Responsive Protein Hydrogels Assembled from Spider Silk Carboxyl-Terminal Domain and Resilin Copolymers

Responsive protein hydrogels are known to respond to target external stimuli that cause changes in their properties, attracting considerable attention for diverse applications. Here we report the design and recombinant biosynthesis of protein copolymers via genetic fusion of repeating units of resilin with spider silk carboxyl-terminal (CT) domain. The resulting copolymers were thermoresponsive in aqueous solutions, and formed reversible hydrogels at low temperatures and irreversible hydrogels at high temperatures within minutes, a peculiar dual thermogelation feature endowed by the CT domain. The incorporation of resilin blocks upshifted the temperature range of reversible gelation and hydrogel stiffness, whereas the temperature of irreversible gelation was differentially affected by the length of the resilin blocks. In addition, sodium chloride and potassium phosphate at moderate concentrations downregulated both the reversible and irreversible gelation temperatures and hydrogel mechanical properties, proving the salts as another level of control over dual thermogelation. Surprisingly, the copolymers were prone to gelate at body temperature in a time-dependent manner, and the resulting hydrogels were pH-responsive to release a highly polar model drug in vitro. The newly developed resilin-CT copolymers and the multistimuli-responsive hydrogels may be potentially useful in biomedicine, such as for drug delivery.