Glowmax multi

Viability of hMSC cells, after 1 and 3 days of treatment with DMEM extracts of the CaP_Si powders, was assessed by using MTT assay (Sigma Aldrich) as previously described in our studies [9 (link),21 (link)]. In brief, the hMSC cells were seeded in a 96-well plate (Sarstedt, Nümbrecht, Germany) at a density of 5 × 10⁴ cells and were allowed to adhere overnight at 37 °C, 5% CO₂. The next day, cell culture medium was removed and cells were treated with DMEM extracts of the CaP_Si powders. Following the incubation period, medium was removed and 40 μL of MTT solution (0.5 mg mL⁻¹) was added to each well. After 3 h of incubation at 37 °C, 170 μL of dimethyl sulfoxide (DMSO, Sigma Aldrich) was added to each well to dissolve formazan crystals. The solution was set for colorimetric detection at 560 nm using a microplate reader (GlowMax-Multi, Promega, Madison, WI, USA). The results are presented as a percentage of cell viability and calculated in reference to negative control (untreated cells).

Viability of HEK 293 and U2OS cells was obtained using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay (Sigma-Aldrich, St. Louis, MO, USA), after 1 and 3 days of cell treatment with the extract of as-prepared powders. After determined time points, the medium was removed and 40 μL of MTT solution was added to each well. After 3 h at 37 °C, 170 μL of dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) was added to each well. When formazan crystals were dissolved, the solution was set for colorimetric detection at 560 nm using a microplate reader (GlowMax-Multi, Promega, Madison, WI, USA). All experiments were performed in triplicates. The percentage of cell viability was calculated from the absorbance readings in reference to untreated cells.