title: The role of helper lipids in the intracellular disposition and transfection efficiency of niosome formulations for gene delivery to retinal pigment epithelial cells.
abstract: In this work, we carried out a comparative study of four different niosome formulations based on the same cationic lipid and non-ionic tensoactive. The niosomes prepared by oil-in-water emulsion technique (o/w) only differed in the helper lipid composition: squalene, cholesterol, squalane or no helper lipid. Niosomes and nioplexes elaborated upon the addition of pCMS-EGFP reporter plasmid were characterized in terms of size, zeta potential and polydispersity index. The capacity of the niosomes to condense, release and protect the DNA against enzymatic degradation was evaluated by agarose gel electrophoresis. In vitro experiments were carried out to evaluate transfection efficiency and cell viability in retinal pigment epithelial cells. Moreover, uptake and intracellular trafficking studies were performed to further understand the role of the helper lipids in the transfection process. Interestingly, among all tested formulations, niosomes elaborated with squalene as helper lipid were the most efficient transfecting cells. Such transfection efficiency could be attributed to their higher cellular uptake and the particular entry pathways used, where macropinocytosis pathway and lysosomal release played an important role. Therefore, these results suggest that helper lipid composition is a crucial step to be considered in the design of niosome formulation for retinal gene delivery applications since clearly modulates the cellular uptake, internalization mechanism and consequently, the final transfection efficiency.
authors.affiliation: NanoBioCel Group, University of Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain.
authors.forename: Edilberto
authors.initials: E
authors.lastname: Ojeda
authors.affiliation: NanoBioCel Group, University of Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Puras
authors.affiliation: NanoBioCel Group, University of Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Agirre
authors.affiliation: NanoBioCel Group, University of Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain.
authors.forename: Jon
authors.initials: J
authors.lastname: Zarate
authors.affiliation: Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain; Institute of Advanced Chemistry of Catalonia, IQAC-CSIC, Barcelona, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Grijalvo
authors.affiliation: Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain; Institute of Advanced Chemistry of Catalonia, IQAC-CSIC, Barcelona, Spain.
authors.forename: Ramon
authors.initials: R
authors.lastname: Eritja
authors.affiliation: Department of Molecular Medicine, "Sapienza" University of Rome, Viale Regina Elena, 324, 00161 Rome, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: DiGiacomo
authors.affiliation: Department of Molecular Medicine, "Sapienza" University of Rome, Viale Regina Elena, 324, 00161 Rome, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Caracciolo
authors.affiliation: NanoBioCel Group, University of Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain. Electronic address: joseluis.pedraz@ehu.es.
authors.forename: Jose-Luis
authors.initials: JL
authors.lastname: Pedraz
mesh_terms: D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage; D004705:Endocytosis; D004848:Epithelium / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes; D010957:Plasmids; D012160:Retina / Q000378:metabolism*
chemical_list: D008055:Lipids; D008081:Liposomes; D004247:DNA
keywords: Cationic lipid; Helper lipid; Intracellular trafficking; Niosomes; Non-viral vector; Transfection