Cytotoxicity Evaluation of Polymer-Based Drug Delivery

EA.hy926 human endothelial cells (CRL-2922^™, ATCC^®, Manassas, VA, USA) were grown in DMEM supplemented with 10% (v/v) FBS and 1 % (v/v) penicillin/streptomycin and incubated at 37 °C and 5% CO₂ until 70% confluency was reached. Then, cells were plated in 4 × 10³ per well. After 24 h, different concentrations (0.125, 0.250, 0.500, and 1.0 mg_BEV/mL) of BEV-loaded PLGA, PCL, and PLA MPs were put in contact with the cells. An equivalent concentration of empty MPs for each type of polymer was used as control. In addition, even the same volume of the solvent (water) in which the MPs were suspended was used as reference for the experimentation (solvent). After 24, 48, and 72 h, MTS assay was assessed as reported by De Negri Atanasio et al. [29 (link)]. Briefly, 20 μL of reagent was added to each well and plates were incubated at 37 °C and 5% CO₂. After 3 h, absorbance at 492 nm was read using a microplate reader (Tecan Spark^® 20M, Tecan, Männendorf, Switzerland). Control cells were considered to be 100% in respect to all the other samples. Experiments were done in triplicate.