Ceramide/PLGA Nanoparticle Fabrication Protocol

Ceramide/PLGA nanoparticles were fabricated following the previous research30 (link): 25 mg poly lactic-co-glycolic acid (PLGA, Sigma-Aldrich, MO, USA) and 10 mg Ceramide (Doosan, South Korea) were dissolved in 0.5 ml chloroform (Sigma-Aldrich, MO, USA), then mixed with 10 ml 2.5% (w/v) polyvinyl alcohol (Junsei, Japan). The resultant sample was subjected to sonication (Sonictopia, South Korea) for 30 s, after which the chloroform was removed by stirring at 4 °C for 24 h. Next, 0.5 ml of 10 mg/ml chitosan (Sigma-Aldrich, MO, USA) dissolved in 1% acetic acid (Daejung, South Korea) was added to each sample and allowed to react for 2 h to develop a chitosan coating on the nanoparticle.