Cck 8 solution

Two days after transfection, we collected cells and plated them in 96-well plates at 2,000 cells/well. After culturing for 0, 1, 2, or 3 d, we added 10 μL CCK-8 solution (Gibco, Logan, Utah, USA) per well and incubated for 1 h. Then, we measured the optical density (OD) value of each well using a microplate reader with 490 nm wavelength.

The Cell Counting Kit‐8 assay was utilized to ascertain the cell viability. The 500 μl of GelMA, GelMA/HA‐E, and GelMA/HA‐E/Ag@MOF hydrogels was coated into 12‐well plates and then irradiated under a UV light (365 nm) for 10 s. Next, a thin layer of the prepared hydrogel was plated as a surface in the plates before 2 ml of 3 T3 cells (2 × 10⁴ cells/ml) were seeded and cultured in the medium consisting of DMEM high‐glucose medium (CGIBO, USA) with 10% fetal bovine serum (FBS, GIBCO, USA) and 1% penicillin/streptomycin at 37°C in 5% CO₂ atmosphere. At given time (first, second, and third days), the culture medium was taken out from the plates which later added a mixed medium with CCK‐8 solution (2% [v/v]， GIBCO). After incubation for 1 h at 37°C, each well was analyzed at 450 nm (Thermo Scientific).

Cell counting kit-8 (CCK-8) was purchased from Solarbio (Beijing, China). To assay the toxicity of the Fy formula, quercetin, luteolin, apigenin, and GT1-7 cells were seeded in 96-well plates at a density of 2.0×10⁵ cells/well. After 24 h of incubation, the cells were pretreated with 100 pmol/L E₂ in SFM overnight. The cells were then incubated with 75, 150, 225, 300, 450, and 525 μg/ml of Fy formula or 5, 10, 15, 20, 30, and 40 μmol/ml of quercetin, luteolin, and apigenin separately in SFM for 24 h. Subsequently, the cells were treated with 10 μL of CCK-8 solution (Gibco) for 1 h at 37°C and 5% CO₂. The optical density (OD) was determined by measuring the absorbance at 450 nm using a microplate reader (BioTek Synergy, United States).