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We report a new class of chitosan-based hybrid nanogels by in-situ immobilization of CdSe quantum dots (QDs) in the chitosan–poly(methacrylic acid) (chitosan–PMAA) networks. The covalently crosslinked hybrid nanogels with chitosan chains semi-interpenetrating in the crosslinked PMAA networks exhibit excellent colloidal and structural stability as well as reversible physical property change in response to a pH variation cross the physiological condition. In contrast, the hybrid nanogels formed by non-covalent physical association exhibit a significant change in the structure and composition upon exposure to physiological pH. This distinction in the structural stability of hybrid nanogels produces very different outcomes for their biomedical applications. The covalently crosslinked hybrid nanogels are low-cytotoxic and could illuminate the B16F10 cells, sense the environmental pH change, and regulate the release …