Abstract
In contemporary research, there is a clear emphasis on the physicochemical characteristics and effectiveness of nanoliposomal (NLs) formulations. However, there has been minimal focus on elucidating nano-bio interactions and understanding the behavior of these formulations at organ and cellular levels. Specifically, it is widely recognized that when exposed to biological fluids, nano-delivery systems, including NLs, rapidly interact with various biomolecules which have a significant impact on the functionality and destiny of the nano-systems but also influence cellular biological functions. Hence, the primary objective of this study was to illuminate the evolution of physicochemical characteristics and surface properties of NLs in biorelevant media. Additionally, in order to point out the influence of specific characteristics on the brain targeting potential of these formulations, we investigated NLs interactions with BBB (hCMEC/D3) and neuroblastoma cells (SH-SY5Y) under different conditions. The results obtained from in vitro comparative cell uptake studies on both cell culture lines after treatment with 3 different concentrations of fluorescently labelled NLs (5, 10 and 100 μg/mL) over a period of 1, 2 and 4 h showed a time- and concentration-dependent internalization pattern, with high impact of the surface characteristics of the different formulations. In addition, transport studies on hCMEC/D3/SH-SY5Y co-culture confirmed the successful transport of NLs across the BBB cells and their subsequent uptake by neurons (ranging from 25.17 to 27.54%). Fluorescence and confocal microscopy micrographs revealed that, once internalized, NLs were concentrated in the perinuclear cell regions.