Protein-Cross-Linked Hydrogels with Tailored Swelling and Bioactivity Performance: A Comparative Study

The design of protein-based hydrogels that include biological activity independent of structural functionality is desirable for many bioengineering applications. Here a general route for construction of protein-based hydrogel is proposed by pretreatment of protein with thiolation agent and succeeding conjugation with 4-arm PEG-acrylate via Michael addition reaction. Different swelling behaviors responding to temperature and ions are comparatively studied for hydrogel cross-linked with hemoglobin (multimeric protein), albumin (monomeric protein), and dithiothreitol (DTT, small molecule). Meanwhile, the microscopic structure change is studied to correlate with the macroscopic hydrogel swelling behavior. Results show that proteins, which function as multisite cross-linkers, affect the gel swelling behaviors, and the effect is more profound for multimeric proteins when exposed to stimulus for protein dissociation. Moreover, the catalytic activity derived from hemoglobin is also preserved in the hydrogel, as demonstrated by the successfully synthesis of the colored product. By taking advantage of each particular protein, a broad range of functional materials can be expected for potential biomedical applications, such as stimuli-responsive hydrogel and immobilized enzyme.