A preliminary study on the interaction between Asn-Gly-Arg (NGR)-modified multifunctional nanoparticles and vascular epithelial cells

Liu, Chunxi; Liu, Tingxian; Yu, Xiaoyue; Gu, Yizhu

To examine the influence of cholesterol depletion, we treated cells with 2 mmol/L MβCD in ECM for 30 min at 37 1C. After being washed three times with PBS, TPIC were added into wells and incubated with cells for another 4 h at 37 1C with 5% CO 2 in an incubator. Then, all cells were harvested and washed, and cellassociated fluorescence was determined by FACS. To exclude the possibility of irreversible damage caused by the above treatment, we further incubated cells with an MβCD-cholesterol complex for 30 min at 37 1C to replenish cholesterol. MβCD-cholesterol was prepared as described previously 36 . Briefly, cholesterol in methanol-chloroform (1:1) was dried and suspended in MβCD in 0.1 mol/L PBS buffer. The solution was sonicated and rotated at 37 1C overnight. The mixture was then filtered and diluted with ECM. The final concentrations were approximately 1 mmol/L MβCD and 100 μg/mL of cholesterol.