Cytocompatibility Evaluation of the PCL/nHA Scaffold

BMSCs-C were cultured in passage 3 and then cultured for cytocompatibility with PCL/nHA scaffold, using cell counting kit 8 (CCK-8) assay and the LIVE/DEAD staining method. BMSCs-C cells from passage 3 were adjusted to the density of 2×10⁶/ml, seeded the PCL/nHA scaffold in a 96-well plate, and then cultured for 1 day, 4 days, and 7 days at 37°C for evaluation of cell proliferation. At the end of each time point, BMSCs-C were incubated with a CCK-8 solution (Sigma, St. Louis, MO, USA) at a concentration of 10% for 2 h. Finally, the optical density (OD) of BMSCs-C was detected at a wavelength of 450 with an ELISA reader (Hiperion MPR4, Germany). For LIVE/DEAD staining, BMSCs-C cells were cultured as in the CCK-8 assay for 28 days and stained with the staining solution (10 mal PBS solution containing 5 μl of AM calcine and 20 μl of ethidium homodimer-1) for 30 min. Finally, the images of LIVE/DEAD stained BMSCs-C cells were captured using laser confocal microscopy (FV1000, Olympus, Tokyo, Japan).