Preparation of thermosensitive polymer nanoparticles by protein-mimetic cross-linking

Thermosensitive nanoparticles were prepared by mimicking protein folding where polymer aggregates were formed by precipitation of thermosensitive polymer chains followed by disulfide formation of their thiol groups. N-Isopropylacrylamide (NIPAM) and methacryloxy succinimide (SuMA) were co-polymerized and then cysteamine was allowed to react with succinimide moieties of the polymer to render thiol moieties. A polymer aqueous solution precipitated to form nano-sized aggregates by increasing temperature above its lower critical solution temperature (LCST), and their sizes were monodispersed and tunable by the polymer concentration. The aggregates were cross-linked to produce nanoparticles by oxidation of thiol groups in a manner similar to formation of a disulfide bond of protein. As a result, the cross-linked nanoparticles exhibited swelling by decreasing temperature below the LCST of the copolymer. Fluorescein and bovine serum albumin (BSA) were chosen as a small and a large substance, respectively, and were encapsulated into the swollen nanoparticles at 25 A degrees C. Fluorescein was rapidly released from both swollen and shrunken nanoparticles. Although BSA exhibited little release at any temperatures, it was released from nanoparticles by adding the reducing agent to dissociate the disulfide cross-linking and incubating below the LCST.