The treatment of diseases, such as cancer, is one of the most significant issues correlated with human beings health. Hydrogels (HGs) prepared from biocompatible and biodegradable materials, especially biopolymers, have been effectively employed for the sort of pharmaceutical and biomedical applications, including drug delivery systems, biosensors, and tissue engineering. Chitosan (CS), one of the most abundant bio-polysaccharide derived from chitin, is an efficient biomaterial in the prognosis, diagnosis, and treatment of diseases. CS-based HGs possess some potential advantages, like high values of bioactive encapsulation, efficient drug delivery to a target site, sustained drug release, good biocompatibility and biodegradability, high serum stability, non-immunogenicity, etc., which made them practical and useful for pharmaceutical and biomedical applications. In this review, we summarize recent achievements and advances associated with CS-based HGs for drug delivery, regenerative medicine, disease detection and therapy.