Journal
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
Volume 65, Issue 4, Pages -Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AAC.01628-20
Keywords
Candida; antifungal activity; phytocompounds; liposomes; carvacrol; cinnamaldehyde; citral; thymol; macrophages
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Funding
- Consejeria de Educacion, Universidades e Investigacion of Gobierno Vasco-Eusko Jaurlaritza [GIC15/78 IT-990-16]
- Fundacion ONCE Oportunidad al Talento
- Fondo Social Europeo
- Federation of European Microbiological Societies (FEMS) by Research and Training grant [FEMS-GO-2017-020]
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The study developed and characterized liposome-based nanoparticles with carvacrol, cinnamaldehyde, citral, and thymol for the therapy of invasive candidiasis. Among the formulations tested, nanoparticles containing cinnamaldehyde, citral, and thymol showed the best characteristics, with high encapsulation efficiencies of citral and carvacrol liposomes. Carvacrol and thymol in liposome-based nanoparticles were non-toxic and maintained their antifungal activity, synergistically working with macrophages to reduce yeast survival.
There is an increasing need for novel drugs and new strategies for the therapy of invasive candidiasis. This study aimed to develop and characterize liposome-based nanoparticles of carvacrol, cinnamaldehyde, citral, and thymol with anti-Candida activities. Dioctadecyldimethylammonium bromide- and monoolein-based liposomes in a 1:2 molar ratio were prepared using a lipid-film hydration method. Liposomes were assembled with equal volumes of liposomal stock dispersion and stock solutions of carvacrol, cinnamaldehyde, citral, or thymol in dimethyl sulfoxide. Cytotoxicity was tested on RAW 264.7 macrophages. In vitro antifungal activity of liposomes with phytocompounds was evaluated according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology using clinical isolates of Candida albicans, Candida auris, Candida dubliniensis, and Candida tropicalis. Finally, the ability of macrophage cells to kill Candida isolates after addition of phytocompounds and their nanoparticles was determined. Nanoparticles with 64 mu g/ml of cinnamaldehyde, 256 mu g/ml of citral, and 128 mu g/ml of thymol had the best characteristics among the formulations tested. The highest encapsulation efficiencies were achieved with citral (78% to 83%) and carvacrol (66% to 71%) liposomes. Carvacrol and thymol in liposome-based nanoparticles were nontoxic regardless of the concentration. Moreover, carvacrol and thymol maintained their antifungal activity after encapsulation, and there was a significant reduction (similar to 41%) of yeast survival when macrophages were incubated with carvacrol or thymol liposomes. In conclusion, carvacrol and thymol liposomes possess high stability, low cytotoxicity, and antifungal activity that act synergistically with macrophages.
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