The anti-inflammatory effect of resveratrol-loaded hydrogel was studied in vitro using lipopolysaccharide (LPS)-induced inflammation on RAW 264.7 macrophage cells. The peptide-hydrogels loaded with/without resveratrol were prepared on the bottom of a 6-well plate. Macrophage cells (1.0 × 106 cells) were added into wells, and LPS was added after 6-h incubation. After 24-h or 48-h treatment, cells were collected for qRT-PCR assay to detect the mRNA expression of inflammatory cytokines TNF-α and IL-1β. The procedures are as follows. Total RNA was extracted by Trizol, and then the cDNA was obtained by using reverse transcription kit (PrimeScript RT reagent Kit, Takara, China). The analysis was performed on the CF96-Real-Time System (Bio-rad, USA) using SYBR Green Premix Ex Taq II (Takara, Beijing, China). The housekeeping gene GAPDH was employed as a control gene. The primers for the target genes are listed in Table 1 .
After 24-h or 48-h treatment, the culture medium was collected to detect the concentration of TNF-α and IL-1β secreted by macrophages using ELISA kit (R&D Systems, USA). The absorbance at 450 nm was measured using a microplate reader, and the concentrations were obtained by measuring the standard curves.
After 24-h or 48-h treatment, the culture medium was collected to detect the concentration of TNF-α and IL-1β secreted by macrophages using ELISA kit (R&D Systems, USA). The absorbance at 450 nm was measured using a microplate reader, and the concentrations were obtained by measuring the standard curves.