The emulsion/solvent evaporation method and miniemulsion technique were combined and applied in the formulation of biodegradable monodisperse nanoparticles at high solid contents using different biocompatible and biodegradable polymers such as poly(L-lactide) (PLLA), poly[(D,L-lactide)-co-glycolide] 50:50 (PLGA), and poly(epsilon-caprolactone) (PCL). Differences between the results of various polymers are found in terms of the particle size and size distribution as well as in the degradation time. An encapsulated hydrophobic fluorescent dye was used as a model marker in order to study the entrapment efficiency and diffusion yield out of the particle. Cellular uptake of the obtained particles was observed in Jurkat and HeLa cells. In the investigated particle size range of 80-200 nm, the surfactant on the particles' surface had a greater influence than the particle size. Uptake kinetics reveals that the PLLA and PCL particles are endocytosed much faster than polystyrene particles.
By using fluorescent polysorbate 80 coated poly(n-butylcyanoacrylate) (PBCA) nanoparticles in an in vivo study, direct evidence was found for the presence of nanoparticles entering the brain and retina of rats. The nanoparticles, prepared with a miniemulsion process, were labeled in situ with a fluorescent dye and coated with polysorbate 80. After preparation the particle size, zeta potential, and the molecular weight distribution were determined. BMEC cells were used as an in vitro model for the BBB. The cells showed significant uptake of the particles, but no transcytosis could be observed in vitro. After applying the particles to the animals at two concentrations, cryosections of the brains and retinas were prepared. Regarding the sections of the rats that received the lower dose, co-localization of the applied fluorescent particles and the stained endothelial cells could be detected in the brain and retina, indicating particle internalization in the endothelial cells. Applied at higher doses, the particles could be detected within the brain and retina with few co-localized signals, suggesting passage through the blood-brain and blood-retina barriers.
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