Enhanced therapeutic action of Trastuzumab loaded ZnxMn1-xFe2O4 nanoparticles using a pre-treatment step for hyperthermia treatment of HER2+ breast cancer.

In this study, Ferrites (Fe3O4, MnFe2O4, ZnFe2O4) and different stoichiometric ratios of ZnxMn1-xFe2O4 (x = 0.2, 0.4, 0.6, and 0.8) nanoparticles (<15 nm) were synthesized by microwave-assisted method and optimised for hyperthermia studies. The selection of the optimised variant of ferrite i.e. Zn0.4Mn0.6Fe2O4 was found to be the best variant based on VSM (38.14 emu g-1) hyperthermia-based temperature rise (maximum ΔT of 38 °C), SAR and ILP values. Trastuzumab, which is known to bind with HER2 receptors of breast cancer was chemically tethered onto Zn0.4Mn0.6Fe2O4 nanoparticles through EDC/NHS coupling with a loading efficiency of 80%. The attached Trastuzumab aided during the pre-treatment step by aiding in the internalisation of Zn0.4Mn0.6Fe2O4 nanoparticles, with cellular uptake of 11% in SK-BR-3 (cancerous HER2+) cells compared to ∼5% for MDA-MB-231 (cancerous HER2-) and RPE-1 (non-cancerous) cells. In the presence of a hyperthermia trigger for 15 mins, ZnxMn1-xFe2O4 -Trastuzumab formulation had a maximum therapeutic effect by reducing the SK-BR-3 cell viability to 14% without adversely affecting the RPE-1 cells. The mechanism of ZnxMn1-xFe2O4-Trastuzumab combination was examined using an internalisation study, MTT-based viability, proliferation study, and ROS generation assay. By utilizing both Trastuzumab and hyperthermia, we achieve their synergistic anticancer properties while minimizing the drug requirement and reducing any effect on non-cancerous cells.

Dielectric spectrum of a ferrofluid layer exposed to a gradient magnetic field.

A low-frequency dielectric response of a ferrofluid based on transformer oil and MnZn ferrite nanoparticles is investigated in a gradient magnetic field. Four ferrofluid samples of various nanoparticle concentrations were introduced into planar micro-capacitors located over a magnetized tip. The dielectric spectra were measured in the frequency range from 0.1 Hz to 200 kHz and in the local magnetic field up to 100 mT. The spectra exhibit a dielectric relaxation ascribed to nanoparticle interfacial polarization. The low-frequency spectrum of each ferrofluid decreases upon application of the magnetic field up to 20 mT. The decrease in dielectric permittivity is caused by a magnetic force acting on larger nanoparticles in the gradient magnetic field. It is assumed that the interfaces of the concentrated nanoparticles in the gradient field do not contribute to the effective dielectric response. This reduces the effective relaxation time and shifts the relaxation toward higher frequencies. The dielectric spectra are well described by a relaxation fit function consisting of one Havriliak-Negami and a conductivity term. The fitting confirms that the only effect of the gradient magnetic field on the dielectric spectra is the shift of the dielectric relaxation and the decrease of the amplitude in the imaginary permittivity. This behavior is evident from a master plot, where all dielectric relaxations are superimposed on a single line. The knowledge of the presented behavior of the ferrofluid may be valuable when applying a ferrofluid to sharply magnetized parts of various electrical equipment (wires, tips, screws, nails, edges) as a liquid dielectric medium.

Optimization of magnetic fluid hyperthermia protocols for the elimination of breast cancer cells MCF7 using Mn-Zn ferrite ferrofluid.

The present study aimed to optimize magnetic fluid hyperthermia (MFH) protocols by standardizing MF incubation time, hyperthermic duration, magnetic field, and MFH sessions to achieve a better hyperthermic response for the profuse killing of human breast cancer cell cells MCF7. Magnetic nanoparticles and MF were characterized using XRD, VSM, and DLS. Induction heating was performed for 30 min at field strengths of 12.5 and 13.3 kA/m at a fixed frequency of 330 kHz with varying concentrations and incubation duration on MCF7 cells. Single and multiple sessions hyperthermia protocols were used to kill MCF7 cells and the cytotoxicity effect was analyzed using MTT assay. Single and multiple sessions MFH protocols were established to kill breast cancer cells utilizing 0.2 mg/mL MF at 13.3 kA/m field and 330 kHz frequency and maintaining the hyperthermic temperature of 43-45 °C for 30 min. The single session MFH revealed severe toxicity of MF leading to more than 75% of cell death after 24 h of MF incubation. Multiple sessions hyperthermia resulted in more than 90% killing of MCF7 cells after two consequent 3 h MF incubation with 3 h gap. Each 3 h of MF incubation was followed by 30 min of induction heating. Multiple sessions hyperthermia was effective in killing a larger cell population compared to the single session protocol. The results may help in optimizing protocols for the profuse killing of cancer cells of multiple origins, and aid in deciding futuristic in vivo MFH-based therapeutic strategies against breast cancer. Variation in MCF7 cells' viability due to HT, MF, and MF + HT in multiple sessions.

In vitro hyperthermic effect of magnetic fluid on cervical and breast cancer cells.

Self-regulating temperature-controlled nanoparticles such as Mn-Zn ferrite nanoparticles based magnetic fluid can be a better choice for magnetic fluid hyperthermia because of its controlled regulation of hyperthermia temperature window of 43-45 °C. To test this hypothesis magnetic fluid with said properties was synthesized, and its effect on cervical and breast cancer cell death was studied. We found that the hyperthermia window of 43-45 °C was maintained for one hour at the smallest possible concentration of 0.35 mg/mL without altering the magnetic field applicator parameters. Their hyperthermic effect on HeLa and MCF7 was investigated at the magnetic field of 15.3 kA/m and frequency 330 kHz, which is close to the upper safety limit of 5 * 109 A/m s. We have tested the cytotoxicity of synthesized Mn-Zn ferrite fluid using MTT assay and the results were validated by trypan blue dye exclusion assay that provides the naked eye microscopic view of actual cell death. Since cancer cells tend to resist treatment and show re-growth, we also looked into the effect of multiple sessions hyperthermia using a 24 h window till 72 h using trypan blue assay. The multiple sessions of hyperthermia showed promising results, and it indicated that a minimum of 3 sessions, each of one-hour duration, is required for the complete killing of cancer cells. Moreover, to simulate an in vivo cellular environment, a phantom consisting of magnetic nanoparticles dispersed in 1 and 5% agarose gel was constituted and studied. These results will help to decide the magnetic fluid based hyperthermic therapeutic strategies using temperature-sensitive magnetic fluid.

Defragmentation of lysozyme derived Amyloid ß fibril using Biocompatible Magnetic fluid.

We present here a modulating effect on lysozyme derived Amyloid ß fibrils by aqueous magnetic fluid. This non-conventional approach of treatment of lysozyme derived Amyloid ß fibrils showed lysing of Amyloid fibrils to its secondary structures which can be seen using optical microscope and scanning electron microscopic image. The size of lysozyme derived amyloid fibrils before and after treatment was measured using dynamic light scattering technique. The mechanism of defragmentation of lysozyme derived Amyloid ß fibrils by magnetic fluid is explained. This is a first report to identify the secondary structure of protein using Fourier Transform Infrared (FTIR) and Circular Dichroism (CD) spectra after lysing. The cyto-toxicity study of this magnetic fluid on neuronal (SH-SY5Y) and non-neuronal (NRK) cell lines shows non-toxicity up to a concentration of 250 µg/mL. The study indicates a novel and unique complementary approach to treat the amyloidogenic brain diseases.

Technique to optimize magnetic response of gelatin coated magnetic nanoparticles.

The paper describes the results of optimization of magnetic response for highly stable bio-functionalize magnetic nanoparticles dispersion. Concentration of gelatin during in situ co-precipitation synthesis was varied from 8, 23 and 48 mg/mL to optimize magnetic properties. This variation results in a change in crystallite size from 10.3 to 7.8 ± 0.1 nm. TEM measurement of G3 sample shows highly crystalline spherical nanoparticles with a mean diameter of 7.2 ± 0.2 nm and diameter distribution (σ) of 0.27. FTIR spectra shows a shift of 22 cm(-1) at C=O stretching with absence of N-H stretching confirming the chemical binding of gelatin on magnetic nanoparticles. The concept of lone pair electron of the amide group explains the mechanism of binding. TGA shows 32.8-25.2% weight loss at 350 °C temperature substantiating decomposition of chemically bind gelatin. The magnetic response shows that for 8 mg/mL concentration of gelatin, the initial susceptibility and saturation magnetization is the maximum. The cytotoxicity of G3 sample was assessed in Normal Rat Kidney Epithelial Cells (NRK Line) by MTT assay. Results show an increase in viability for all concentrations, the indicative probability of a stimulating action of these particles in the nontoxic range. This shows the potential of this technique for biological applications as the coated particles are (i) superparamagnetic (ii) highly stable in physiological media (iii) possibility of attaching other drug with free functional group of gelatin and (iv) non-toxic.

Ultrasonic propagation: a technique to reveal field induced structures in magnetic nanofluids.

The paper reports the study of magnetic field induced structures in magnetic nanofluid investigated through ultrasonic wave propagation. Modified Tarapov's theory is used to study variation in velocity anisotropy with magnetic field. The types of field induced structures depend upon the chemical structure of the carrier in which magnetic nanoparticles are dispersed. Our study indicates formation of fractals and chain respectively, in transformer oil and kerosene based fluid. This difference is explained on the basis of particle-particle interaction and particle-medium interaction.

Effect of carrier and particle concentration on ultrasound properties of magnetic nanofluids.

Ultrasound wave propagation in nanofluids and its rheological behavior has been studied as a function of solid volume fraction, temperature and magnetic field for magnetic nanofluids synthesized in kerosene and transformer oil. Ultrasonic velocity decreases while viscosity increases with increasing volume fraction. The attenuation of ultrasonic wave is explained using dipolar coupling co-efficient which favors oligomer structures with increasing number density of particles. The structure formation increases further with increase in magnetic field which is prominent in transformer oil compared to kerosene. This difference can be due to the structural difference between these two carriers.

Magnetization dynamics in rare earth Gd3+ doped Mn(0.5)Zn(0.5)Fe2O4 magnetic fluid: electron spin resonance study.

The electron spin resonance (ESR) technique has been applied to study the spin dynamics in broad temperature range for rare earth doped Mn(0.5)Zn(0.5)Fe(1.9)Gd(0.1)O(4) (MZG5) magnetic fluid. Zero field cooled (ZFC) ESR spectra of MZG5 fluid exhibit an isotropic shift in the resonance field below 40 K, while the field cooled (FC) ESR spectra show a deviation from sin(2)θ behavior and an angle dependent hysteresis, this unambiguously points to the dominating unidirectional freezing of surface spins below 40 K. Above 60 K, the resonance field exhibits sin(2)θ behavior, indicating the uniaxial anisotropy contribution of core spin. This indicates that surface spin freezing temperature is around 40 K. The presence of surface spin freezing and the coupling between core and surface spins are further supported by cycle dependent FC ESR spectra measured at 20 K, which show the systematic increase in resonance field (H(res)) and intensity. The double peak behavior of blocking temperature distribution retrieved from ZFC-FC magnetization measurement is an additional corroboration of the existence of surface spin glass like layer.

Monodispersed superparamagnetic FeO4 nanoparticles: synthesis and characterization.

Monodispersed Fe3O4 (magnetite) nanoparticles are synthesized by non-aqueous thermal decomposition route. The powder XRD reveals single phase spinel structure with 60 angstroms particle size. The TEM image shows nearly monodispersed spherical particles with log-normal median diameter of 49 A and size distribution in In(D) 0.12. The monodispersivity is also confirmed by SANS experiment. TGA pattern shows a sharp single step transition at 606 K indicating a monolayer coating of the oleic acid on particle surface. Temperature dependent low field dc magnetization study suggest that particles are superparamagnetic above 25 K with an effective blocking temperature 12 K @ 50 Oe field, indicating the dominance of particle-particle interaction in the system. The saturation magnetization of the nanocrystals, M(s), at 5 K is very close to that of bulk materials. This technique of synthesis gives high M(s) of particles compared to other chemical routes.

Maghemite nanocrystal impregnation by hydrophobic surface modification of mesoporous silica.

Here, we report the design of a hybrid inorganic/organic mesoporous material through simultaneous pore engineering and hydrophobic surface modification of the intramesochannels to improve the uptake of superparamagnetic maghemite nanocrystals via impregnation techniques. The mesoporous material of the SBA-15 type was functionalized in situ with thiol organo-siloxane groups. Restricting the addition of the thiol organo-siloxane to 2 mol % yielded an inorganic/organic hybrid material characterized by large pores and a well-ordered hexagonal p6mm mesophase. The hydrophobic surface modification promoted the incorporation of 7.5 nm maghemite (gamma-Fe2O3) nanocrystals, prepared through temperature-controlled decomposition of iron pentacarbonyl in organic solvents. The hydrophobic, oleic acid capped superparamagnetic maghemite nanocrystals were incorporated into the porous network via wet impregnation from organic suspensions. Combining diffraction, microscopy, and adsorption data confirmed the uptake of the nanocrystals within the intramesochannels of the silica host. Magnetization dependencies on magnetic field at different temperatures show a constriction in the loop around the origin, which indicates immobilization of maghemite nanocrystals inside the thiol-functionalized silica host.

Spin-glass-like magnetic ordering in Zn substituted magnetite magnetic fluids.

Electron spin resonance (ESR) spectra of magnetic fluids involving polydispersed Zn(0.5)Fe(0. 5)Fe(2)O(4) (FZ5) and Zn(0.7)Fe(0. 3)Fe(2)O(4) (FZ7) nanomagnetic particles are scanned from 4.2 to 300K. The FZ7 fluid exhibits certain distinct features below 40K which are different from FZ5 fluid. These include (i) an isotropic shift in resonance field in zero-field-cooled ESR study, (ii) deviation of resonance field from sin(2)theta behavior (where theta is the angle between axis of the particle and field) in field cooled (FC) sample and (iii) abrupt increase in anisotropy field for FC sample. The results are analyzed in light of the core-shell model for nanomagnetic particles.

Experimental evidence of zero forward scattering by magnetic spheres.

Magnetically induced diffraction patterns by micron sized magnetic spheres dispersed in a ferrofluid disappear at a certain critical magnetic field. This critical field is found to depend on the concentration of the ferrofluid and on the volume of the magnetic spheres. We attribute this effect to the zero forward scattering by magnetic spheres as predicted by Kerker, Wang, and Giles [J. Opt. Soc. Am. 73, 765 (1983)]. We suggest that such a dispersion can be used to study the optical analogues of localization of electrons in condensed matter, the Hall effect, and the anisotropic diffusion, etc. The combination of the micron sized magnetic spheres and the ferrofluid will also be useful to design magnetically tunable photonic devices.

Magneto-optical effects in temperature-sensitive ferrofluids.

A temperature-dependent magneto-optical study of three different temperature-sensitive fluids was carried out. The changes in transmitted intensity as a function of applied magnetic field and temperature are recorded. The study provides an alternative technique for determining the Curie temperature of such fluids, and it is also feasible for the monitoring of temperature changes of such fluids optically within a limited time span.