Fig. 3

Fig. 3 Nanosystem affinity towards macrophages and effect on bacterial uptake.

a Schematic representation of the protocol used for the affinity assay. b Percentages of FITC-positive non-infected and Mmar-infected THP-1 cells after treatment with 200 µg/mL of each nanosystem for 2 h. The experiment was performed in 3 biological replicates and 3 technical replicates each. Data are shown as mean ± SD (n = 3). Comparisons were done using two-way ANOVA with Tukey’s multiple comparisons test. Asterisks above each bar correspond to comparison to MSN within the condition. Adjusted p values (from left to right): Non-infected: ****p < 0.0001; M. marinum: ****p < 0.0001, *p = 0.0171 (TPP vs AVA-TPP), *p = 0.0152 (AVA-TPP vs AVA₂-TPP). c Representative images of non-infected and Mmar-infected THP-1 cells after treatment with 200 µg/mL of each nanosystem. First and third line correspond to xy orthogonal projection of z-stack confocal images, with a more detailed 3D reconstruction (second and fourth line) of the white squares. Three independent experiments were performed. d Schematic representation of the protocol used for the internalization assay. e Percentage of infected monocyte-derivate macrophages (MDM) (left panel) and monocyte-derivate dendritic cells (MoDC) (right panel) measured by flow cytometry. Both cell types were infected with mScarlet-Mmar pre-incubated with control (gray), 50 or 200 µg/mL of MSN (blue), or MSN-AVA-TPP (orange). The experiment was performed with 3 donors, and 3 technical replicates for each donor. Each dot corresponds to the mean of 3 technical replicates for one donor, normalized to the control. Data are shown as median and interquartile range (n = 3). Comparisons were done using ordinary one-way ANOVA with Šídák’s multiple comparisons test. Asterisks above each bar correspond to comparison to non-treated condition (gray). Adjusted p values (from left to right): MDM: ns p = 0.7783 (MSN 50), ns p = 0.1575 (MSN 200), ***p = 0.0008, ****p < 0.0001, **p = 0.0065 (MSN 50 vs AVA-TPP 50), **p = 0.0015 (MSN 200 vs AVA-TPP 200); MoDC: ns p = 0.5958 (MSN 50), ns p = 0.3277 (MSN 200), **p = 0.0049, ***p = 0.0006, ns p = 0.0814 (MSN 50 vs AVA-TPP 50), *p = 0.016 (MSN 200 vs AVA-TPP 200). Created in BioRender. Kremer, L. (2025) https://BioRender.com/1qn1m1o. Source data are provided as a SourceData file.