title: Novel non-viral gene delivery systems composed of carbosilane dendron functionalized nanoparticles prepared from nano-emulsions as non-viral carriers for antisense oligonucleotides.
abstract: The development of novel and efficient delivery systems is often the limiting step in fields such as antisense therapies. In this context, poly(d,l-lactide-co-glycolide) acid (PLGA) nanoparticles have been obtained by a versatile and simple technology based on nano-emulsion templating and low-energy emulsification methods, performed in mild conditions, providing good size control. O/W polymeric nano-emulsions were prepared by the phase inversion composition method at 25°C using the aqueous solution/polysorbate80/[4 wt% PLGA in ethyl acetate] system. Nano-emulsions formed at oil-to-surfactant (O/S) ratios between 10/90-90/10 and aqueous contents above 70 wt%. Nano-emulsion with 90 wt% of aqueous solution and O/S ratio of 70/30 was chosen for further studies, since they showed the appropriate characteristics to be used as nanoparticle template: hydrodynamic radii lower than 50 nm and enough kinetic stability. Nanoparticles, prepared from nano-emulsions by solvent evaporation, showed spherical shape, sizes about 40 nm, negative surface charges and high stability. The as-prepared nanoparticles were functionalized with carbosilane cationic dendrons through a carbodiimide-mediated reaction achieving positively charged surfaces. Antisense oligonucleotides were electrostatically attached to nanoparticles surface to perform gene-silencing studies. These complexes were non-haemolytic and non-cytotoxic at the concentrations required. The ability of the complexes to impart cellular uptake was also promising. Therefore, these novel nanoparticulate complexes might be considered as potential non-viral carriers in antisense therapy.
authors.affiliation: Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), C/Jordi Girona, 18-26, 08034, Barcelona, Spain; CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Fornaguera
authors.affiliation: Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), C/Jordi Girona, 18-26, 08034, Barcelona, Spain; CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Grijalvo
authors.affiliation: CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain; Group of Dendrimers for Biomedical Applications, University of Alcalá (GDAB-UAH), Alcalá de Henares, Madrid, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Galán
authors.affiliation: CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain; Group of Dendrimers for Biomedical Applications, University of Alcalá (GDAB-UAH), Alcalá de Henares, Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Fuentes-Paniagua
authors.affiliation: CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain; Group of Dendrimers for Biomedical Applications, University of Alcalá (GDAB-UAH), Alcalá de Henares, Madrid, Spain.
authors.forename: Francisco Javier
authors.initials: FJ
authors.lastname: de la Mata
authors.affiliation: CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain; Group of Dendrimers for Biomedical Applications, University of Alcalá (GDAB-UAH), Alcalá de Henares, Madrid, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Gómez
authors.affiliation: Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), C/Jordi Girona, 18-26, 08034, Barcelona, Spain; CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain.
authors.forename: Ramon
authors.initials: R
authors.lastname: Eritja
authors.affiliation: Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), C/Jordi Girona, 18-26, 08034, Barcelona, Spain; CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Calderó
authors.affiliation: Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), C/Jordi Girona, 18-26, 08034, Barcelona, Spain; CIBER-BBN Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain. Electronic address: conxita.solans@iqac.csic.es.
authors.forename: Conxita
authors.initials: C
authors.lastname: Solans
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D050091:Dendrimers / Q000008:administration & dosage / Q000737:chemistry; D004655:Emulsions; D004912:Erythrocytes / Q000187:drug effects / Q000473:pathology; D018014:Gene Transfer Techniques; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry; D049410:Luciferases, Renilla / Q000235:genetics; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000737:chemistry; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012821:Silanes / Q000737:chemistry; D055672:Static Electricity
chemical_list: D050091:Dendrimers; D004655:Emulsions; D016376:Oligonucleotides, Antisense; D012821:Silanes; C504072:carbosilane; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D049410:Luciferases, Renilla
keywords: Antisense therapy; Cationic dendron; Nano-emulsion; Non-viral gene delivery systems; PIC emulsification method; PLGA nanoparticles