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.