Exploiting Niosomes as Efficient Non-Viral Vectors for Enhanced Gene Transfer to Human Mesenchymal Stem Cells

Abstract

[Abstract] Gene therapy of mesenchymal stem cells (MSCs) represents a powerful tool to enhance their therapeutic potential for regenerative medicine approaches. Among current non-viral vectors used for MSCs transfection, niosomes are preferred over classical liposomes due to their reduced cytotoxicity. However, the use of niosomes is hindered by the lack of optimization in its composition, precluding their efficiency, especially when using in hard-to-transfect cells as MSCs. To develop effective niosome formulations for gene delivery to human MSCs, this study explores, for the first time, a broad range of compositions and mole ratios and their impact on transfection efficiency. Formulations containing the cationic lipid DOTMA, the non-ionic surfactants polysorbate 20 or polysorbate 80, and the helper lipids cholesterol (DP80CH) or chloroquine (DP20CQ), at a 1:2:2 M ratio and DOTMA/DNA ratio of 5/1, achieved the highest lacZ and GFP reporter transgene expressions in MSCs. Endocytosis pathway analysis revealed that niosome composition strongly influenced cellular uptake, with DP80CH niosomes being internalized via macropinocytosis and caveolae-dependent endocytosis, while DP20CQ niosomes primarily utilized the latter mechanism. Physicochemical characterization confirmed their suitability for gene delivery applications. These findings demonstrate the potential of niosomes as effective vectors for MSCs genetic modification, offering new strategies for the prevention and treatment of various diseases.