Preparation and application of polymer-grafted magnetic nanoparticles for lipase immobilization

Functionalized superparamagnetic particles were prepared by graft polymerization of glycidyl methacrylate and methacryloxyethyl trimethyl ammonium chloride onto the surface of modified-Fe₃O₄ nanoparticles. The resultant particles were characterized by X-ray powder diffraction, transmission electron microscopy, Fourier transform infrared spectroscopy, and vibrating sample magnetometry. The results indicate that the polymer chains had been effectively grafted onto the surface of Fe₃O₄ nanoparticles. The functionalized particles remained dispersive and superparamagnetic. Lipase was immobilized on the magnetic particles under mild conditions by electrostatic adsorption and covalent binding with the activity recovery up to 70.4%. The immobilized lipase had better thermal stability compared to free lipase.     

Surface functionalization of silica-coated magnetic nanoparticles for covalent attachment of cholesterol oxidase

A systematic approach towards the fabrication of highly functionalized silica shell magnetic nanoparticles, presently used for enzyme immobilization, is herein fully presented. The synthesis of bare maghemite (γ-Fe₂O₃) nanoparticles was accomplished by thermal co-precipitation of iron ions in ammonia alkaline solution at harsh reaction conditions, respectively. Primary surface engineering of maghemite nanoparticles was successfully performed by the proper deposition of silica onto nanoparticles surface under strictly regulated reaction conditions. Next, the secondary surface functionalization of the particles was achieved by coating the particles with organosilane followed by glutaraldehyde activation in order to enhance protein immobilization. Covalent immobilization of cholesterol oxidase was attempted afterwards. The structural and magnetic properties of magnetic silica nanocomposites were characterized by TEM and vibrating sample magnetometer (VSM) instruments. X-ray diffraction measurements confirmed the spinel structure and average size of uncoated maghemite nanoparticles to be around 20 nm in diameter. SEM-EDS spectra indicated a strong signal for Si, implying the coating procedure of silica onto the particles surface to be successfully accomplished. Fourier transform infrared (FT-IR) spectra analysis confirmed the binding of amino silane molecules onto the surface of the maghemite nanoparticles mediated Si-O-Si chemical bonds. Compared to the free enzyme, the covalently bound cholesterol oxidase retained 50% of its activity. Binding of enzyme onto chemically modified magnetic nanoparticles via glutaraldehyde activation is a promising method for developing biosensing components in biomedicine.     

Applications of polymeric micelles with tumor targeted in chemotherapy

The chitosan-coated magnetic nanoparticles (CS MNPs) were in situ synthesized by cross-linking method. In this method; during the adsorption of cationic chitosan molecules onto the surface of anionic magnetic nanoparticles (MNPs) with electrostatic interactions, tripolyphosphate (TPP) is added for ionic cross-linking of the chitosan molecules with each other. The characterization of synthesized nanoparticles was performed by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS/ESCA), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), dynamic light scattering (DLS), thermal gravimetric analysis (TGA), and vibrating sample magnetometry (VSM) analyses. The XRD and XPS analyses proved that the synthesized iron oxide was magnetite (Fe₃O₄). The layer of chitosan on the magnetite surface was confirmed by FTIR. TEM results demonstrated a spherical morphology. In the synthesis, at higher NH₄OH concentrations, smaller sized nanoparticles were obtained. The average diameters were generally between 2 and 8?nm for CS MNPs in TEM and between 58 and 103?nm in DLS. The average diameters of bare MNPs were found as around 18?nm both in TEM and DLS. TGA results indicated that the chitosan content of CS MNPs were between 15 and 23?% by weight. Bare and CS MNPs were superparamagnetic. These nanoparticles were found non-cytotoxic on cancer cell lines (SiHa, HeLa). The synthesized MNPs have many potential applications in biomedicine including targeted drug delivery, magnetic resonance imaging?(MRI), and magnetic hyperthermia.     

Synthesis and functionalization of magnetite nanoparticles with different amino-functional alkoxysilanes

Superparamagnetic iron oxide (SPIO) nanoparticles show great promise for many biotechnological applications. This paper addresses the synthesis and characterization of SPIO nanoparticles grafted with three different alkoxysilanes: 3-aminopropyl-triethoxysilane (APTES), 3-aminopropyl-ethyl-diethoxysilane (APDES) and 3-aminopropyl-diethy-ethoxysilane (APES). SPIO nanoparticles with an average particle diameter of 10 nm were prepared by chemical sonoprecipitation. As confirmed by Fourier transform infrared (FTIR) spectroscopy, silylation of these nanoparticles occurs through a two-step process. Decreasing the number of alkoxide groups reduced the concentration of free amino groups on the SPIO surface ([SPIO-NH₂]—APTES>APDES>APES). This phenomenon results from steric contributions and the formation of H-bonded amines provided by the ethyl groups present in the APDES and APES molecules. A simulation of SPIO nanoparticles in a saline physiologic solution shows that the ethyl groups impart larger steric stability onto the ferrofluids, which reduces aggregation. The magnetization (M) versus magnetic field (H) curves show that the synthesized iron oxide nanoparticles display superparamagnetic behavior. The zero-field cooling (ZFC) and field cooling (FC) curves show that the changes in the blocking temperature depend on the alkoxysilane-functionalized particle surface.     

Synthesis,characterization, and cytotoxicity of glutathione-PEG-iron oxide magnetic nanoparticles

Recently, increasing interest is spent on the synthesis of superparamagnetic iron oxide nanoparticles, followed by their characterization and evaluation of cytotoxicity towards tumorigenic cell lines. In this work, magnetite (Fe₃O₄) nanoparticles were synthesized by the polyol method and coated with polyethylene glycol (PEG) and glutathione (GSH), leading to the formation of PEG-Fe₃O₄ and GSH-PEG-Fe₃O₄ nanoparticles. The nanoparticles were characterized by state-of-the-art techniques: dynamic light scattering (DLS), atomic force microscopy (AFM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and superconducting quantum interference device (SQUID) magnetic measurements. PEG-Fe₃O₄ and GSH-PEG-Fe₃O₄ nanoparticles have crystallite sizes of 10 and 5 nm, respectively, indicating compression in crystalline lattice upon addition of GSH on the nanoparticle surface. Both nanoparticles presented superparamagnetic behavior at room temperature, and AFM images revealed the regular spherical shape of the nanomaterials and the absence of particle aggregation. The average hydrodynamic sizes of PEG-Fe₃O₄ and GSH-PEG-Fe₃O₄ nanoparticles were 69 ± 37 and 124 nm ± 75 nm, respectively. The cytotoxicity of both nanoparticles was screened towards human prostatic carcinoma cells (PC-3). The results demonstrated a decrease in PC-3 viability upon treatment with PEG-Fe₃O₄ or GSH-PEG-Fe₃O₄ nanoparticles in a concentration-dependent manner. However, the cytotoxicity was not time-dependent. Due to the superparamagnetic behavior of PEG-Fe₃O₄ or GSH-PEG-Fe₃O₄ nanoparticles, upon the application of an external magnetic field, those nanoparticles can be guided to the target site yielding local toxic effects to tumor cells with minimal side effects to normal tissues, highlighting the promising uses of iron oxide nanoparticles in biomedical applications.     

Surface functionalization of chitosan-coated magnetic nanoparticles for covalent immobilization of yeast alcohol dehydrogenase from Saccharomyces cerevisiae

A novel and efficient immobilization of yeast alcohol dehydrogenase (YADH, EC1.1.1.1) from Saccharomyces cerevisiae has been developed by using the surface functionalization of chitosan-coated magnetic nanoparticles (Fe₃O₄/KCTS) as support. The magnetic Fe₃O₄/KCTS nanoparticles were prepared by binding chitosan alpha-ketoglutaric acid (KCTS) onto the surface of magnetic Fe₃O₄ nanoparticles. Later, covalent immobilization of YADH was attempted onto the Fe₃O₄/KCTS nanoparticles. The effect of various preparation conditions on the immobilized YADH process such as immobilization time, enzyme concentration and pH was investigated. The influence of pH and temperature on the activity of the free and immobilized YADH using phenylglyoxylic acid as substrate has also been studied. The optimum reaction temperature and pH value for the enzymatic conversion catalyzed by the immobilized YADH were 30 °C and 7.4, respectively. Compared to the free enzyme, the immobilized YADH retained 65% of its original activity and exhibited significant thermal stability and good durability.     

Magnetic Study of Nanocrystalline Ferrites and the Effect of Swift Heavy Ion Irradiation

100 MeV Si⁺⁷ irradiation induced modifications in the structural and magnetic properties of Mg_0.95Mn_0.05Fe₂O₄ nanoparticles have been studied by using X-ray diffraction, Mössbauer spectroscopy and a SQUID magnetometer. The X-ray diffraction patterns indicate the presence of single-phase cubic spinel structure of the samples. The particle size was estimated from the broadened (311) X-ray diffraction peak using the well-known Scherrer equation. The milling process reduced the average particle size to the nanometer range. After irradiation a slight increase in the particle size was observed. With the room temperature Mössbauer spectroscopy, superparamagnetic relaxation effects were observed in the pristine as well as in the irradiated samples. No appreciable changes were observed in the room temperature Mössbauer spectra after ion irradiation. Mössbauer spectroscopy performed on a 12 h milled pristine sample (6 nm) confirmed the transition to a magnetically ordered state for temperatures less than 140 K. All the samples showed well-defined magnetic ordering at 5 K, whereas, at room temperature they were in a superparamagnetic state. From the magnetization studies performed on the irradiated samples, it was concluded that the saturation magnetization was enhanced. This was explained on the basis of SHI irradiation induced modifications in surface states of the nanoparticles.     

Effects of surfactant and polymerization method on the synthesis of magnetic colloidal polymeric nanoparticles

The addition of superparamagnetic iron nanoparticles into polystyrene matrix allows for the modification of the physical properties as well as the implementation of new features in the hybrid nanomaterials. These materials have excellent potential for biomedical and bioengineering applications. Nevertheless, it is necessary to achieve a good dispersion of magnetic nanoparticles for its successful incorporation into polymer particles. This can be obtained through the use of a stabilizer, which provides stability against aggregation. In this work, magnetic nanoparticles were dispersed using different stabilizers. Subsequently, ferrofluids stabilized using the mixture of ABEX/IGEPAL and acrylic acid (AA) were used to synthesize PS-Fe₃O₄ nanocomposites, through miniemulsion and emulsion polymerization conventional techniques. Semicontinuous and batch processes were compared, by varying surfactants and their concentrations. The PS-Fe₃O₄ nanoparticles were characterized by dynamic light scattering, scanning electron microscopy, Raman spectroscopy, and vibrating sample magnetometer. Magnetic nanoparticle dispersions show better results when the anionic and nonionic surfactants are used as a mixture rather than when used alone. Results of DLS showed that the semicontinuous process allowed obtaining monodisperse materials, whereas polidisperse systems are generated in batch process. Raman spectroscopy confirmed the presence of magnetite and polystyrene in the nanocomposites. PS-Fe₃O₄ nanoparticles showed superparamagnetic behavior with final magnetization of around 0.01 emu/g and low coercivity, properties that make them suitable for applications in wide fields of technology. Particle size (Dz), was lower than 300 nm in all cases. Moreover, the use of AA as stabilizer allows enhancing the PS-Fe₃O₄ composite properties. These findings showed that particle size, morphology, and agglomeration are directly influenced by the concentration and the type of surfactant employed.     

Synthesis and characterization of uncoated and gold-coated magnetite nanoparticles

We report on the synthesis and characterization of uncoated and gold coated magnetite nanoparticles. Structural characterizations, carried out using X-ray diffraction, confirm the formation of magnetite phase with a mean size of ~7 and ~8 nm for the uncoated and gold covered magnetite nanoparticles, respectively. The value of the gold coated Fe₃O₄ nanoparticles is consistent with the mean physical size determined from transmission electron microscopy images. Mössbauer spectra at room temperature are consistent with the thermal relaxation of magnetic moments mediated by particle-particle interactions. The 77 K Mössbauer spectra are modeled with four sextets. Those sextets are assigned to the signal of iron ions occupying the tetrahedral and octahedral sites in the core and shell parts of the particle. The room-temperature saturation magnetization value determined for the uncoated Fe₃O₄ nanoparticles is roughly ~60 emu/g and suggests the occurrence of surface effects such as magnetic disorder or the partial surface oxidation. These surface effects are reduced in the gold-coated Fe₃O₄ nanoparticles. Zero-field–cooled and field-cooled curves of both samples show irreversibilities which are consistent with a superparamagnetic behavior of interacting nanoparticles.     

Magnetic and Mössbauer studies of fucan-coated magnetite nanoparticles for application on antitumoral activity

Fucan-coated magnetite (Fe₃O₄) nanoparticles were synthesized by the co-precipitation method and studied by Mössbauer spectroscopy and magnetic measurements. The sizes of the nanoparticles were 8–9 nm. Magnetization measurements and Mössbauer spectroscopy at 300 K revealed superparamagnetic behavior. The magnetic moment of the Fe₃O₄ is partly screened by the Fucan coating aggregation. When the magnetite nanoparticles are capped with oleic acid or fucan, reduced particle-particle interaction is observed by Mössbauer and TEM studies. The antitumoral activity of the fucan-coated nanoparticles were tested in Sarcoma 180, showing an effective reduction of the tumor size.     

海藻酸改性的空心Fe3O4纳米颗粒的制备与应用

不使用任何模板一步制得空心Fe₃O₄纳米颗粒,然后将海藻酸钠嫁接在氨基化的空心Fe₃O₄表面,再利用海藻酸盐与钙离子的作用,在空心Fe₃O₄表面形成一个凝胶化层,制得海藻酸盐凝胶化的空心Fe₃O₄纳米颗粒,粒径约为400～500 nm．采用TEM、XRD、XPS、VSM等手段对纳米微球进行表征．VSM表征结果表明在室温下样品磁性材料为超顺磁性．改性Fe₃O₄纳米颗粒成功地用于柔红霉素的载负和缓释,最大载负率和载药量分别为28.4%和14.2%．缓释结果表明,海藻酸盐凝胶化层的存在,能更有效控制柔红霉素缓慢地释放．     

Immobilization of cholesterol oxidase to finely dispersed silica-coated maghemite nanoparticles based magnetic fluid

In the recent years, the potential applicability of magnetic nanoparticles (MNPs) has witnessed a significant increase in interest towards the medical field, in particular, towards the usage of novel nanoparticles in diagnostics and disease treatment, respectively. In a present study, cholesterol oxidase (ChOx) was covalently immobilized to magnetic nanoparticles of maghemite (γ-Fe₂O₃) and further functionalized by silica (SiO₂) and amino-silane molecules. The activity of the bound enzyme was retained up to 60%, respectively. The binding of cholesterol oxidase was confirmed using FT-IR spectrophotometer. SEM analysis showed uniformly dispersed functional magnetic nanoparticles, which ranged in size from 22.5 to 50.8 nm, surrounded by amorphous silica. In this paper, the potential applications of chemically modified magnetic nanoparticles as carriers for cholesterol oxidase and other enzymes are discussed.     

Characterization of magnetic core-shell nanoparticles by fluxgate magnetorelaxometry, ac susceptibility, transmission electron microscopy and photon correlation spectroscopy—A comparative study

We have compared the structure parameters of magnetic core-shell nanoparticles determined from fluxgate magnetorelaxometry measurements applying the moment superposition model with the results from other methods. For the characterization of the magnetic cores, the nanoparticles are immobilized by freeze-drying. The core size distribution estimated for superparamagnetic Fe₃O₄ magnetic nanoparticles (MNPs) with polyacrylic acid shell agrees well with that from transmission electron microscopy measurements. The distribution of hydrodynamic diameters of nanoparticle suspensions estimated from magnetorelaxometry measurements is in good agreement with that obtained from ac susceptibility and photon correlation spectroscopy measurements. Advantages of magnetorelaxometry compared to the other two integral techniques are that it is fast and the signal is less dominated by larger particles.     

Magnetic properties and the mechanism of formation of the uncompensated magnetic moment of antiferromagnetic ferrihydrite nanoparticles of a bacterial origin

The magnetic properties of the superparamagnetic ferrihydrite nanoparticles that form as a result of the vital activity of Klebsiella oxytoca bacteria are studied. Both an initial powder with an average number of iron atoms N _Fe ～ 2000–2500 in a particle and this powder after annealing at 140°C for 3 h in air are investigated. The following substantial modifications of the magnetic properties of the ferrihydrite nanoparticles are detected after annealing: the superparamagnetic blocking temperature increases from 23 to 49.5 K, and the average magnetic moment of a particle increases (as follows from the results of processing of magnetization curves). The particles have antiferromagnetic ordering, and the magnetic moment resulting in the superparamagnetism of the system appears due to random spin decompensation inside the particle. For this mechanism, the number of uncompensated spins is proportional to the number of magnetically active atoms raised to the one-half power, and this relation holds true for the samples under study at a good accuracy. The possible causes of the detected shift of magnetic hysteresis loops at low temperatures upon field cooling are discussed.     

Preparation and Characterization of Magnetic Microspheres with an Epoxy Group Coating and Their Applications for Lipase Immobilization

Magnetic microspheres coated with epoxy groups (Ep) were prepared by suspension polymerization of glycidyl methacrylate (GMA) and styrene (St) in the presence of oleic acid-coated Fe₃O₄ (OA-Fe₃O₄) nanoparticles, using ethylene glycol dimethacrylate (EGDMA) as a cross-link reagent. The resulting magnetic microspheres were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and vibrating sample magnetometry (VSM). It was verified that the magnetic microspheres had small diameters of 10–30 μm, as well as having super paramagnetism with a saturation magnetization of 10.5 emu/g. Porcine pancreas lipase (PPL) was selected as a model enzyme to be covalently immobilized (IPPL) on the magnetic microspheres via the reactive Ep; they had an activity yield up to 58.6%, when the protein loading reached 55.4 mg/g supports. In addition, the resulting immobilized lipases exhibited a better thermal stability, temperature and pH endurance than that of the free ones. In particular, an excellent reusability of IPPL, which could meet industrial requirements, was achieved.     

Magnetic nanoparticles coated with polysaccharide polymers for potential biomedical applications

This study reports a two-steps route for obtaining magnetic nanoparticles–polysaccharide hybrid materials consisting of Fe₃O₄, NiFe₂O₄ and CuFe₂O₄ nanoparticles synthesis by coprecipitation method in the presence of a soft template followed by coating of ferrite nanoparticles of 8–10-nm size range with polysaccharide type polymers—sodium alginate or chitosan. Magnetic oxide nanoparticles and the corresponding hybrid materials were characterized by X-ray diffraction (XRD), Mössbauer spectroscopy, atomic absorption spectroscopy (AAS), FTIR spectroscopy, scanning and transmission electron microscopy (SEM and TEM) and specific surface area measurements. The vibrating sample magnetometry confirms the superparamagnetic properties of the synthesized ferrites and hybrids. Using this route, the percent of magnetic nanoparticles retained in chitosan-based hybrid materials is nearly double in comparison with that of sodium alginate–based materials. The biological activity tests on Escherichia coli ATCC 25922, Pseudomonas aeroginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Candida scotti microorganisms show the non-toxic properties of prepared hybrid materials.     

Synthesis of immunomagnetic nanoparticles and their application in the separation and purification of CD34 hematopoietic stem cells

The silica-coated superparamagnetic nanoparticles with the uniform diameter of about 60 nm were synthesized by reverse microemulsions method. And the magnetic nanoparticles were modified with N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPS). The immunomagnetic nanoparticles were then successfully prepared by covalently immobilizing anti-CD34⁺ monoclonal antibodies to the surface of amino silane modified magnetic particles. The cell separation results showed that the synthesized immunomagnetic nanoparticles could rapidly and conveniently separate the CD34⁺ cells with high efficiency and specificity than normal ones. The surface morphology of separated target cells was examined by scanning electron microscope (SEM). Atomic force microscope (AFM) also characterized the magnetic materials on the surface of the separated target cells for the first time, which further confirmed that the target cells were separated by the immunomagnetic nanoparticles. The viability of the separated cells was studied by culturing method and Beckman Vi-cell viability analyst. Therefore, our experiments provided a new, direct, rapid mode to separate target cells.     

Structural characterization and magnetic properties of superparamagnetic zinc ferrite nanoparticles synthesized by the coprecipitation method

Single phase zinc ferrite (ZnFe₂O₄) nanoparticles have been prepared by the coprecipitation method without any subsequent calcination. The effects of precipitation temperature in the range 20–80 °C on the structural and the magnetic properties of zinc ferrite nanoparticles were investigated. The crystallite size, microstructure and magnetic properties of the prepared nanoparticles were studied using X-ray diffraction (XRD), Fourier transmission infrared spectrum, transmission electron microscope (TEM), energy dispersive X-ray spectrometer and vibrating sample magnetometer. The XRD results showed that the coprecipitated nanoparticles were single phase zinc ferrite with mixture of normal and inverse spinel structures. Furthermore, ZnFe₂O₄ nanoparticles have the crystallite size in the range 5–10 nm, as confirmed by TEM. The magnetic measurements exhibited that the zinc ferrite nanoparticles synthesized at 40 °C were superparamagnetic with the maximum magnetization of 7.3 emu/g at 10 kOe.     

Implications of nanoparticle concentration and size distribution in the superparamagnetic behaviour of aging-improved maghemite xerogels

Evolution of static magnetic properties of a set of enhanced γ-Fe₂O₃/SiO₂ nanocomposites with different iron concentration has been studied on the basis of their corresponding hysteresis loops, zero-field/field-cooled (ZFC/FC) magnetization curves and transmission electron microscopy images. The lack of coercivity in all compositions, as well as the fulfillment of the H/T scaling law by the magnetization above the blocking temperature of each system under study, evidence a superparamagnetic behaviour in the iron oxide nanoparticles. In order to study the influence of iron content in the unblocking processes of nanoparticles, ZFC curves under different applied magnetic fields have been fitted to a model considering the systems under study as a distribution of energy barriers. Depart from the superparamagnetic model is discussed considering interparticle interactions.     

Mechanosynthesis,magnetic and Mössbauer characterization of pure and Ti4+-doped cubic phase BiFeO3 nanocrystalline particles

The mechanosynthesis of cubic γ-phase pure BiFeO₃ and Ti⁴⁺-doped BiFeO₃ nanocrystalline particles and their preliminary characterization with magnetic measurements and Mössbauer spectroscopy are reported. The BiFeO₃ nanoparticles (5–40 nm) were prepared by heating a 48 h pre-milled 1:1 molar mixture of α-Bi₂O₃ and α-Fe₂O₃ at 400 °C for (1 h). Doping α-Fe₂O₃ in the initial mixture of reactants with Ti⁴⁺ was found to lead to the formation of Ti⁴⁺-doped BiFeO₃ nanoparticles by milling the reactants for 32 h. The magnetization of the BiFeO₃ nanoparticles is found to be tripled under a maximum external field of 1.35 T and their magnetic hardness increases by ～15 times relative to those of the corresponding bulk. The Ti⁴⁺-doped BiFeO₃ nanoparticles exhibit higher magnetization relative to the pure ones. These observations are related to the spiral modulated spin structure of the compound. The Mössbauer data show ～12 % of the BiFeO₃ nanocrystalline particles to be superparamagnetic having blocking temperatures of less than 78 K. The quadrupole shift values of the magnetic spectral component favor the cubic structural symmetry. These observations were mainly associated with possible collective magnetic excitations as well as transverse relaxation of canted surface spins. The Ti-doped BiFeO₃ nanoparticles gave statistically-poor Mössbauer spectra with no signs of a superparamagnetic behavior.