e-Posters - Nano Drug Delivery 2018
Fatemeh Radmanesh
Shahid Beheshti University
Optimizing trnsfection of primary human umbilical vein endothelial cells using facial
amphipathic deoxycholic acid-conjugated polyethyleneimine
Fatemeh Radmanesh (Biography)
Fatemeh Radmanesh (Abstract)
Introduction: Currently, RNA interference (RNAi)-\r\nbased gene therapy has been investigated for treating\r\nvarious disease conditions. However, successful\r\napplication of RNAi including siRNA or miRNA has\r\nbeen limited by several factors in vitro and in vivo. To\r\novercome these challenges, various non-viral carriers\r\nhave been developing for efficient RNAi-mediated\r\ngene silencing. However, it is necessary to improve\r\nthe efficacy of these non-viral strategies to achieve\r\ndesired therapeutic effect. In this study, we carried out\r\nsynthesis, characterization, and optimization of a\r\npolymeric conjugate based on low molecular weight\r\npolyethylenimine which was modified by high\r\nmembrane permeable deoxycholic acid (PEI-DA) for\r\nefficient delivery of RNAi-based therapeutics into\r\nprimary human umbilical vein endothelial cells\r\n(HUVECs)\r\nMethods: Herein, DA-PEI conjugate was\r\nsynthesized based on DCC/NHS chemistry at various\r\nDA to PEI molar ratios of 2 to 4 and used for delivery\r\nof a fluorescent labeled siRNA into HUVECs.\r\nConjugates were characterized for chemical structure,\r\nsize, and cell cytotoxicity. The effect of various\r\nparameters including DA/PEI molar ratio,\r\npolymer/siRNA weight ratio, and different buffer\r\nsolutions was investigated on transfection efficacy of\r\nconjugates. Results: DA was conjugated to the\r\nterminal amine groups of the PEI1.8 via amide bonds.\r\nThe polyplexes had smaller sizes (about 130~150 nm)\r\nthan the parent PEI1.8 at different weight ratios. MTS\r\nassay revealed that the conjugates were non-toxic at\r\npolymer concentrations used in transfection\r\nexperiments. The higher intracellular uptake and\r\ntransfection efficiency were achieved by the\r\nconjugates synthesized in DA/PEI molar ratio of 3 or 4,\r\npolymer/siRNA weight ratio of 5 when they were\r\nprepared in salty buffers. Conclusion: These results\r\nsuggest that the DA-PEI1.8 conjugate can be applied\r\nas a promising candidate to enhance delivery of RNAi\r\ntherapeutics into primary endothelial cells under the\r\noptimized transfection conditions.\r\n