Self-assembled biomaterials are capable of spontaneously organizing their constituent monomers into well-defined structures like nanofibers, vesicles, and hydrogels, among others. The assembly is driven primarily by molecular interactions, including hydrogen bonds and ionic, hydrophobic, and van der Waals forces. Moreover, environmental factors like pH, temperature, solvent properties, and concentration significantly influence the process. Biomaterials have been extensively studied for various biomedical applications. This chapter provides an overview of the advancements in self-assembled biomaterials and the non-covalent interactions involved in their formation. It highlights their evolution from traditional implants to cutting-edge platforms for tissue engineering and biomimetic environments. Various types of self-assembled materials based on peptides, nucleic acids, amphiphilic blocks, proteins, lipids, and hybrid materials have also been covered. Their physicochemical and functional properties are analyzed using structure elucidation and characterization techniques such as microscopy, spectroscopy, and scattering, alongside others. These materials find extensive applications in drug delivery systems, tissue engineering, biosensing and diagnostics, wound healing, and bioprinting, which have been summarized in the chapter. Despite their significant potential, challenges persist, including optimizing the tolerance and response of the human body, achieving long-term bioavailability, and cost-effective mass production. Still, biomaterials are a promising platform for future healthcare solutions. Interdisciplinary research collaboration among researchers across materials science, biology, chemistry, and biomedical engineering can lead to the development of cost-effective, sustainable, biocompatible, and biodegradable self-assembled systems.