Thermoresponsive Injectable Hydrogels Cross-Linked by Native Chemical Ligation

Abstract

Temperature-induced physical gelation was combined with native chemical ligation (NCL) as a chemical cross-linking mechanism to yield rapid network formation and mechanically strong hydrogels. To this end, a novel monomer <i>N</i>-(2-hydroxypropyl)­methacrylamide-cysteine (HPMA-Cys) was synthesized that copolymerizes with <i>N</i>-isopropylacrylamide (NIPAAm) to yield thermoresponsive polymers decorated with cysteine functionalities. Triblock copolymers consisting of a poly­(ethylene glycol) (PEG) middle block flanked by random blocks of NIPAAm and HPMA-Cys were successfully synthesized and characterized. Additionally, thioester cross-linkers were synthesized based on PEG and hyaluronic acid, respectively. Upon mixing the thermoresponsive polymer with PEG or hyaluronic acid cross-linker, cysteine and thioester functionalities react under physiological conditions to generate a native peptide bond. An immediate physical network was formed after elevation of the temperature to 37 °C due to the self-assembly of the pNIPAAm chains. This network was stabilized in time by covalent cross-linking due to NCL reaction between thioester and cysteine functionalities, resulting in hydrogels with up to 10 times higher storage moduli than without chemical cross-links. Finally, a collagen mimicking peptide sequence was successfully ligated to this hydrogel using the same reaction mechanism, showing the potential of this hydrogel for tissue engineering applications