Effect on in vitro cell response of the statistical insertion of N-(2-hydroxypropyl) methacrylamide on linear pro-dendronic polyamine's gene carriers.
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 300 nm. 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.
Institute of Polymer Science and Technology, ICTP-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain. Electronic address: jaredondo@ictp.csic.es.
Juan Alfonso
JA
Redondo
Institute of Biofunctional Studies (IEB), Tissue Engineering Group, (UCM), Associated Unit to the Institute of Polymer Science and Technology (CSIC), Paseo de Juan XXIII 1, 28040 Madrid, Spain.
Enrique
E
Martínez-Campos
Institute of Polymer Science and Technology, ICTP-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
Rodrigo
R
Navarro
Institute of Polymer Science and Technology, ICTP-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
Helmut
H
Reinecke
Institute of Polymer Science and Technology, ICTP-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
Carlos
C
Elvira
Institute of Biofunctional Studies (IEB), Tissue Engineering Group, (UCM), Associated Unit to the Institute of Polymer Science and Technology (CSIC), Paseo de Juan XXIII 1, 28040 Madrid, Spain.
José Luis
JL
López-Lacomba
Institute of Polymer Science and Technology, ICTP-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
Alberto
A
Gallardo
D000178:Acrylamides / Q000138:chemical synthesis* / Q000633:toxicity; D000818:Animals; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D050091:Dendrimers / Q000138:chemical synthesis* / Q000633:toxicity; D004587:Electrophoresis, Agar Gel; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008027:Light; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D015233:Models, Statistical; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D010316:Particle Size; D011094:Polyethyleneimine / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000633:toxicity; D012542:Scattering, Radiation; D041701:Swiss 3T3 Cells; D014162:Transfection / Q000379:methods*
D000178:Acrylamides; D050091:Dendrimers; D011094:Polyethyleneimine; D004247:DNA; D008156:Luciferases; C032976:N-(2-hydroxypropyl)methacrylamide
Cationic polymers; Cytocompatibility; Dendronic; HPMA; Statistical copolymers; Transfection