Nanomedicine Journal

Nanomedicine Journal

Voriconazole-Loaded Nanoliposomes against Cryptococcus neoformans: Antifungal Activity, Biofilm Biomass Reduction, and Expression of CXT1 and LAC1

Articles in Press

Document Type : Research Paper

Authors
sanam Nami 1
Hamed Hamishekar 2
Behzad Baradaran 3
Zahra Ramezanalipour 4
Parviz Hassanpour 1, 4
1 Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
2 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
3 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
4 Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
10.22038/nmj.2026.97952.2468
Abstract
Objective(s): Cryptococcus neoformans is an opportunistic fungal pathogen whose virulence is associated with capsule formation, melanin production, and biofilm development. This pilot study evaluated the biological activity of voriconazole-loaded nanoliposomes (VCZ-NLs), with emphasis on biofilm biomass reduction and potential modulation of the virulence-associated genes CXT1 and LAC1.
Materials and Methods: A reference strain of C. neoformans was used in this pilot study. VCZ-NLs were prepared and characterized using dynamic light scattering, zeta potential analysis, and transmission electron microscopy. Antifungal susceptibility of yeast-phase cells was determined by minimum inhibitory concentration (MIC) testing. Biofilm biomass was quantified using the crystal violet microtiter assay. Urease activity was qualitatively assessed using Christensen’s urea agar. Relative expression of CXT1 and LAC1 was evaluated by quantitative real-time PCR using the 2^−ΔΔCt method, with untreated cells as the calibrator. Experiments were performed in technical triplicate as part of a pilot design.
Results: C. neoformans showed a VCZ MIC of 4 µg/mL, and liposomal encapsulation did not alter the observed yeast-phase MIC. Urease activity showed no apparent change, while CXT1 expression showed a modest downward trend and LAC1 expression remained largely unchanged under the tested conditions. VCZ-NLs produced a greater reduction in crystal violet-measured biofilm biomass than free VCZ under the conditions tested.
Conclusion: VCZ-NLs preserved planktonic antifungal activity and produced a greater reduction in crystal violet-measured biofilm biomass under the tested conditions. Because crystal violet measures biomass rather than viability, these findings do not establish a biofilm-specific or enhanced antifungal effect. Further studies with biological replication and viability-based assays are warranted.
Keywords
Cryptococcus neoformans
Voriconazole
Biofilms
Urease
Nanoliposomes
Subjects
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Nanomedicine Journal

Articles in Press, Accepted Manuscript
Available Online from 24 October 2026