Glycol chitosan as a stabilizer for protein encapsulated into poly(lactide-co-glycolide) microparticle

Glycol chitosan (GC), a chitosan derivative conjugated with ethylene glycol, is soluble in water at a neutral/acidic pH and is viscous. This GC was incorporated into poly(lactide-co-glycolide) (PLGA) microparticles (prepared by the multi-emulsion W-1/O/W-2 (water-in-oil-in-water) method) to stabilize lysozyme (Lys) used as a model protein. Herein, GC's viscous property helped to improve Lys encapsulation efficacy and reduce Lys denaturaton at the water/organic solvent interface. When the GC concentration in the W-1 phase increased, the formation of non-covalent Lys aggregates decreased. This may be because the aqueous microdroplets surrounded by the firm viscous interface protect Lys from the degrading environment formed by the water/organic solvent interface. In an in vitro Lys release test, 40 mg incorporation of GC led to continuous Lys release of up to 78 wt.% for 1 month and presented bioactivity of more than 95% for Lys released from microparticles. In addition, there was negligible immune response in the tissue treated with the GC-incorporated PLGA microparticles, whereas there was a moderate foreign body reaction in the muscle layer and many configurations of neutrophils in the tissue treated with the PLGA microparticles without GC. It is expected that GC facilitates a decrease in immune responses exacerbated as a consequence of PLGA degradation. (c) 2007 Elsevier B.V. All rights reserved.