Skeleton-Controlled pDNA Delivery of Renewable Steroid-Based Cationic Lipids, the Endocytosis Pathway Analysis and Intracellular Localization

Abstract

Using renewable and biocompatible natural-based resources to construct functional biomaterials has attracted great attention in recent years. In this work, we successfully prepared a series of steroid-based cationic lipids by integrating various steroid skeletons/hydrophobes with (l-)-arginine headgroups via facile and efficient synthetic approach. The plasmid DNA (pDNA) binding affinity of the steroid-based cationic lipids, average particle sizes, surface potentials, morphologies and stability of the steroid-based cationic lipids/pDNA lipoplexes were disclosed to depend largely on the steroid skeletons. Cellular evaluation results revealed that cytotoxicity and gene transfection efficiency of the steroid-based cationic lipids in H1299 and HeLa cells strongly relied on the steroid hydrophobes. Interestingly, the steroid lipids/pDNA lipoplexes inclined to enter H1299 cells mainly through caveolae and lipid-raft mediated endocytosis pathways, and an intracellular trafficking route of “lipid-raft-mediated endocytosis→lysosome→cell nucleic localization” was accordingly proposed. The study provided possible approach for developing high-performance steroid-based lipid gene carriers, in which the cytotoxicity, gene transfection capability, endocytosis pathways, and intracellular trafficking/localization manners could be tuned/controlled by introducing proper steroid skeletons/hydrophobes. Noteworthy, among the lipids, Cho-Arg showed remarkably high gene transfection efficacy, even under high serum concentration (50% fetal bovine serum), making it an efficient gene transfection agent for practical application.

Self‐Assembly of Cholesterol‐Doxorubicin and TPGS into Prodrug‐Based Nanoparticles with Enhanced Cellular Uptake and Lysosome‐Dependent Pathway in Breast Cancer Cells

“…For the cells incubated with Chol‐Dox/TPGS 1:1 and 1:2, stronger red fluorescence inside the cells could be observed. Notably, lysosome co‐localization effects [ 33 ] of these assemblies were observed, indicating that the introduction of TPGS could remarkably enhance the intracellular uptake of Chol‐Dox and lead to lysosome localization. Similar intracellular localization of the Chol‐Dox/TPGS nanoassemblies was also observed in MDA‐MB‐231 cells (Figure S4, Supporting Information), indicating that intracellular trafficking of the nanoassemblies was not cell type‐dependent.…”

Lipophilic Polyamines as Promising Components of Liposomal Gene Delivery Systems

“…Amphiphiles 62a – d are based on l -arginine and certain sterols linked by amide and ester bonds ( Figure 23 ) [ 85 ]. Cholesterol-based compound 62а exhibited the highest TA, forming lipoplexes with a particle size of approximately 100 nm.…”

“…When sterol derivatives are used as hydrophobic domains, one should prefer natural compounds, which do not cause significant toxicity. In this case, it is optimal to use a common and widely available sterol such as cholesterol [ 38 , 78 , 85 ], although diosgenin derivatives are also capable of efficient NA delivery [ 75 , 76 , 77 , 83 ]. Tocopherol is also employed as a hydrophobic domain [ 80 , 92 , 101 ].…”

“…The positively charged CA domain is responsible for the electrostatic interactions of amphiphiles and/or liposomes with NAs, the formation of stable lipoplexes, and the interaction of complexes with cell membranes. An increase in the number of amino groups in the structure of the cationic domain leads to an increase in the TA [ 31 , 56 , 60 , 81 , 82 , 85 ]. The most effective are CAs with domains based on polyamines, with the number (more than two [ 74 ]) and distribution of amino groups in the polyamine chain [ 31 , 53 , 93 , 102 ] playing important roles.…”

Steroid-Based Supramolecular Systems and their Biomedical Applications: Biomolecular Recognition and Transportation

“…The study provided possible approach for developing high-performance steroid-based lipid gene carriers, in which the cytotoxicity, gene transfection capability, endocytosis pathways, as well as intracellular trafficking/localization manners could be tuned/controlled by introducing proper steroid skeletons/hydrophobes. Noteworthy, among the lipids, Cho-Arg showed remarkably high gene transfection efficacy even under high serum concentration (50% FBS), making it an efficient gene transfection agent for practical application [65].…”