Abstract format for biomechanica iv

European Cells and Materials Vol. 20. Suppl. 3, 2010 (page 124) ISSN 1473-2262 Self-Assembled Nanostructures from Organic Bolaamphiphiles for
Gene Delivery
N. Jain, Y. Arntz, V. Goldschmidt, G. Duportail, Y. Mely, Andrey S. Klymchenko LBP, UMR 7213 CNRS, Faculté de Pharmacie, Université de Strasbourg, France helping agents. Finally, all bolas showed low INTRODUCTION: The success in gene therapy
relies strongly on new efficient gene delivery vectors. Nonviral vectors based on lipids and polymers constitute an important alternative to the viral vectors. However, the key problem with these vectors is poor structural control of their DNA complexes. (bolas) bearing positively charged and neutral head groups could be an attractive alternative. These molecules can generate asymmetric membranes (in form of vesicles or nanotubes) having positively charged inner and neutral outer surfaces, where the inner membrane surface can be used to wrap the DNA molecule. The current work presents new bolaamphiphiles, characterization of their assembly with DNA and application for gene delivery. METHODS: The design methodology is based on
Fig. 1: Schematic presentation of bolaamphiphile bola molecules bearing neutral sugar (gluconic or and its assembly with DNA (A). Gel lactonic) and ornithine di-cationic residues as head electrophoresis (B) and AFM images (C) of bola- groups, connected by a long hydrophobic spacer (Fig. 1). Gel Electrophoresis in 0.9% agarose was used to study interaction of bolas with DNA. DNA DISCUSSION & CONCLUSIONS: Obtained data
condensation within the bola-DNA complexes suggests that the size and shape of the bolaplexes (bolaplexes) was followed by ethidium bromide depends on bola structure and the formulation used. The increase in size at higher N/P ratio is probably related to (EtBr) exclusion assay. Size and charge of the charge neutralization which is in line with zeta potential bolaplexes at different N/P ratios was examined by analysis. Increase in transfection efficiency with the use Dynamic Light Scattering (DLS) and Zetasizer of DOPE or chloroquine suggests that the key barrier respectively. Atomic Force Microscopy (AFM) in for their internalization can be endosomal escape. Thus, liquid phase with tapping mode was performed to the new bola molecules show the potential for study the morphology of bolaplexes. Transfection construction of nonviral vectors featuring controlled efficiency of the bolaplexes was tested for different small size, high efficiency and low cytotoxicity. formulations by luciferase assay. DOPE, a helper lipid, or an endosmolytic reagent chloroquine was REFERENCES: 1Zuber G., Daut, E., Nothisen M.,
used in the formulations. Cytotoxicity of the bolas Belguise P. & Behr J.P. Adv Drug Deliv Rev 52, 245-
253 (2001).2Shimizu T., Masuda M. & Minamikawa H.
Chemical Reviews 105, 1401-1443 (2005).3Fuhrhop
RESULTS: New unsymmetrical bolaamphiphiles
J.H. & Tank H. Chemistry and Physics of Lipids 43,
were synthesized. Strong interaction of bolas with 193-213 (1987). 4Masuda M. & Shimizu T. Langmuir DNA was observed with particle size of ~200-300 20, 5969-5977 (2004).
nm. Moreover, an increase in size and surface ACKNOWLEDGEMENTS:
charge of bolaplexes was noticed at higher N/P financial support of FRM. We thank to the groups of J.- ratios. AFM studies reveals nano-structural rod S. Remy and B. Frisch for help with luminometry and shaped or spherical morphology of bolaplexes. Transfection efficiency of the bolaplexes improves considerably using DOPE or chloroquine, as

Source: http://www.ecmjournal.org/journal/supplements/vol020supp03/pdf/v020supp03a124.pdf

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