Influence of Lipid Thioether Modification and Formulation Method on Lipoplex Structure, Stability and Transfection Efficiency in HEK293T Cells
Abstract
Lipoplexes, formed by electrostatic interactions between nucleic acids and cationic lipids, must protect their cargo, enable cellular uptake and promote intracellular release for non-viral gene delivery. Although traditionally produced by bulk complexation, scalable microfluidic methods are now available. However, how lipid chemistry and formulation jointly influence physicochemical properties and biological performance remains poorly understood. To address this gap, we compared two structurally matched cationic lipids differing only in their hydrophobic chains: BSV107-S containing two thioether-functionalized chains, and BSV107, containing two mono-unsaturated chains. Each lipid was formulated either by complexing DNA with preformed liposomes obtained by thin-film hydration or by one-step DNA/lipid complexation through microfluidics assembly. We hypothesized that thioether incorportation would increase membrane fluidity and enhance intracellular plasmid release. Both lipids formed nanosized lipoplexes, with hydrodynamic diameters ranging 70-100 nm for microfluidic formulations and 100-140 nm for bulk-prepared lipopexes at R ≥ 2. Cryo-TEM and SAXS revealed a multilamellar organization, with BSV107-S microfluidic lipoplexes showing larger interlamellar spacings than BSV107 or bulk-prepared systems. In HEK293T cells, transfection efficiency depended on by the interplay between lipid composition and formulation, with BSV107-S consistently outperforming BSV107 regardless of formulation method. This improved performance correlated with increased membrane fluidity, measured by FRAP, supporting the role of thioether groups in modulating membrane dynamics. Together, these results establish structure-formulation-property relationships linking lipid chemical structure, formulation, physicochemical properties and biological performance, providing insights for the rational design of more efficient non-viral gene delivery systems.
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BibTeX
@article{rochedy2026influence,
title = {{Influence of Lipid Thioether Modification and Formulation Method on Lipoplex Structure, Stability and Transfection Efficiency in HEK293T Cells}},
author = {Manon Rochedy and Laurie Dechamboux and Aurélien Dupont and Mathieu Berchel and Paul-Alain Jaffrès and Jean-Paul Chapel and Thomas Vivès and Patricia Leroyer and Sylvain Tranchimand and Fabienne Gauffre and Frédéric Nallet and Christophe Schatz and Pascal Loyer and Lourdes Mónica Bravo-Anaya},
journal = {Journal of materials chemistry B},
year = {2026},
month = aug,
doi = {10.1039/d6tb00356g},
url = {https://hal.science/hal-05733778v2},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Influence of Lipid Thioether Modification and Formulation Method on Lipoplex Structure, Stability and Transfection Efficiency in HEK293T Cells
AU - Manon Rochedy
AU - Laurie Dechamboux
AU - Aurélien Dupont
AU - Mathieu Berchel
AU - Paul-Alain Jaffrès
AU - Jean-Paul Chapel
AU - Thomas Vivès
AU - Patricia Leroyer
AU - Sylvain Tranchimand
AU - Fabienne Gauffre
AU - Frédéric Nallet
AU - Christophe Schatz
AU - Pascal Loyer
AU - Lourdes Mónica Bravo-Anaya
PY - 2026
DA - 2026/08/31
JO - Journal of materials chemistry B
DO - 10.1039/d6tb00356g
UR - https://hal.science/hal-05733778v2
AB - Lipoplexes, formed by electrostatic interactions between nucleic acids and cationic lipids, must protect their cargo, enable cellular uptake and promote intracellular release for non-viral gene delivery. Although traditionally produced by bulk complexation, scalable microfluidic methods are now available. However, how lipid chemistry and formulation jointly influence physicochemical properties and biological performance remains poorly understood. To address this gap, we compared two structurally matched cationic lipids differing only in their hydrophobic chains: BSV107-S containing two thioether-functionalized chains, and BSV107, containing two mono-unsaturated chains. Each lipid was formulated either by complexing DNA with preformed liposomes obtained by thin-film hydration or by one-step DNA/lipid complexation through microfluidics assembly. We hypothesized that thioether incorportation would increase membrane fluidity and enhance intracellular plasmid release. Both lipids formed nanosized lipoplexes, with hydrodynamic diameters ranging 70-100 nm for microfluidic formulations and 100-140 nm for bulk-prepared lipopexes at R ≥ 2. Cryo-TEM and SAXS revealed a multilamellar organization, with BSV107-S microfluidic lipoplexes showing larger interlamellar spacings than BSV107 or bulk-prepared systems. In HEK293T cells, transfection efficiency depended on by the interplay between lipid composition and formulation, with BSV107-S consistently outperforming BSV107 regardless of formulation method. This improved performance correlated with increased membrane fluidity, measured by FRAP, supporting the role of thioether groups in modulating membrane dynamics. Together, these results establish structure-formulation-property relationships linking lipid chemical structure, formulation, physicochemical properties and biological performance, providing insights for the rational design of more efficient non-viral gene delivery systems.
SN - 2050-750X
ER -