In vitro photodynamic inactivation of Candida spp. growth and adhesion to buccal epithelial cells

In this study, photodynamic inactivation (PDI) was used to inhibit in vitro growth and adhesion of different Candida isolates to buccal epithelial cells (BEC). Experimental conditions were optimized and 25 μM toluidine blue O (TBO) and 15 min of irradiation time by light emitting diode (LED) (energy density of 180 J/cm²) were selected due to higher reductions in cellular viability obtained after treatment. Reduction media of Log₁₀ 3.41 in viable cellular growth and media of 55% in the inhibition of adhesion to buccal epithelial cells were obtained. Two fluconazole resistant isolates were susceptible to PDI (Log₁₀ 3.54 in IB05 and Log₁₀ 1.95 in CG09) and a second session of this treatment for CG09 isolate inhibited cellular viability in 100%, without producing heat. The results permit to conclude that photodynamic inactivation under these experimental conditions would be a possible alternative approach to inhibit Candida spp. cellular growth and adhesion to buccal epithelial cells.     

Ultra-high-performance liquid chromatography using a fused-core particle column for fast analysis of propolis phenolic compounds

Propolis is a bee product with a complex chemical composition formed by several species from different geographical origins. The complex propolis composition requires an accurate and reproducible characterization of samples to standardize the quality of the material sold to consumers. This work developed an ultra-high-performance liquid chromatography with a photodiode array detector method to analyze propolis phenolic compounds based on the two key propolis biomarkers, Artepillin C and p-Coumaric acid. This choice was made due to the complexity of the sample with the presence of several compounds. The optimized method was hyphenated with mass spectrometry detection allowing the detection of 23 different compounds. A step-by-step strategy was used to optimize temperature, flow rate, mobile phase composition, and re-equilibration time. Reverse-phase separation was achieved with a C18 fused-core column packed with the commercially available smallest particles (1.3 nm). Using a fused-core column with ultra-high-performance liquid chromatography allows highly efficient, sensitive, accurate, and reproducible determination of compounds extracted from propolis with an outstanding sample throughput and resolution. Optimized conditions permitted the separation of the compounds in 5.50 min with a total analysis time (sample-to-sample) of 6.50 min.