Preparation and properties of an injectable thermo-sensitive double crosslinking hydrogel based on thiolated chitosan/beta-glycerophosphate

An injectable thermo-sensitive double crosslinking hydrogel based on thiolated chitosan (CS-TGA)/beta-glycerophosphate (β-GP) was prepared by combining physical and chemical crosslinking. The effect of the concentrations of CS-TGA and β-GP on gelation temperature of CS-TGA-GP hydrogel was investigated. The gelation and mechanical properties of CS-TGA-GP in situ gel system were studied by oscillatory rheology and unconfined compression testing. By the physical interaction of CS-TGA and β-GP, CS-TGA-GP system undergoes a fast gelation at body temperature within 2 min. In addition, CS-TGA-GP hydrogel contains a low concentration of β-GP, which significantly decreases the toxicity of the gel. Owing to the chemical crosslinking of disulfide bonds, the gel is durable and possesses high-mechanical strength without introducing any potential cytotoxicity. The integrity of CS-TGA-GP hydrogel maintains for more than 30 days both in vitro and in vivo, and the interior morphology visualized by scanning electron microscopy reveals that the gel has interconnected porous network structure. In vitro release behavior of protein from CS-TGA-GP hydrogel was investigated using BSA as model protein. There is a sustained protein release from the gel without any initial burst. In vitro cytotoxicity, hemolysis, and histopathological analysis reveal that the gel is biocompatible. These features indicate that CS-TGA-GP hydrogel is a promising candidate for injectable protein delivery system and biomedical applications.