Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane

Soo-Yong Park; Yang H Yun; Bum-Joon Park; Hyung-Il Seo; Ildoo Chung

doi:10.3390/ph15010017

Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane

Pharmaceuticals (Basel). 2021 Dec 24;15(1):17. doi: 10.3390/ph15010017.

Authors

Soo-Yong Park¹, Yang H Yun², Bum-Joon Park³, Hyung-Il Seo⁴, Ildoo Chung¹

Affiliations

¹ Department of Polymer Science and Engineering, Pusan National University, Busan 46241, Korea.
² Department of Biomedical Engineering, College of Engineering, The University of Akron, Akron, OH 44325, USA.
³ Department of Molecular Biology, College of Natural Science, Pusan National University, Busan 46241, Korea.
⁴ Department of Surgery, Biomedical Research Institute, Pusan National University Hospital, Busan 49241, Korea.

Abstract

Gene therapy is a suitable alternative to chemotherapy due to the complications of drug resistance and toxicity of drugs, and is also known to reduce the occurrence of cellular mutation through the use of gene carriers. In this study, gene carrier nanoparticles with minimal toxicity and high transfection efficiency were fabricated from a biocompatible and biodegradable polymer, l-tyrosine polyurethane (LTU), which was polymerized from presynthesized desaminotyrosyl tyrosine hexyl ester (DTH) and polyethylene glycol (PEG), by using double emulsion and solvent evaporation techniques, resulting in the formation of porous nanoparticles, and then used to evaluate their potential biological activities through molecular controlled release and transfection studies. To assess cellular uptake and transfection efficiency, two model drugs, fluorescently labeled bovine serum albumin (FITC-BSA) and plasmid DNA-linear polyethylenimine (LPEI) complex, were successfully encapsulated in nanoparticles, and their transfection properties and cytotoxicities were evaluated in LX2 as a normal cell and in HepG2 and MCF7 as cancer cells. The morphology and average diameter of the LTU nanoparticles were confirmed using light microscopy, transmission electron microscopy, and dynamic light scattering, while confocal microscopy was used to validate the cellular uptake of FITC-BSA-encapsulated LTU nanoparticles. Moreover, the successful cellular uptake of LTU nanoparticles encapsulated with pDNA-LPEI and the high transfection efficiency, confirmed by gel electrophoresis and X-gal assay transfection, indicated that LTU nanoparticles had excellent cell adsorption ability, facilitated gene encapsulation, and showed the sustained release tendency of genes through transfection experiments, with an optimal concentration ratio of pDNA and LPEI of 1:10. All the above characteristics are ideal for gene carriers designed to transport and release drugs into the cytoplasm, thus facilitating effective gene therapy.

Keywords: DNA-LPEI complex; LTU; biodegradable; double emulsion; gene carrier; transfection.

Grants and funding

2017R1D1A1B04032465/National Research Foundation of Korea