Cell survival rate

Three groups of cells were extracted from the hydrogel at the time of the assay so that each group was kept in the same culture environment. All three groups were maintained in equal volumes of a cell and medium mixture. The quantity of the mixture was determined based on the blank control. Because the blank control group had the most comprehensive cell counts, changes in cell counts could be clearly observed when cells were injured. The mixtures were diluted to a density of 1×10⁴ cells/well, uniformly inoculated in a 96-well plate (Thermo Fisher Scientific, Shanghai, China), and incubated for 12 h. The cell survival rate was then measured using an enhanced CCK-8 kit (Beyotime, Shanghai, China). The cells were incubated for an additional 4 h after adding CCK-8, and the absorbance was measured at a wavelength of 450 nm using a fully automated enzyme labeling instrument (ELX800, Biotek, Vermont, USA), which was operated while being protected from light. The cell survival rates of the normal control and injured groups were determined by comparing their optical density with that of the blank control group. Then, by subtracting the cell survival rate of the control group from that of the injured group, we calculated the impact of mechanical injury on the cell survival rate while eliminating the influence of the hydrogel.