Biocompatibility Evaluation of AgNPs, PVP, and AgNPs-PVP
PC-12 cell line (ATCC CRL-1721.1) was cultured with F-12K medium supplemented (Corning, USA) with 2.5% fetal bovine serum (Gibco, USA), 15% horse serum (Gibco, USA) and 1% penicillin and streptomycin (Gibco, USA); and maintained in a humidified 5% CO2-containing balanced-air incubator at 37°C.
PC-12 cells used in the assays were seeded in each well in the 96-well plates at the density of \(1\times 10^{4}\) 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 and 48h dissolved in fresh medium. Viability of PC-12 cells was evaluated using the CCK-8 kit for mammalian cells (Dojindo Molecular Technologies, Japan). The absorbance at 450 nm was measured after incubating cells with the CCK-8 assay for 1-2h. Empty wells with CCK-8 assay only were used as blanks and were subtracted from the final reading. Relative cell viability was calculated by normalizing the absorbance readings by that of untreated cells . All assays were carried out in quadruplicates.