Effect on in vitro cell response of the statistical insertion of N-(2-hydroxypropyl) methacrylamide on linear pro-dendronic polyamine’s gene carriers

• Published in: European journal of pharmaceutics and biopharmaceutics 2015-06, Vol.93, p.303-310
• Main Authors: Redondo, Juan Alfonso, Martínez-Campos, Enrique, Navarro, Rodrigo, Reinecke, Helmut, Elvira, Carlos, López-Lacomba, José Luis, Gallardo, Alberto
• Format: Article
• Language: English
• Subjects: Acrylamides - chemical synthesis; Acrylamides - toxicity; Animals; Cationic polymers; Cytocompatibility; Dendrimers - chemical synthesis; Dendrimers - toxicity; Dendronic; DNA - biosynthesis; DNA - chemistry; Electrophoresis, Agar Gel; Fibroblasts - drug effects; Fibroblasts - metabolism; Gene Expression Regulation; Genes, Reporter; HPMA; Light; Luciferases - biosynthesis; Luciferases - genetics; Mice; Models, Statistical; Molecular Structure; Nucleic Acid Conformation; Particle Size; Polyethyleneimine - analogs & derivatives; Polyethyleneimine - chemical synthesis; Polyethyleneimine - toxicity; Scattering, Radiation; Statistical copolymers; Swiss 3T3 Cells; Transfection; Transfection - methods
• ISSN: 0939-6411; 1873-3441
• DOI: 10.1016/j.ejpb.2015.04.014

[Display omitted] Statistical copolymers of N-(2-hydroxypropyl) methacrylamide (HPMA) and the dendronic methacrylic monomer 2-(3-(Bis(2-(diethylamino)ethyl)amino)propanamido)ethyl methacrylate (TEDETAMA, derived from N,N,N′,N′-tetraethyldiethylenetriamine, TEDETA), were synthesized through radical copolymerization and evaluated in vitro as non-viral gene carriers. Three copolymers with nominal molar percentages of HPMA of 25%, 50% and 75% were prepared and studied comparatively to the positive controls poly-TEDETAMA and hyperbranched polyethyleneimine (PEI, 25kDa). Their ability to complex DNA at different N/P molar ratios, from 1/1 up to 8/1, was determined through agarose gel electrophoresis and Dynamic Light Scattering. The resulting complexes (polyplexes) were characterized and evaluated in vitro as possible non-viral gene carriers for Swiss-3T3 fibroblasts, using luciferase as reporter gene and a calcein cytocompatibility assay. All the copolymers, except the one with highest HPMA proportion (75 molar %) at the lowest N/P ratio, condensed DNA to a particle size between 100 and 300nm. The copolymers with 25 and 50 molar % of HPMA displayed higher transfection efficiency and cytocompatibility than the positive controls poly-TEDETAMA and PEI. A higher proportion of HPMA (75 molar %) led to copolymers that displayed very low transfection efficiency, despite their full cytocompatibility even at the highest N/P ratio. These results indicate that the statistical combination of TEDETAMA and HPMA and its fine compositional tuning in the copolymers may fulfill the fine balance of transfection efficiency and cytocompatibility in a superior way to the control poly-TEDETAMA and PEI.