Nanoscience and nanotechnology have a historical background that dates back to ancient times, with evidence suggesting their use in Mesopotamian religious temple windows and in the production of royal artifacts such as crowns and ceremonial cups. However, the modern advancement and widespread integration of nanotechnology began following Richard Feynman’s groundbreaking lecture at the California Institute of Technology (CALTECH) in December 1959, which laid the foundation for its transformative impact on various scientific disciplines. Over the decades, nanotechnology has revolutionized fields such as medicine, chemistry, biology, and materials science, leading to unprecedented advancements in healthcare and biotechnology. Among the key innovations emerging from nanotechnology, ionic liquids (ILs) have garnered significant attention due to their remarkable physicochemical properties. ILs, characterized by their tuneability, thermal stability, and biocompatibility, have become essential components in biomedical, biotechnological, and nano-biotechnological applications. Their ability to function as non-volatile, customizable solvents has enabled significant progress in pharmaceutical formulations, drug delivery systems, and biosensing technologies. The biomedical applications of ILs span diverse areas, including targeted drug delivery systems, tissue engineering, regenerative medicine, and diagnostic imaging. By modulating ILs composition, researchers can design systems that enhance drug solubility, improve bioavailability, and enable controlled release, thereby optimizing therapeutic outcomes. Nanofluids (NFs) are a mixture of a base fluid, as water, oil, or ethylene glycol, with an adequate amount of nanoparticles (NPs) dispersed in suspension along this fluid, enabled by advancements in nanotechnology. The synergetic effect between ILs and nanotechnology has led to innovative solutions for cancer therapy, antimicrobial coatings, and advanced bioimaging techniques. Despite these promising applications, concerns about toxicity, biodegradability, and large-scale production must be addressed to enable their safe and long-term integration into clinical practice. This chapter provides a comprehensive review of the latest advancements, applications, and challenges in the biomedical use of ILs, NFs, and NPs, by exploring recent breakthroughs and evaluating their translational potential.