EVALUATION OF MAGNETIC-HYDROXYAPATITE NANOPARTICLES FOR GENE DELIVERY CARRIER

2010 ◽  
Vol 22 (01) ◽  
pp. 33-39 ◽  
Author(s):  
Hsi-Chin Wu ◽  
Feng-Huei Lin
Keyword(s):  
Magnetic Field ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Binding Capacity ◽  
Fold Increase ◽  
Great Promise ◽  
Precipitation Process ◽  
Hydroxyapatite Nanoparticles ◽  
Enzyme Link Immunosorbent Assay ◽  
Transfection Complex

With the advancement of nanotechnology, nano-sized bioceramics have been dedicated much attention to biomedical applications in the past decades. In this study, the magnetic-hydroxyapatite nanoparticles (mHap NPs) were synthesized and magnetized by the addition of irons into hydroxyapatite using precipitation process. The mHap NPs were with good biocompatibility and superparamagnetic property which would have great promise in biological applications. We have evaluated the essential properties including plasmid DNA (pDNA) binding capacity by electrophoresis, qualitative transfection efficiency by fluorescence microscopy and quantitative GDNF expression by enzyme-link immunosorbent assay (ELISA). The results indicated that mHap NPs could be successfully binding with pDNA to form pDNA-mHap NPs transfection complex. The endocytosis of pDNA-mHap NPs transfection complex was mediated into cells and facilitated if magnetic field was applied. The transfection efficiency could also be significantly improved by magnetofection. The gene expression had been four-fold increase in high pDNA loading (2 μg/well) in transfection procedure with magnetic field compared with traditional transfection method. In brief, the mHap NPs have great potential as non-viral inorganic delivery carriers for gene delivery system.

scholarly journals Dopamine-Grafted Hyaluronic Acid Coated Hyperbranched Poly(β-Amino Esters)/DNA Nano-Complexes for Enhanced Gene Delivery and Biosafety

Crystals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 347
Author(s):  
Man Guo ◽  
Yingcai Meng ◽  
Xiaoqun Qin ◽  
Wenhu Zhou
Keyword(s):  
Hyaluronic Acid ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Genetic Diseases ◽  
Great Promise ◽  
Cationic Polymers ◽  
Nano Structure ◽  
Robust Solution ◽  
Amino Esters

Gene therapy has attracted particular attention for the treatment of various genetic diseases, and the development of gene delivery vectors is of utmost importance for in vivo applications of gene drugs. Various cationic polymers with high nucleic acid loading and intracellular transfection efficiency have been reported, however, their biological applications are limited by potential toxicity. Surface modification is a robust solution to detoxify the cationic vectors, but this can inevitably weaken the transfection efficiency. To address this dilemma, we reported the ability of a dopamine (DA)-grafted hyaluronic acid (HA) to modify gene vectors for enhanced gene delivery and biosafety. The nano-vector was formed by using branched poly(β-amino esters) (PAEs), and surface coating with HA-DA to form a core-shell nano-structure via electrostatic attraction. Upon HA-DA modification, the biosafety of the gene delivery vehicle was improved, as demonstrated by the cell cytotoxicity assay and hemolysis test. Notably, the nano-system displayed a DA-dependent transfection efficiency, in which a higher DA grafting degree resulted in better efficacy. This can be explained by the adhesive nature of DA, facilitating cell membrane interaction, as well as DA receptor mediated active targeting. At the optimal DA grafting ratio, the nano-system achieved a transfection efficiency even better than that of commonly used polyethylenimine (PEI) vectors. Together with its excellent biocompatibility, the vector presented here holds great promise for gene delivery applications.

scholarly journals Application of Ferriferous Oxide Modified by Chitosan in Gene Delivery

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Yu Kuang ◽  
Tun Yuan ◽  
Zhongwei Zhang ◽  
Mingyuan Li ◽  
Yuan Yang
Keyword(s):  
Magnetic Field ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Spherical Surface ◽  
Cytotoxicity Test ◽  
The Novel ◽  
Permanent Magnetic Field ◽  
In Vivo Test

New approaches to improve the traditional gene carriers are still required. Here we explore Fe3O4 modified with degradable polymers that enhances gene delivery and target delivery using permanent magnetic field. Two magnetic Fe3O4 nanoparticles coated with chitosan (CTS) and polyethylene glycol (PEG) were synthesized by means of controlled chemical coprecipitation. Plasmid pEGFP was encapsulated as a reported gene. The ferriferous oxide complexes were approximately spherical; surface charge of CTS-Fe3O4 and PEG-Fe3O4 was about 20 mv and 0 mv, respectively. The controlled release of DNA from the CTS-Fe3O4 nanoparticles was observed. Concurrently, a desired Fe3O4 concentration of less than 2 mM was verified as safe by means of a cytotoxicity test in vitro. Presence of the permanent magnetic field significantly increased the transfection efficiency. Furthermore, the passive target property and safety of magnetic nanoparticles were also demonstrated in an in vivo test. The novel gene delivery system was proved to be an effective tool required for future target expression and gene therapy in vivo.

scholarly journals ULTRASOUND-ASSISTED MAGNETIC NANOPARTICLE-BASED GENE DELIVERY

2020 ◽  
Author(s):  
Wei Zhang ◽  
Gaser N. Abdelrasoul ◽  
Oleksandra Savchenko ◽  
Abdalla Abdrabou ◽  
Zhixiang Wang ◽  
...  
Keyword(s):  
Magnetic Field ◽  
External Magnetic Field ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Genetic Material ◽  
Positive Control ◽  
Superparamagnetic Nanoparticles ◽  
Cell Permeability ◽  
Hek 293 ◽  
Delivery Technique

AbstractLow-intensity pulsed ultrasound (LIPUS), a special type of ultrasonic stimulation, is attracting a lot of attention for both clinical and scientific research. In this paper, we report a concept of a new method using magnetic nanoparticles (MNPs) for LIPUS-assisted gene delivery. The MNPs are iron oxide superparamagnetic nanoparticles, coated with polyethyleneimine (PEI), which introduces a high positive surface charge, favorable for the binding of genetic material. Due to the paramagnetic properties of the MNPs, the application of an external magnetic field increases transfection efficiency; meanwhile, LIPUS stimulation enhances cell permeability. We found out that stimulation at the intensity of 30 mW/cm2 for 10 minutes yields optimal results with a minimal adverse effect on the cells. Combining the effect of the external magnetic field and LIPUS, the genetic material (GFP or Cherry Red plasmid in our case) can enter the cells. The flow cytometry results showed that by using just a magnetic field to direct the genetic material, the transfection efficiency of HEK 293 cells that were treated with our MNPs was 56.1%. Coupled with LIPUS stimulation, it increased to 61.5% or 19% higher than the positive control (Lipofectamine 2000). In addition, compared with the positive control, our method showed less toxicity. Cell viability after transfection was 63.61%, 19% higher than with the standard transfection technique. In conclusion, we designed a new gene-delivery technique that is affordable, targeted, shows low-toxicity, yet high transfection efficiency, compared to other conventional approaches.The Graphical Abstract

scholarly journals Mannan-Modified PLGA Nanoparticles for Targeted Gene Delivery

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Fansheng Kong ◽  
Linfu Ge ◽  
Ximin Liu ◽  
Ning Huang ◽  
Fang Zhou
Keyword(s):  
Flow Cytometry ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Binding Capacity ◽  
Plga Nanoparticles ◽  
Gene Expressions ◽  
Targeted Gene Delivery ◽  
Modified Dna ◽  
Lipofectamine 2000

The studies of targeted gene delivery nanocarriers have gained increasing attention during the past decades. In this study, mannan modified DNA loaded bioadhesive PLGA nanoparticles (MAN-DNA-NPs) were investigated for targeted gene delivery to the Kupffer cells (KCs). Bioadhesive PLGA nanoparticles were prepared and subsequently bound withpEGFP. Following the coupling of the mannan-based PE-grafted ligands (MAN-PE) with the DNA-NPs, the MAN-DNA-NPs were delivered intravenously to rats. The transfection efficiency was determined from the isolated KCs and flow cytometry was applied for the quantitation of gene expression after 48 h post transfection. The size of the MAN-DNA-NPs was found to be around 190 nm and the Zeta potential was determined to be −15.46mV. ThepEGFPbinding capacity of MAN-DNA-NPs was ()% and thein vitrorelease profiles of the MAN-DNA-NPs follow the Higuchi model. When compared with non-modified DNA-NPs and Lipofectamine 2000-DNA, MAN-DNA-NPs produced the highest gene expressions, especiallyin vivo. Thein vivodata from flow cytometry analysis showed that MAN-DNA-NPs displayed a remarkably higher transfection efficiency (39%) than non-modified DNA-NPs (25%) and Lipofectamine 2000-DNA (23%) in KCs. The results illustrate that MAN-DNA-NPs have the ability to target liver KCs and could function as promising active targeting drug delivery vectors.

scholarly journals The transition from linear to highly branched poly(β-amino ester)s: Branching matters for gene delivery

2016 ◽  
Vol 2 (6) ◽  
pp. e1600102 ◽  
Author(s):  
Dezhong Zhou ◽  
Lara Cutlar ◽  
Yongsheng Gao ◽  
Wei Wang ◽  
Jonathan O’Keeffe-Ahern ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Branched Polymers ◽  
Nonviral Gene Delivery ◽  
Great Promise ◽  
Amino Ester ◽  
Design And Synthesis ◽  
Gene Delivery Vectors

Nonviral gene therapy holds great promise but has not delivered treatments for clinical application to date. Lack of safe and efficient gene delivery vectors is the major hurdle. Among nonviral gene delivery vectors, poly(β-amino ester)s are one of the most versatile candidates because of their wide monomer availability, high polymer flexibility, and superior gene transfection performance both in vitro and in vivo. However, to date, all research has been focused on vectors with a linear structure. A well-accepted view is that dendritic or branched polymers have greater potential as gene delivery vectors because of their three-dimensional structure and multiple terminal groups. Nevertheless, to date, the synthesis of dendritic or branched polymers has been proven to be a well-known challenge. We report the design and synthesis of highly branched poly(β-amino ester)s (HPAEs) via a one-pot “A2 + B3 + C2”–type Michael addition approach and evaluate their potential as gene delivery vectors. We find that the branched structure can significantly enhance the transfection efficiency of poly(β-amino ester)s: Up to an 8521-fold enhancement in transfection efficiency was observed across 12 cell types ranging from cell lines, primary cells, to stem cells, over their corresponding linear poly(β-amino ester)s (LPAEs) and the commercial transfection reagents polyethyleneimine, SuperFect, and Lipofectamine 2000. Moreover, we further demonstrate that HPAEs can correct genetic defects in vivo using a recessive dystrophic epidermolysis bullosa graft mouse model. Our findings prove that the A2 + B3 + C2 approach is highly generalizable and flexible for the design and synthesis of HPAEs, which cannot be achieved by the conventional polymerization approach; HPAEs are more efficient vectors in gene transfection than the corresponding LPAEs. This provides valuable insight into the development and applications of nonviral gene delivery and demonstrates great prospect for their translation to a clinical environment.

High Resolution Current Imaging by Direct Magnetic Field Sensing

2003 ◽  
Author(s):  
S.I. Woods ◽  
Nesco M. Lettsome ◽  
A.B. Cawthorne ◽  
L.A. Knauss ◽  
R.H. Koch
Keyword(s):  
Magnetic Field ◽  
High Resolution ◽  
Spatial Resolution ◽  
Great Promise ◽  
Magnetoresistive Sensor ◽  
Magnetic Sensitivity ◽  
Current Lines ◽  
Scanning Squid ◽  
Scanning Fiber ◽  
Ferromagnetic Probe

Abstract Two types of magnetic microscopes have been investigated for use in high resolution current mapping. The scanning fiber/SQUID microscope uses a SQUID sensor coupled to a nanoscale ferromagnetic probe, and the GMR microscope employs a nanoscale giant magnetoresistive sensor. Initial scans demonstrate that these microscopes can resolve current lines less than 10 µm apart with edge resolution of 1 µm. These types of microscopes are compared with the performance of a standard scanning SQUID microscope and with each other with respect to spatial resolution and magnetic sensitivity. Both microscopes show great promise for identifying current defects in die level devices.

Stepwise Development of Biomimetic Chimeric Peptides for Gene Delivery

2020 ◽  
Vol 27 (8) ◽  
pp. 698-710
Author(s):  
Roya Cheraghi ◽  
Mahboobeh Nazari ◽  
Mohsen Alipour ◽  
Saman Hosseinkhani
Keyword(s):  
Gene Delivery ◽  
Targeted Delivery ◽  
Viral Vectors ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Cationic Lipids ◽  
Target Cells ◽  
Scale Production ◽  
Stepwise Development ◽  
Chimeric Peptides

Gene-based therapy largely relies on the vector type that allows a selective and efficient transfection into the target cells with maximum efficacy and minimal toxicity. Although, genes delivered utilizing modified viruses transfect efficiently and precisely, these vectors can cause severe immunological responses and are potentially carcinogenic. A promising method of overcoming this limitation is the use of non-viral vectors, including cationic lipids, polymers, dendrimers, and peptides, which offer potential routes for compacting DNA for targeted delivery. Although non-viral vectors exhibit reduced transfection efficiency compared to their viral counterpart, their superior biocompatibility, non-immunogenicity and potential for large-scale production make them increasingly attractive for modern therapy. There has been a great deal of interest in the development of biomimetic chimeric peptides. Biomimetic chimeric peptides contain different motifs for gene translocation into the nucleus of the desired cells. They have motifs for gene targeting into the desired cell, condense DNA into nanosize particles, translocate the gene into the nucleus and enhance the release of the particle into the cytoplasm. These carriers were developed in recent years. This review highlights the stepwise development of the biomimetic chimeric peptides currently being used in gene delivery.

scholarly journals Highly Osmotic Oxidized Sucrose-Crosslinked Polyethylenimine for Gene Delivery Systems

Pharmaceutics ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 87
Author(s):  
Jaehong Park ◽  
Kyusik Kim ◽  
Sohee Jeong ◽  
Migyeom Lee ◽  
Tae-il Kim
Keyword(s):  
Gene Delivery ◽  
Reductive Amination ◽  
Transfection Efficiency ◽  
Delivery Systems ◽  
Buffering Capacity ◽  
Serum Stability ◽  
High Osmolality ◽  
Cox 2 ◽  
Serum Free Conditions ◽  
Positively Charged

In this work, highly osmotic oxidized sucrose-crosslinked polyethylenimine (SP2K) polymers were developed for gene delivery systems, and the transfection mechanism is examined. First, periodate-oxidized sucrose and polyethylenimine 2K (PEI2K) were crosslinked with various feed ratios via reductive amination. The synthesis was confirmed by 1H NMR and FTIR. The synthesized SP2K polymers could form positively charged (~40 mV zeta-potential) and nano-sized (150–200 nm) spherical polyplexes with plasmid DNA (pDNA). They showed lower cytotoxicity than PEI25K but concentration-dependent cytotoxicity. Among them, SP2K7 and SP2K10 showed higher transfection efficiency than PEI25K in both serum and serum-free conditions, revealing the good serum stability. It was found that SP2K polymers possessed high osmolality and endosome buffering capacity. The transfection experiments with cellular uptake inhibitors suggest that the transfection of SP2K polymers would progress by multiple pathways, including caveolae-mediated endocytosis. It was also thought that caveolae-mediated endocytosis of SP2K polyplexes would be facilitated through cyclooxygenase-2 (COX-2) expression induced by high osmotic pressure of SP2K polymers. Confocal microscopy results also supported that SP2K polyplexes would be internalized into cells via multiple pathways and escape endosomes efficiently via high osmolality and endosome buffering capacity. These results demonstrate the potential of SP2K polymers for gene delivery systems.

Boosting transfection efficiency: A systematic study using layer-by-layer based gene delivery platform

2021 ◽  
pp. 112161
Author(s):  
Yana V. Tarakanchikova ◽  
Dmitrii S. Linnik ◽  
Tatiana Mashel ◽  
Albert R. Muslimov ◽  
Sergey Pavlov ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Systematic Study ◽  
Transfection Efficiency ◽  
Layer By Layer ◽  
Delivery Platform

scholarly journals Electrical Field-Assisted Gene Delivery from Polyelectrolyte Multilayers

Polymers ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 133
Author(s):  
Yu-Che Cheng ◽  
Shu-Lin Guo ◽  
Kun-Da Chung ◽  
Wei-Wen Hu
Keyword(s):  
Gene Delivery ◽  
Electric Field ◽  
Treatment Time ◽  
Transfection Efficiency ◽  
Polyelectrolyte Multilayers ◽  
Aqueous Environment ◽  
Layer By Layer ◽  
Electrical Field ◽  
Lbl Assembly ◽  
Electric Field Treatment

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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