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

Park, SY; Yun, YH; Park, BJ; Seo, HI; Chung, I

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

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DNA gene carrier nanoparticles represent a revolutionary breakthrough in the realm of biotechnology and medicine. These minuscule particles, typically composed of biocompatible materials such as lipids, polymers, or inorganic substances, serve as highly efficient vehicles for the delivery of therapeutic genes into target cells. Their small size and customizable surface properties enable precise targeting of specific tissues or cell types, offering immense potential for personalized medicine. By encapsulating DNA or RNA molecules encoding therapeutic proteins or regulatory factors, gene carrier nanoparticles can modulate cellular functions, correct genetic defects, or induce desired biological responses. Moreover, their ability to protect genetic cargo from degradation and deliver it directly into the cell cytoplasm or nucleus enhances the efficiency and specificity of gene therapy approaches. This technology holds promise for treating a wide range of diseases, including genetic disorders, cancer, infectious diseases, and neurological disorders. However, challenges such as off-target effects, immune responses, and scalability need to be addressed to fully harness the therapeutic potential of DNA gene carrier nanoparticles. As research continues to advance, these nanoparticles are poised to revolutionize the landscape of medicine, offering new avenues for targeted therapy, disease prevention, and personalized healthcare.