Fig. 2: Synthesize and characterization of novel multifunctional nanostructure FSGG/siGal-9.
From: Tumor-targeted nanodrug FSGG/siGal-9 for transdermal photothermal immunotherapy of melanoma
a, b TEM images of FSG and FSGG. FSG and FSGG were synthesized as described in Materials and Methods. c Zeta potential and (d) Normalized UV-Vis absorption spectra of FS, FSG, GNR-MUA, FSGG, which were synthesized as described in Materials and Methods. e Gel shift assay for assessing the siRNA-loading capacity of FSGG. Equal volumes containing 1 μg of siRNA and the desired weights (0 μg, 1 μg, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg) of FSGG were mixed and incubated at the indicated weight ratios for 30 min. Gels were visualized with Ethidium Bromide. f Temperature increase induced by photothermal effects of GNR-MUA or FSGG in vitro. 1 ml of GNR-MUA (40 μg/ml) or FSGG (40 μg/ml), or deionized water was irradiated by 808 NIR laser at 2 W/cm2 for 10 min. The temperatures were measured using infrared radiation thermometer and plotted at indicated time points. g, h Cellular transfection efficiency of siRNA by FSGG in vitro. After 24 h incubation, the intracellular fluorescence intensity was observed under fluorescence microscope (g) (The cell nuclei were stained with DAPI, scale bar = 100 μm) and the Cy3 positive population of B16-F10 was determined by flow cytometry (h). i Assessment of cytotoxicity of FSGG in vitro. B16-F10 cells were incubated with a cell culture medium containing various concentrations (10 µg/ml, 20 µg/ml, 30 µg/ml, 40 µg/ml, 50 µg/ml, 60 µg/ml, 70 µg/ml) of FSGG or PBS as control for 24 h or 48 h. The cell viabilities were measured by the MTT assay. j, k In vitro photothermal effects of FSGG killing B16-F10 tumor cells. B16-F10 cells were incubated with 40 μg/ml FSGG or PBS overnight. Subsequently, the cells were irradiated by 808 nm NIR laser for 5 min at various densities (0 W/cm2, 1 W/cm2, 2 W/cm2, 3 W/cm2 and 4 W/cm2) (j), or at 2 W/cm2 for various times (0 s, 100 s, 200 s, 300 s, 400 s, 500 s) (k). After 24 h, the cell viabilities were measured by MTT assays and the percentages of dead cells = (Cell viabilities before laser irradiation - cell viabilities after laser irradiation)/ (cell viabilities before laser irradiation) were calculated. Error bars represent the standard deviation of 3 experiments (*P ≤ 0.05; **P ≤ 0.01; ****P ≤ 0.0001; Adding the magnet field is denoted with “+M”).
