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The highly intricate structures of biological systems make the precise probing of biological behaviors at the cellular‐level particularly difficult. As an advanced toolset capable of exploring diverse biointerfaces, high‐aspect‐ratio nanowires stand out with their unique mechanical, optical, and electrical properties. Specifically, semiconductor nanowires show much promise in their tunability and feasibility for synthesis and fabrication. Thus far, semiconductor nanowires have shown favorable results in deciphering biological communications and translating this cellular language through the nanowire‐based biointerfaces. In this perspective, the synthesis and fabrication methods for different kinds of nanowires and nanowire‐based structures are first surveyed. Next, several cellular‐level nanowire‐enabled applications in biophysical dynamics probing, physiological or biochemical sensing, and biological activity modulation are highlighted. Then, the progress of functionalized nanowires in drug delivery and bioenergy production is reviewed. Finally, the current limitations of nanowires and an outlook into the next generation of nanowire‐based devices at the biointerfaces are concluded.