Fig 1.
Chemical structures of CIS and VOR.
Fig 2.
Schematic illustration of Niosomes and their components.
Fig 3.
Characterization of CIS and VOR formulations.
(A) Calibration curve of vorinostat showing linearity between concentration and peak area. (B) Representative HPLC chromatogram of vorinostat. (C) Calibration curve of cisplatin showing linearity between concentration and peak area. (D) Representative HPLC chromatogram of cisplatin. (E) Encapsulation efficiency (EE%) of CIS and VOR in single- and co-loaded niosomal formulations (NIO-CIS, NIO-VOR, NIO-CIS-VOR). (F) In vitro cumulative release profile of free drugs (CIS, VOR) and drug-loaded niosomal formulations (NIO-CIS-VOR)(Data are presented as mean ± SD (n = 3).
Fig 4.
Characterization and Stability of Niosomal Formulation Encapsulating Cisplatin and Vorinostat (NIO-CIS-VOR).
(A) Size distribution by intensity showing a narrow peak around ~150 nm, indicating uniform particle size. (B) Zeta potential distribution demonstrating surface charge stability with values near −10 mV. (C) Transmission electron microscopy (TEM) image confirming spherical morphology of niosomes with sizes ranging from ~120–173 nm. (D) Stability profile at 4°C over 60 days, showing consistent hydrodynamic diameter (blue) and zeta potential (red), indicating good physical stability of the formulation.
Fig 5.
FTIR spectral analysis of cisplatin (CIS), vorinostat (VOR), and their formulations.
(A) FTIR spectra of pure CIS, pure VOR, and physical mixture (CIS–VOR), showing characteristic functional group peaks including NH₂ bending, OH stretching, and C = O vibrations. (B) FTIR spectra of CIS–VOR, plain niosomes (NIO), and drug-loaded niosomes (NIO–CIS–VOR), demonstrating characteristic peak shifts and changes in intensity, confirming successful encapsulation of CIS and VOR within the niosomal carrier system.
Fig 6.
Dose–response curves showing cell viability after 72 h treatment with cisplatin (CIS), vorinostat (VOR), and niosomal formulations (NIO–CIS, NIO–VOR, NIO–CIS–VOR) on different cancer cell lines.
(A & D) Cytotoxic effects on HT 29 cells. (B & F) Cytotoxic effects on A549 cells. (C & F) Cytotoxic effects on PANC-1 cells. Data are presented as mean ± SD (n = 3). (G) Table summarizing IC50 for the (NIO–CIS, NIO–VOR, NIO–CIS–VOR) against the three cell lines HT29, A549 and PANC 1.
Fig 7.
Caspase 3/7 activity in cells following treatment with CIS, VOR, CIS-VOR, NIO-CIC, NIO-VOR, and NIO-CIS-VOR (A) and (D) HT 29. (B) and (E) A549 cells. (C0 and (F) PANC 1 cells. Caspase 3/7 activity was measured and expressed as fold change relative to untreated control. Data are presented as mean ± SD (n = 3). Statistical significance was assessed using one-way ANOVA followed by post hoc analysis. *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 8.
Quantitative analysis of wound closure (%) after 48 hours of Cell migration assay showing the effects of free and niosomal formulations of CIS and VOR on the migration of HT-29, A549, and PANC-1 cancer cells. (A, B, G, H) (n = 3).
Fig 9.
Dose–Effect and Combination Analysis of Niosomal Formulations in Different Cancer CellC Lines.
(A, D, G) Dose–response curves for HCT29, A549, and PANC-1 cells treated with NIO-VOR, NIO-CIS, and NIO-CIS-VOR formulations. (B, E, H) Combination Index (CI) plots for NIO-CIS-VOR at varying fractional effects (Fa), indicating synergism at lower Fa values. (C, F, I) Dose Reduction Index (DRI) plots for cisplatin and vorinostat in combination, showing significant dose reduction potential, especially in HCT29 cells. Table K: Combination Index (CI) and Dose Reduction Index (DRI) for NIO-CIS-VOR in Different Cancer Cell Lines Values represent the estimated CI at fractional effects (Fa ≈ 0.5, 0.75, and 0.9) and DRI for cisplatin and vorinostat at Fa ≈ 0.5. CI < 1 indicates synergism, CI = 1 indicates additive effect, and CI > 1 indicates antagonism. DRI > 1 suggests the potential for dose reduction when drugs are combined compared to single-agent treatment.