Rhodamine phalloidin (Solarbio, China) was used to stain microfilaments. rBMSCs were incubated at 37 °C and 5% CO2 for 2 h and then rinsed with PBS followed by fixation in 4% paraformaldehyde for 15 min. After rinsing with PBS three times, the cells were dyed with Rhodamine phalloidin for 30 min and 4’,6-diamidino-2-phenylindole (DAPI) for 3 min. The samples were kept moist, and the cell conditions were observed and imaged under the red and blue channels using a laser confocal microscope.
After rBMSCs were cultured on the surface of the samples (10 mm × 10 mm × 1 mm) for 1, 3, 5 and 7 d, cell proliferation was tested using CCK-8 (APE x BIO, USA). For each group (n=3), 2×104 viable cells were seeded per sample, with α-minimal essential medium (α-MEM) as the blank control. At each predetermined time interval, samples were rinsed 3 times with PBS to remove unattached cells and transferred to new 24-well plates containing 500 μL of 10% CCK-8 solution (APE x BIO, USA) per well. The plates were incubated in the dark at 37 °C for 1 h, and 100 μL of each solution was transferred into 96-well plates in triplicate and read at 450 nm using a microplate reader (Multiskan FC, Thermo Scientific, USA).
After rBMSCs were cultured on the surface of the samples (10 mm × 10 mm × 1 mm) for 1, 3, 5 and 7 d, cell proliferation was tested using CCK-8 (APE x BIO, USA). For each group (n=3), 2×104 viable cells were seeded per sample, with α-minimal essential medium (α-MEM) as the blank control. At each predetermined time interval, samples were rinsed 3 times with PBS to remove unattached cells and transferred to new 24-well plates containing 500 μL of 10% CCK-8 solution (APE x BIO, USA) per well. The plates were incubated in the dark at 37 °C for 1 h, and 100 μL of each solution was transferred into 96-well plates in triplicate and read at 450 nm using a microplate reader (Multiskan FC, Thermo Scientific, USA).