Nowadays, great attention has been paid to the design and development of novel macromolecular constructions in drug delivery. Hyperbranched polymers (HBPs) are amongst various types of polymeric structures with exceptional physicochemical properties and biomedical applications relevant to their dendritic structure. Unlike perfect dendrimers, HBPs can be produced via one-step polymerization reactions. Moreover, they can be applied as building blocks in preparing dendronized, dendrigraft or dendritic nanostructures. Hyperbranched polyglycerols (HPGs) and their derivatives are widely regarded as promising biomaterials in a variety of diagnostic and therapeutic applications through solubilization, microencapsulation or conjugation with diagnostic agents, drugs, genetic materials, proteins, peptides, etc. HPGs have good chemical stability, inertness under biological conditions, water-solubility with low viscosity and multi-functionalization, which prompt their application as ideal or interesting nanocarriers or modifying agents. Herein, it was aimed to review recent developments and targeted approaches in utilizing HPGs for delivery of drugs, proteins and nucleic acids, as well as tissue engineering, diagnostic and theranostic applications.