Evaluating A. cinnamomea Extracts' Cytotoxicity

Cell viability was measured using Cell Proliferation Reagent WST-1 (Roche, Basel, BS, Switzerland). In brief, Meg-01 cells were seeded at a density of 1000 cells per well in 100 μL culture medium containing gradient concentrations of A. cinnamomea extracts in 96-well plates. Cells cultured in a medium without A. cinnamomea extract were used as control cells. The plates were incubated at 37 °C in an incubator with 5% CO₂ for 7 days. After incubation, 10 μL of WST-1 assay reagent was added to each well and incubated for another 2 h. Absorbance at 440 nm was determined using theenzyme-linked immunosorbent assay (ELISA) reader (Multiskan SkyHigh Microplate Spectrophotometer, Thermo Fisher Scientific, Waltham, MA, USA). All experiments were performed in duplicate three times. According previous studies described [64 (link),65 (link),66 (link)], the values of CC₁₀ and CC₅₀ (cytotoxicity concentration that causes a 10% and 50% reduction in cell numbers compared to the control) were calculated a 4-parameter logistic (4PL) model and were regarded as the working concentration of the A. cinnamomea extracts for the virus inhibition assay.