Calcein am pi cytotoxicity assay kit

Alpha-modified Eagle’s Medium (α-MEM), penicillin-streptomycin (PS), 10% foetal bovine serum (FBS) and 0.25% trypsin-ethylenediaminetetraacetic acid (EDTA) were purchased from Gibco Company (United States). 4% paraformaldehyde fixative (4% PFA) and 2-(4-aminophenyl) - 6-indolyl methylamino dihydro ether (DAPI) were purchased from Shanghai BTS Biotechnology Co. Ltd. Calcein AM/PI Cytotoxicity Assay Kit was purchased from Solarbio (China). Staphylococcus aureus (Staphylococcus aureus) and E. coli (Escherichia coli) required for the present study were purchased through the American Type Culture Collection (ATCC) purchased from the Liaoning Provincial Center for Disease Control and Prevention. ICA, PVA (polyvinyl alcohol) and Alg (sodium alginate) were obtained from Aladdin Reagents Ltd. (China).

The effect of ICA + CNF@H on the cell vitality was determined by CCK-8 method. BMSCs cells were inoculated in 96-well plates at a concentration of 5,000 cells/well with a medium volume of 100 μL and cultured at 37°C and 5% CO₂ for 24 h. Cells were processed for 1, 3 and 7 days with blank control and different materials, respectively. 10 μL of CCK-8 solution was added to each hole, the incubation was carried out for 2 h, and the absorbance was measured at 450 nm using an enzyme marker.
The biosafety of ICA + CNF@H was tested by live-dead staining of cells. ICA + CNF@H and ICA + CNF@H (NIR) were co-cultured with BMSCs for 1, 3 and 5 days, and the effect of composites on BMSCs was examined employing the Calcein AM/PI Cytotoxicity Assay Kit (Solarbio, China). The red fluorescent nucleic acid dye PI is able to penetrate impaired cell walls for labeling dead cells. In addition, Calcein green gets into living cells and is broken down to Calcein by intracellular esterase cleavage, emits intense green fluorescence and stays inside the living cells. Cells were incubated in the staining solution for 30 min and photographed under a fluorescent microscope for observation.