Stability of poly(ethylene glycol)-graft-polyethylenimine copolymer/DNA complexes: influences of PEG molecular weight and PEGylation degree

Polyethylenimine (PEI) is one of the most widely investigated cationic polymers for gene delivery. However, PEI/DNA complexes are unstable and tend to aggregate. PEGylation was used to improve the stability. The stability of polymer/DNA complexes was investigated including complexation stability, aggregation stability, sedimentation stability, and nuclease stability. PEI25K/DNA complexes were liable to aggregate to large particles (500–700 nm). The aggregation was proved to be induced by phosphate anion. In the medium without phosphate anion, aggregation was prevented by electrostatic repulsion. Owing to more efficient steric repulsion, PEG2 and PEG5K excelled PEG750 in facilitating copolymers to form stable small polyplexes (below 100 nm) without aggregation regardless of phosphate anion. The steric repulsion predominated over electrostatic repulsion in stabilization.     

Decaarginine-PEG-artificial lipid/DNA complex for gene delivery: nanostructure and transfection efficiency

Oligoarginine conjugates are highly efficient vectors for the delivery of plasmid DNA into cells. Decaarginine-conjugated lipid (Arg10-PEG-lipid) was synthesized and the effects of Arg10-PEG-lipid concentration at a fixed DNA concentration on transfection efficiency and the structure of the complexes were studied below and above critical micelle concentration (CMC), and at the lipid nitrogen/DNA phosphate (N/P) ratio corresponding to transfection, respectively. Arg10-PEG-lipid at the concentration below CMC showed stronger interaction with DNA by fluorescence intensity distribution analysis, and significantly higher luciferase and green fluorescent protein expression than that above CMC. A phase-contrast cryo-transmission electron microscope (cryo-TEM) experiment showed that the morphology of the complexes depended on the N/P ratio. At a low N/P ratio corresponding to that in transfection at a lipid concentration below CMC, a net-like structure developed in which plasmid DNA was involved. A further increase in the N/P ratio, a large fibrous nanostructure of complexes, was also observed. Without DNA, these structures were not obtained. The cellular uptake mechanism of complexes using flow cytometry with inhibitors suggested that complexes with two different morphologies showed similar cellular uptake and uptake mechanism, macropinocytosis. Differences in transfection efficiency of the complexes may be explained by a large fibrous nanostructure inhibiting the cellular internalization of complexes or the release of DNA from macropinosomes into cytoplasm. Arg10-PEG-lipid/DNA complexes formed a favorable nanostructure for gene delivery, depending on the N/P ratio in water.     

镀Ni-Cu硅酸钙镁晶须/镍粉/环氧树脂电磁屏蔽涂料研究

以一种新型镀Ni-Cu硅酸钙镁矿物晶须和羰基镍粉作为屏蔽复合填料,研制出了一种新型镀Ni-Cu硅酸钙镁晶须/镍粉/环氧树脂电磁波屏蔽复合涂料,并研究了其导电和电磁波屏蔽性能.结果表明,添加4%(wt)的镀Ni-Cu硅酸钙镁晶须能显著改善涂层的电磁性能,膜厚300μm,涂层电阻率为1.09Ω·cm,涂层屏蔽效能在0.3-...     

The use of myristic acid as a ligand of polyethylenimine/DNA nanoparticles for targeted gene therapy of glioblastoma

To establish a gene delivery system for brain targeting, a low molecular weight polyethylenimine (PEI(10?K)) was modified with myristic acid (MC), and complexed with DNA, yielding MC-PEI(10?K)/DNA nanoparticles successfully. The nanoparticles were observed to be successfully taken up by the brains of mice. The transfection efficiency of the nanoparticles was then investigated, and both the in?vitro and in?vivo gene expression of MC-PEI(10?K)/DNA nanoparticles is significantly higher than that of unmodified PEI(10?K)/DNA nanoparticles. The anti-glioblastoma effect of MC-PEI(10?K)/pORF-hTRAIL was demonstrated by the survival time of intracranial U87 glioblastoma-bearing mice. The median survival time of the MC-PEI(10?K)/pORF-hTRAIL group (28 days) was significantly longer than that of the PEI(10?K)/pORF-hTRAIL group (24 days), the MC-PEI(10?K)/pGL(3) group (21 days) and the saline group (22 days). Therefore, our results suggested that MC-PEI(10?K) could be potentially used for brain-targeted gene delivery and in the treatment of glioblastoma.     

Cationic liposome-DNA complexes: from liquid crystal science to gene delivery applications

At present, there is an unprecedented level of interest in the properties and structures of complexes consisting of DNA mixed with oppositely charged cationic liposomes (CLs). The interest arises because the complexes mimic natural viruses as chemical carriers of DNA into cells in worldwide human gene therapy clinical trials. However, since our understanding of the mechanisms of action of CL-DNA complexes interacting with cells remains poor, significant additional insights and discoveries will be required before the development of efficient chemical carriers suitable for long-term therapeutic applications. Recent studies describe synchrotron X-ray diffraction, which has revealed the liquid crystalline nature of CL-DNA complexes, and three-dimensional laser-scanning confocal microscopy, which reveals CL-DNA pathways and interactions with cells. The importance of the liquid crystalline structures in biological function is revealed in the application of these modern techniques in combination with functional transfection efficiency measurements, which shows that the mechanism of gene release from complexes in the cell cytoplasm is dependent on their precise liquid crystalline nature and the physical and chemical parameters (for example, the membrane charge density) of the complexes. In [section sign] 5, we describe some recent new results aimed at developing bionanotube vectors for gene delivery.     

电刷镀电磁屏蔽涂层制备工艺与性能研究

采用电刷镀方法在环氧树脂表面制备镍铜层状电磁屏蔽复合镀层。分析了工作电压对复合镀层表面形貌、沉积速率、复合镀层与基体的结合力的影响,最后对不同工艺参数下层状复合电磁屏蔽镀层的电磁屏蔽效能进行了测试。结果表明,电刷镀铜的最佳刷镀电压是5V,电刷镀镍的最佳电压是4V,镀笔运动最佳速度为6m/min。在此工艺条件下制备的复合镀层与基体的结合力可达20.44N,电磁屏蔽效能达到74dB以上。     

碳纳米管/环氧丙烯酸酯紫外光固化涂层的制备及光阻隔性能

利用碳纳米管(CNTs)表面官能团与环氧丙烯酸酯(EA)齐聚物的反应, 制备了碳纳米管/环氧丙烯酸酯(CNTs/EA)光敏性树脂, 通过紫外光(UV)固化交联制备出新型复合涂层。研究了涂层的阻隔红外/紫外线性能, 在近红外区(4000~12000 cm-1)的阻隔率为68.55%, 对可见光(380~780 nm)的透过率大于85％, 对于＜300 nm的紫外线完全阻隔。同时复合涂层力学性能与EA涂层相比也得到提高, 在光阻隔涂层方面具有潜在的应用前景。      

Receptor-mediated gene delivery into antigen presenting cells using mannosylated chitosan/DNA nanoparticles

Dendritic cells (DCs) are professional antigen presenting cells that induce, sustain, and regulate immune responses. Gene modification of DCs is of particular interest for immunotherapy of diseases where the immunes system has failed or is abnormally regulated, such as in cancer or autoimmune disease. Gene transfer using non-viral vectors is a promising approach for the safe delivery of therapeutic DNA. Among various non-viral vectors, chitosan is considered to be a good candidate for gene delivery system, however, lack of cell specificity and low transfection of chitosan need to be overcome prior to clinical use. In this study, mannosylated chitosan (MC) was prepared to induce the receptor-mediated endocytosis and targeting into antigen presenting cells (APCs), especially DCs having mannose receptors. MC showed great ability to form complexes with DNA and showed suitable physicochemical properties for gene delivery system. It had low cytotoxicity and exhibited much enhanced gene transfer efficiency on the macrophage cell line than chitosan itself. Also, MC/DNA complex was more efficient for transferring IL-12 gene into DCs rather than water-soluble chitosan (WSC)/DNA one, which resulted in better induction of INF-gamma from DCs. Therefore, MC is a promising gene delivery system for repeated administration to maintain sustained gene expression, thereby opening the possibility for immunotherapy.     

不同基体对复合涂料的微结构及电磁屏蔽效能的影响

采用镍粉(Ni)与樟脑磺酸二次掺杂聚苯胺(PAn-CSA)为导电填料,以KH-550硅烷为偶联剂,分别以丙烯酸树脂、醇酸油漆、酚醛油漆为基体制备成各种电磁屏蔽涂料,分别对其电导率(ρ)和电磁屏蔽效能(SE)的测试与分析。结果表明,ρ丙烯酸树脂ρ醇酸油漆ρ酚醛油漆,SE丙烯酸树脂SE醇酸油漆SE酚醛油漆;采用扫描电子显微镜对其表面形貌进行观察与分析,观测的结果,导电填料在基体中分散的均匀性大小:丙烯酸树脂醇酸油漆酚醛油漆,这与电导率和电磁屏蔽效能测量的结果是一致的。     

电磁屏蔽涂料用改性短碳纤维的研究

利用阻抗匹配原理和电容器通交流断直流的频率特性,提出了在导电填料表面包覆不导电层设计的构想,来实现对表面DC电导率随填料含量的提高而增大、从而形成回路而短路等问题的改善.采用原位聚合法在短碳纤维(SCF)的表面均匀包覆一层不导电的聚苯胺(PANI),使用扫描电子显微镜(SEM),热分析(TG)以及傅里叶变换红外光谱(FTIR)等表征了改性SCF的表观形貌和结构,分析并探讨了SCF微结构与电磁波的交互作用机制.研究表明:在聚合温度为5℃,聚合反应时间为12h,苯胺与引发剂的摩尔比为1:1,碱性溶液摩尔浓度为1mol/L时,可制得表面均匀包覆不导电PANI的改性SCF;应用改性填料制得电磁屏蔽涂料,通过同轴法兰装置测试,在频率＜1.5GHz下,相对使用碳纤维体系而言,其屏蔽效能(最大值)提高了3倍多.     

Stealth dendrimers for drug delivery: correlation between PEGylation,cytocompatibility, and drug payload

It is advantageous to utilize low generation polyamidoamine (PAMAM) dendrimers for drug delivery because low generations (generation 4.0 or below) have more biologically favorable properties as compared to high generations. Nevertheless, modification of low generation dendrimers with PEG to create stealth dendrimers is still necessary to avoid potential side effects by long term accumulation. However, low generation dendrimers have much fewer surface sites for drug loading as compared to higher generations. To efficiently utilize low generation dendrimer-based stealth dendrimers for drug loading, PEGylation needs to be optimized. In this study, we synthesized a series of stealth dendrimers based on low generation Starburst™ PAMAM dendrimers (i.e., G2.5, G3.0, G3.5, and G4.0) and quantitatively assessed PEGylation efficacy in modulating cytocompatibility of low generation PAMAM dendrimers. Cytocompatibility of stealth dendrimers was examined using endothelial cells. The results showed that PEGylation degree on low generation dendrimers could be dramatically reduced to leave as many unoccupied surface groups as possible for drug loading, while maintaining the drug carrier cytocompatibility. 3PEGs-G3.0 and 10PEGs-G4.0 were considered initially optimized stealth dendrimers that would be further modified to deliver drugs of interest. Correlation of PEGylation, cytocompatibility, and drug payload allowed us to optimize low generation dendrimer-based stealth dendrimers for drug delivery and advance the understanding of structure-property relationship of stealth dendrimers.     

Ultra-small graphene oxide functionalized with polyethylenimine (PEI) for very efficient gene delivery in cell and zebrafish embryos

Efficient DNA delivery is essential for introducing new genes into living cells. However, effective virus-based systems carry risks and efficient synthetic systems that are non-toxic remain to be discovered. The bottle-neck in synthetic systems is cytotoxicity, caused by the high concentration of DNA-condensing compounds required for efficient uptake of DNA. Here we report a polyethyleneimine (PEI) grafted ultra-small graphene oxide (PEI-g-USGO) for transfection. By removing the free PEI and ensuring a high PEI density on small sized graphene, we obtained very high transfection efficiencies combined with very low cytotoxicity. Plasmid DNA could be transfected into mammalian cell lines with up to 95% efficiency and 90% viability. Transfection in zebrafish embryos was 90%, with high viability, compared to efficiencies of 30% or lower for established transfection technologies. This result suggests a novel approach to the design of synthetic gene delivery vehicles for research and therapy.      

Influence of aspect ratio and skin effect on EMI shielding of coating materials fabricated with carbon nanofiber/PVDF

The effectiveness of electromagnetic interference (EMI) shielding was measured for poly vinylidene fluoride (PVDF) coating materials containing carbon nanofibers. Carbon nanofibers produced from the C₂H₄/NC73 system exhibited higher shielding effectiveness (SE) by relatively large specific surface area and high aspect ratio than those from others. When the thickness of carbon nanofibers filled PVDF coating materials varied from 25 to 50 m, the electrical conductivity of coating materials increased sharply from 1.34 to 1.91 S/cm. However, the electrical conductivity approached a certain value with further raise of the thickness. This phenomenon denotes that a critical thickness of coating materials exists around 50 m. The electrical conductivity and SE of coating materials decreased dramatically as the carbon nanofiber fillers were milled. It could be concluded that the decrease of the shielding effectiveness of carbon nanofiber filled composite was due to the decrease of the filler's aspect ratio by ball milling.     

Synergistic effects of micro-/nano-fillers on conductive and electromagnetic shielding properties of polypropylene nanocomposites

This study presents the synergistic effects of graphene nanosheets (GNSs) and carbon fibers (CFs) additions on the electrical and electromagnetic shielding properties of GNS/CF/polypropylene (PP) composites. These composites were fabricated by the melt blending of different ratios of GNSs and CFs (20:0, 15:5, 10:10, 5:15 and 0:20 wt/wt%) into a PP polymer matrix using a Brabender mixer. Besides, the chemical and crystalline structures and the thermal stability of the resultant GNS/CF/PP composites were characterized by Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD) and thermogravimetric analysis (TGA). FT-IR and XRD showed that with the addition of GNSs content, transmittances at 1373.4?cm^?1 and 1454.4?cm^?1 became smaller and the characteristic peak at 26.82° became stronger. TGA showed that the GNS/CF/PP composite can be used at high temperature below 456°C. Blending 10?wt% CFs and 10?wt% GNSs into the PP polymer resulted in excellent conductivity (0.397 S/cm), which indicated the occurrence of the critical percolation threshold phenomenon, and also reached the maximum electromagnetic shielding effectiveness (EMSE) of 20?dB at 1.28–2.00?GHz. Laminated with five layers of composites, its EMSE achieved 25–38?dB at 0.3–3.0?GHz, corresponding to blocking of 94.38–98.74% electromagnetic waves.     

膨胀石墨/金属网/ABS复合材料电磁屏蔽性能的研究

以膨胀石墨(EG)和金属网(MN)作为电磁屏蔽基元材料与ABS树脂采用共混、挤出、热压等成型工艺制备了电磁屏蔽复合材料,研究了膨胀石墨的含量、处理方式、复合材料的厚度和金属网的目数对电磁屏蔽复合材料屏蔽性能的影响。结果表明,在膨胀石墨/ABS电磁屏蔽复合材料中,其电磁屏蔽效能随着膨胀石墨含量增加及复合材料厚度增加而增大,膨胀石墨经超声处理后,可以提高复合材料的屏蔽效能。在两种单层金属网/ABS电磁屏蔽夹层复合材料中,屏蔽效能并不随着金属网目数增加而增大。在30MHz～1.8GHz频率范围内,200目不锈钢网/ABS复合材料和100目铜网/ABS复合材料的屏蔽性能最好,最大屏蔽效能分别为76.1和70dB。在多相电磁屏蔽复合材料中,膨胀石墨/不锈钢网/ABS复合材料的屏蔽效能比不锈钢网/ABS复合材料高约5dB。     

镀银织物基Fe3O4/WPU发泡涂层复合材料电磁屏蔽性能研究

为实现材料与结构协同增强织物电磁屏蔽性能,以镀银织物为基布,引入磁损耗材料Fe₃O₄,利用多孔结构,采用发泡涂层法制备镀银导电布基Fe₃O₄/水性聚氨酯（WPU）发泡涂层材料,探究Fe₃O₄添加量对涂层表观结构以及电磁材料协同和泡孔结构协同作用下复合材料的电磁屏蔽性能的影响。实验结果表明,当Fe₃O₄添加量不超过20%的情况下,Fe₃O₄在水性聚氨酯基体中分散性相对较好,泡孔孔径分布均匀,复合材料整体协同的电磁屏蔽性能在8.2～12.4GHz内高达85dB,较原基布提高了18%,体现较好的电磁屏蔽性能。     

Anionic solid lipid nanoparticles supported on protamine/DNA complexes

The objective of this study was to design novel anionic ternary nanoparticles for gene delivery. These ternary nanoparticles were equipped with protamine/DNA binary complexes (150-200?nm) as the support, and the anionic formation was achieved by absorption of anionic solid lipid nanoparticles (≤20?nm) onto the surface of the binary complexes. The small solid lipid nanoparticles (SLNs) were prepared by a modified film dispersion-ultrasonication method, and adsorption of the anionic SLNs onto the binary complexes was typically carried out in water via electrostatic interaction. The formulated ternary nanoparticles were found to be relatively uniform in size (257.7 ± 10.6?nm) with a 'bumpy' surface, and the surface charge inversion from 19.28 ± 1.14?mV to -17.16 ± 1.92?mV could be considered as evidence of the formation of the ternary nanoparticles. The fluorescence intensity measurements from three batches of the ternary nanoparticles gave a mean adsorption efficiency of 96.75 ± 1.13%. Circular dichroism spectra analysis showed that the protamine/DNA complexes had been coated by small SLNs, and that the anionic ternary nanoparticles formed did not disturb the construction of the binary complexes. SYBR Green I analysis suggested that the ternary nanoparticles could protect the DNA from nuclease degradation, and cell viability assay results showed that they exhibit lower cytotoxicity to A549 cells compared with the binary complexes and lipofectamine. The transfection efficiency of the ternary nanoparticles was better than that of naked DNA and the binary complexes, and almost equal to that of lipofectamine/DNA complexes, as revealed by inversion fluorescence microscope observation. These results indicated that the anionic ternary nanoparticles could facilitate gene transfer in cultured cells, and might alleviate the drawbacks of the conventional cationic vector/DNA complexes for gene delivery in vivo.     

Transferrin-mediated PEGylated nanoparticles for delivery of DNA/PLL

The purpose of this work was to determine the stability of pDNA/poly(L-lysine) complex (DNA/PLL) during microencapsulation, prepare transferrin (TF) conjugated PEGylated nanoparticles (TF-PEG-NP) loading DNA/PLL, and assess its physicochemical characteristics and in vitro transfection efficiency. The DNA/PLL was prepared by mixing plasmid DNA (pDNA) in deionized water with various amounts of PLL. PEGylated nanoparticles (PEG-NP) loading DNA/PLL were prepared by a water-oil-water double emulsion solvent evaporation technique. TF-PEG-NP was prepared by coupling TF with PEG-NP. The physicochemical characteristics of TF-PEG-NP and in vitro transfection efficiency on K562 cells were measured. The results showed that free pDNA reserved its double supercoiled form (dsDNA) for only on average 25.7% after sonification, but over 70% of dsDNA was reserved after pDNA was contracted with PLL. The particle size range of TF-PEG-NP loading DNA/PLL was 150-450?nm with entrapment efficiency over 70%. TF-PEG-NP exhibited the low burst effect (<10%) within 1 day. After the first phase, DNA/PLL displayed a sustained release. The amount of cumulated DNA/PLL release from TF-PEG-NP with 2% polymer over 7 days was 85.4% for DNA/PLL (1:0.3 mass ratio), 59.8% and 43.1% for DNA/PLL (1:0.6) and DNA/PLL (1:1.0), respectively. To TF-PEG-NP loading DNA/PLL without chloroquine, the percentage of EGFP expressing cells was 28.9% for complexes consisting of DNA/PLL (1:0.3), 38.5% and 39.7% for DNA/PLL (1:0.6) and DNA/PLL (1:1.0), respectively. In TF-PEG-NP loading DNA/PLL with chloroquine, more cells were transfected, the percentage of positive cells was 37.6% (DNA/PLL, 1:0.3), 47.1% (DNA/PLL, 1:0.6) and 45.8% (DNA/PLL, 1:1.0), which meant that the transfection efficiency of pDNA was increased by over 50 times when PLL and TF-PEG-NP were jointly used as a plasmid DNA carrier, in particular, the maximal percentage of positive cells (47.1%) from TF-PEG-NP loading DNA/PLL (1:0.6) was about 70 times the transfection efficiency of free plasmid DNA. The average cell viability of TF-PEG-NP loading DNA/PLL was about 90%, which meant that TF-PEG-NP appeared to be safer than PLL alone. As a result, TF-PEG-NP loading DNA/PLL could be a more effective non-viral vector for the delivery of pDNA.     

高分子超/微滤膜的亲水化改性:从PEG化到离子化

众多高分子超/微滤(UF/MF)膜材料存在疏水性强、抗污染能力低下、生物相容性不佳的缺点,限制了UF/MF膜的推广应用,膜材料的亲水化改性被认为是解决这一难题的有效方法,也是近年来膜材料领域的研究热点之一.高分子UF/MF膜的亲水化改性方法可分为物理改性和化学改性两大类,在这些方法中,膜表面的聚乙二醇(PEG)化和离子...