Facile formation of dynamic hydrogel microspheres for triggered growth factor delivery

Dynamic hydrogels have emerged as an important class of biomaterials for temporal control over growth factor delivery. In this study we formed dynamic hydrogel microspheres from protein–polymer conjugates using an aqueous two-phase suspension polymerization process. This polymerization process enabled rapid microsphere formation without the use of an organic phase, surfactants, mechanical strain or toxic radical initiators. The microspheres’ size distribution was modulated by varying the protein–polymer conformation in the pre-polymer solution. Notably, the protein’s ligand-induced, nanometer-scale conformational change translated to maximum hydrogel volume changes of 76 ± 10%. The magnitude of the microspheres’ volume change was tuned by varying the crosslinking time and ligand identity. After characterizing the microspheres’ dynamic properties, we encapsulated two important therapeutic proteins, vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2), in the hydrogel microspheres and characterized how the microspheres’ dynamic properties controlled their release. Significantly, the aqueous two-phase suspension polymerization process enabled high encapsulation efficiencies (65.8 ± 4.8% and 79.5 ± 3.0% for VEGF and BMP-2, respectively). Also, the microspheres’ ligand-induced volume change triggered VEGF and BMP-2 release at specific, predetermined times. There are hundreds of proteins that undergo well-characterized conformational changes that could be processed into hydrogel microspheres via aqueous two-phase suspension polymerizations. Therefore, this approach could be used to form dynamic, growth-factor-releasing hydrogel microspheres that respond to a broad range of specific biochemical ligands.