4.3.4. Nitric Oxide Release

We performed the nitric oxide (NO) release assessment by seeding the cells with various concentrations of EGCG-loaded liposomes in one hand, and on the other hand, cells were seeded without EGCG. After 24 h incubation, cells were treated with 50 ng/mL LPS for both conditions and incubated for 24 h. Next, we collected 50 μL of culture media from each culture condition and transferred them into 96-well plates. 50 μL of Griess reagent was used to evaluate NO concentration in cells and was then added into different wells. The disposal was protected from light and incubated at 25 ◦C for 20 min. ELISA reader assessed the optic density (OD) values at 540 nm wavelength for determining the concentration of NO released by cells.
