Ethylcellulose nanoparticles as a new 'in vitro' transfection tool for antisense oligonucleotide delivery
| Autor: | S. Leitner, Conxita Solans, Ramon Eritja, María José García-Celma, Gabriela Calderó, Santiago Grijalvo |
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| Přispěvatelé: | Ministerio de Economía y Competitividad (España), Eritja Casadellà, Ramón, Eritja Casadellà, Ramón [0000-0001-5383-9334] |
| Rok vydání: | 2020 |
| Předmět: |
Renilla
Polymers and Plastics Static Electricity Oligonucleotides 02 engineering and technology “in vitro” transfection 010402 general chemistry 01 natural sciences HeLa chemistry.chemical_compound Nanoparticle Materials Chemistry Zeta potential Animals Humans Nanotechnology Gene Silencing RNA Messenger Cellulose Luciferases HEPES Antisense oligonucleotides Aqueous solution biology Nanopartícules Chemistry Nanotecnologia Organic Chemistry Cationic polymerization Gene Transfer Techniques Oligonucleòtids Serum Albumin Bovine Transfection Buffer solution Oligonucleotides Antisense 021001 nanoscience & nanotechnology biology.organism_classification 0104 chemical sciences Nano-emulsion Ethylcellulose Nanoparticles Cattle 0210 nano-technology Fetal bovine serum Nuclear chemistry HeLa Cells |
| Zdroj: | Dipòsit Digital de la UB Universidad de Barcelona Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Oil-in-water nano-emulsions have been obtained in the HEPES 20 mM buffer solution / [Alkylamidoammonium:Kolliphor EL = 1:1] / [6 weight% ethylcellulose in ethyl acetate] system over a wide oil-to-surfactant range and above 35 weight% aqueous component at 25 °C. The nano-emulsion with an oil-to-surfactant ratio of 70/30 and 95 weight% aqueous component was used for nanoparticles preparation. These nanoparticles (mean diameter around 90 nm and zeta potential of +22 mV) were non-toxic to HeLa cells up to a concentration of 3 mM of cationic species. Successful complexation with an antisense phosphorothioate oligonucleotide targeting Renilla luciferase mRNA was achieved at cationic/anionic charge ratios above 16, as confirmed by zeta potential measurements and an electrophoretic mobility shift assay, provided that no Fetal Bovine Serum is present in the cell culture medium. Importantly, Renilla luciferase gene inhibition shows an optimum efficiency (40%) for the cationic/anionic ratio 28, which makes these complexes promising for “in vitro” cell transfection. The DLS, zeta potential and Turbiscan analyses have been performed at the Nanostructured Liquid Characterization Unit, located at the Institute of Advanced Chemistry of Catalonia (IQAC), belonging to the Spanish National Research Council (CSIC) and affiliated to the NANBIOSIS ICTS of the Biomedical Networking Center (CIBER-BBN). CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011. Financial support from MINECO (grant CTQ2017-84998-P (AEI/FEDER,UE) and Generalitat de Catalunya (grant 2009SGR-961) is acknowledged. S.L. is grateful to CIBER-BBN for a research scholarship. Colorcon and Bonderalia S.A./Quimivita S.A. and BASF are gratefully acknowledged for the gift of ethylcellulose, ricinoleamidopropyltrimonium methosulfate and Kolliphor EL respectively. |
| Databáze: | OpenAIRE |
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