Cytotoxicity was
evaluated according
to the ISO10993-5-2009 regulation, where the L929 cell line was chosen,
and an MTT cell viability assay was performed. To obtain extraction
solutions, samples were cut into square pieces (2 × 2 cm2) and sterilized by UV irradiation for 40 min. Then, samples
were placed in six-well tissue culture plates and filled with a culture
medium (DMEM) with an extraction ratio of 4 cm2/0.666 mL
at 37 °C in a 5% CO2 incubator for 24 h. L929 cells
were seeded into 96-well culture plates at a density of 6 × 103 cells/well in 100 μL of a culture medium and incubated
for 24 h at 37 °C in a humidified atmosphere containing 5% CO2 in air. After this time, culture media were replaced with
100 μL of the sample and control group extracts (the extraction
medium without the sample and only a glass slide were used as control
groups). Cells were examined microscopically after 24 h of incubation
to assess the general morphology of cells. Then, the cells were incubated
with 10 μL of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide solution (MTT, 5 mg/mL, Sigma-Aldrich, Germany) for 3 h. Formazan
crystals were dissolved in 100 μL of dimethyl sulfoxide, and
the absorbance (OD) was measured with a microplate reader (Promega
Multireader Glomax) at 560 nm. Cell viability was calculated by the
following equation
Experiments
were performed in triplicate,
and mean OD values were normalized to the control group and represented
as cell viability (%).
evaluated according
to the ISO10993-5-2009 regulation, where the L929 cell line was chosen,
and an MTT cell viability assay was performed. To obtain extraction
solutions, samples were cut into square pieces (2 × 2 cm2) and sterilized by UV irradiation for 40 min. Then, samples
were placed in six-well tissue culture plates and filled with a culture
medium (DMEM) with an extraction ratio of 4 cm2/0.666 mL
at 37 °C in a 5% CO2 incubator for 24 h. L929 cells
were seeded into 96-well culture plates at a density of 6 × 103 cells/well in 100 μL of a culture medium and incubated
for 24 h at 37 °C in a humidified atmosphere containing 5% CO2 in air. After this time, culture media were replaced with
100 μL of the sample and control group extracts (the extraction
medium without the sample and only a glass slide were used as control
groups). Cells were examined microscopically after 24 h of incubation
to assess the general morphology of cells. Then, the cells were incubated
with 10 μL of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide solution (MTT, 5 mg/mL, Sigma-Aldrich, Germany) for 3 h. Formazan
crystals were dissolved in 100 μL of dimethyl sulfoxide, and
the absorbance (OD) was measured with a microplate reader (Promega
Multireader Glomax) at 560 nm. Cell viability was calculated by the
following equation
Experiments
were performed in triplicate,
and mean OD values were normalized to the control group and represented
as cell viability (%).
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