Rosa canina L. seeds are rich in bioactive components that can add value to the various formulations. The focus of the study was the development of liposomes for R. canina oil to protect its sensitive compounds and prolong their shelf-life. Oil-loaded liposomes were characterized via the determination of the particle size, polydispersity index (PDI), zeta potential, conductivity, mobility, density, surface tension, viscosity, and stability. Raman and FT-IR spectroscopy were employed to investigate the chemical composition of the non-treated and UV-treated samples, and the presence of different interactions. Antioxidant and antimicrobial activities were examined as well. The liposome size was 970.4 ± 37.4 nm, the PDI 0.438 ± 0.038, the zeta potential -32.9 ± 0.8 mV, the conductivity 0.068 ± 0.002 mS/cm, the mobility -2.58 ± 0.06 µmcm/Vs, the density 0.974 ± 0.004 g/cm3, the surface tension 17.2 ± 1.4 mN/m, and the viscosity 13.5 ± 0.2 mPa•s. The Raman and FT-IR spectra showed the presence of lipids, fatty acids, polyphenols, and carotenoids. It was approved that the oil compounds were distributed inside the phospholipid bilayer and were combined with the membrane interface of the bilayer. The UV irradiation did not cause any chemical changes. However, neither the pure oil nor the oil-loaded liposomes showed any antimicrobial potential, while the antioxidant capacity of the oil-loaded liposomes was significantly low. The sizes of the liposomes did not change significantly during 60 days of storage. Due to the proven stability of the oil-loaded liposomes, as well as the liposome's ability to protect the sensitive oil compounds, their potential application in the pharmaceutical and cosmetic formulations could be investigated with a focus on the skin regeneration effects.