Abstract: The construction methods of chiral nanostructured materials and their latest progress in biological applications was reviewed. Firstly， the basic concepts and classification of chiral nanostructured materials were introduced， including inorganic chiral nanomaterials and chiral supramolecular assemblies. Subsequently， various construction strategies of chiral nanostructured materials were elaborated in details， such as surface-assisted self-assembly， self-assembly with chiral transfer， wet chemical method， chiral ligand modification， and chiral doping. The advantages and limitations of these strategies in regulating the morphology and properties of chiral nanostructures were discussed. In terms of biological applications， the application examples of chiral nanostructured materials in the fields of biomedicine （drug synthesis and delivery， disease diagnosis and treatment）， tissue engineering （stem cell differentiation regulation， tissue repair and regeneration） and biocatalysis were mainly discussed. In addition， the chiral nanostructured materials also showed unique application prospects in the fields of chiral recognition， nanoreactors， etc. Finally， the challenges faced by chiral nanomaterials in biological applications were presented， such as the stability of chiral nanostructures， biocompatibility， and large-scale preparation. In response to these issues， the future research directions and development trends were proposed.