# Influence of surface characteristics on the in vitro stability and cell uptake of nanoliposomes for brain delivery

## TL;DR

This study explores how the surface properties of nanoliposomes affect their stability and uptake by brain cells in laboratory conditions.

## Contribution

The novel aspect is the detailed investigation of how surface characteristics influence nanoliposome behavior at the blood-brain barrier and neuronal cells.

## Key findings

- Nanoliposome uptake by cells was time- and concentration-dependent.
- Surface characteristics strongly influenced the internalization and transport of nanoliposomes across the blood-brain barrier.
- Internalized nanoliposomes were concentrated in perinuclear regions of cells.

## 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, nanodelivery systems, including NLs, rapidly interact with various biomolecules which have a significant impact on the functionality and fate of the nanosystems but also influence cellular biological functions. Hence, the primary objective of this study was to elucidate 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 interactions between NLs and blood–brain barrier (BBB, hCMEC/D3) and neuroblastoma cells (SH-SY5Y) under different conditions. The results obtained from comparative in vitro cell uptake studies on both cell culture lines after treatment with three 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-cultures 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.