The dye was applied according to the manufacturer's protocol (General Method for Measuring Cytotoxicity or Proliferation Using alamarBlue® by Fluorescence, Bio-Rad). Five controls were performed: (a) cells with alamarBlue® (positive control), (b) cell with 1% Triton-X 100 (negative control), (c) medium alone with dye (blank), (d) dead cells with alamarBlue®, (e) a pure medium without dye. 20-30 μl of specific compounds of layer were dropped on the each well and incubate at 37°C for 48 hours. After this time cells were seeded into a 96-well plate (Greiner Bio-One) with approximately 7500 cells number per well (controlled with Countess II Cell Counter) and were incubated at 37°C for 24 hours. Plates was read after incubation with 10% alamarBlue® dye at 37°C in phenol red-free culture medium (to eliminate the background fluorescence), lasted for 4 hours. The fluorescence was measured at 590 nm (590/20 filter) for excitation at 560 nm (560/20 filter). Plot of cell viability was made by taking averages of the six repeats.