Cell scratch assay

To assess cell migration, RWPE-1 and DU145 cells were treated with 1000 pulses of Li-ESWT at the G2 and G3 phases, using a nonadherent in vitro model. Following Li-ESWT, cells were resuspended in complete culture medium and counted. Cells were seeded in 6-well plates at a density of approximately 400 000 cells per well and incubated at 37°C for 24 h to establish a monolayer. Subsequently, scratches (wounds) were created across the cell monolayer. Detached cells and debris were carefully removed. RWPE-1 cells were supplemented with complete culture medium, whereas DU145 cells were supplemented with serum-free medium. These plates were incubated at 37°C for 24 h and 48 h in the presence of 5% CO₂. Images were captured using phase-contrast microscopy (AxioVert.A1; Carl Zeiss AG, Oberkochen, Germany) to monitor cell migration into the scratched areas.