| Sumario: | 13 pages. -- SI Figure 1: Characterization of rGO. (a) Raman spectra and (b) FTIR patterns of rGO. functionalized-rGO (green), rGO (red) and GO (black). -- SI Figure 2: Characterization of functionalized rGO.(a) XPS spectra of the functionalized rGO. Deconvolution of (b) C1s and (c) O1s.-- SI Figure 3: TGA of functionalized rGO. -- SI Figure 4: TEM micrographs of rGO after the APTES functionalization. -- SI Figure 5: Physicochemical characterization of rGO and abraded composite materials. (a) AFM images of rGO and PA6-rGO . Lateral dimension and thickness distribution analysis for rGO and PA6-rGO. (b) Optical microscopy images of rGO, PA6-rGO and PA6. -- SI Figure 6: (a) EPR spectra of the powder of PA6, rGO and PA6-rGO at room temperature. (b)Assessment of ROS. Fluorescent spectra of PA6, rGO and PA6-rGO in 0.1 % BSA at 40 µg/mL in the presence of DHR123. Fenton reaction (Fe2+ + H2O2) corresponds to the positive control. -- EPR analysis of PA6, rGO and PA6-rGO. -- Fluorescence analysis of PA6, rGO and PA6-rGO. -- SI Figure 7: Particle size distributions of abraded (a) PA6-rGO and (b) PA6 particles in the nanometer range (13–573 nm) measured, measured by SMPS. The results show mean ± SD from at least three-independent measurements. -- SI Figure 8: TNF-α Expression Test (TET) for endotoxin detection in HMDMs exposed to PA6, PA6-rGO, and rGO (20 g/mL) or to bacterial LPS (0.01 µg/mL) in the presence or absence of the specific LPS inhibitor, polymyxin B (Poly-B; 10 µM) for 24h. . * indicates statistical significance compared to the negative control in absence of Poly-B (p < 0.05). # shows a statistically significant response (p < 0.05) in presence of Poly-B. -- SI Figure 9: (a) SEM images of the crack surface of the freeze-fractured samples of neat PA6 and PA6-rGO composite. Red line and red arrows indicate pulled-out rGOs. (b) TEM image of the abraded particles from the PA6-rGO composite showing the protruding rGOs from PA6 matrix marked with a dashed line. Layered structure of rGO can be observed at higher magnification. -- SI Figure 10: Pro-inflammatory response of A549 epithelial cells after treatment with the inflammogenic material DQ (100 µg/mL). (a) IL-6 and (b) IL-8 release measured after 24 h. Results are shown as mean ± standard error of the mean (SEM) from three independent experiments. * indicates statistical significance compared to the negative control at 24 h of exposure (p < 0.05). -- SI Figure 10: Pro-inflammatory response of A549 epithelial cells after treatment with the inflammogenic material DQ (100 µg/mL). (a) IL-6 and (b) IL-8 release measured after 24 h. Results are shown as mean ± standard error of the mean (SEM) from three independent experiments. * indicates statistical significance compared to the negative control at 24 h of exposure (p < 0.05). -- SI Figure 12: (a) Cytokine release by THP-1 macrophages. IFN-γ, TNF-α and IL-1β levels were determined after incubation with 20 µg/mL of PA6, PA6-rGO and rGO. Data are the mean ± SEM of 3 independent experiments. The * symbol represents p<0.05 as compared to the negative control. (b) THP-1 cells incubated with rGO (20 µg/mL) in the presence or absence of the pan-caspase inhibitor, zVAD-fmk (20 µM) or the NLRP3 inhibitor MCC950 (10 µM). * = p < 0.05, ** = p < 0.01, *** = p < 0.001. -- SI Figure 13: Cell morphology of RAW 264.7 macrophages exposed to different concentrations of rGO and abraded PA6 and PA6-rGO composites after 24 h. Scale bar: 50 μm. -- SI Figure 14: Cytokine profiling and hierarchical cluster analysis of inflammatory mediators released in differentiated HL-60 neutrophils exposed to rGO, abraded PA6-rGO and PA6. -- SI Figure 15: Interference assessment of the abraded particles (PA6 and PA6-rGO) and rGO with the MTS assay. -- SI Figure 16: : TNF-α and IL-6 measurements in the Bronchoalveolar lavage fluids.
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