Fig. 3.

Isolation and characterization of neutrophils. a Flow cytometry data of neutrophils treated with culture media and D-MMSNs for 1 h, respectively. Dox fluorescence signal was monitored at 488 nm via the PE-A channel. b CLSM images of neutrophils after coincubation with D-FMMSNs for 1 h ([Dox] = 10 μg ml⁻¹). Blue fluorescence represents the polymorphonuclear of neutrophils stained with DAPI. Green fluorescence represents FITC-labeled MMSNs. Red fluorescence represents Dox encapsulated in the D-MMSNs. Scale bar: 2.5 μm. c T₂-weighted MRI of neutrophils after coincubation with D-MMSNs of varied concentrations (from left to right: 0, 12.5, 25, and 50 μg ml⁻¹, respectively) for 1 h. d Schematic illustration of in vitro inflammation response evaluation. e Representative images of migration of ND-MMSNs towards fresh serum-free media and fresh serum-free media + TNF-α for 30 min, 1, and 2 h. Scale bar: 100 μm. f CLSM images of NETs after ND-FMMSNs incubated with U87 glioma cells for 2 h. Neutrophil-derived DNA networks were stained with DAPI emitting blue fluorescence. Green fluorescence: FITC-labeled MMSNs. Red fluorescence: Dox. Scale bar: 5 μm. g CLSM images of U87 glioma cells after incubation with ND-FMMSNs for 4 h ([Dox] = 10 μg ml⁻¹). After incubation, U87 cell nuclei were stained with DAPI, which emits blue fluorescence. Green fluorescence: FITC-labeled MMSNs. Red fluorescence: Dox. Scale bar: 10 μm