Sh sy5y human

SH-SY5Y human neuroblastoma cells were obtained from the American Type Culture Collection and were maintained in DMEM supplemented with 2 mM glutamine, 10% FBS, and 1% antibiotic-antimycotic solution. Cells were grown to confluence at 37 °C in a 5% CO₂ atmosphere. All experiments were performed in 96-well plates and 100-mm cell culture dishes based on requirements. The cells were prepared in 100-µL aliquots at a density of 1 × 10⁵ mL and plated in 96-well plates. After the cells were cultured for 24 h, the medium was replaced with a medium containing various concentrations or required concentrations of GO or AgNPs or GO-AgNPs. After incubation for an additional 24 h, the cells were analyzed for viability. The cells that were not exposed to GO, AgNPs or GO-AgNPs served as controls. For all differentiation experiments, SH-SY5Y cells were grown in the same culture media supplemented with 1% FBS for 24 h and then washed with fresh media and further incubated with GO (10 µg/mL), AgNPs (5 µg/mL) or GO-AgNPs (1 µg/mL) for another 24 h.

The SH-SY5Y human neuroblastoma cell line was obtained from American type culture collection (ATCC) and maintained in high glucose modified Eagle’s medium (DMEM) supplemented with 15% fetal bovine serum and penicillin/streptomycin (100 μg/ml) at 37 °C with 5% CO₂. The medium was refreshed once a day during cell growth. When cells reached 80~ 90% confluence, cell suspension was sub-cultured on flask according to 1:3 proportion. Cells were plated in a 96-well plate at a concentration of 1 × 10⁴ cells in per well and cultured for 24 h. Then, the medium with the concentration of Aβ25–35 (5 μM/L, 10 μM/L, 20 μM/L, 40 μM/L) was added into the culture plate for 48 h. And the physiological saline acted as the negative control.