Additional file 1 of Different coatings on magnetic nanoparticles dictate their degradation kinetics in vivo for 15 months after intravenous administration in mice
Creators: name: Portilla, Yadileiny
name: Fernández-Afonso, Yilian
name: Pérez-Yagüe, Sonia
name: Mulens-Arias, Vladimir
name: Morales, M. Puerto
name: Gutiérrez, Lucía
name: Barber, Domingo F.
Publisher: name: figshare
Year: 2024
Subjects: subject: Space Science
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1: Fig. S1. Physicochemical characterization of MNPs. (a) TEM images of iron oxide coated APS-, DEX-and DMSA-MNPs. (b) Nanoparticle size distributions after coating with APS, DEX and DMSA, as determined by TEM. (c, d) The hydrodynamic size and Z potential of the APS, DEX and DMSA coated MNPs at 24 h as determined by DLS. The data shown for means ± SD corresponding to the hydrodynamic size of the MNPs were obtained by calculating the mean value of hydrodynamic size measured in intensity distribution for three measurements made with this batch of MNPs. (e) Magnetization curve at RT for the MNPs showing their superparamagnetic behavior. (f) Magnetic susceptibility of APS-, DEX- and DMSA-MNPs diluted in agar. Scale bars 50 nm and 20 nm. Fig. S2. Evaluation of the blood circulation time of MNPs. (a) ICP-OES analysis of iron content in blood samples from PBS (Control), APS-, DEX- or DMSA-MNP-treated mice 7 days post-administration. The data are shown as the mean ± SD (n = 3). (b, c) Analysis of the presence of APS-MNPs in blood determined by AC magnetic susceptibility 7 (b) and 14 (c) days after the last administration of the APS-MNPs. The in-phase (real, χ′) and out-of-phase (imaginary, χ′′- inset inside of the χ′ profile) components of the AC magnetic susceptibility measurements are shown. Fig. S3. AC magnetic susceptibility of the organs of PBS-treated mice 7 days after they received the last dose of PBS. Temperature dependence of the AC magnetic susceptibility: a) in-phase and b) out-of-phase components of tissues from mice treated with PBS at 7 days post-administration. Fig. S4. Analysis of protein corona formation dynamics on APS-, DEX- and DMSA-MNPs in DMEM with 10% MS. Hydrodynamic size and Z-Potential over time as determined by DLS: (a) APS-MNPs, (b) DEX-MNPs and (c) DMSA-MNPs. Control sample, MNPs incubated in DMEM without MS; Corona sample, MNPs incubated in DMEM with 10% MS to allow PC formation. In both cases, the DLS measurements were performed in triplicate. Fig. S5. Magnetically isolated endolysosomes containing MNPs. (a) Endolysosomal Lamp 1 marker found in endolysosomes containing MNPs assessed in Western blots. (b) Quantification of Lamp1 levels by densitometry of the bands obtained in Western blot using the Image J software. Fig. S6. Viability of macrophage cells treated with different concentrations of APS-, DEX- and DMSA-MNPs. Cell viability of RAW264.7 (a) and NCTC1469 cells (b). Concentration dependent cytotoxic effects of MNPs evaluated with the PrestoBlue assay after a 24 h incubation. The data are the mean ± SD of three independent experiments in both analyses. Fig. S7. TEM images of MNPs before and after their internalization in endolysosomes in RAW 264.7 and NCTC1469 macrophagic cells. These images were analyzed to determine the decrease in the size of the iron oxide core as indicative of intracellular degradation. Scale bar 20 nm. Fig. S8. Cellular iron concentrations in macrophage cells after APS-, DEX- and DMSA-MNP uptake. (a) Quantification of the iron concentration in RAW 264.7 cells by ICP-OES. (b) (a) Quantification of the iron concentration in NCTC1469 cells by ICP-OES. The data are shown as the mean ± SD (n = 3) and the iron concentration in both cell types was compared by a one-way analysis of variance (ANOVA) and Tukey’s multiple tests. The asterisks indicate statistically significant differences in iron concentration: ns – no significant differences, *p &lt;0.05, **p &lt;0.01, ***p &lt;0.001, and **** p &lt;0.0001. Fig. S9. Temperature dependence of the in-phase and out-of-phase susceptibility scaled to the maximum associated with the presence of particles in the spleen. These plots help visualize the change in position of the AC magnetic susceptibility maximum over time, indicating particle transformation. (a) APS-MNPs, (b) DEX-MNPs and (c) DMSA-MNPs. The in-phase (real, χ′ - Top) and out-of-phase (imaginary, χ′′ - Bottom) component of the AC magnetic susceptibility measurements are shown. Fig. S10. Temperature dependence of the in-phase and out-of-phase susceptibility scaled to the maximum associated with the presence of particles in the liver. These plots help visualize the change in position of the AC magnetic susceptibility maximum over time indicating particle transformation. (a) APS-MNPs, (b) DEX-MNPs and (c) DMSA-MNPs. The in-phase (real, χ′ - Top) and out-of-phase (imaginary, χ′′ - Bottom) component of the AC magnetic susceptibility measurements are shown. Fig. S11. Evolution of the ferritin and paramagnetic ion signals in PBS-treated mice measured by AC magnetic susceptibility at different times. The magnetic susceptibility showed a paramagnetic signal in the in-phase magnetic susceptibility component in the liver, although no paramagnetic contribution was observed in the spleen. In addition, a ferritin signal in the out-of-phase magnetic susceptibility component that increased over time was observed in the spleen but not in the liver. The in-phase (real, χ′ - Top) and out-of-phase (imaginary, χ′′ - Bottom) component of the AC magnetic susceptibility measurements of the liver (a) and spleen (b) are shown.