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Designed synthesis and assembly of nanoparticles assisted by their surface ligands can create “smart” materials with programmed responses to external stimuli for biomedical applications. These assemblies can be designed to respond either exogenously (for example, to magnetic field, temperature, ultrasound, light, or electric pulses) or endogenously (to pH, enzymatic activity, or redox gradients) and play an increasingly important role in a diverse range of biomedical applications, such as biosensors, drug delivery, molecular imaging, and novel theranostic systems. In this review, the recent advances and challenges in the development of stimuli‐responsive nanoparticle assemblies are summarized; in particular, the application‐driven design of surface ligands for stimuli‐responsive nanoparticle assemblies that are capable of sensing small changes in the disease microenvironment, which induce the related …