In 96-well culture plates (Thermo-Fisher Scientific, Waltham, MA), 2 × 105 cells in 1 mL of DMEM per well were cultured and grown to subconfluent monolayers for 24 hours. The culture medium was then altered with equal volumes (25 µL) of sealer extracts (conditioning medium), using the culture medium itself as a negative control. The assessment of the cytotoxic activity was substantiated by the colorimetric method bromide (3-{4,5-dimethylthiazol-2-yl}-2,5-diphenyl tetrazolium bromide) (MTT). After 24 hours of incubation, 25 µL (5 mg/mL) of MTT solution was inserted to each well, and the plates were incubated for 3 hours. The MTT was then detached and 25 µL per well dimethyl sulfoxide (Sigma Chemical Co.) was put into each well to dissolve the formazan crystals.
Based on ISO 10993–12,
16
a reduction in the number of alive cells leads to a decline in the metabolism in the sample. Such reduction is directly associated with the quantity of blue-violet formazan created as observed by the optical density at 570 nm using enzyme-linked immunosorbent assay reader (Tecan Spark, Tecan Trading AG, Switzerland). The percentage of viable cells in each well was calculated as below
18 (link)
:
Based on ISO 10993–12,
16
a reduction in the number of alive cells leads to a decline in the metabolism in the sample. Such reduction is directly associated with the quantity of blue-violet formazan created as observed by the optical density at 570 nm using enzyme-linked immunosorbent assay reader (Tecan Spark, Tecan Trading AG, Switzerland). The percentage of viable cells in each well was calculated as below
18 (link)
:
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