Fig. 8: Antioxidant bioactivity of nanoalgosomes in cellular systems and in C. elegans.

a Percentage of ROS production in 1-7 HB2 and MDA-MB 231 cells treated with nanoalgosomes (0.5–1-2 µg/mL) for 24 h, with/without oxidant agent (250 µM TBH), normalized to untreated cells. Values are means ± SEM of three independent experiments. One-way ANOVA was used to assess the statistical significance of the differences: MDA-MB 231 vs MDA-MB 231 2 µg/mL nanoalgosome, *p < 0.05; 1-7HB2 vs 1-7 HB2 2 µg/mL nanoalgosome, *p < 0.05; MDA-MB 231 + oxidant agent vs MDA-MB 231 0.5-1-2 µg/mL nanoalgosome + oxidant agent, ****p < 0.0001; 1-7 HB2 + oxidant agent vs 1-7 HB2 0.5-1-2 µg/mL nanoalgosome + oxidant agent, ****p < 0.0001. b–d Real-time PCR quantification of antioxidant enzyme expression levels after exposure to nanoalgosomes (2 µg/mL) for 24 h, with/without oxidant agent (250 µM H₂O₂) in 1-7 HB2 cells. Each experiment was performed in triplicate (n = 3 biologically independent experiments). Three-way ANOVA statistical test was used to assess the statistical significance of the differences: Control vs 2 µg/mL nanoalgosome; Control vs Oxidant agent; Oxidant agent vs Oxidant agent + 2 µg/mL nanoalgosome, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns=not significant (p > 0.9). e In vivo response to exogenous oxidative stress. Animals have been pre-treated with mock, NAC (2 mM) and nanoalgosome (20 µg/mL). Animal movement was assessed right after acute exposure with water (untreated) or H₂O₂ 5 mM. One-way ANOVA statistical test was used to assess the statistical significance of the differences: H₂O₂ mock vs H₂O₂ nanoalgosome ***p = 0.0009; H₂O₂ nanoalgosome vs H₂O₂ NAC ns=not significant (p > 0.5). f Quantification of the fluorescence in gst-4p::GFP expressing animals after chronic treatment with mock or nanoalgosomes (20 µg/mL). Each dot represents the total fluorescence in a picture corrected for the background. Bars represent the mean; error bar is the SEM. n is the total number of animals analyzed. Mann Whitney t-test was used for comparing: mock vs nanoalgosome *p = 0.0435. g Thrashing assay on young and old animals after acute treatments with mock and nanoalgosomes (10 µg/mL). Violin plots show the distribution of thrashes performed by the animals in a minute. Bold dashed lines in the center correspond to the medians, while upper and lower dashed lines correspond to the quartiles. n is the number of animals analyzed. Mann Whitney t-test was used for comparing: mock vs nanoalgosome treatment of young animals, ns=not significant (p > 0.3); mock vs nanoalgosome treatment of old animals *p = 0.0362.