纳米铂颗粒在酶生物传感器中的应用研究

采用硼氢化钠作为还原剂制备纳米铂颗粒,并分别用羧甲基纤维素(CMC)、羟丙基纤维素(HPC)、聚丙烯酸(PAA)为保护剂,提高纳米铂溶胶的稳定性。将制备的纳米铂颗粒与聚乙烯醇缩丁醛构成复合固酶膜基质,用溶胶-凝胶法固定葡萄糖氧化酶,构建葡萄糖生物传感器。实验表明,纳米铂颗粒可以大幅度提高固定化酶的催化活性。在葡萄糖浓度为10mmol／L的溶液中,响应电流从318nA／cm^2提高到13657nA／cm^2。探讨纳米颗粒效应在固定化酶中所起的作用,并分析不同条件对酶电极响应灵敏度的影响。     

Conductometric biosensor based on glucose oxidase and beta-galactosidase for specific lactose determination in milk

This paper investigates the development of a biosensor associating two distinct enzymatic activities, that of the beta-galactosidase and that of the glucose oxidase, in order to apply it for the quantitative detection of lactose in milk. To eliminate interferences with glucose, a differential mode of measurement was used. Results show a linear calibration curve for lactose concentration between 60 and 800 μM (0.03 to 0.3 g/L). Tests with real commercial milk samples were carried out to validate the conductometric biosensor.     

Fabrication of silica/platinum core-shell particles by electroless metal plating

An electroless metal plating method was used to form Pt shells on sub-micrometer-sized silica (SiO₂) particles fabricated by a sol-gel method. The electroless metal plating method was comprised of three steps: (1) surface-modification of SiO₂ particles with polyvinylpyrrolidone (PVP) (SiO₂/PVP) or poly-diallyldimethylammonium chloride (PDADMAC) (SiO₂/PDADMAC), (2) pre-deposition of Pt nuclei or Pt fine particles on the SiO₂ particles by reducing Pt ions in the presence of SiO₂/PVP particles (SiO₂/PVP-Pt) or SiO₂/PDADMAC particles (SiO₂/PDADMAC-Pt), and (3) growth of the pre-deposited Pt by immersing the SiO₂/PVP-Pt or SiO₂/PDADMAC-Pt particles in a Pt-plating solution. The pre-deposition of Pt nanoparticles was successfully performed for the surface-modified SiO₂ particles since the surface modification possibly strengthened the affinity between the SiO₂ particle surfaces and Pt ions. The Pt nanoparticles were pre-deposited more uniformly in the case of PVP because the pre-deposition took place more slowly for the PVP, which provided uniform surface-modification followed by the uniform pre-deposition of Pt nanoparticles. The formation of Pt shells was successfully performed on the SiO₂/PVP-Pt particles in the electroless metal plating process because Pt nuclei were generated by the reduction of H₂PtCl₆ and then further deposited on the Pt particle surfaces on the SiO₂/PVP-Pt particles.     

Growth of anisotropic platinum nanostructures catalyzed by gold seed nanoparticles

This paper reports an effective method for the synthesis of platinum nanostructures with anisotropic morphologies by decomposition of platinum dichloride in oleylamine at intermediate temperatures catalyzed by gold seed nanoparticles. A small quantity of spherical gold nanoparticles formed in situ was used to trigger the nucleation and anisotropic growth of the Pt nanocrystals. By varying the amount of gold seed nanoparticles, porous flower-like, irregular polyhedron-shaped, multi-branched rod shaped, and caterpillar-like Pt nanostructures were produced in high yields at 190–240 °C in reaction times of a few minutes. Control of morphology under different conditions has been systematically studied and a kinetically controlled induced growth mechanism has been proposed. Electronic Supplementary Material  Supplementary material is available for this article at and is accessible for authorized users. This article is published with open access at Springerlink.com     

Cell membrane disruption induced by amorphous silica nanoparticles in erythrocytes,lymphocytes, malignant melanocytes,and macrophages

It is of critical importance to examine carefully the potential adverse effects of engineered nanoparticles (NPs) on human health and environments. In the present study, we have investigated the disruption of cell membranes induced by amorphous silica NPs in erythrocytes, lymphocytes (Jurkat), malignant melanocytes (B16F10), and macrophages (J774.1); these four types of mammalian cells have distinctive characteristics in terms of nucleated/non-nucleated cells, adherent/non-adherent cells, endocytosis, and phagocytosis. The silica-induced membranolysis was examined by exposing these different cells to serum-free culture media containing the amorphous silica NPs of different diameters (28, 50, 55, 156, and 461 nm) under similar conditions. We investigated how the silica-induced membranolysis of the cells of different origins is influenced by the size and dose of the silica NPs. Additionally, the interaction forces of a silica microsphere with a living cell or a giant unilamellar vesicle composed of zwitterionic phosphatidylcholine lipids were measured by colloid-probe atomic force microcopy, whereby the affinities of silica surface for plasma membranes and protein-free phospholipid membranes were estimated. Possible mechanism of the silica-induced membranolysis was discussed.     

Fabrication and characterization of platinum black and mesoporous platinum electrodes for in-vivo and continuously monitoring electrochemical sensor applications

Most electrochemical biosensors are disposable due to enzymes that are living creatures. Thus, these are limited to use in in-vivo and continuously monitoring biosensor system applications. The mesoporous (pores with a size of 2-50 nm) platinum (Pt) structure formed on a rod-shaped Pt microelectrode was reported for developments glucose sensors without any enzymes. In this paper, plane Pt electrode (non-treated), Pt black electrode, and mesoporous Pt electrode are fabricated and characterized on a silicon substrate in order to check their usability as enzymeless sensing electrodes for in-vivo and continuously monitoring electrochemical biosensors integrated with silicon CMOS read-out circuitry. The Pt black electrode with rough surface was fabricated by using an electrodeposition technique with hexachloroplatinic acid hydrate (HCPA) solutions. The proposed mesoporous Pt electrode with approximately 3 nm in pore diameter was fabricated by using an electrodeposition technique with nonionic surfactant octaethylene glycol monohexadecyl ether (C₁₆EO₈) and HCPA. The measured current responses at 40 mM glucose solution of the fabricated plane Pt, Pt black, and mesoporous Pt electrodes are approximately 12.4 nA/mm², 2.1 μA/mm², and 2.8 μA/mm², respectively. These data indicate that the mesoporous Pt electrode is much more sensitive than the other Pt electrodes and has strong potential for enzymeless electrochemical sensor applications.     

金属与非金属纳米颗粒增强葡萄糖生物传感器

为了提高葡萄糖传感器的灵敏度和抗干扰性,利用纳米增强效应,以Au、Ag、Pt、SiO2纳米颗粒及金属-无机复合纳米颗粒与聚乙烯醇缩丁醛(PVB)构成复合固定酶膜基质,采用溶胶-凝胶法固定葡萄糖氧化酶(GOD),组成葡萄糖生物传感器.研究表明,纳米颗粒可以大幅度地提高固定化酶的催化活性,增加电极的电流响应灵敏度,改进生物传感器的抗干扰性能,使信噪比提高了32倍.     

Reagentless biosensor based on glucose oxidase wired by the mediator freely diffusing in enzyme containing membrane

Wiring glucose oxidase in the membrane with an immobilized mediator is possible due to the diffusion ability of the latter, if the enzyme containing membrane is formed according to the proposed protocol, including exposing proteins to water-organic mixtures with the high content of organic solvent. In the course of the study, the new glucose oxidase mediator, unsubstituted phenothiazine, was discovered. The diffusion coefficient of the mediator in the resulting membrane is independent of the presence of enzyme. The cyclic voltammograms of the enzyme electrode after appearance of the only glucose in solution obtain a well-defined catalytic shape, which is normally observed for both the enzyme and the mediator in solution. Analytical performances of the resulting biosensor are comparable to the advanced second generation ones, which, however, require covalent linking of the mediator either to the membrane forming polymer or to the enzyme. Even without such covalent linking, the reported biosensor is characterized by an appropriate long-term operational stability allowing reagentless sensing.     

Application of fractional factorial design for green synthesis of cyano-modified silica nanoparticles: Chemometrics and multifarious response optimization

Cyano-functionalized spherical silica nanoparticles (SNPs) were synthesized via Stöber method. A 2?^k-p_IV–fractional factorial design (2^k-p_IV–FFD) was used to smartly prepare monodispersed evenly distributed SNPs. Six factors were considered; concentrations of tetraethylorthosilicate (TEOS), 3-Cyanopropyltriethoxysilane (CPTS), water, and ammonia, reaction time (RT) and stirring time (ST). Two responses; particle size (PS, measured by SEM) and particle-size distribution (PSD, calculated as standard deviation, ±SD) were measured. Control charts were used to decide on impacts of linear and two-way interactions on both responses. Derringer’s function was used to consolidate these multifarious responses into a uniform execution characteristic. Both screening and optimization were always accompanied by ANOVA testing at a 95.0% confidence interval (CI). The ideal synthetic conditions were obtained from the composite desirability plots. Cyano-functionalized SNPs with an average PS of 474.04?±?86.71?nm were produced. Raman spectroscopy and FTIR were used to confirm the functionalization process. Thermogravimetric analysis (TGA) was used to evaluate the thermal behavior of synthesized particles.     

Enhancing the grafting of poly(2-hydroxyethyl methacrylate) on silica nanoparticles (SiO2-g-PHEMA) by the sequential UV-induced graft polymerization with a multiple-UV irradiation

Grafting of poly(2-hydroxyethyl methacrylate) on silica nanoparticles was accomplished via the sequential UV-induced graft polymerization. Under UV-irradiation, the silica was functionalized with the surface initiator, benzophenone (BP) and subsequently graft-polymerized with 2-hydroxyethyl methacrylate (HEMA). The grafting on the silica particles was confirmed by DSC analysis which revealed a shift of the glass transition temperature (T_g) of grafted PHEMA to higher temperature than T_g of ungrafted PHEMA. A significant improvement in the grafting efficiency and the grafting percentage was achieved when a sequential grafting approach was taken, employing multiple UV exposures. Using this approach, the efficient chain extension from the grafted-PHEMA was possible without producing significant amounts of ungrafted PHEMA when low HEMA concentrations were used during each UV-exposure.     

Ag-Cu离子注入SiO2玻璃后形成纳米颗粒的研究

Ag、Cu离子经200和110keV加速后分别以5×1016和1.5×1017ions /cm2的剂量在室温下先后注入到非晶SiO2玻璃中.注入后样品的光学吸收谱显示两个吸收峰,其峰位为407和569nm,分别对应单独Ag和单独Cu纳米颗粒的等离子体共振吸收峰,样品在还原-保护气氛下退火后吸收峰峰强明显增加.样品的透射电镜选区电子衍射花样含有Ag、Cu两套衍射环,透射电镜的明场像观察到大量的纳米颗粒呈现出中心亮斑特征.在样品倾转过程中,中心亮斑特征依然存在,证实这种现象是离子辐照产生的纳米空位团簇.扫描透射电子显微镜高角环形暗场像进一步证实了这一点.综上所述,样品中形成了单Ag和单Cu包裹空位团簇的纳米颗粒.     

Effect of annealing on the structural and optical properties of non-coated and silica-coated ZnS:Mn nanoparticles

The Mn²⁺-doped ZnS nanoparticles stabilized by sodium citrate were synthesized through a simple chemical route. Using the ZnS:Mn nanoparticles as seeds, the silica-coated ZnS:Mn nanocomposites were formed in isopropanol by the controlled hydrolysis of tetraethyl orthosilicate. The photoluminescence spectra confirmed that the Mn²⁺ ions were incorporated into the ZnS nanoparticles. The annealing effect on the structural and optical properties of these particles was studied over a range of 100–400 °C. The results of X-ray diffraction and photoluminescence showed that the silica shell not only improved the thermal stability but also resisted the lattice-deformation and oxidation of the particles. The thermal analysis further confirmed that the non-coated ZnS:Mn nanoparticles were unstable beyond 200 °C.     

TEM and HRTEM evidence for the role of ligands in the formation of shape-controlled platinum nanoparticles

The morphology of platinum nanoparticles synthesized using an organometallic approach from PtMe(2) (C(8) H(12) ) is influenced by the nature of the ligands used as stabilizing agents. The use of long alkyl chain amines leads to the formation of multipodal nanoparticles that transform into compact nano-objects, adopting cubic, truncated cubic, or cuboctahedral shapes. In contrast, the use of diamine ligands allows the growth of compact (111) arrowlike faces, forming polycrystalline nanoparticles of an overall desert-rose aspect. Different reaction parameters are studied ([ligand]/[metal] ratio, temperature, solvent identity) in order to optimize the various shapes.     

Investigation into the synergistic effects in ternary cementitious systems containing portland cement, fly ash and silica fume

This research was primarily conducted to verify the presence of synergistic effects in ternary cementitious systems containing portland cement (OPC), class C fly ash (FA) and silica fume (SF). A subsequent objective of the study was to quantify the magnitude of the synergy and to determine its source. For a ternary mixture containing 20% FA and 5% SF by mass, the synergistic effect was observed mostly at later ages (7 days onward) and it resulted in an increased compressive strength and resistance to chloride ion penetration as well as a reduced rate of water absorption (sorptivity) compared to predictions based on individual effects of FA and SF in respective binary systems. The observed synergy was attributed to both chemical and physical effects. The chemical effect manifested itself in the form of an increased amount of hydration products. The physical effect associated with packing density was, somewhat contrary to general belief, not due to an optimized particle size distribution of the binder components of the ternary cementitious system. Instead, it was the result of smaller initial inter-particle spacing caused by lower specific gravities of both FA and SF which, in turn, led to a lower volumetric w/cm. If the mixture design was adjusted to account for these differences, the physical effect would be diminished.     

Optimization of PLGA nanoparticles formulation containing L-DOPA by applying the central composite design

The aim of this work was to prepare L-DOPA loaded poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles by a modified water-in-oil-in-water (W₁/O/W₂) emulsification solvent evaporation method. A central composite design was applied for optimization of the formulation parameters and for studying the effects of three independent variables: PLGA concentration, polyvinyl alcohol (PVA) concentration and organic solvent removal rate on the particle size and the entrapment efficiency (response variables). Second-order models were obtained to adequately describe the influence of the independent variables on the selected responses. The analysis of variance showed that the three independent variables had significant effects (p < 0.05) on the responses. The experimental results were in perfect accordance with the predictions estimated by the models. Using the desirability approach and overlay contour plots, the optimal preparation area can be highlighted. It was found that the optimum values of the responses could be obtained at higher concentration of PLGA (5%, w/v) and PVA (6%, w/v); and faster organic solvent removal rate (700 rpm). The corresponding particle size was 256.2 nm and the entrapment efficiency was 62.19%. FTIR investigation confirmed that the L-DOPA and PLGA polymer maintained its backbone structure in the fabrication of nanoparticles. The scanning electron microscopic images of nanoparticles showed that all particles had spherical shape with porous outer skin. The results suggested that PLGA nanoparticles might represent a promising formulation for brain delivery of L-DOPA. The preparation of L-DOPA loaded PLGA nanoparticles can be optimized by the central composite design.     

Ag nanoparticles capped by a nontoxic polymer: Electrochemical and spectroscopic characterization of a novel nanomaterial for glucose detection

Modified electrodes with metal or metal oxides nanoparticles are particularly appealing to improve sensor performances and fabricate miniaturized devices, as required also in glucose detection. A Pt electrode modified by drop casting of a novel nanostructured film based on silver nanoparticles (Ag-NPs) capped in a commercial nontoxic polyvinyl alcohol (PVA) matrix is proposed here as a valid alternative to classical glucose (bio)sensors. The extensive electrochemical and spectroscopic characterization by X-ray Photoelectron Spectroscopy (XPS) of this advanced nanomaterial is presented to study its response to glucose and to investigate the chemical nature of deposited Ag.     

Design of high payload PLGA nanoparticles containing melittin/sodium dodecyl sulfate complex by the hydrophobic ion-pairing technique

The water-soluble peptide, melittin, was modified with an anionic agent, sodium dodecyl sulfate by hydrophobic ion-pairing. Investigations showed that the formed complex was very soluble in organic solvent, especially, in dimethylsulfoxide and dehydrated alcohol. Furthermore, the physiochemical properties of the complex in the solid state or in an aqueous medium were characterized using octanol/water partition measurement, Fourier transform infrared spectroscopy, and differential scanning calorimetry. The complex was formulated into poly(d,l-lactide–co-glycolide acid) nanoparticles by an emulsion solvent diffusion method. It was found that the nanoparticles of about 130 nm in size can be produced with a high encapsulation efficiency, and the entrapment of nanoparticles prepared with the formed complex increased from about 50% to nearly 100% compared with that for pure melittin. Moreover, the growth inhibitory effects of modified melittin and melittin-loaded nanoparticles in breast cancer MCF-7 cells were not changed comparing with free melittin as determined by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay.     

MGF纳米微球静电纺丝材料及对hMSC增殖和迁移的影响

力生长因子(MGF,mechano-growth factor)是由Yang等发现并命名,是细胞对力刺激响应的产物,对人骨髓间充质干细胞(hMSC,human mesenchymal stem cells)有重要的调控作用。利用静电纺丝技术(electrospinning)及相分离法制备了基于聚已内酯(PCL,polycaprolactone)的含MGF的葡聚糖纳米微球的纤维支架,间接性地保护了MGF活性,采用扫描电镜和透射电镜对微球和纤维支架进行了基础表征。同时,进一步采用激光共聚焦显微镜和酶标仪分别考察了纳米微球中释放出来的MGF对细胞的作用。研究结果表明含MGF的纳米微球支架能够显著促进hMSC增殖和迁移。此静电纺丝纤维支架能够较好地保护MGF的活性,有望在组织工程领域得到应用。     

Sharp cubic and octahedral morphologies of poly(vinylpyrrolidone)-stabilised platinum nanoparticles by polyol method in ethylene glycol: their nucleation,growth and formation mechanisms

Pt nanoparticles (NPs) were synthesised by a modified polyol method with the addition of silver nitrate. The results showed that the specific shapes of Pt NPs were influenced by the relevant factors, which are the contents of silver nitrate, synthetic time and temperature. A small content of silver nitrate has played an important role in determining their final shapes of platinum NPs. We observed that Pt NPs in the forms of very sharp shapes such as Pt cubes, octahedrons, cuboctahedrons and tetrahedrons have been obtained. In addition, the shape growth mechanisms and formation of Pt NPs have been studied. They exist in both cubic and octahedral shapes. Importantly, Pt nanocrystals can grow into main cubic and octahedral shapes for a short time less than 15?min. Moreover, Pt nanocrystals can also grow into different shapes from cubic and octahedral into spherical ones for several hours. Especially, they exhibited interesting shapes of multiple-branched Pt nanostructures because of their overgrowth and aggregations. Clearly, large cubic and octahedral Pt NPs of 160?nm diameter were observed. The growth and formation of large cubic and octahedral Pt NPs were due to the aggregation of Pt clusters or initial Pt seeds, even small Pt nanocrystals.