title: Cationic liposomes produced via ethanol injection method for dendritic cell therapy.
abstract: Cationic liposomes can be designed and developed in order to be an efficient gene delivery system for mammalian cells. Dendritic cell (DC) vaccines can be used to treat cancer, as cationic liposomes can deliver tumor antigens to cells while cells remain active. However, most methods used for liposome production are not able to reproduce in large scale the physicochemical and biological properties of liposomes produced in laboratory scale. In this context, ethanol injection method achieved promising results, although requiring post-treatment for size reduction and/or to remove residual ethanol. Thus, the purpose of this study was to generate cationic liposomes suitable for gene therapies via ethanol injection method in only one step (VEI) and compared to those submitted to a size reduction processes by microfluidization (MFV). For this, the method to produce cationic liposomes composed of egg phosphatidylcholine (EPC), 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and 1,2-dioleoylphosphatidylethanolamine (DOPE) was optimized using a statistical design approach. As a result, the size of VEI decreased from 290 nm to 110 nm and the polydispersity from 0.54 to 0.17. In the case of MFV, size decreased from 128 nm to 107 nm and polydispersity from 0.40 to 0.18. ST and MFV before and after optimization were also characterized in terms of morphology by transmission electron microscopy (TEM) and structure by differential scanning calorimetry (DSC). Finally, to show their potential in gene/immune therapies applications, DCs were stimulated by such liposomes. Cells internalized liposomes, increasing expression of the costimulatory molecule CD86 and inducing T lymphocyte proliferation.
authors.affiliation: a Department of Materials and Bioprocesses Engineering , School of Chemical Engineering, University of Campinas (Unicamp) , Campinas , Brazil and.
authors.forename: Micaela Tamara
authors.initials: MT
authors.lastname: Vitor
authors.affiliation: b Department of Immunology , Institute of Biomedical Sciences, University of São Paulo (USP) , São Paulo , Brazil.
authors.forename: Patrícia Cruz
authors.initials: PC
authors.lastname: Bergami-Santos
authors.affiliation: a Department of Materials and Bioprocesses Engineering , School of Chemical Engineering, University of Campinas (Unicamp) , Campinas , Brazil and.
authors.forename: Rafael Henrique Freitas
authors.initials: RHF
authors.lastname: Zômpero
authors.affiliation: b Department of Immunology , Institute of Biomedical Sciences, University of São Paulo (USP) , São Paulo , Brazil.
authors.forename: Karen Steponavicius Piedade
authors.initials: KSP
authors.lastname: Cruz
authors.affiliation: b Department of Immunology , Institute of Biomedical Sciences, University of São Paulo (USP) , São Paulo , Brazil.
authors.forename: Mariana Pereira
authors.initials: MP
authors.lastname: Pinho
authors.affiliation: b Department of Immunology , Institute of Biomedical Sciences, University of São Paulo (USP) , São Paulo , Brazil.
authors.forename: José Alexandre Marzagão
authors.initials: JAM
authors.lastname: Barbuto
authors.affiliation: a Department of Materials and Bioprocesses Engineering , School of Chemical Engineering, University of Campinas (Unicamp) , Campinas , Brazil and.
authors.forename: Lucimara Gaziola
authors.initials: LG
authors.lastname: de la Torre
mesh_terms: D000818:Animals; D051940:B7-2 Antigen / Q000378:metabolism; D002412:Cations; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000276:immunology; D000431:Ethanol / Q000737:chemistry*; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D007167:Immunotherapy; D008081:Liposomes / Q000737:chemistry*; D010316:Particle Size; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013499:Surface Properties; D013601:T-Lymphocytes / Q000166:cytology
chemical_list: D051940:B7-2 Antigen; D002412:Cations; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C094877:dioleoyl phosphatidylethanolamine; D000431:Ethanol; C020888:1,2-dielaidoylphosphatidylethanolamine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Cationic liposomes; dendritic cells; ethanol injection method; experimental design; immunotherapy