Synthetic poly(ester amine) and poly(amido amine) nanoparticles for efficient DNA and siRNA delivery to human endothelial cells

Abstract

Biodegradable poly(ester amine) (PEA)-based and poly(amido amine) (PAA)-based nanoparticles were developed for efficient in vitro siRNA delivery to human umbilical vein endothelial cells (HUVECs). They were screened, characterized, and compared with traditionally studied DNA-containing particles. Several of the polymeric nanoparticles tested were found to be effective for delivering functional siRNA to green fluorescent protein (GFP) + HUVECs, achieving 60%–75% GFP knockdown while maintaining high viability. While PEAs have been used previously to form polyplexes or nanoparticles for DNA delivery, highly effective siRNA delivery in hard-to-transfect human cell types has not been previously reported. PEAs and linear nondendrimeric PAAs were also found to be effective for DNA delivery to HUVECs using GFP-encoding plasmid DNA (up to 50%–60% transfection efficiency). PEAs and PAAs can be separated into groups that form polymeric nanoparticles effective for siRNA delivery, for DNA delivery, or for both.

Keywords:

• nanoparticles
• siRNA
• DNA
• gene delivery
• human umbilical vein endothelial cells

Supporting figure

Figure S1 GFP⁺ HUVECs were seeded in 96-well plates at known densities and allowed to settle and adhere for 1 hr before measuring fluorescence with a microplate reader. Linear regression showed a positive, linear correlation following the relation RFU = 33073*(#cells)+6599 with correlation coefficient r² = 0.985. The gray, shaded region shows the 95% confidence interval of the regression line.

Note: Error bars represent standard deviation of multiple replicates at each cell density.

Abbreviations: GFP, green fluorescent protein; HUVECs, human umbilical vein endothelial cells; RFU, relative fluorescence units.

Acknowledgments

This work was supported in part by the Johns Hopkins University Institute for NanoBioTechnology, Maryland Technology Development Corporation and Maryland Stem Cell Research Fund (2009-MSCRFE-0098-00), and the National Institutes of Health (R21CA152473). Preliminary studies were funded by the Provost’s Undergraduate Research Award at Johns Hopkins. SYT thanks the National Science Foundation for fellowship support. GFP-positive HUVECs were transduced by Brian Goh using a viral vector provided by Dr David Yue. The authors thank Joel C Sunshine for synthesizing some of the polymers used in this study, Ron B Shmueli for assistance with HUVEC culture and transfection, and Corey J Bishop and the Johns Hopkins School of Medicine Microscope Facility for assistance with TEM.

Disclosure

The authors report no conflicts of interest in this work.