Biocompatibility Evaluation of Hydrogel Formulations
The primary dermal fibroblast cell line (ATCC PCS-201-012) was cultured
using the Fibroblast Growth Kit-Low Serum (ATCC PCS-201-041) in a
humidified 5% CO2-containing balanced-air incubator at
37°C.
Fibroblast cells used in the assays were seeded in each well in the
24-well plates at the density of 4×104 cells per well
overnight at 37 °C in a humidified 5% CO2-containing atmosphere. After
that, the media in each well was discarded. And cells were exposed to
the AgNPs-PVP at concentrations of 3.125 µM, 6.25 µM, 12.5 µM, 25 µM, 50
µM, and 100 µM for 24h dissolved in fresh medium. Viability of
fibroblast cells was evaluated using a LIVE/DEAD cytotoxicity kit
(Invitrogen, USA) which was diluted 1000-fold into PBS. The live/dead
image was obtained by Confocal (Zeiss LSM800) after incubating cells
with the LIVE/DEAD staining assay for about 30 mins. Untreated cells
with the assay were used as control. Relative cell viability was
calculated by normalizing the reading of number of live cells by that of
untreated cells via ImageJ. All assays were carried out in
quadruplicates.