Figure 4.

Apoptosis and necrosis induced by encapsulated Fe₃O₄ NPs in breast cancer cells. Cells were treated with 100 μg/mL NPs for 4 h, and apoptotic and necrotic cell death was revealed using Annexin V (phosphatidylserine externalization, an apoptotic marker) and 7-AAD (rupture of the plasma membrane, a necrotic marker) dual staining and flow cytometry. (a) Representative dot plots are shown. (b) Bars indicate SD, n = 3, and ***p < 0.001 compared to untreated control (ANOVA and Dunnett’s a posteriori test). Four subpopulations are shown, namely, live cells (Annexin V (AV)-negative, 7-AAD-negative), early apoptotic cells (Annexin V (AV)-positive, 7-AAD-negative), late apoptotic cells (Annexin V (AV)-positive, 7-AAD-positive), and necrotic cells (Annexin V (AV)-negative, 7-AAD-positive). CTRL, untreated control; Fe₃O₄@Dex, dextran-based coated iron oxide nanoparticles; and Fe₃O₄@aC, glucosamine-based amorphous carbon-coated iron oxide nanoparticles.