The cytotoxicity of the naproxen-based
ILs of L929 cells in vitro was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) assay. Cells were seeded at a density of 1 × 104 cells·mL–1 cells/well (200 μL)
in DMEM containing 10% fetal bovine serum and 1% antibiotic solution
in 96-well plates for 24–48 h at 37 °C. In serum-free
DMEM, wells were rinsed with sterile PBS and treated with various
concentrations of the API-IL samples. Each sample was reproduced three
times, and the cells were cultured for 24 h at 37 °C. Twenty
microliters of MTT (5 mg·mL–1) was added to
each well, and the cells were incubated for 4 h until purple precipitates
were visible under a microscope. Finally, the medium was gently aspirated
from the wells and rinsed with 1× PBS (200 μL). Then we
added dimethylsulfoxide (DMSO) (100 μL) to dissolve the formazan
crystals and shook the plate for 5 min. Using a microplate reader
(Thermo Fisher Scientific, USA), the absorbance value of each well
was measured at 490 nm, and the cell viability and IC50 value were calculated using GraphPad Prism 5.0 (USA).
ILs of L929 cells in vitro was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) assay. Cells were seeded at a density of 1 × 104 cells·mL–1 cells/well (200 μL)
in DMEM containing 10% fetal bovine serum and 1% antibiotic solution
in 96-well plates for 24–48 h at 37 °C. In serum-free
DMEM, wells were rinsed with sterile PBS and treated with various
concentrations of the API-IL samples. Each sample was reproduced three
times, and the cells were cultured for 24 h at 37 °C. Twenty
microliters of MTT (5 mg·mL–1) was added to
each well, and the cells were incubated for 4 h until purple precipitates
were visible under a microscope. Finally, the medium was gently aspirated
from the wells and rinsed with 1× PBS (200 μL). Then we
added dimethylsulfoxide (DMSO) (100 μL) to dissolve the formazan
crystals and shook the plate for 5 min. Using a microplate reader
(Thermo Fisher Scientific, USA), the absorbance value of each well
was measured at 490 nm, and the cell viability and IC50 value were calculated using GraphPad Prism 5.0 (USA).
