Evaluation of siRNA and cationic liposomes complexes as a model for in vitro siRNA delivery to cancer cells

Author(s)

Es, Ismail ^[1] ; Ok, Meryem Tyrrasch ^[2] ; Puentes-Martinez, Ximena E. ^[3] ; Szymanski de Toledo, Marcelo Augusto ^[4] ; de Pinho Favaro, Marianna Teixeira ^[4] ; Cavalcanti, Leide Passos ^[1] ; Cassago, Alexandre ^[3] ; Portugal, Rodrigo Villares ^[3] ; Azzoni, Adriano Rodrigues ^[5] ; de la Torre, Lucimara Gaziola ^[1]

Affiliation

^[1] Univ Estadual Campinas, Sch Chem Engn, BR-13083852 Campinas, SP - Brazil

^[2] MIT, Dept Biol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 - USA

^[3] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Nanotechnol Natl Lab LNNano, Campinas, SP - Brazil

^[4] Univ Estadual Campinas, Mol Biol & Genet Engn Ctr, Lab Genet & Mol Anal, Campinas, SP - Brazil

^[5] Univ Sao Paulo, Chem Engn Dept, Polytech Sch, Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS; v. 555, p. 280-289, OCT 20 2018.
• Web of Science Citations: 1

Abstract

Controlled release of genetic material such as small interfering RNA (siRNA) using lipid-based non-viral vectors has gained substantial importance in gene therapy applications. Therefore, the interaction between siRNA and these vectors must be well understood. This study aims to investigate the effect of different molar charge ratios (R+/-) between positive charges from microfluidics-produced cationic liposomes (CL) (egg phosphatidylcholine, 1,2-dioleoyl-3-trimethylammonium-propane and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and negative charges from siRNA and on physico-chemical and morphological properties of the lipoplexes (CL/siRNA) as well as their in vitro luciferase silencing effect in HeLa cells. R+/- 3.27 was found to be the optimum point for complexation. This finding was confirmed by gel retardation and siRNA accessibility assays. According to Cryo-TEM analysis, the lipoplexes had multi-lamellarity. In vitro transfection efficiency of lipoplexes in HeLa cells was tested at three different siRNA concentrations (10, 25, and 35 nM). Significant knockdown of luciferase by siRNA lipoplexes was observed based on reduced luciferase activity of transfected HeLa cells. Our findings were comparable with the silencing effect of siRNA complexed with Lipofecamine (R). No cytotoxicity of lipoplexes was detected at the tested concentrations. This study was essential for further complexation studies which will be performed using microfluidic systems to formulate next-generation lipid-based controlled release systems. (AU)

• FAPESP's process: 15/14468-0 - MICROFLUIDIC SYSTEMS FOR INCORPORATION OF SMALL INTERFERING RNA (siRNA) IN CATIONIC LIPOSOMES AND FOR IN VITRO ANIMAL CELL TRANSFECTION TARGETING GENE THERAPY
• Grantee: Ismail Es
• Support Opportunities: Scholarships in Brazil - Doctorate