For the migration studies, HaCaT cells were seeded into 24-well plates at a concentration of 500,000 cells per well and allowed to adhere in standard conditions (37 °C, 5% CO 2 ) for 24 h. Then, the scratches were prepared in each well. Cells were washed thrice with PBS buffer, and a new medium containing tested compounds at the final concentration of 100 μM was added. Cells were then cultured with tested compounds in a cell culture incubator under standard conditions (37 °C, 5% CO 2 ). The microscopic observations and measurements of the sizes of wounds were performed immediately after adding tested compounds (time 0), and subsequently after 6 h and 24 h of incubation using a Nis Eclipse Ti microscope (Nikon, Tokyo, Japan). Data were recorded and analyzed using Nis-Elements BR 4.30.00 software (Nikon).
Analysis for individual samples after incubation with the tested compounds was performed based on the initial scratch size in each well. The data obtained in this way were then compared with the control sample (untreated cells). The mean size of cell free gap in the wells without tested compounds was used as a control and set as 100%. The influence on the migration rate was estimated based on the size of the wound area after compound treatment related to the control sample.
Analysis for individual samples after incubation with the tested compounds was performed based on the initial scratch size in each well. The data obtained in this way were then compared with the control sample (untreated cells). The mean size of cell free gap in the wells without tested compounds was used as a control and set as 100%. The influence on the migration rate was estimated based on the size of the wound area after compound treatment related to the control sample.