Synthesis of Injectable, Thermally Responsive, Chondroitin Sulfate-Cross-Linked Poly(N-isopropylacrylamide) Hydrogels

In this study, we describe the synthesis and characterization of a biosynthetic hydrogel system that consists of a thermally responsive macromer and biological cross-linkers. By combining a poly(N-isopropylacrylamide)-based thermogelling macromer with epoxy pendant groups and chondroitin sulfate cross-linkers that are modified to contain either hydrazide or N-hydroxysuccinimide pendant groups, we successfully fabricated a system that undergoes gelation when the temperature is raised from room temperature to 37 °C and is further stabilized via covalent links between the macromers. The anionic charge on chondroitin sulfate contributed to a high degree of gel swelling, while the cross-linking reaction between the macromers prevented post-formation syneresis. The rate of degradation of CS-cross-linked hydrogels was dependent on the degree of substitution of hydrazide-modified chondroitin sulfate cross-linkers. A higher molar content of chondroitin sulfate led to a greater osmotic pressure within the hydrogel and thus a higher compressive modulus. On the other hand, excessive amounts of chondroitin sulfate caused time-dependent cytotoxicity, as confirmed by a leachables cytocompatibility study. Overall, the system described in this study provides a versatile platform to synthesize hydrogels with differing combinations of compressive moduli and rates of degradation, which is achievable by varying the degree of substitution of hydrazide groups on CS-based cross-linkers.