Surface immobilization of poly(ethyleneimine) and plasmid DNA on electrospun poly(L‐lactic acid) fibrous mats using a layer‐by‐layer approach for gene delivery

Abstract

The method of coating electrospun ultrafine poly(L-lactic acid) fibers with DNA, by building up polyelectrolyte layer(s) of poly(ethyleneimine) (PEI) and plasmid DNA using an electrostatic layer-by-layer deposition method, for gene delivery is presented. The pGL3 encoding luciferase was applied as plasmid DNA. The quantity of pGL3 immobilized on individual fibers increased with increasing pGL3 concentration in the immersion solution (0.017-0.870 mg/mL) and increasing bilayer number of PEI/pGL3 (single-triple). With the exception of one specimen prepared under the condition 0.870 mg/mL pGL3 solution and double PEI/pGL3 layers, the transfection efficiency of COS-7 cells, defined by the ratio of fluorescence intensity (resulting from the presence of luciferase) with respect to the quantity of cellular protein on the fibrous mat increased with increasing quantity of pGL3 on the fibers. In addition to the ease of controlling the quality of polyelectrolyte bilayer(s) by simply changing the concentrations of substances and number of immersing cycles, the features of the electrospun fibrous mat such as a very large surface-to-volume ratio and flexibility, could potentially be employed as a strategy for gene therapy combined with tissue engineering technology.

Core–sheath structured fibers with pDNA polyplex loadings for the optimal release profile and transfection efficiency as potential tissue engineering scaffolds

“…The use of DNA condensation techniques in combination with electrospinning should have the potential to achieve localized transfection of cells and induce sufficient protein production to stimulate new tissue formation on electrospun fibrous scaffolds, however, few attempts have been made to this end. Sakai et al coated electrospun fibers with DNA by building up polyelectrolyte layers of polycations and pDNA through electrostatic layer by layer deposition [16]. pDNA release and transgene expression were due to biodegradation of the fiber matrix and competitive binding of the proteins in serum.…”

Growth factor-eluting technologies for bone tissue engineering

“…LbL films can be generated from a wide variety of synthetic and natural polymers, and bioactive proteins; DNA and gene delivery vectors can be incorporated within PE multilayers without any need for covalent attachment [74] and without any significant changes to their native conformation. The deposition of PE multilayers onto 3D scaffolds has enabled scaffold-based delivery of both genes and proteins [75][76][77]. For example, 3D printed β-tricalcium phosphate/ polycaprolactone scaffolds coated with LbL films consisting of a poly (β-aminoester) (Bpoly 2^), chondroitin sulphate (CS), and BMP-2 resulted in a system that successfully induced in vivo bone formation when implanted intramuscularly in rats [76].…”

Localized, non-viral delivery of nucleic acids: Opportunities, challenges and current strategies

“…The average transfection efficiency was affected by PEG molecular weight and concentration and lasted for up to 60 days under optimized conditions in vitro. An alternative approach to encapsulation of NA into polymer fibers was presented by Sakai et al, who used a layer-by-layer assembly to load NA onto the fiber surface [104]. In this study, plasmid DNA and PEI were employed as polyanion and polycation, respectively to achieve successive deposition and Interestingly, several studies reported that significant cytotoxicity of polyplexes was observed despite encapsulation or adsorption of polyplexes to electrospun nonwovens, which was at least in part attributed to significant burst release [100e102].…”