Propolis is a natural substance and consists of bioactive compounds, which gives it antioxidant and antimicrobial properties. However, the use of propolis is limited by the low solubility in aqueous solutions. Thus, nanoparticles may be likely to accomplish enhanced delivery of poorly water-soluble phytomedicine. The aim of the present study was to fabricate and evaluate the biological activity of ethanolic extract of propolis-loaded poly(lactic-co-glycolic acid) nanoparticles (EEP-NPs). The EEP-NPs were prepared using the oil-in-water (o/w) single-emulsion solvent evaporation technique. The physicochemical properties of EEP-NPs were characterized and tested on their cytotoxicity, antifungal activity, and impact on key virulence factors that contribute to pathogenesis of C. albicans. EEP-NPs were successfully synthesized and demonstrated higher antifungal activity than EEP in free form. Moreover, EEP-NPs exhibited less cytotoxicity on Vero cells and suppressed the virulence factors of C. albicans, including adhesion, hyphal germination, biofilm formation, and invasion. Importantly, EEP-NPs exhibited a statistical decrease in the expression of hyphal adhesion-related genes, ALS3 and HWP1, of C. albicans. The results of this study revealed that EEP-NPs mediates a potent anticandidal activity and key virulence factors by reducing the gene-encoding virulence-associated hyphal- adhesion proteins of C. albicans and, thereby, disrupting the morphologic presence and attenuating their virulence.
Cryptococcus neoformans, a life-threatening human yeast pathogen, has the ability to produce melanin, which is one of the common virulence factors contributing to cryptococcal pathogenesis. This virulence factor is closely associated with the cryptococcal cell wall, specifically chitin and chitosan polysaccharides, a complex structure that is essential for maintaining cellular structure and integrity. In this study, we aim to investigate the effects of two stingless bee (SLB) propolis from Tetragonula laeviceps and Tetrigona melanoleuca against cell wall-associated melanin in C. neoformans, and its immune response in RAW 264.7 macrophage. The ethanolic extract of SLB propolis (EEP) has strongly exhibited anti-cryptococcal activity. Moreover, EEP from both sources reduced chitin/chitosan and melanin production against C. neoformans in a dose-dependent manner. Likewise, the mRNA expression level of CDA1, IPC1-PKC1 and LAC1 genes involved in the cryptococcal melanization pathway was significantly decreased at 2 mg/mL in EEP treatment. Additionally, pretreatment with EEP prior to yeast infection dramatically reduced intracellular replication of C. neoformans in RAW 264.7 macrophages in a dose-dependent manner. This study might be a new insight to use a natural powerful source, not only acting to target cell wall-associated molecules, but also being capable to explore a novel strategy by which dysregulation of these molecules leads to promote immunomodulatory activity.
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