Light-triggered release from dye-loaded fluorescent lipid nanocarriers in vitro and in vivo

Light is an attractive trigger for release of active molecules from nanocarriers in biological systems. Here, we describe a phenomenon of light-induced release of a fluorescent dye from lipid nano-droplets under visible light conditions. Using auto-emulsification process we prepared nanoemulsion droplets of 32nm size encapsulating the hydrophobic analogue of Nile Red, NR668. While these nano-droplets cannot spontaneously enter the cells on the time scale of hours, after illumination for 30s under the microscope at the wavelength of NR668 absorption (535nm), the dye showed fast accumulation inside the cells. The same phenomenon was observed in zebrafish, where nano-droplets initially staining the blood circulation were released into endothelial cells and tissues after illumination. Fluorescence correlation spectroscopy revealed that laser illumination at relatively low power (60mW/cm²) could trigger the release of the dye into recipient media, such as 10% serum or blank lipid nanocarriers. The photo-release can be inhibited by deoxygenation with sodium sulfite, suggesting that at least in part the release could be related to a photochemical process involving oxygen, though a photo-thermal effect could also take place. Finally, we showed that illumination of NR668 can provoke the release into the cells of another highly hydrophobic dye co-encapsulated into the lipid nanocarriers. These results suggest dye-loaded lipid nano-droplets as a prospective platform for preparation of light-triggered nanocarriers of active molecules.