Fig 1.
Histograms overlay and cellular uptake for 4,24h for 2D cells by flow cytometry.
(A) Histogram overlay illustrating the 2D cellular uptake for 4h. (B) Histogram overlay illustrating the 2D cellular uptake for 24h. X-axis represents fluorescence intensity, while the Y-axis represents cell count. (C) Fold increase in fluorescence intensity for both con (1 µM, 2 µM) over time. * P < 0.05, **** P < 0.0001 (two-way ANOVA). (D) Positive cells percentage for both con (1 µM, 2 µM) over time. **P < 0.01*** P < 0.001 (two-way ANOVA). Error bars indicate SD (n = 3).
Fig 2.
Histograms overlay and cellular uptake for 4,24h for 3D spheroids by flow cytometry.
(A) Histogram overlay illustrating the 3D cellular uptake for 4h. (B) Histogram overlay illustrating the 3D cellular uptake for 24h. Fluorescence intensity is represented by the X-axis, while the Y-axis represents cell count. (C) Fold increase in fluorescence intensity for both con (1 µM, 2 µM) over time. * P < 0.05, **** P < 0.0001 (two-way ANOVA). (D) Positive cells percentage for both con (1 µM, 2 µM) over time. **P < 0.01*** P < 0.001 (two-way ANOVA). Error bars indicate SD (n = 3).
Fig 3.
2D and 3D cellular uptake for 24h.
(A) Histogram overlay for 3D cells by flow cytometry. The X-axis represents fluorescent intensity, while the Y-axis represents cell count. (B) Fold increase in fluorescence intensity for 2 µM con over time. ** P < 0.01, *** P < 0.001, **** P < 0.0001 (two-way ANOVA). (C) Positive cells percentage for 2 µM con over time. **** P < 0.0001 (two-way ANOVA). Error bars indicate SD (n = 3).
Fig 4.
U87MG cells after 4 and 24h cellular interaction and uptake of FD by Confocal Microscopy.
Localization of Dox in 1 µM and 2 µM FD treated cells. Blue fluorescence is for DAPI nuclear stain, and red fluorescence is for Dox localization. Untreated cells were used as a negative control.
Fig 5.
U87MG cells after 4 and 24h cellular interaction and uptake of Doxil® by Confocal Microscopy.
Localization of Dox in 1 µM and 2 µM FD treated cells. Blue fluorescence is for DAPI nuclear stain, and red fluorescence is for Dox localization. Untreated cells were used as a negative control.
Fig 6.
Z-stacking of U87-MG spheroids after 24h in (A) Untreated cell, (B) 2 µM FD treated spheroid (C) 1 µM FD treated spheroid, (D) 2 µM Doxil® treated spheroid (E) 1 µM Doxil® treated spheroid.
Blue fluorescence is for DAPI nuclear stain, and red fluorescence is for Dox localization.
Fig 7.
(A) Comparison between FD and Doxil® treatment on U87-MG cells after 72h of incubation, as measured by MTT assay. (B) Dose- response curve of U87MG cells in FD and Doxil®. *** P < 0.001, **** P < 0.0001 (two-way ANOVA). Error bars indicate SD (n = 3).
Fig 8.
FD and Doxil® treated spheroids in 3D collagen model.
(A) The invasion area (represented by red area) after (62.5nM and 125nM) FD treatment for 4 days. (B) The invasion area after (250nmM and 400nM) Doxil® treatment for 4 days. (C) Comparison of FD and Doxil® invasion areas. (D)The diagram shows the relationship between the invasion area and time. The X-axis represents TIA, and the Y-axis represents times (days). * P < 0.05, **** P < 0.0001 (two-way ANOVA). Error bars indicate SD (n = 3).
Fig 9.
FD and Doxil® treated cells in wound healing-2D model.
(A) The effect of (250nM, 400nM) Doxil® concentrations on the migration area vs time. (B) The effect of (62.5nM, 125nM) FD concentrations on the migration area vs time. (C) The relation between migration area and time, and (D) the diagram demonstrates the TMA vs time comparison for FD and Doxil®. **** P < 0.0001 * P < 0.05 (Two-way ANOVA). Error bars indicate SD (n = 3).