研究目的
To evaluate the antimicrobial effect of curcumin-ethylenediaminetetraacetic acid (EDTA)-mediated antimicrobial photodynamic therapy (aPDT) on the vitality of intact biofilms of dentin caries microcosms.
研究成果
The combination of curcumin and EDTA enhances the antimicrobial effect of aPDT on dentin caries microcosms, particularly with lower light densities and in deeper biofilm layers, rejecting the null hypotheses that it has no effect on vitality.
研究不足
The study used non-fluorescent glass slabs instead of dentin to avoid interference in CLSM, which may not fully replicate natural dental conditions. The LED light source had low power, leading to long irradiation times that are not clinically feasible; high-power LEDs could overcome this. The biofilm model may not capture all aspects of in vivo biofilms, and the sample size was small (n=4 per group).
1:Experimental Design and Method Selection:
The study aimed to assess the effect of curcumin-EDTA-mediated aPDT on dental biofilms. Biofilms were grown on glass slabs under microaerophilic conditions, treated with curcumin and EDTA combinations, irradiated with LED light, and vitality was measured using confocal laser scanning microscopy (CLSM). Statistical analysis used non-parametric tests due to non-normal data distribution.
2:Sample Selection and Data Sources:
Dentin caries samples were collected from three children aged 7-11 years with dentin caries lesions in primary molars, excluding those with systemic diseases or recent antibiotic use. Samples were stored and used to grow biofilms.
3:List of Experimental Equipment and Materials:
Glass slabs, McBain medium, curcumin, EDTA, LED light source (Biotable RGB), optical power meter, CLSM, staining reagents (fluorescein diacetate and ethidium bromide), and statistical software (SPSS).
4:Experimental Procedures and Operational Workflow:
Biofilms were grown for 5 days, treated with curcumin and/or EDTA for 2 minutes in dark, irradiated with LED at specified energies, stained, and imaged with CLSM to determine vitality based on live/dead cell ratios.
5:Data Analysis Methods:
Data were analyzed using Kruskal-Wallis and post hoc Dunn's tests in SPSS version 21.0, with significance set at P < 0.05.
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Confocal laser scanning microscope
TCS-SPE
Leica
Used to determine biofilm vitality
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Glass ionomer cement
Ketac Fil Plus
3M Espe
Used for restoring teeth after dentin collection
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Brain Heart Infusion broth
Used for storing dentin samples
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Glass slabs
Menzel
Used to grow biofilms to avoid fluorescence interference
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Sonicator
Single Ultra-Sonic Cell Disruptor
Merse
Used for sonicating dentin samples
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Mucin
Sigma Chemical Co.
Component of McBain medium
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Peptone
Becton, Dickinson & Co.
Component of McBain medium
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Casein peptone
Becton, Dickinson & Co.
Component of McBain medium
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Yeast extract
Becton, Dickinson & Co.
Component of McBain medium
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PIPES buffer
Sigma Chemical Co.
Component of McBain medium
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Curcumin
Sigma-Aldrich
Used as a photosensitizing agent
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EDTA
Sigma-Aldrich
Used as a chelating agent
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LED light source
Biotable RGB
Institute of Physics of S?o Carlos
Used for irradiation in aPDT
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Optical power meter
1916-C Optical Power Meter
Newport
Used to adjust irradiation time
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Fluorescein diacetate
Sigma Chemicals Co.
Staining reagent for live cells
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Ethidium bromide
Sigma Chemicals Co.
Staining reagent for dead cells
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Statistical software
SPSS version 21.0
IBM
Used for statistical analysis
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