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Antifungal, cytotoxic, and immunomodulatory properties of tea tree oil and its derivative components: potential role in management of oral candidosis in cancer patients.

Ramage G, Milligan S, Lappin DF, Sherry L, Sweeney P, Williams C, Bagg J, Culshaw S - Front Microbiol (2012)

Bottom Line: The aims of the study were to evaluate the antifungal efficacy of TTO and key derivatives against C. albicans biofilms, to assess the toxicological effects of TTO on a clinically relevant oral cell line, and to investigate its impact on inflammation.Transcript and protein analysis showed a reduction of IL-8 when treated with TTO and T-4-ol.These data provide further in vitro evidence that TTO and its derivative components, specifically T-4-ol, exhibit strong antimicrobial properties against fungal biofilms.

View Article: PubMed Central - PubMed

Affiliation: Infection and Immunity Research Group, Glasgow Dental School, School of Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow Glasgow, UK.

ABSTRACT
Candida albicans forms oral biofilms that cause disease and are difficult to treat with conventional antifungal agents. Tea tree oil (TTO) is a natural compound with reported antimicrobial and immunomodulatory activities. The aims of the study were to evaluate the antifungal efficacy of TTO and key derivatives against C. albicans biofilms, to assess the toxicological effects of TTO on a clinically relevant oral cell line, and to investigate its impact on inflammation. TTO and its derivatives were examined against 100 clinical strains of C. albicans. Planktonic minimum inhibitory concentrations (MICs) were determined using the CLSI M-27A broth microdilution method. Sessile MICs were determined using an XTT reduction assay. Inhibition, time-kill, and mode of action studies were performed. OKF6-TERT2 epithelial cells were used for cytotoxicity and cytokine expression assays. Planktonic C. albicans isolates were susceptible to TTO, terpinen-4-ol (T-4-ol), and α-terpineol, with an MIC(50) of 0.5, 0.25, and 0.25%, respectively. These three compounds also displayed potent activity against the 69 biofilm-forming strains, of which T-4-ol and α-terpineol displayed rapid kill kinetics. For all three compounds, 1 × MIC(50) effectively inhibited biofilm growth when C. albicans were treated at 0, 1, and 2 h post adhesion. By scanning electron microscopy analysis and PI uptake, TTO and derivative components were shown to be cell membrane active. TTO and T-4-ol were cytotoxic at 1 × MIC(50), whereas at 0.5 × MIC(50) T-4-ol displayed no significant toxicity. Transcript and protein analysis showed a reduction of IL-8 when treated with TTO and T-4-ol. These data provide further in vitro evidence that TTO and its derivative components, specifically T-4-ol, exhibit strong antimicrobial properties against fungal biofilms. T-4-ol has safety advantages over the complete essential oil and may be suitable for prophylaxis and treatment of established oropharyngeal candidosis. A clinical trial of T-4-ol is worthy of consideration.

No MeSH data available.


Related in: MedlinePlus

Candida albicans biofilm killing by TTO, T-4-ol, and α-terpineol is time dependent. Standardized C. albicans (1 × 106 cells/ml) were incubated in flat-bottomed 96-well plates for 24 h, washed in PBS, and treated with 2 × SMIC50 at 2, 5, 10, 15, and 60 min. Metabolic activity of treated biofilms was then quantified using the XTT assay by reading at 492 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Three isolates were used for each assay, and this was performed on two independent occasions in triplicate. **p < 0.001. Error bars represent the ±standard error of the mean.
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Figure 1: Candida albicans biofilm killing by TTO, T-4-ol, and α-terpineol is time dependent. Standardized C. albicans (1 × 106 cells/ml) were incubated in flat-bottomed 96-well plates for 24 h, washed in PBS, and treated with 2 × SMIC50 at 2, 5, 10, 15, and 60 min. Metabolic activity of treated biofilms was then quantified using the XTT assay by reading at 492 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Three isolates were used for each assay, and this was performed on two independent occasions in triplicate. **p < 0.001. Error bars represent the ±standard error of the mean.

Mentions: The biofilm rate of kill for the three most effective components (TTO, T-4-ol, and α-terpineol) at 2 × SMIC50 was performed, where time dependent killing was observed (Figure 1). TTO was shown to reduce the viability of the biofilms within 2 min by 44%, but after 5, 15, and 60 min this had declined only slightly to 46, 56, and 65%, respectively. However, both T-4-ol and α-terpineol displayed comparable and more effective killing, reducing the viability rapidly after 2 min by approximately 55%. This anti-biofilm activity remained, reducing biofilm viability by approximately 61, 75, and 94% after 5, 15, and 60 min, respectively. Both T-4-ol and α-terpineol showed significantly better activity than TTO only at 60 min (p < 0.001).


Antifungal, cytotoxic, and immunomodulatory properties of tea tree oil and its derivative components: potential role in management of oral candidosis in cancer patients.

Ramage G, Milligan S, Lappin DF, Sherry L, Sweeney P, Williams C, Bagg J, Culshaw S - Front Microbiol (2012)

Candida albicans biofilm killing by TTO, T-4-ol, and α-terpineol is time dependent. Standardized C. albicans (1 × 106 cells/ml) were incubated in flat-bottomed 96-well plates for 24 h, washed in PBS, and treated with 2 × SMIC50 at 2, 5, 10, 15, and 60 min. Metabolic activity of treated biofilms was then quantified using the XTT assay by reading at 492 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Three isolates were used for each assay, and this was performed on two independent occasions in triplicate. **p < 0.001. Error bars represent the ±standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376416&req=5

Figure 1: Candida albicans biofilm killing by TTO, T-4-ol, and α-terpineol is time dependent. Standardized C. albicans (1 × 106 cells/ml) were incubated in flat-bottomed 96-well plates for 24 h, washed in PBS, and treated with 2 × SMIC50 at 2, 5, 10, 15, and 60 min. Metabolic activity of treated biofilms was then quantified using the XTT assay by reading at 492 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Three isolates were used for each assay, and this was performed on two independent occasions in triplicate. **p < 0.001. Error bars represent the ±standard error of the mean.
Mentions: The biofilm rate of kill for the three most effective components (TTO, T-4-ol, and α-terpineol) at 2 × SMIC50 was performed, where time dependent killing was observed (Figure 1). TTO was shown to reduce the viability of the biofilms within 2 min by 44%, but after 5, 15, and 60 min this had declined only slightly to 46, 56, and 65%, respectively. However, both T-4-ol and α-terpineol displayed comparable and more effective killing, reducing the viability rapidly after 2 min by approximately 55%. This anti-biofilm activity remained, reducing biofilm viability by approximately 61, 75, and 94% after 5, 15, and 60 min, respectively. Both T-4-ol and α-terpineol showed significantly better activity than TTO only at 60 min (p < 0.001).

Bottom Line: The aims of the study were to evaluate the antifungal efficacy of TTO and key derivatives against C. albicans biofilms, to assess the toxicological effects of TTO on a clinically relevant oral cell line, and to investigate its impact on inflammation.Transcript and protein analysis showed a reduction of IL-8 when treated with TTO and T-4-ol.These data provide further in vitro evidence that TTO and its derivative components, specifically T-4-ol, exhibit strong antimicrobial properties against fungal biofilms.

View Article: PubMed Central - PubMed

Affiliation: Infection and Immunity Research Group, Glasgow Dental School, School of Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow Glasgow, UK.

ABSTRACT
Candida albicans forms oral biofilms that cause disease and are difficult to treat with conventional antifungal agents. Tea tree oil (TTO) is a natural compound with reported antimicrobial and immunomodulatory activities. The aims of the study were to evaluate the antifungal efficacy of TTO and key derivatives against C. albicans biofilms, to assess the toxicological effects of TTO on a clinically relevant oral cell line, and to investigate its impact on inflammation. TTO and its derivatives were examined against 100 clinical strains of C. albicans. Planktonic minimum inhibitory concentrations (MICs) were determined using the CLSI M-27A broth microdilution method. Sessile MICs were determined using an XTT reduction assay. Inhibition, time-kill, and mode of action studies were performed. OKF6-TERT2 epithelial cells were used for cytotoxicity and cytokine expression assays. Planktonic C. albicans isolates were susceptible to TTO, terpinen-4-ol (T-4-ol), and α-terpineol, with an MIC(50) of 0.5, 0.25, and 0.25%, respectively. These three compounds also displayed potent activity against the 69 biofilm-forming strains, of which T-4-ol and α-terpineol displayed rapid kill kinetics. For all three compounds, 1 × MIC(50) effectively inhibited biofilm growth when C. albicans were treated at 0, 1, and 2 h post adhesion. By scanning electron microscopy analysis and PI uptake, TTO and derivative components were shown to be cell membrane active. TTO and T-4-ol were cytotoxic at 1 × MIC(50), whereas at 0.5 × MIC(50) T-4-ol displayed no significant toxicity. Transcript and protein analysis showed a reduction of IL-8 when treated with TTO and T-4-ol. These data provide further in vitro evidence that TTO and its derivative components, specifically T-4-ol, exhibit strong antimicrobial properties against fungal biofilms. T-4-ol has safety advantages over the complete essential oil and may be suitable for prophylaxis and treatment of established oropharyngeal candidosis. A clinical trial of T-4-ol is worthy of consideration.

No MeSH data available.


Related in: MedlinePlus