Cytotoxicity Assessment of β-CDs/CeO2 NPs

The potential cytotoxicity of β-CDs/CeO₂ NPs was investigated in MTT assay as we reported earlier.46 (link) The HaCaT human keratinocyte cells line was selected to analyze cell viability in MTT assay (Sigma, MO, USA). HaCaT human keratinocyte cells were seeded into 96-well plates at a concentration of 1×10⁴ cells/well and then incubated at 37°C and 5% CO₂ for 24 h to ensure proper stability and adherence. Then, the culture medium was removed. The cells were incubated with fresh medium containing 100 µg/mL β-CDs/CeO₂ NPs for 24 h before 20 µM H₂O₂ was added. After 24 h of cell culture, 10 µL of MTT (5 mg/mL) was added into each well. Incubation was continued for 4 h, and then 100 µL of DMSO was appended to each well to dissolve MTT crystals. The reactions were monitored using an iMark microplate reader (Bio-rad, CA, USA) at a wavelength of 490 nm. The experiment was repeated three times.