Limited cellular uptake of liposomes: Might thiolated phospholipids hold the key?

Aim: It was the aim of this study to evaluate the impact of surface thiolation on cellular uptake of liposomes.

Methods: Liposomes were prepared via the thin lipid film method, incorporating cholesterol, dipalmitoylphosphatidylcholin (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol). The characterization of liposomes included size, polydispersity index, surface morphology, zeta potential and stability in simulated gastric and intestinal fluid. Hemocompatibility and cytotoxicity of liposomes were investigated. Cellular uptake studies were performed on Caco-2, HEK, HeLa and SW620 cells, involving both quantitative analysis through flow cytometry and qualitative evaluation via confocal microscopy. Additionally, we investigated the impact of an oxidizing agent on thiol-dependent uptake.

Results: Non-thiolated and thiolated liposomes exhibited a size of 149 nm to 274 nm and a PDI between 0.3 and 0.45. Liposomes were stable in simulated intestinal and gastric fluid. Hemocompatibility studies and cytocompatibility studies of liposomes showed negligible toxic effects of liposomes. Cellular uptake of thiolated liposomes was 1.8-, 2.1-, 5.4- and 1.4-fold enhanced in comparison to non-thiolated liposomes on Caco-2, HEK, HELA and SW620 cells, respectively. The results were qualitatively verified by confocal microscopy. Thiol dependent uptake was influenced by oxidizing agents on HeLa cells.

Conclusion: Surface thiolation represents a promising approach to enhance cellular uptake of liposomes.