Development and characterization of new insulin containing polysaccharide nanoparticles

A nanoparticle insulin delivery system was prepared by complexation of dextran sulfate and chitosan in aqueous solution. Parameters of the formulation such as the final mass of polysaccharides, the mass ratio of the two polysaccharides, pH of polysaccharides solution, and insulin theorical loading were identified as the modulating factors of nanoparticle physical properties. Particles with a mean diameter of 500 nm and a zeta potential of approximately −15 mV were produced under optimal conditions of DS:chitosan mass ratio of 1.5:1 at pH 4.8. Nanoparticles showed spherical shape, uniform size and good shelf-life stability. Polysaccharides complexation was confirmed by differential scanning calorimetry and Fourier transformed infra-red spectroscopy. An association efficiency of 85% was obtained. Insulin release at pH below 5.2 was almost prevented up to 24 h and at pH 6.8 the release was characterized by a controlled profile. This suggests that release of insulin is ruled by a dissociation mechanism and DS/chitosan nanoparticles are pH-sensitive delivery systems. Furthermore, the released insulin entirely maintained its immunogenic bioactivity evaluated by ELISA, confirming that this new formulation shows promising properties towards the development of an oral delivery system for insulin.     

Preparation and properties of ionically cross‐linked chitosan nanoparticles

Chitosan nanoparticles were fabricated by a method of tripolyphosphate (TPP) cross‐linking. The influence of fabrication conditions on the physical properties and drug loading and release properties was investigated by transmission electron microscopy (TEM), dynamic light scattering (DLS), and UV–vis spectroscopy. The nanoparticles could be prepared only within a zone of appropriate chitosan and TPP concentrations. The particle size and surface zeta potential can be manipulated by variation of the fabrication conditions such as chitosan/TPP ratio and concentration, solution pH and salt addition. TEM observation revealed a core–shell structure for the as‐prepared nanoparticles, but a filled structure for the ciprofloxacin (CH) loaded particles. Results show that the chitosan nanoparticles were rather stable and no cytotoxicity of the chitosan nanoparticles was found in an in vitro cell culture experiment. Loading and release of CH can be modulated by the environmental factors such as solution pH and medium quality. Copyright © 2008 John Wiley & Sons, Ltd.     

烷基壳聚糖纳米微球的制备及其紫杉醇负载研究

在碱性条件下通过卤代烃的N-烷基取代反应制备得到烷基壳聚糖衍生物。X射线光电子能谱计算结果表明烷基取代反应主要发生在壳聚糖的氨基上。动态光散射研究表明，该衍生物在水中可自动形成粒径在10—200nm范围的纳米微粒，负载紫杉醇后微球的粒径增大，在磷酸缓冲液（pH=7．4）中体外释放研究表明，随着烷基链长的增加，紫杉醇在磷酸缓冲液中达到平衡时的浓度降低。     

Immunonanogold-catalytic resonance scattering spectral assay of trace human chorionic gonadotrophin

Gold nanoparticles in diameter of 10 nm were used to label rabbit anti-human chorionic gonadotrophin (RhCG) antiserum to obtain a resonance scattering spectral probe (AuRhCG) for human chorionic gonadotrophin (hCG). The immunoreaction between AuRhCG and hCG take place to form hCG–AuRhCG immunocomplex in pH 5.0 citric acid–Na₂HPO₄ buffer solution. The immunocomplex solutions were centrifuged to obtain the supernatant solution. The AuRhCG in the supernatant solution exhibited strong catalytic effect on the particle reaction between Ag⁺ and hydroquinone to produce gold–silver composite particles in pH 3.4 citric acid–trisodium citrate buffer solution. There is a stronger resonance scattering (RS) peak at 423 nm for the particles. With the addition of hCG, the AuRhCG in the supernatant solution decreased, and the RS intensity at 423 nm decreased. The decreased RS intensity ΔI_423 nm was proportional to the concentration of hCG in the range of 2.5–208.3 mIU/mL with a detection limit of 0.83 mIU/mL. This method has been applied to the determination of hCG in urine samples, with satisfactory results.     

Nanoparticles prepared by self-assembly of Chitosan and poly-γ-glutamic acid

The present paper describes the preparation and characterization of novel biodegradable nanoparticles based on self-assembly of poly-gamma-glutamic acid (γ-PGA) and chitosan (CH). The nanosystems were stable in aqueous media at low pH conditions. Solubility of the systems was determined by turbidity measurements. Surface charge and mobility were measured electrophoretically. The particle size and the size distribution of the polyelectrolyte complexes were identified by dynamic light scattering and transmission electron microscopy (TEM). It was found that the size and size distribution of the nanosystems depends on the concentrations of γ-PGA and CH solutions and their ratio as well as on the pH of the mixture and the order of addition. The diameter of individual particles was in the range of 20–285 nm measured by TEM, and the average hydrodynamic diameters were between 150 and 330 nm. These biodegradable, self-assembling stable nanocomplexes might be useful for several biomedical applications.     

Self-aggregated nanoparticles of cholesterol-modified glycol chitosan conjugate: Preparation, characterization, and preliminary assessment as a new drug delivery carrier

Cholesterol-modified glycol chitosan (CHGC) conjugate was synthesized and characterized by FTIR and ¹H NMR. The degree of substitution (DS) was 6.7 cholesterol groups per 100 sugar residues of glycol chitosan. CHGC formed self-aggregated nanoparticles with a roughly spherical shape and a mean diameter of 228 nm by probe sonication in aqueous medium. The physicochemical properties of the self-aggregated nanoparticles were studied using dynamic light scattering (DLS), transmission electron microscopy (TEM) and fluorescence spectroscopy. The critical aggregation concentration (CAC) of self-aggregated nanoparticles in aqueous solution was 0.1223 mg/mL. Indomethacin (IND), as a model drug, was physically entrapped into the CHGC nanoparticles by dialysis method. The characteristics of IND-loaded CHGC (IND-CHGC) nanoparticles was analyzed using DLS, TEM and high performance liquid chromatography (HPLC). The IND-CHGC nanoparticles were almost spherical in shape and their size increased from 275 to 384 nm with the IND-loading content increasing from 7.14% to 16.2%. The in vitro release behavior of IND from CHGC nanoparticles was studied by a dialysis method in phosphate buffered saline (PBS, pH 7.4). IND was released in a biphasic way. The initial rapid release in 2 h and slower release for up to 12 h were observed. The results indicated that CHGC nanoparticles had a potential as a drug delivery carrier.     

Synthesis,Characterization of Copper-Loaded Carboxymethyl-Chitosan Nanoparticles with Effective Antibacterial Activity

Summary: Copper-loaded carboxymethyl-chitosan (CMCS-Cu) nanoparticles were successfully prepared by chelation under aqueous conditions. The effect of degree of deacetylation and substitution, the molecular weight of CMCS, CMCS concentration, Cu(II) ions concentration, pH value of the solution, as well as temperature, on the morphology of the yielded particles were systematically investigated. The physicochemical properties of the particles were determined by size and zeta potential analysis, FTIR analysis, DLS, TEM, SEM and XRD pattern. FTIR and XRD revealed that Cu (II) ions and CMCS formed a chelate complex. The size of CMCS-Cu particles shows a good consistency by DLS, TEM, and SEM. The nanoparticles with the size of about 70 nm have been prepared at 0.13 wt% CMCS, 16 mmol/L Cu(II) ions, pH value 4.56 at 25 °C. The antibacterial activity of CMCS, CMCS-Cu normal particles with the size of about 1000 nm and CMCS-Cu nanoparticles with the size of less than 100 nm against Staphylococcus aureus was evaluated by vibration method. Results show that the antibacterial efficiency of nanoparticles reached 99%, which is much more efficient than 68.9% of the normal one and 6.1% of CMCS. CMCS-Cu nanoparticles were proved to be a good novel antibacterial material.     

Improving Lanthanide Nanocrystal Colloidal Stability in Competitive Aqueous Buffer Solutions using Multivalent PEG-Phosphonate Ligands

The range of properties available in the lanthanide series has inspired research into the use of lanthanide nanoparticles for numerous applications. We aim to use NaLnF(4) nanoparticles for isotopic tags in mass cytometry. This application requires nanoparticles of narrow size distribution, diameters preferably less than 15 nm, and robust surface chemistry to avoid nonspecific interactions and to facilitate bioconjugation. Nanoparticles (NaHoF(4), NaEuF(4), NaGdF(4), and NaTbF(4)) were synthesized with diameters from 9 to 11 nm with oleic acid surface stabilization. The surface ligands were replaced by a series of mono-, di-, and tetraphosphonate PEG ligands, whose synthesis is reported here. The colloidal stability of the resulting particles was monitored over a range of pH values and in phosphate containing solutions. All of the PEG-phosphonate ligands were found to produce non-aggregated colloidally stable suspensions of the nanoparticles in water as judged by DLS and TEM measurements. However, in more aggressive solutions, at high pH and in phosphate buffers, the mono- and diphosphonate PEG ligands did not stabilize the particles and aggregation as well as flocculation was observed. However, the tetraphosphonate ligand was able to stabilize the particles at high pH and in phosphate buffers for extended periods of time.     

Nanoparticles from Chitosan

Novel biodegradable nanoparticles were synthesized by chemical modification of the chitosan linear chain. A natural dicarboxylic acid (malic acid) was used as a crosslinking agent for intramolecular covalent condensation reaction to obtain hydrophilic nanoparticles based on chitosan. A variety of methods including, solubility studies, laser light scattering (DLS), transmission electron microscopy (TEM) and nuclear magnetic resonance (NMR) was used to characterize the crosslinked macromolecules. The prepared biodegradable chitosan nanoparticles, soluble in aqueous media, might be useful for various biomedical applications, like injectable drug- or gene-delivery systems.     

Submicrometer intermediates in the citrate synthesis of gold nanoparticles: new insights into the nucleation and crystal growth mechanisms

The reduction of tetrachloroaurate by citrate ions in aqueous solutions yielding gold nanoparticles (GNPs) has been studied using in situ tapping mode atomic force microscopy (AFM), UV-vis absorption and dynamic light scattering (DLS) spectroscopies, small-angle X-ray scattering (SAXS) along with ex situ TEM, EDX and XPS. Special attention is given to mesoscale intermediates responsible for the intense coloring of the transient solutions and their role in nucleation and crystal growth. AFM detects liquid droplet-like domains, globules 30-50 nm in diameter arranged in submicrometer aggregates in the gray and blue solutions, and well separated individual particles in the final red sols. DLS shows abrupt appearance of species about 30 nm and larger but not growing Au nanoparticles, while SAXS reveals gradually increasing nanoparticles and no aggregates. The mesoscale structures observed in TEM become looser as the reaction proceeds; they contain signatures of oxidized Au and other solutes. The results are interpreted in terms of decomposition of supersaturated solutions to afford domains ("dense droplets") enriched by gold, and then, after nucleation and coalescence of Au nuclei inside them, rather slow growth of gold nanoparticles within the associated globules; the color changes of the transient solutions are due to increasing interparticle distances.     

Preparation and properties of inhalable nanocomposite particles: effects of the size, weight ratio of the primary nanoparticles in nanocomposite particles and temperature at a spray-dryer inlet upon properties of nanocomposite particles

Nanoparticles are expected to be applicable to inhalation as carrier but there exist disadvantages because of their size. Their deposition dose to the lung will be small. To overcome this problem and utilize nanoparticles for inhalation, we have prepared nanocomposite particles as drug carriers targeting lungs. The nanocomposite particles are prepared as drug-loaded nanoparticles–additive complex to reach deep in the lungs and to be decomposed into nanoparticles when they deposit into lung. In this study, we examined the effect of preparation condition – inlet temperature, size of primary nanoparticles and weight ratio of primary nanoparticles – on the property of nanocomposite particles.

When the size of primary nanoparticles was 400 nm and inlet temperature was 90 °C, only the nanocomposite particles containing between 45 and 55% of primary nanoparticles could be decomposed into nanoparticles in water. On the other hand, when the inlet temperature was 80 °C, nanocomposite particles were decomposed into nanoparticles independent of the weight ratio of primary nanoparticles. Also, the aerodynamic diameter of the nanocomposite particles was between 1.5 and 2.5 μm, independent of the weight ratio of primary nanoparticles.

When the size of primary nanoparticles was 200 nm and inlet temperature was 70 °C, nanocomposite particles were decomposed into nanoparticles independent of the weight ratio of primary nanoparticles. Also, the aerodynamic diameters of them were almost 2.0 μm independent of the weight ratio of primary nanoparticles. When the nanocomposite particles containing nanoparticles with the size of 200 nm are prepared at 80 °C, no decomposition into nanoparticles was observed in water.

Fine particle values, FPF, of the nanocomposite particles were not affected by the weight ratio of primary nanoparticles when they were prepared at optimum inlet temperature.     

Green Synthesis and Characterization of Silver Nanoparticles Using Ascomycota Fungi Penicillium nalgiovense AJ12

A new method for green synthesis of silver nanoparticles using the cell-free filtrate of Penicillium nalgiovense AJ12 as reducing and protecting agent was described. The pathway is based on the reduction of Ag¹⁺ by protein(s). Various techniques such as UV–Vis spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier transform infrared and Zeta potential measurements were used to characterize the silver nanoparticle obtained. The results revealed synthesis of the spherical silver nanoparticles coated with protein(s). The average size of the particles obtained from TEM was 15.2 ± 2.6 nm. DLS measurements showed that the particle size was higher than that estimated from TEM measurements and was 25.2 ± 2.8 nm. Studies on the role of the cell-free filtrate proteins in the synthesis of silver nanoparticles indicated that the process is non-enzymatic but involves amino acids interactions with silver ions. It was found that the aqueous silver nanoparticles suspensions exhibited excellent stability over a wide range of ionic strength, pH and temperature.     

壳聚糖纳米粒子荧光探针的制备和表征

通过低分子量的壳聚糖(LCS)聚阳离子与三聚磷酸钠(TPP)的静电作用制备纳米级壳聚糖微球,并利用壳聚糖链上丰富的氨基与荧光素异硫氰酸酯(FITC)反应从而制备纳米壳聚糖微球荧光探针(NFCS)。结果表明,当壳聚糖分子量为60000,LCS与TPP的质量比为6∶1时,可得到粒度均一的球形纳米粒子,平均粒径为40±3 nm。荧光倒置显微镜观察证实FITC结合到壳聚糖微球上。荧光光谱分析显示NFCS的最大激发波长、最大发射波长与游离态FITC无显著差异。光漂白实验证实NFCS的稳定性比游离态FITC有显著提高。     

Preparation and characterization of poly(acrylic acid)-based nanoparticles

The present investigation describes the synthesis and characterization of nanoparticles based on poly(acrylic acid) (PAA) intramolecularly cross-linked with diamine, 2,2′-(ethylenedioxy)bis(ethylamine), using water-soluble carbodiimide. The aqueous colloid dispersions of nanoparticles were clear or mildly opalescent depending on the ratio of cross-linking, pH of the solution, and the molecular weight of PAA, finding consistent with values of transmittance between 3% and 99%. The structure was determined by nuclear magnetic resonance spectroscopy, and the particle size was identified by dynamic light scattering (DLS) and transmission electron microscopy (TEM) measurements. It was found that particle size depends on the pH, and at a given pH, it was caused by the ratio of cross-linking and the molecular weight of PAA. Particle size measured by TEM varied in the range of 20 and 80 nm. In the swollen state, the average size of the particles measured by DLS was in the range of 35–160 nm.     

Comparison of nanoparticle size and electrophoretic mobility measurements using a carbon-nanotube-based coulter counter, dynamic light scattering, transmission electron microscopy, and phase analysis light scattering

The precision and accuracy of measurements of the diameter and electrophoretic mobility (mu) of polymeric nanoparticles is compared using four different analytical approaches: carbon-nanotube-based Coulter counting, dynamic light scattering (DLS), transmission electron microscopy (TEM), and phase analysis light scattering (PALS). Carbon-nanotube-based Coulter counters (CNCCs) use a 132 nm diameter channel to simultaneously determine the diameter (28-90 nm) and mu value for individual nanoparticles. These measurements are made without calibration of the CNCC and without labeling the sample. Moreover, because CNCCs measure the properties of individual particles, they provide true averages and polydispersities that are not convoluted into the intrinsic instrumental response function of the CNCC. CNCCs can be used to measure the size of individual nanoparticles dispersed in aqueous solutions, which contrasts with the TEM-measured size of individual dehydrated particles and the ensemble size averages of dispersed particles provided by DLS. CNCCs provide more precise values of mu than PALS.     

A novel nonaqueous route to V2O3 and Nb2O5 nanocrystals

The reaction between transition metal alkoxides and benzyl alcohol provides a novel soft chemistry route to metal oxide nanoparticles. The method allows the preparation of nanocrystals of two important transition metal oxides, namely V₂O₃ and Nb₂O₅. Although the reaction temperatures of 200–220 °C are comparably low, the obtained particles are highly crystalline. According to TEM investigations, the V₂O₃ crystals exhibit particle sizes between 20 and 50 nm, and the Nb₂O₅ crystals display platelet-like particle shapes with sizes of 50–80 nm, without any indications of amorphous character.     

In situ synthesis of polysaccharide nanoparticles via polyion complex of carboxymethyl cellulose and chitosan

Biocompatible polymer-magnetite hybrid nanoparticles were prepared by means of in situ synthesis of magnetite within polysaccharide hydrogel nanoparticles. Hydrogel nanoparticles were first fabricated by blending high-molecular-weight carboxymethyl cellulose as an anionic polymer, and low-molecular-weight chitosan as a cationic polymer to form polyion complexes (CC particles). These polyion complexes were then chemically crosslinked using genipin, a bio-based cross-linker, to form stable nanoparticles having a semi-IPN structure (CCG particles). Magnetite was lastly synthesized within CCG particles by the coprecipitation method to obtain polymer-magnetite hybrid nanoparticles (CCGM particles). The formations of CC, CCG and CCGM particles were mainly observed by transmittance, absorbance of genipin and TEM, respectively, and their hydrodynamic diameters and zeta-potentials were analyzed. It was confirmed that the hydrodynamic diameters and the zeta-potentials of these particles were significantly influenced by pH of the suspension, which was attributed to the charges of polymers. The diameters of CCGM particles were smaller than 200 nm at any pH conditions, suggesting the possibility to apply them as drug delivery carriers. CCGM particles exhibited the responsiveness to a magnetic field in addition to their high dispersion stability, indicating their potential to be utilized as a biomaterial for hyperthermia.     

Preparation and characterization of core-shell nanoparticles hardened by gamma-ray

Core-shell nanoparticles have been prepared by irradiation of gamma-ray on block copolymer micelles consisting of hydrophilic polyacrylic acid and hydrophobic polyisoprene with each 40 monomer units. The structure was determined by means of dynamic light scattering (DLS), small angle X-ray scattering (SAXS) and atomic force microscopy (AFM). The size distribution of the core-shell nanoparticles determined by DLS and AFM was very narrow. The average diameter of the particles decreased from 48 nm for the original micelles to 26 nm by the irradiation of 30 kGy. The core size determined by SAXS combined with DLS was roughly constant of 10 nm, irrespective of irradiation dose, whereas the shell thickness of the micelles was twice as large as the core size, and decreased with increasing irradiation dose.     

一种便捷方法制备表面氨基化的超顺磁Fe3O4纳米粒子

提出了一种简便易行的对磁性纳米粒子表面进行氨基化的方法. 首先使用化学共沉淀法合成了粒径为10 nm左右的Fe3O4纳米粒子, 然后用阿仑膦酸钠对其表面进行修饰, 使其表面具有了功能化的氨基. 利用透射电子显微镜(TEM)、X射线衍射(XRD)、振动样品磁强计(VSM)、动态光散射(DLS)仪、热重分析(TGA)仪、傅里叶变换红外(FT-IR)光谱仪、X射线光电子能谱(XPS)仪等对其进行表征. 结果显示磁性纳米粒子表面被成功地修饰了一层双膦酸分子. 所制备的纳米粒子可在pH=6.3稳定存在4周以上.     

Redispersible and stable amorphous calcium phosphate nanoparticles functionalized by an organic bisphosphate

Although much effort has been focused on the preparation of stable amorphous calcium phosphate （ACP） nanoparticles in aqueous solution, the redispersibility and long-term stability of ACP nanoparticles in aqueous solution remains an unresolved problem. In this work, stable colloidal ACPs were prepared by using an organic bisphosphonate （BP） as a sterically hindered agent in aqueous solution. The harvested calcium phosphate nanoparticles were characterized by inductively coupled plasma atomic emission spectrometry （ICP-AES）, Fourier transform infrared （FTIR）, X-ray diffraction （XRD）, dynamic light scattering （DLS） and transmission electron microscopy （TEM）. ICP-AES, FTIR and XRD results suggested the particles were ACP. DLS and TEM results indicated that the size of the ACP nanoparticles were in the range of 60 nm with a spherical morphology. The resulting calcium phosphate nanoparticles retained its amorphous nature in aqueous solution for at least 6 months at room temperature due to the stabilizing effect of the organic bisphosphonate. Moreover, the surface of the ACP nanoparticles adsorbed with the organic bisphosphate used showed good redispersibility and high colloid stability both in organic and aqueous solutions.