Introducing Electrospray As a Potent Technique to Deliver Chitosan/Pdna Nanoparticles to Eukaryotic Cells Publisher



Landi FS¹ ; Negahdari B² ; Esmaeili F¹ ; Kolivand S² ; Amani A^{3, 4} Show Affiliations
Source: Journal of Nano Research Published:2021

Abstract

Electrospray technique has received increasing attention for intracellular gene delivery as well as the production of nanoparticles. In this study, chitosan-pDNA nanoparticles with an N/P ratio of 5 were prepared and transferred to HEK293T cells by electrospray. Physicochemical characterization of prepared nanoparticles, including size, zeta potential, and entrapment efficiency was performed. Gel electrophoresis was employed to confirm pDNA attachment to chitosan. Moreover, transfection efficiency was investigated using flow cytometry. MTT assay was performed for cell viability studies. Nanoparticles were prepared at three pDNA concentrations of 10, 55, and 100 μg/ml at a fixed N/P ratio. The size of nanoparticles was obtained as 110, 188, and 240 nm, using DLS. SEM showed a size of 102.3 ± 10.7 nm for samples having 55 μg/ml of pDNA. Zeta potential and entrapment efficiency were +25 mV and 85±4%, respectively. The effect of pDNA concentration, electrospray time, and incubation time on transfection efficiency was investigated using Box- Behnken design. Percent of GFP-positive cells was 41.05 ± 3.04% which was taken as an indicator of transfection efficiency. The transfection efficiency of this method was then compared with that of calcium phosphate (31.1 ± 2.4%), showing improved efficiency. Because electrospray is an easy, low cost, one-step process which makes low damage to cells and produces monodispersed nanoparticles, the method is introduced as a fascinating approach in gene transfection. © 2021 Trans Tech Publications Ltd, Switzerland.