Customizing biohybrid cryogels to serve as ready-to-use delivery systems of signaling proteins.

Macroporous cryogels have recently gained increasing interest for the controlled administration of signaling proteins in tissue engineering due to an advantageous combination of material properties. However, most of the previously reported cryogel systems did not allow for tunable, sustained protein release. We therefore designed a set of ready-to-use multi-armed polyethylene glycol (starPEG)-heparin cryogel systems containing different amounts of the protein-affine glycosaminoglycan component heparin to enable systematically tunable long-term delivery of different signaling proteins without affecting other cell-instructive properties. Experimental data and mathematical modeling indicate that the macroporous structure causes local differences in the concentration of proteins released into the pores and in the surrounding of the cryogels. As a proof-of-concept for their ready-to-use potential, cryogels pre-functionalized with signaling proteins and cell adhesion-peptides were demonstrated to induce the neuronal differentiation of colonizing pheochromocytoma cells. The elaborated approach opens up new perspectives for cryogels as easily storable and applicable systems for the precision delivery of signaling proteins.