Recent studies demonstrated the potential therapeutic use of newly synthesized omega-3 (omega-3) polyunsaturated fatty acid (PUFA) monoglycerides owing to their beneficial health effects in various disorders including cancer and inflammation diseases. To date, the research was mainly focused on exploring the biological effects of these functional lipids. However, to the best of our knowledge, there is no report on the hydration-mediated self assembly of these lipids that leads to the formation. of nanostructures, which are attractive for use as vehicles for the delivery of drugs and functional foods. In the present study, we investigated the temperature-composition phase 29 behaviour of eicosapentaenoic acid monoglyceride (MAG-EPA), which is one of the most investigated w-3 PUFA monoglycerides, during a heating cooling cycle in the temperature range of 5-60 degrees C. Experimental synchrotron small-angle X-ray scattering (SAXS) evidence on the formation of a dominant inverse hexagonal (H-2) lyotropic liquid crystalline phase and its temperature-induced transition to an inverse micellar solution (L-2 phase) is presented for the fully hydrated bulk MAC-EPA system and its corresponding dispersion. We produced colloidal MAG-EPA hexosomes with an internal inverse hexagonal (H2) lyotropic crystalline phase in the presence of F127, a well-known polymeric stabilizer, or citrem, which is a negatively charged food-grade emulsifier. In this work, we report also on the formation of MAG-EPA hexosomes by vortexing MAG-EPA in excess aqueous medium containing F127 at room temperature. This low-energy emulsification method is different than most reported studies in the literature that have demonstrated the need for using a high-energy input during the emulsification step or adding an organic solvent for the formation of such colloidal nonlamellar liquid crystalline dispersions. The designed nanoparticles hold promise for future drug and functional food delivery applications due to their unique structural properties and the potential health-promoting effects of MAG-EPA.