In Vitro Assessment of Cellular and Genetic Safety for a DSPCar-Based Amphotericin B Nanoformulation Using MTT and Ames Assays
Archives of Advances in Biosciences,
Vol. 17 No. 1 (2026),
28 Ordibehesht 2026
,
Page 1-12
https://doi.org/10.22037/aab.v17i1.52478
Abstract
Background: Despite the potent antifungal properties of amphotericin B (AmB), its clinical deployment remains restricted by adverse effects whose severity rises proportionally with administered dose. Our laboratory had previously engineered a DSPCar-containing nanoliposomal carrier for AmB that demonstrated superior antifungal action. The current investigation sought to characterize the cellular and genetic harmlessness profile of this DSPCar-AmB nanosystem.
Methods: The fabricated nanocarrier underwent physicochemical profiling to establish its mean diameter, surface charge, and medication entrapment percentage. We evaluated cytotoxicity via the MTT method using HEK293 cells (concentration range: 0.01–100 µM; duration: 24 hours). Mutagenicity screening employed the bacterial reverse mutation (Ames) procedure on Salmonella typhimurium TA98 alongside TA100 strains, both with and without S9 metabolic supplementation, using three dosage levels (50, 500, and 5000 µg/plate).
Results: The DSPCar-AmB preparation exhibited a hydrodynamic size of 89.4 ± 6.2 nm, a polydispersity index of 0.18, and a zeta potential measuring -32.5 mV. Relative to a reference liposomal AmB product, the DSPCar-AmB carrier induced significantly less cellular death, with viability percentages remaining above 85% for each examined concentration (p < 0.05). The half-maximal inhibitory concentration (IC₅₀) for DSPCar-AmB exceeded 100 µM, whereas the reference formulation gave an IC₅₀ of 8.2 ± 1.4 µM. Within the Ames assay, no condition produced a significant rise in revertant colony numbers (fold increase consistently below 1.5 relative to negative controls), confirming a lack of mutation-inducing potential.
Conclusion: The DSPCar-AmB nanocarrier presents a beneficial in vitro safety profile featuring substantially diminished cytotoxicity alongside undetectable genotoxicity. When integrated with previously documented antifungal effectiveness, this formulation implies a potentially enhanced safety-efficacy equilibrium and merits continued preclinical advancement.
- Nanoliposome
- DSPCar AmB
- MTT assay
- Bacterial reverse mutation test
- Cytotoxicity
- Genotoxicity
- Amphotericin B
How to Cite
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