Abstract

Targeted gene delivery is important in biomedical research and applications. In this paper, we synergistically combine non-viral chemical materials, magnetic nanoparticles (MNPs), and a physical technique, low-intensity pulsed ultrasound (LIPUS), to achieve efficient and targeted gene delivery. The MNPs are iron oxide super-paramagnetic nanoparticles, coated with polyethyleneimine (PEI), which makes a high positive surface charge and is favorable for the binding of genetic materials. Due to the paramagnetic properties of the MNPs, the application of an external magnetic field increases transfection efficiency while LIPUS stimulation enhances cell viability and permeability. We found that stimulation at the intensity of 30 mW/cm2 for 10 minutes yields optimal results with a minimal adverse effect on the cells. By combining the effect of the external magnetic field and LIPUS, the genetic material (GFP or Cherry Red plasmid) can enter the cells. The flow cytometry results showed that by using just a magnetic field to direct the genetic material, the transfection efficiency on HEK 293 cells that were treated by our MNPs was 56.1%. Coupled with LIPUS stimulation, it increased to 61.5% or 19% higher than the positive control (Lipofectamine 2000). Besides, compared with the positive control, our method showed less toxicity. Cell viability after transfection was 63.61%, which is 19% higher than the standard transfection technique. In conclusion, we designed a new gene-delivery method that is affordable, targeted, shows low-toxicity, yet high transfection efficiency, compared to other conventional approaches.

Highlights

  • Gene delivery is a popular research area with high demand on the market, and applications in both clinical and scientific biomedical research [1, 2]

  • From the previous work done in our group as well as by other groups and reported in the literature, we knew that low-intensity pulsed ultrasound (LIPUS) could transiently increase cell membrane permeability [31] as well as can be beneficial for cell viability [35, 43]

  • We combined the application of an external magnetic field with magnetic nanoparticles (MNPs) and LIPUS stimulation for gene delivery

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Summary

Introduction

Gene delivery is a popular research area with high demand on the market, and applications in both clinical and scientific biomedical research [1, 2]. The applications include, but are not limited to, treating cancers, immune-deficient diseases, and genetic diseases [3]. Mammalian cells have a selectively permeable plasma membrane that protects them from the external environment. Effective methods to transfect cells are needed.

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