Electrophoretic deposition to promote layer-by-layer assembly for in situ gene delivery application

Hu, Wei Wen; Zheng, Yan Rong

doi:10.1016/j.colsurfb.2015.05.046

Cited by 8 publications

(4 citation statements)

References 28 publications

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“…The objective of this work was to investigate the immobilization of DNA complexes to substrates functionalized with polymer brushes, taking advantage of the high negative surface charge of the brushes to attract and load cationic complexes, while also presenting cell-binding ligands to potentially influence the cellular response. Previous studies have indicated that the chemical properties of the substrate (e.g., self-assembly monolayers, polymer films, protein coatings) affect DNA complex binding and the efficiency of SMD (Segura et al, 2003; Bengali et al, 2005, 2007, 2009; Pannier et al, 2005, 2008; Li et al, 2009; Rea et al, 2009b; Zhang et al, 2009; Holmes and Tabrizian, 2013; Hu and Zheng, 2015), but many of those studies have focused on substrates like TCPS or glass, rather than biomaterials with possible clinical applications such as Ti. In this paper, we investigated the ability of chemically modified Ti substrates (with PAA brushes with or without peptide modifications) to support SMD.…”

Section: Discussionmentioning

confidence: 99%

Free Polyethylenimine Enhances Substrate-Mediated Gene Delivery on Titanium Substrates Modified With RGD-Functionalized Poly(acrylic acid) Brushes

et al. 2019

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Substrate mediated gene delivery (SMD) is a method of immobilizing DNA complexes to a substrate via covalent attachment or nonspecific adsorption, which allows for increased transgene expression with less DNA compared to traditional bolus delivery. It may also increase cells receptivity to transfection via cell-material interactions. Substrate modifications with poly(acrylic) acid (PAA) brushes may improve SMD by enhancing substrate interactions with DNA complexes via tailored surface chemistry and increasing cellular adhesion via moieties covalently bound to the brushes. Previously, we described a simple method to graft PAA brushes to Ti and further demonstrated conjugation of cell adhesion peptides (i.e., RGD) to the PAA brushes to improve biocompatibility. The objective of this work was to investigate the ability of Ti substrates modified with PAA-RGD brushes (PAA-RGD) to immobilize complexes composed of branched polyethyleneimine and DNA plasmids (bPEI-DNA) and support SMD in NIH/3T3 fibroblasts. Transfection in NIH/3T3 cells cultured on bPEI-DNA complexes immobilized onto PAA-RGD substrates was measured and compared to transfection in cells cultured on control surfaces with immobilized complexes including Flat Ti, PAA brushes modified with a control peptide (RGE), and unmodified PAA. Transfection was two-fold higher in cells cultured on PAA-RGD compared to those cultured on all control substrates. While DNA immobilization measured with radiolabeled DNA indicated that all substrates (PAA-RGD, unmodified PAA, Flat Ti) contained nearly equivalent amounts of loaded DNA, ellipsometric measurements showed that more total mass (i.e., DNA and bPEI, both complexed and free) was immobilized to PAA and PAA-RGD compared to Flat Ti. The increase in adsorbed mass may be attributed to free bPEI, which has been shown to improve transfection. Further transfection investigations showed that removing free bPEI from the immobilized complexes decreased SMD transfection and negated any differences in transfection success between cells cultured on PAA-RGD and on control substrates, suggesting that free bPEI may be beneficial for SMD in cells cultured on bPEI-DNA complexes immobilized on PAA-RGD grafted to Ti. This work demonstrates that substrate modification with PAA-RGD is a feasible method to enhance SMD outcomes on Ti and may be used for future applications such as tissue engineering, gene therapy, and diagnostics.

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Section: Discussionmentioning

confidence: 99%

Free Polyethylenimine Enhances Substrate-Mediated Gene Delivery on Titanium Substrates Modified With RGD-Functionalized Poly(acrylic acid) Brushes

et al. 2019

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“…Because the assembly of PEM is based on the electrostatic interaction between polyelectroelectolytes, ions in aqueous solutions may facilitate the dissociation of PEM from substrates [28]. However, the passive dissociation is limited due to not only electrostatic interaction but also the entanglement between polymer chains [12,24]. Therefore, an external force may be beneficial to destabilize the PEM structure and promote the following release.…”

Section: The Effects Of Electrical Field On the Dissociation Of Lbl Mmentioning

confidence: 99%

“…However, this switch-type manipulation exhibits quick DNA release, which is unsuitable to long-term delivery for regenerative medicine purposes [23]. On the other hand, our previous study also applied electrical fields to promote the deposition of DNA/chitosan multilayers, which suggested that electrical fields can lead the electrophoresis of polyelectrolytes, by which the loading efficiency can thus be improved [24]. These studies indicated that electrical fields can manipulate the stability of PEMs through electrochemical reaction and electrophoresis of polyelectrolytes, which should be a promising strategy to enhance the dissolution of polyelectrolytes from PEMs.…”

Section: Introductionmentioning

confidence: 99%

Electrical Field-Assisted Gene Delivery from Polyelectrolyte Multilayers

et al. 2020

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To sustain gene delivery and elongate transgene expression, plasmid DNA and cationic nonviral vectors can be deposited through layer-by-layer (LbL) assembly to form polyelectrolyte multilayers (PEMs). Although these macromolecules can be released for transfection purposes, their entanglement only allows partial delivery. Therefore, how to efficiently deliver immobilized genes from PEMs remains a challenge. In this study, we attempt to facilitate their delivery through the pretreatment of the external electrical field. Multilayers of polyethylenimine (PEI) and DNA were deposited onto conductive polypyrrole (PPy), which were placed in an aqueous environment to examine their release after electric field pretreatment. Only the electric field perpendicular to the substrate with constant voltage efficiently promoted the release of PEI and DNA from PEMs, and the higher potential resulted in the more releases which were enhanced with treatment time. The roughness of PEMs also increased after electric field treatment because the electrical field not only caused electrophoresis of polyelectrolytes and but also allowed electrochemical reaction on the PPy electrode. Finally, the released DNA and PEI were used for transfection. Polyplexes were successfully formed after electric field treatment, and the transfection efficiency was also improved, suggesting that this electric field pretreatment effectively assists gene delivery from PEMs and should be beneficial to regenerative medicine application.

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“…Using the electric-field-directed layer-by-layer deposition, CdTe light-emitting devices that emit green and red colors 16 and two different types of enzymes 17,18 were fabricated on an indium-tin-oxide substrate. The process of the electrophoretic deposition promoted the efficiency of DNA delivery, increased the duration of the release of DNA from polyelectrolyte multilayers, 19 enhanced the pervaporation performances of polyelectrolyte multilayers for separating isopropanol-water mixtures, 20,21 and rapidly fabricated functional protein nanoarrays. 22 The adsorption of polyelectrolytes was weakened at an intermediate electric field strength because of the electrolysis of the water at the electrode surface.…”

Section: Introductionmentioning

confidence: 99%

The fast deposition of antireflection coating

Wang

Peng

et al. 2017

J Coat Technol Res

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In this investigation, the effect of deposition time (5, 60, or 300 s) on a double-sided antireflection (AR) coating done by layer-by-layer deposition of polyelectrolyte was determined. The different structures and transmittance of an AR coating induced by electric-field-assisted layer-by-layer deposition (ELBLD) (with voltage of 0.5, 5, 50, or 500 V) and normal deposition (with voltage of 0 V) were compared. The electric field was perpendicular to the glass surface. After 5.5 bilayers of PAH/PAA were fabricated, the film was exposed to an acidic treatment of HCl solution (pH 2.4) for 65 s, followed by a brief rinse ($15 s) in deionized water, blown dry with air, and heat treated at 220°C for 2 h. Regardless of whether the electric field was applied, the average transmittance of visible light of the AR coating was more than 96% except for the reduced transmittance because of the excessive deposition of polyelectrolyte at an applied voltage of 500 V and a deposition time of 300 s. Average transmittance of 97.68 ± 0.31% was obtained at normal deposition of 5.5 bilayers with deposition time of 60 s. The polyelectrolytes had a propensity to form larger particles or accumulate into bundles under ELBLD, which was detrimental to the transmittance of an AR coating. Therefore, at the same deposition time, transmittance values of ELBLD samples were slightly lower than those of normal deposition. In other words, in the overall performance, the AR films of normal deposition are superior to those of ELBLD. The blowing pressure applied to dry the coating had significant impact on the quality of the film.

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Electrophoretic deposition to promote layer-by-layer assembly for in situ gene delivery application

Cited by 8 publications

References 28 publications

Free Polyethylenimine Enhances Substrate-Mediated Gene Delivery on Titanium Substrates Modified With RGD-Functionalized Poly(acrylic acid) Brushes

Free Polyethylenimine Enhances Substrate-Mediated Gene Delivery on Titanium Substrates Modified With RGD-Functionalized Poly(acrylic acid) Brushes

Electrical Field-Assisted Gene Delivery from Polyelectrolyte Multilayers

The fast deposition of antireflection coating

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