Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis,Characterization, and Evaluation

There is an increasing interest in cationic polymers as important constituents of non-viral gene delivery vectors. In the present study, we developed a versatile synthetic route for the production of covalent polymeric conjugates consisting of water-soluble depolymerized chitosan (dCS; M_W 6–9 kDa) and low molecular weight polyethylenimine (PEI; 2.5 kDa linear, 1.8 kDa branched). dCS-PEI derivatives were evaluated based on their physicochemical properties, including purity, covalent bonding, solubility in aqueous media, ability for DNA condensation, and colloidal stability of the resulting polyplexes. They were complexed with non-integrating DNA vectors coding for reporter genes by simple admixing and assessed in vitro using liver-derived HuH-7 cells for their transfection efficiency and cytotoxicity. Using a rational screening cascade, a lead compound was selected (dCS-Suc-LPEI-14) displaying the best balance of biocompatibility, cytotoxicity, and transfection efficiency. Scale-up and in vivo evaluation in wild-type mice allowed for a direct comparison with a commercially available non-viral delivery vector (in vivo-jetPEI). Hepatic expression of the reporter gene luciferase resulted in liver-specific bioluminescence, upon intrabiliary infusion of the chitosan-based polyplexes, which exceeded the signal of the in vivo jetPEI reference formulation by a factor of 10. We conclude that the novel chitosan-derivative dCS-Suc-LPEI-14 shows promise and potential as an efficient polymeric conjugate for non-viral in vivo gene therapy.     

硅疏水改性聚胺阻垢剂的制备及其性能

用γ-缩水甘油醚氧丙基三甲氧基硅烷(KH560)、1-氯辛烷对低相对分子质量聚乙烯亚胺(PEI)进行疏水接枝改性,得到了含硅疏水改性聚胺阻垢剂。通过FTIR、TG、粒径分析、Zeta电位测试等分析改性后聚胺阻垢剂的结构与性能。结果表明,改性后产物热稳定性提高、黏度增大、表面张力降低、溶液粒径增大。将改性聚胺阻垢剂用于分散铝硅酸钠颗粒,对处理前后的颗粒进行红外光谱表征,并通过对悬浮分散液进行稳定性、颗粒沉降等测试,评价聚胺阻垢剂的阻垢分散性能并初步讨论其分散机理。当m(PEI, M_w=10000)∶m(KH560)∶m(1-氯辛烷)=10∶2∶0.5,反应时间6 h,反应温度75℃时,合成的阻垢剂分散性能最佳。当结疤的质量浓度为30 g/L时,悬浮液颗粒沉降速度达到最低值0.047 cm/d,沉降速率减小程度达到94.8%,分散性显著提高。经过改性后的PEI在氧化铝工业生产中具有高效的阻垢作用,可通过在工厂进行条件优化,进一步替代传统的强酸型阻垢剂,减小管道腐蚀程度,降低能耗。     

尾静脉注射方法考察Stearic-PEI/SPIO 纳米基因载体的基因转染性能及磁共振可视化效果研究

实验应用硬脂酸-聚乙烯亚胺包裹超顺磁氧化铁(Superparamagnetic Iron Oxide,SPIO)装载目的基因构建可视化 纳米复合物,进行 BABL/c 雌性小鼠体内磁共振成像实验、组织学检测,探讨其是否能够有效地介导体内基因表达以及 产生磁共振成像(Magnetic Resonance Imaging,MRI)效果。MRI 实验结果显示,对小鼠尾静脉注射复合物 0.5 小时后进 行磁共振扫描,与注射生理盐水的对照组相比实验组 T2 加权信号明显降低,信号强度仅为对照组的 35%;48 小时后, 实验组 T2 加权信号强度略有回升,但与对照组信号强度相比仍存在显著差异。组织学结果显示可视化基因纳米复合物 能够穿过血管内皮细胞进入肝脏组织,并释放 DNA 在细胞内成功表达,但基因表达效果较差。综合 MRI 成像及组织检 查实验结果可知,该纳米颗粒具有优良的 MRI 成像性能,但通过尾静脉注射方法基因转染效率较低。     

Low molecular weight PEI-grafted carboxyl-modified soybean protein as gene carriers with reduced cytotoxicity and greatly improved transfection in vitro

SA-SP-PEI were synthesized via acylation reaction between carboxyl-modified soybean protein (SA-SP) and branched polyethylenimine (PEI) with molecular weight of 600, 1200, and 1800?Da, and designed as SA-SP-PEI600, SA-SP-PEI1200, and SA-SP-PEI1800, respectively. SA-SP-PEI could effectively condense plasmid DNA into nanoscale polyplexes and protect them from enzymatic digestion. MTT assay revealed that SA-SP-PEI exhibited reduced cytotoxicity on 293?T and SH-SY5Y cells. SA-SP-PEI1800/DNA complexes hold highest transfection efficiency on 293?T and SH-SY5Y cells with or without 10% serum, which was owing to its better serum stable and improved biocompatibility. Such polycationic soybean proteins have great potentials as gene carriers by further optimization.     

醇脱氢酶在聚乙烯膜表面的固定化研究

以聚丙烯酸（PAA）改性的聚乙烯（PE）膜为载体，研究了醇脱氢酶（ADH）的两种固定化路线，并以甲醛为底物考察了固定化酶的催化性能。路线1用聚乙烯亚胺（PEI）进一步改性，使用戊二醛（GA）固定化ADH。最优固定化pH为6.0，温度为5～15℃，酶浓度为1.0 mg/ml，GA浓度为0.01%(质量)；固定化酶的最适反应pH为6.5，温度为15～30℃，反应速率最高为9.6 μmol/(L·min)；重复利用10次后可保持47.3%的活性。路线2以PAA-PE为载体，用1-(3-二甲氨基丙基)-2-乙基碳二亚胺盐酸盐（EDC）和N-羟基琥珀酰亚胺（NHS）为活化剂，固定化ADH。EDC和NHS最优摩尔比为1∶0.5，固定化时间为24 h；固定化酶的最适反应pH为6.5，温度为20～37℃，反应速率为15.58 μmol/(L·min)；重复利用10次后可保持53.8%的活性。     

Dendrimer as nanocarrier for drug delivery

Dendrimers are novel three dimensional, hyperbranched globular nanopolymeric architectures. Attractive features like nanoscopic size, narrow polydispersity index, excellent control over molecular structure, availability of multiple functional groups at the periphery and cavities in the interior distinguish them amongst the available polymers. Applications of dendrimers in a large variety of fields have been explored. Drug delivery scientists are especially enthusiastic about possible utility of dendrimers as drug delivery tool. Terminal functionalities provide a platform for conjugation of the drug and targeting moieties. In addition, these peripheral functional groups can be employed to tailor-make the properties of dendrimers, enhancing their versatility. The present review highlights the contribution of dendrimers in the field of nanotechnology with intent to aid the researchers in exploring dendrimers in the field of drug delivery.     

Electrochemical regeneration of partially ethoxylated polyethylenimine used in the polymer-supported ultrafiltration of copper

This work is focussed on the application of an electrochemical technology to regenerate the bonding agent of a polymer-supported ultrafiltration process, technique commonly used to remove metal ions from wastewaters. To do this, a batch rotating-electrode electrochemical cell has been set up to recover the copper bind to partially ethoxylated polyethyleneimine by electrodeposition. The influence of the main parameters (current efficiency, stirring rate, pH, electrode material and nature of the counterion) on the performance of the process has been studied. Current efficiency is clearly enhanced with the stirring rate, reaching values as high as 0.93 for the optimum working conditions. Regarding pH, this variable has been observed to play an important role in process efficiency. Thus, it has been found that the reactor can operate at pH 4 without affecting process performance. This is a clear advantage with respect to other regeneration techniques and also to previous works of electrochemical regeneration. The electrode material seems to have a clear influence on process behaviour and especially on the appearance of a first transitory stage with lower current efficiency. Finally, the electrolyte salt also influences significantly on the results and the presence of sulphate as counterion leads to the best system performance.     

Facile fabrication of graphene oxide-polyethylenimine composite and its application for the Cr(VI) removal

A binary composite consisting of graphene oxide (GO) and polyethylenimine (PEI) was fabricated by a facile physical mixing. Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscope (FE-SEM), thermo-gravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS) and Zeta potential were used to characterize the prepared graphene oxide-polyethylenimine composite (GOPC). A series of experiments were carried out to investigate the effects of some important parameters, such as molecular weight of PEI, pH, time and temperature, on the adsorption efficiency of GOPC. Due to the high amine density of GOPC, its adsorption for Cr(VI) occurred more easily at lower pH mainly via electrostatic interaction. The adsorption process matched well with the Langmuir isotherm model and the pseudo-second-order kinetic model. The maximum adsorption capacity from the Langmuir model was 370.37 mg/g at pH 2.0 and 45°C for GOPC. Thermodynamic parameters revealed spontaneous and endothermic nature of the Cr(VI) adsorption onto GOPC. The main adsorption mechanism of GOPC toward Cr(VI) was electrostatic interaction. The adsorption-desorption experiments suggested GOPC was easily recycled and its stable adsorption capacity endowed it great potential as an adsorbent of Cr(VI) from wastewater.