Triton x 100

HUVECs in logarithmic growth phase were used for further study. The cells were digested by trypsin and separately seeded on the AD scaffold and AD/PRP composite scaffold in a 96-well plate with a density of 10⁴ cells/mL. On day 7 of culture, the samples were divided into 2 parts for further study. One part was fixed with 4% paraformaldehyde at room temperature for 1 h. After the fixative samples were washed twice with PBS every 10 min, the cells were sequentially penetrated with 0.1% Triton X-100 (Coolaber, China) for 10 min and blocked with 1% bovine serum albumin (Sigma-Aldrich, America) for 30 min. Subsequently, 5 μg/mL phalloidin-rhodamine (Invitrogen, America) and 4′, 6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich) solution were utilized to stain the cytoskeletons and the cell nuclei, respectively, in a dark room. Finally, a laser scanning confocal microscope (LSCM, Leica, SP8) was used to obtain images of cell adhesion and proliferation at 405/552 nm. For the other part of the samples, gradient dehydration was performed with 50%, 70%, 90%, and 100% ethanol. After freeze-drying, the samples were coated with a thin layer of gold and observed using a scanning electron microscope (SEM) (ZEISS Merlin) at 5 kV with a Schottky Thermal Field Emission (TFE) filament.