PLGA microspheres by Supercritical Emulsion Extraction: a study on insulin release in myoblast culture

Supercritical Emulsion Extraction in a Continuous operation layout is proposed for the production of poly-lactic-co-glycolic acid (PLGA) microspheres loaded with insulin, selected as a model of bioactive signal. Microspheres with different mean sizes of 2m (+/- 0.9m) and 3m (+/- 2.2m) and insulin loadings of 3 and 6mg/g were obtained by processing different water-oil-water emulsions; an encapsulation efficiency of about 60% w/w was measured in all cases. Insulin release profiles from PLGA microspheres were also characterized in two different media (Phosphate-Buffered Saline and Dulbecco's Modified Eagle Medium) and kinetic constants were estimated by using a model proposed in literature. The produced microspheres were, then, used for the cultivation of rat embryonic ventricular myoblasts in a serum-free medium to monitor the biological effect of the released insulin. The best cell viability and proliferation, supported by released insulin, was monitored when microspheres with mean size of 3m loaded with 3mg/g of insulin were added.