilm attached to the enamel and dentin block surfaces with significant terpinen-4-ol effects against these microorganisms. The expression of the gbpA and slpA genes involved in adherence and biofilm formation was investigated using RT-PCR. Expression of these genes decreased after 15 min with 0.24% and 0.95% terpinen-4-ol in S. mutans and L. acidophilus, respectively. These findings demonstrate the antimicrobial activity of terpinen-4-ol and its ability to modulate the expression of gbpA and slpA genes, emphasizing the therapeutic capacity of terpinen-4-ol as an alternative to inhibit adherence in biofilm.

In vitro antibacterial and cytotoxic activities of carvacrol and terpinen-4-ol against biofilm formation on titanium implant surfaces.

This study evaluated the antibacterial properties of carvacrol and terpinen-4-ol against Porphyromonas gingivalis and Fusobacterium nucleatum and its cytotoxic effects on fibroblast cells. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were examined. The minimum biofilm inhibition concentration (MBIC) was evaluated by XTT assay. Biofilm decontamination on titanium surfaces was quantified (CFU ml-1), evaluated by confocal laser scanning microscopy (CLSM) and cytotoxic activity by MTT. The MIC and MBC for carvacrol were 0.007% and 0.002% for P. gingivalis and F. nucleatum, and 0.06% for terpinen-4-ol for both microorganisms. The MBIC for carvacrol was 0.03% and 0.06% for P. gingivalis and F. nucleatum, and for terpinen-4-ol was 0.06% and 0.24%. The results indicated anti-biofilm activity using carvacrol (0.26%, 0.06%) and terpinen-4-ol (0.95%, 0.24%) and showed cytotoxic activity similar to chlorohexidine (CHX). However, terpinen-4-ol (0.24%) showed higher cell viability than other treatments. Carvacrol and terpinen-4-ol showed antibacterial activity in respect of reducing biofilms. Moreover, CHX-like cytotoxicity was observed.

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.

Antibacterial activity of glass ionomer cement modified by zinc oxide nanoparticles.

This study evaluated the antibacterial activity of zinc oxide nanoparticles incorporated into self-cured glass ionomer cement (GIC) and light-cured resin-reinforced GIC on Streptococcus mutans biofilm. The GICs, Fuji II (GC America) and Fuji II LC (GC America), were incorporated with nanoparticles at concentrations of 0%, 1%, and 2% by weight, and the biofilm maturation time was one and seven days. Circular specimens of each GIC type were prepared. The antibacterial activity was evaluated by determining the number of colony forming units of S. mutans strain per milliliter. Morphology of the biofilm was analyzed by scanning electron microscopy (SEM). The data obtained for each GIC were analyzed by two-way ANOVA (α = 5%). For chemically activated GIC, no significant difference was observed in relation to the time of biofilm maturation (p = 0.744), concentration of nanoparticles (p = 0.966), and their interaction (p = 0.800). The results from analysis of GIC modified by light-polymerized resin showed that only of the maturing time significantly affected the number of adhered cells on the biofilm (p = 0.034, F = 4.778). The more mature the biofilm, higher the number of cells. SEM analysis showed no change in cell morphology in relation to the type of GIC, maturation time, and nanoparticles concentration. We conclude that the inclusion of zinc oxide nanoparticles at concentrations of 1% and 2% by weight into the GICs evaluated here, did not promote their antimicrobial activity against S. mutans.

Tratamento de manchas brancas associadas ao tratamento ortodôntico / Treatment of white spots during orthodontic treatment

O aparecimento de mancha branca durante o tratamento ortodôntico é um problema clínico relevante. O objetivo deste trabalho foi destacar possíveis tratamentos da mancha branca durante e após o tratamento ortodôntico. Analisando a literatura encontrada tem-se que os principais tratamentos são: o controle do biofilme dental, microabrasão, aplicação de flúor (verniz, gel, dentifrício e enxaguatório), laser de argônio e fosfopeptídeo caseína-fosfato de cálcio amorfo (CPP-ACP). A condição dentária, principalmente o controle do biofilme dental, deve ser rigorosamente avaliada pelo ortodontista, para evitar o aparecimento das manchas brancas.

The white spot appearance during orthodontic treatment is an important clinical problem. The objective of this study was to highlight possible treatments of white spot during and after orthodontic treatment. The literature review has demonstrated that the main treatments are biofilm control, sandblasting fluoride application (varnish, gel, toothpaste and mouthwash), argon laser and amorphous calcium phosphate-casein phosphopeptide (CPP-ACP). The dental health, particularly the control of the biofi lm must be rigorously evaluated by the orthodontist to prevent the appearance of white spots.

Efeito do óleo essencial de Melaleuca alternifolia e de seu principal componente Terpinen-4-ol sobre isolados clínicos de Candida albicans resistentes / Effect of Melaleuca alternifolia essential oil and its main component Terpinen-4-ol against clinical isolates of Candida albicans resistant

O óleo essencial de Melaleuca alternifolia (TTO) é um extrato de ação antifúngica e preventiva em escala farmacêutica ou cosmética. O objetivo deste estudo foi avaliar a eficácia do TTO e Terpinen-4-ol sobre isolados clínicos de Candida albicans. Este estudo foi realizado em três fases: 1- Identificação da CIM (Concentração Inibitória Mínima) e CFM (Concentração Fungicida Mínima) do TTO (0,25 a 2%) e Terpinen-4- ol (0,11 a 0,95%) sobre C. albicans na forma planctônica. 2- Análise das diferentes concentrações do óleo sobre biofilme monoespécie de C. albicans por meio da contagem das UFC/mL e avaliação da atividade metabólica das células pelo método colorimétrico de XTT. 3- Análise da ação inibitória das soluções de TTO e Terpinen-4- ol sobre biofilmes de C.albicans (cepa padrão e isolados clínicos) formados em corpos de prova de resina acrílica termopolimerizável previamente cobertos com saliva humana, por meio do teste de XTT e por microscopia confocal de varredura à laser (MCVL). A nistatina (Sigma) foi utilizada como controle positivo. Os resultados mostraram que isolados de C. albicans planctônicos foram sensíveis ao TTO 1%, Terpinen-4-ol 0,47% e Nistatina 8 ?g/mL. As menores concentrações fungicidas para os isolados foram TTO 2 %, Terpinen-4-ol 0,95 % e Nistatina 16 ?g/mL. Quando analisado em biofilme através da quantificação (UFC/mL) e teste de XTT as concentrações de TTO 2 % e Terpinen-4-ol 0,95 % foram eficazes quando comparado ao controle, sendo que as amostras da genotipagem A2 e B1 foram as mais resistentes. Os resultados de MCVL mostraram que todos os biofilmes desenvolvidos em corpos de resina acrílica apresentaram-se semelhantes à ação da Nistatina. Os extratos avaliados apresentaram ação antifúngica para os isolados clínicos e podem ser considerados tratamento alternativo para paciente com candidíase.

The essential oil of Melaleuca alternifolia (TTO) is an extract of antifungal action and preventive in pharmaceutical or cosmetic scale. The aim of this study was to evaluate the efficacy of TTO and Terpinen-4-ol against clinical isolates of Candida albicans. This study was conducted in three phases: 1- Identification of MIC (Minimum Inhibitory Concentration) and CFM (Minimum Fungicidal Concentration) of TTO (0.25 to 2%) and Terpinen-4-ol (0.11 to 0.95 %) on C. albicans in planktonic form . 2- Analysis of different concentrations of oil on C. albicans biofilm single species by counting CFU/mL and evaluation of the metabolic activity of cells by XTT colorimetric method. 3- Analysis of the inhibitory action of TTO and Terpinen-4-ol solutions on C. albicans biofilms (standard strain and clinical isolates) formed in specimens of polymerizable resin acrylic microwave previously coated with human saliva , by means of the XTT test and confocal laser scanning microscopy (CLSM) . The nystatin (Sigma) was used as a positive control. The results showed that isolates of C. albicans planktonic were sensitive to TTO 1%, Terpinen-4-ol 0.47% and Nystatin 8 mg/mL. The smaller fungicidal concentrations for the isolates were TTO 2%, Terpinen-4-ol 0.95% and Nystatin 16 mg / mL. When analyzed by quantifying biofilm (CFU / ml) and XTT test, the concentrations TTO 2% and Terpinen-4-ol 0.95% were effective when compared to the control, and genotyping of samples A2 and B1 were more resistant. The CLSM results showed that all biofilms developed in bodies of acrylic resin were similar to the action of Nystatin. The extracts showed antifungal action for the clinical isolates and can be considered an alternative treatment for patients with candidiasis