An in vitro anti-inflammatory effect of Thai propolis in human dental pulp cells

Objective: To explore the potential for development of Thai propolis extract as a pulp capping agent to suppress pulpal inflammation from dental pulp infections. This study aimed to examine the anti-inflammatory effect of the propolis extract on the arachidonic acid pathway, activated by interleukin (IL)-1β, in cultured human dental pulp cells.  Methodology: Dental pulp cells, isolated from three freshly extracted third molars, were first characterized for their mesenchymal origin and treated with 10 ng/ml of IL-1β in the presence or absence of non-toxic concentrations of the extract from 0.08 to 1.25 mg/ml, as determined by the PrestoBlue cytotoxic assay. Total RNA was harvested and analyzed for mRNA expressions of 5-lipoxygenase (5-LOX) and cyclooxygenase-2 (COX-2). Western blot hybridization was performed to investigate COX-2 protein expression. Culture supernatants were assayed for released prostaglandin E2 levels. Immunofluorescence was conducted to determine involvement of nuclear factor-kappaB (NF-kB) in the inhibitory effect of the extract. Results: Stimulation of the pulp cells with IL-1β resulted in the activation of arachidonic acid metabolism via COX-2, but not 5-LOX. Incubation with various non-toxic concentrations of the propolis extract significantly inhibited upregulated COX-2 mRNA and protein expressions upon treatment with IL-1β (p<0.05), resulting in a significant decrease in elevated PGE2 levels (p<0.05). Nuclear translocation of the p50 and the p65 subunits of NF-kB upon treatment with IL-1β was also blocked by incubation with the extract. Conclusions: Upregulated COX-2 expression and enhanced PGE2 synthesis upon treatment with IL-1β in human dental pulp cells were suppressed by incubation with non-toxic doses of Thai propolis extract via involvement of the NF-kB activation. This extract could be therapeutically used as a pulp capping material due to its anti-inflammatory properties

Combined bisdemethoxycurcumin and potassium iodide-mediated antimicrobial photodynamic therapy

Antimicrobial photodynamic therapy is emerging as a promising way to treat infections with minimal side effects. Typically, a single photosensitizer used in photodynamic therapy is capable of generating only one type of reactive oxygen species, which may have inadequate capability to eradicate certain types of microbes, especially Candida species. Thus, the use of combined photosensitizers is examined as a means of achieving superior antimicrobial results. We postulate that bisdemethoxycurcumin, a type I reactive oxygen species generator, combined with potassium iodide, an antimicrobial iodide molecule, might exhibit superior antimicrobial effects compared to a single photosensitizer-mediated photodynamic therapy. The effects of bisdemethoxycurcumin + potassium iodide + dental blue light on Candida albicans reduction were examined. Candida biofilms were treated with 20, 40 or 80 μM bisdemethoxycurcumin, 100 mM potassium iodide or a combination of these species for 20 min before irradiation with a dental blue light (90 J/cm2). The negative and positive controls were phosphate buffer saline and nystatin at 1 : 100,000 units/ml, respectively. Candidal numbers were quantified at 0, 1, 6 and 24 h. Hydroxyl radicals were spectrophotometrically measured using 2-[6-(4′amino phynoxyl-3H-xanthen-3-on-9-yl)] benzoic acid or APF probe-mediated fluorescence intensity (Varioskan) at 490/515 nm (excitation/emission). Candidal counts and hydroxyl radical comparisons were performed using the Kruskal-Wallis test and one-way ANOVA, respectively. Correlations between candidal numbers and hydroxyl radical levels were done with a Pearson correlation test. Forty μM bisdemethoxycurcumin+100 mM KI could provide a 3.5 log10 CFU/ml reduction after 6 h. Bisdemethoxycurcumin alone generated OH levels that were strongly correlated with candidal reduction. In conclusion, 40 μM bisdemethoxycurcumin+100 mM KI could reduce C. albicans biofilm

Inhibitory Effects of Erythrosine/Curcumin Derivatives/Nano-Titanium Dioxide-Mediated Photodynamic Therapy on Candida albicans

This study focuses on the role of photosensitizers in photodynamic therapy. The photosensitizers were prepared in combinations of 110/220 µM erythrosine and/or 10/20 µM demethoxy/bisdemethoxy curcumin with/without 10% (w/w) nano-titanium dioxide. Irradiation was performed with a dental blue light in the 395–480 nm wavelength range, with a power density of 3200 mW/cm2 and yield of 72 J/cm2. The production of ROS and hydroxyl radical was investigated using an electron paramagnetic resonance spectrometer for each individual photosensitizer or in photosensitizer combinations. Subsequently, a PrestoBlue® toxicity test of the gingival fibroblast cells was performed at 6 and 24 h on the eight highest ROS-generating photosensitizers containing curcumin derivatives and erythrosine 220 µM. Finally, the antifungal ability of 22 test photosensitizers, Candida albicans (ATCC 10231), were cultured in biofilm form at 37 °C for 48 h, then the colonies were counted in colony-forming units (CFU/mL) via the drop plate technique, and then the log reduction was calculated. The results showed that at 48 h the test photosensitizers could simultaneously produce both ROS types. All test photosensitizers demonstrated no toxicity on the fibroblast cells. In total, 18 test photosensitizers were able to inhibit Candida albicans similarly to nystatin. Conclusively, 20 µM bisdemethoxy curcumin + 220 µM erythrosine + 10% (w/w) nano-titanium dioxide exerted the highest inhibitory effect on Candida albicans