[en] The aim of this study was to improve the colloidal stability, decrease unspecific interactions with cells and blood components of a novel gene delivery system composed of epsilon-caprolactone and quaternized epsilon-caprolactone. For this purpose, diblock 50/50 copolymer was used to generate complexes-with DNA by either the solvent evaporation technique and by dialysis. The size, surface charge and degree of interaction of the plasmid-loaded formulations were measured. Then, polyplexes were combined with a poly(CL)-b-PEG copolymer to create a hydrophilic corona on the surface of the complexes. The cytotoxicity, transfection efficiency and cellular uptake of polyplexes and their association with PEG were evaluated on HeLa cells. The dialysis method did not allow to reduce the size of complexes as compared to the solvent evaporation method. The zeta potential of polyplexes became positive from a charge ratio of 4. The degree of interaction of copolymer with plasmid DNA was very high. Cytotoxicity and transfection efficiency were found to be comparable to polyethylenimine 50 kDa. Association of polyplexes with poly(CL)-b-PEG copolymer led to a small increase in particle size and a sharp decrease of charge surface. Cytotoxicity, transfection efficiency and cellular uptake were significantly reduced relative to unshielded copolymer/DNA complexes. The PEGylated formulations may be an attractive approach for an in vivo application.
Research center :
Center for Education and Research on Macromolecules (CERM)
Boussif O., Lezoualch F., Zanta M.A., Mergny M.D., Scherman D., Demeneix B., and Behr J.P. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc. Natl. Acad. Sci. U.S.A. 92 (1995) 7297-7301
De Smedt S.C., Demeester J., and Hennink W.E. Cationic polymer based gene delivery systems. Pharm. Res. 17 (2000) 113-126
De Wolf H.K., Luten J., Snel C.J., Oussoren C., Hennink W.E., and Storm G. In vivo tumor transfection mediated by polyplexes based on biodegradable poly(DMAEA)-phosphazene. J. Contr. Release 109 (2005) 275-287
Deshpande M.C., Davies M.C., Garnett M.C., Williams P.M., Armitage D., Bailey L., Vamvakaki M., Armes S.P., and Stolnik S. The effect of poly(ethylene glycol) molecular architecture on cellular interaction and uptake of DNA complexes. J. Contr. Release 97 (2004) 143-156
Detrembleur C., Mazza M., Halleux O., Lecomte Ph., Mecerreyes D., Hedrick J.L., and Jérôme R. Ring-opening polymerization of γ-bromo-ε-caprolactone: a novel route to functionalized aliphatic polyesters. Macromolecules 33 (2000) 14-18
Detrembleur C., Mazza M., Lou X., Halleux O., Lecomte Ph., Mecerreyes D., Hedrick J.L., and Jérôme R. New functional aliphatic polyesters by chemical modification of copolymers of ε-caprolactone with γ-(2-bromo-2-methylpropionate)-ε-caprolactone, γ-bromo-ε-caprolactone and a mixture of β and γ-ene-ε-caprolactone. Macromolecules 33 (2000) 7751-7760
Fischer D., Bieber T., Li Y., Elsasser H.P., and Kissel T. A novel non-viral vector for DNA delivery based on low molecular weight, branched polyethylenimine: effect of molecular weight on transfection efficiency and cytotoxicity. Pharm. Res. 16 (1999) 1273-1279
Funhoff A.M., Monge S., Teeuwen R., Koning G.A., Schuurmans-Nieuwenbroek N.M., Crommelin D.J., Haddleton D.M., Hennink W.E., and van Nostrum C.F. PEG shielded polymeric double-layered micelles for gene delivery. J. Contr. Release 102 (2005) 711-724
Garinot M., Fievez V., Pourcelle V., Stoffelbach F., des Rieux A., Theate I., Freichels H., Jérôme C., Marchand-Brynaert J., Schneider Y.-J., and Préat V. PEGylated PLGA-based nanoparticles targeting M cells for oral vaccination. J. Contr. Release 120 (2007) 195-204
Gaucher G., Dufresne M.-H., Sant V.P., Kang N., Maysinger D., and Leroux J.-C. Block copolymer micelles: preparation, characterization and application in drug delivery. J. Contr. Release 109 (2005) 169-188
Hans M.L., and Lowman A.M. Biodegradable nanoparticles for drug delivery and targeting. Curr. Opin. Solid State Mater. Sci. 6 (2002) 319-327
Jeong J.H., Song S.H., Lim D.W., Lee H., and Park T.G. DNA transfection using linear poly(ethylenimine) prepared by controlled acid hydrolysis of poly(2-ethyl-2-oxazoline). J. Contr. Release 73 (2001) 391-399
Jeong J.H., and Park T.G. Poly(l-lysine)-g-poly(d,l-lactic-co-glycolic acid) micelles for low cytotoxic biodegradable gene delivery carriers. J. Contr. Release 82 (2002) 159-166
Kichler A. Gene transfer with modified polyethylenimines. J. Gene Med. 6 (2004) S3-S10
Kursa M., Walker G.F., Roessler V., Ogris M., Roedl W., Kircheis R., and Wagner E. Novel shielded transferrin-polyethylene glycol-polyethylenimine/DNA complexes for systemic tumor-targeted gene transfer. Bioconjug. Chem. 14 (2003) 222-231
Lam J.K.W., Ma Y., Armes S.P., Lewis A.L., Baldwin T., and Stolnik S. Phosphorylcholine-polycation diblock copolymers as synthetic vectors for gene delivery. J. Contr. Release 100 (2004) 293-312
Lee H., Jeong J.H., and Park T.G. A new gene delivery formulation of polyethylenimine/DNA complexes coated with PEG conjugated fusogenic peptide. J. Contr. Release 76 (2001) 183-192
Lee M., and Kim S.W. Polyethylene glycol-conjugated copolymers for plasmid DNA delivery. Pharm. Res. 22 (2005) 1-10
Merdan T., Callahan J., Petersen H., Kunath K., Bakowsky U., Kopeckova P., Kissel T., and Kopecek J. Pegylated polyethylenimine-Fab' antibody fragment conjugates for targeted gene delivery to human ovarian carcinoma cells. Bioconjug. Chem. 14 (2003) 989-996
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65 (1983) 55-63
Nimesh S., Goyal A., Pawar V., Jayaraman S., Kumar P., Chandra R., Singh Y., and Gupta K.C. Polyethylenimine nanoparticles as efficient transfecting agents for mammalian cells. J. Contr. Release 110 (2006) 457-468
Perez C., Sanchez A., Putnam D., Ting D., Langer R., and Alonso M.J. Poly(lactic acid)-poly(ethylene glycol) nanoparticles as new carriers for the delivery of plasmid DNA. J. Contr. Release 75 (2001) 211-224
Soppimath K.S., Aminabhavi T.M., Kulkarni A.R., and Rudzinski W.E. Biodegradable polymeric nanoparticles as drug delivery devices. J. Contr. Release 70 (2001) 1-20
Vangeyte P., and Jérôme R. Amphiphilic block copolymers of high-molecular-weight poly(ethylene oxide) and either ε-caprolactone or γ-methyl-ε-caprolactone: synthesis and characterization. J. Polym. Sci. Part A: Polym. Chem. 42 (2004) 1132-1142
Vroman B., Mazza M., Fernandez M.R., Jérôme R., and Préat V. Copolymers of ε-caprolactone and quaternized ε-caprolactone as gene carriers. J. Contr. Release 118 (2007) 136-144
Zaric V., Weltin D., Erbacher P., Remy J.-S., Behr J.-P., and Stephan D. Effective polyethylenimine-mediated gene transfer into human endothelial cells. J. Gene Med. 6 (2004) 176-184
