Rutin is an herbal polyphenolic compound recognized for its numerous therapeutic benefits, including antioxidant, anticancer, and antimicrobial properties. However, its application in biomedical fields encounters significant challenges, such as low solubility, poor absorption, low bioavailability, short half-life, and rapid metabolism. In recent years, advancements in nanotechnology have presented promising solutions to these limitations. Consequently, various nano-formulation strategies have been developed to enhance rutin's solubility, absorption, and overall efficacy. These strategies can be broadly categorized into two approaches. The first involves transforming rutin into nanocrystals without the use of any secondary compounds. The second approach entails nano-formulating rutin with other compounds, including proteins, polysaccharides, lipids, polymers, and metals. This study offers a review of these approaches and their applications in biomedical sciences, focusing on their categories, preparation methods, and biomedical properties. Initially, the pharmacological potential of rutin, its application in recent clinical trials, and its mechanisms of action are outlined. Next, it explores how nano-carriers can enhance rutin's bioavailability. Subsequently, the types of nanostructures employed are categorized, along with a discussion of their fabrication methods and benefits. In some instances, certain drawbacks are also reported. The data provided can guide the selection of optimal strategies for rutin nano-formulation, based on the intended biomedical application.