The layer-by-layer (LbL) technique was introduced in the early 1990s. Since then, it has undergone a series of technological developments, making it possible to engineer various theranostic platforms, such as fi lms and capsules, with precise control at the nanometer and micrometer scales. Recent progress in the applications of LbL assemblies in the fi eld of cancer therapy, diagnosis, and fundamental biological study are highlighted here. The potential of LbL-based systems as drug carriers is discussed, especially with regard to the engineering of innovative stimuli-responsive systems, and their advantageous multifunctionality in the development of new therapeutic tools. Then, the diagnostic functions of LbL assemblies are illustrated for detection and capture of rare cancer cells. Finally, LbL-mimicking extracellular environments demonstrate the emerging potential for the study of cancer cell behavior in vitro. The advantages of LbL systems, important challenges that need to be overcome, and future perspectives in clinical practice are then highlighted.
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