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Clay nanomaterials are an emerging class of two-dimensional (2D) biomaterials of interest due to their atomically thin layered structure, discotic charged characteristics and well-defined composition. Synthetic nanoclays are plate-like polyions composed of simple or complex salts of silicic acids with a heterogeneous charge distribution and patchy interactions. Due to their biocompatible characteristics, unique shape, high surface-to-volume ratio and charge distribution, nanoclays are investigated for various biomedical applications. This review article will provide a critical overview of the physical, chemical and physiological interactions of nanoclays with biological moieties including cells, proteins and polymers. The state-of-the-art biomedical applications of 2D nanoclay in regenerative medicine, therapeutic delivery and additive manufacturing are reviewed. In addition, recent developments that are shaping this emerging field are discussed and promising new research directions for 2D nanoclay-based biomaterials are identified.