Epirubicin‐Loaded Superparamagnetic Iron‐Oxide Nanoparticles for Transdermal Delivery: Cancer Therapy by Circumventing the Skin Barrier

The transdermal administration of chemotherapeutic agents is a persistent challenge for tumor treatments. A model anticancer agent, epirubicin (EPI), is attached to functionalized superparamagnetic iron‐oxide nanoparticles (SPION). The covalent modification of the SPION results in EPI–SPION, a potential drug delivery vector that uses magnetism for the targeted transdermal chemotherapy of skin tumors. The spherical EPI–SPION composite exhibits excellent magnetic responsiveness with a saturation magnetization intensity of 77.8 emu g^?1. They feature specific pH‐sensitive drug release, targeting the acidic microenvironment typical in common tumor tissues or endosomes/lysosomes. Cellular uptake studies using human keratinocyte HaCaT cells and melanoma WM266 cells demonstrate that SPION have good biocompatibility. After conjugation with EPI, the nanoparticles can inhibit WM266 cell proliferation; its inhibitory effect on tumor proliferation is determined to be dose‐dependent. In vitro transdermal studies demonstrate that the EPI–SPION composites can penetrate deep inside the skin driven by an external magnetic field. The magnetic‐field‐assisted SPION transdermal vector can circumvent the stratum corneum via follicular pathways. The study indicates the potential of a SPION‐based vector for feasible transdermal therapy of skin cancer.     

Characterization of superparamagnetic iron oxide nanoparticles and its application in protein purification

The application of surface modified magnetic adsorbent particles in combination with magnetic separation techniques has received considerable awareness in recent years. There is a particular need in protein purification and analysis for specific, functional and generic methods of protein binding on solid supports. Nanoscale superparamagnetic iron oxide particles have been used to purify a natural coagulant protein extracted from Moringa oleifera seeds. Spectrophotometric analysis of the coagulant protein was performed using synthetic clay solution as substrate. Protein binding with carboxyl and silica surface modified superparamagnetic iron oxide nanoparticles (SPION) were compared with the known carboxyl methyl cellulose (CMC) beads of approximately 1 microm. SPION modified with carboxyl surface showed higher binding capacity towards the coagulant protein compared to the CMC beads. The high surface area to volume ratio of the carboxyl-coated SPION resulted in high binding capacity and rapid adsorption kinetics of the crude protein extract. The purification and molecular weight of coagulant protein is analyzed by SDS-PAGE. This approach utilizes the most efficient, feasible and economical method of coagulant protein purification and it can also be applicable to other proteins that possess similar properties.     

Rapid magnetic cell delivery for large tubular bioengineered constructs

Delivery of cells into tubular tissue constructs with large diameters poses significant spatial and temporal challenges. This study describes preliminary findings for a novel process for rapid and uniform seeding of cells onto the luminal surface of large tubular constructs. Fibroblasts, tagged with superparamagnetic iron oxide nanoparticles (SPION), were directed onto the luminal surface of tubular constructs by a magnetic field generated by a k4-type Halbach cylinder device. The spatial distribution of attached cells, as measured by the mean number of cells, was compared with a conventional, dynamic, rotational cell-delivery technique. Cell loading onto the constructs was measured by microscopy and magnetic resonance imaging. The different seeding techniques employed had a significant effect on the spatial distribution of the cells (p < 0.0001). The number of attached cells at defined positions within the same construct was significantly different for the dynamic rotation technique (p < 0.05). In contrast, no significant differences in the number of cells attached to the luminal surface were found between the defined positions on the construct loaded with the Halbach cylinder. The technique described overcomes limitations associated with existing cell-delivery techniques and is amenable to a variety of tubular organs where rapid loading and uniform distribution of cells for therapeutic applications are required.     

Superparamagnetic iron oxide nanoparticles-loaded polymersome-mediated gene delivery guided by enhanced magnetic resonance signal

Positively charged superparamagnetic iron oxide nanoparticle (SPION)-loaded polymersome was prepared in order to deliver genes to the target sites, which was monitored by magnetic resonance imaging (MRI), concomitantly. The transfection efficiency in vitro was tested by treating CT-26 colon cancer cell line with luciferase-expressing plasmids/SPION complex. MRI was also used to check the detectability of SPION in vitro and in vivo. SPION-loaded polymersome, carrying genetic materials, was delivered and then accumulated at the tumor site of the murine colon cancer xenograft model after intravenous injection, possibly through a passive targeting mechanism. Clinical MRI monitored this accumulation. This result indicates that the SPION-loaded polymersomecan be applied to MR image-guided gene therapy.     

Direct visualization and identification of biofunctionalized nanoparticles using a magnetic atomic force microscope

Because of its outstanding ability to image and manipulate single molecules, atomic force microscopy (AFM) established itself as a fundamental technique in nanobiotechnology. (1) We present a new modality that distinguishes single nanoparticles by the surrounding magnetic field gradient. Diamagnetic gold and superparamagnetic iron oxide nanoparticles become discernible under ambient conditions. Images of proteins, magnetolabeled with nanoparticles, demonstrate the first steps toward a magnetic analogue to fluorescence microscopy, which combines nanoscale lateral resolution of AFM with unambiguous detection of magnetic markers.     

Folate encapsulation in PEG‐diamine grafted mesoporous Fe3 O4 nanoparticles for hyperthermia and in vitro assessment

Effective and targeted delivery of the antitumour drugs towards the specific cancer spot is the major motive of drug delivery. In this direction, suitably functionalised magnetic iron oxide nanoparticles (NPs) have been utilised as a theranostic agent for imaging, hyperthermia and drug delivery applications. Herein, the authors reported the preparation of multifunctional polyethyleneglycol‐diamine functionalised mesoporous superparamagnetic iron oxide NPs (SPION) prepared by a facile solvothermal method for biomedical applications. To endow targeting ability towards tumour site, folic acid (FA) is attached to the amine groups which are present on the NPs surface by 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride/N‐hydroxysuccinimide chemistry. FA attached SPION shows good colloidal stability and possesses high drug‐loading efficiency of ∼ 96% owing to its mesoporous nature and the electrostatic attachment of daunosamine (NH₃ ⁺) group of doxorubicin (DOX) towards the negative surface charge of carboxyl and hydroxyl group. The NPs possess superior magnetic properties in result endowed with high hyperthermic ability under alternating magnetic field reaching the hyperthermic temperature of 43°C within 223 s at NP''s concentration of 1 mg/ml. The functionalised NPs possess non‐appreciable toxicity in breast cancer cells (MCF‐7) which is triggered under DOX‐loaded SPION.Inspec keywords: nanoparticles, nanocomposites, mesoporous materials, colloids, biochemistry, nanomagnetics, molecular biophysics, tumours, superparamagnetism, drugs, toxicology, biomedical materials, nanofabrication, hyperthermia, cancer, magnetic particles, cellular biophysics, nanomedicine, iron compounds, drug delivery systems, filled polymers, biological organs, liquid phase depositionOther keywords: NP surface, colloidal stability, drug‐loading efficiency, hydroxyl group, magnetic properties, high hyperthermic ability, magnetic field, DOX‐loaded SPION, folate encapsulation, targeted delivery, antitumour drugs, specific cancer spot, magnetic iron oxide nanoparticles, theranostic agent, drug delivery applications, multifunctional polyethyleneglycol‐diamine, facile solvothermal method, biomedical applications, tumour site, amine groups, mesoporous superparamagnetic nanoparticles, PEG‐diamine grafted mesoporous nanoparticles, 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride‐N‐hydroxysuccinimide chemistry, daunosamine group, carboxyl group, breast cancer cells, temperature 43.0 degC, Fe₃ O₄      

Differential internalization of superparamagnetic iron oxide nanoparticles in different types of cells

Superparamagnetic iron oxide nanoparticles (SPION) have attracted great attention for nanomedical applications, but the mechanisms underlying the transmembrane transport of SPION in variant cells has not been fully defined. The present study investigated the internalization of SPION in three cell models with different phagocytic capacity using transmission electron microscopy (TEM) and energy dispersive spectrometer (EDS) analyses. The EDS study aimed to further confirm if the suspected internalized particles were iron-containing SPION. SPION could be taken up quickly by macrophage-like cell line RAW264.7 (with strong phagocytic capacity) and slowly by the 3T3-L1 cells (with weak phagocytic capacity), but not by red blood cells (with no phagocytic capacity). The internalized SPION were mainly found in the cytoplasmic vesicles, with no localization in the endoplasmic reticulum, mitochondria and nucleus. We conclude that the internalization of SPION in the three types of mammalian cells was mediated by phagocytosis, not by direct membrane penetration.     

Superparamagnetic Iron Oxide Nanoparticles (SPION) for the Treatment of Antibiotic‐Resistant Biofilms

Bacterial infections caused by antibiotic‐resistant strains are of deep concern due to an increasing prevalence, and are a major cause of morbidity in the United States of America. In particular, medical device failures, and thus human lives, are greatly impacted by infections, where the treatments required are further complicated by the tendency of pathogenic bacteria, such as Staphylococcus aureus, to produce antibiotic resistant biofilms. In this study, a panel of relevant antibiotics used clinically including penicillin, oxacillin, gentamicin, streptomycin, and vancomycin are tested, and although antibiotics are effective against free‐floating planktonic S. aureus, either no change in biofilm function is observed, or, more frequently, biofilm function is enhanced. As an alternative, superparamagnetic iron oxide nanoparticles (SPION) are synthesized through a two‐step process with dimercaptosuccinic acid as a chelator, followed by the conjugation of metals including iron, zinc, and silver; thus, the antibacterial properties of the metals are coupled to the superparamagnetic properties of SPION. SPION might be the ideal antibacterial treatment, with a superior ability to decrease multiple bacterial functions, target infections in a magnetic field, and had activity better than antibiotics or metal salts alone, as is required for the treatment of medical device infections for which no treatment exists today.     

Mesoporous silica nanoparticles improve magnetic labeling efficiency in human stem cells

Tumblerlike magnetic/fluorescein isothiocyanate (FITC)-labeled mesoporous silica nanoparticles, Mag-Dye@MSNs, have been developed, which are composed of silica-coated core-shell superparamagnetic iron oxide (SPIO@SiO(2)) nanoparticles co-condensed with FITC-incorporated mesoporous silica. Mag-Dye@MSNs can label human mesenchymal stem cells (hMSCs) through endocytosis efficiently for magnetic resonance imaging (MRI) in vitro and in vivo, as manifested by using a clinical 1.5-T MRI system with requirements of simultaneous low incubation dosage of iron, low detection cell numbers, and short incubation time. Labeled hMSCs are unaffected in their viability, proliferation, and differentiation capacities into adipocytes and osteocytes, which can still be readily detected by MRI. Moreover, a higher MRI signal intensity decrease is observed in Mag-Dye@MSN-treated cells than in SPIO@SiO(2)-treated cells. This is the first report that MCM-41-type MSNs are advantageous to cellular uptake, as manifested by a higher labeling efficiency of Mag-Dye@MSNs than SPIO@SiO(2).     

Systemic distribution and elimination of plain and with Cy3.5 functionalized poly(vinyl alcohol) coated superparamagnetic maghemite nanoparticles after intraarticular injection in sheep in vivo

PVA coated and fluorescent dye (Cy3.5) functionalized vinyl alcohol/vinyl amine copolymer coated superparamagnetic iron oxide nanoparticles (SPION) were evaluated for systemic distribution and elimination after intraarticular injection in sheep. Observation was done at 3, 24, 72, and 120 hours after injection using light microscopy, fluorescent microscopy, and confocal microscopy. No pathologic influence of SPION on the tissue harvested could be seen. A significantly increased iron content could be identified in the kidneys, lymph nodes, and spleen after injection of SPION. No particles were detected in the liver, the urinary, and the gall bladder. No positive fluorescent signal could be attributed to SPION throughout the organs. Our results indicated that the iron component of the SPION is possible to be incorporated into the physiologic iron metabolism after reabsorption in the proximal tubule system of the kidney and that concentration levels of Cy3.5 are too low to be detected throughout the body.     

Surface modified superparamagnetic iron oxide nanoparticles: as a new carrier for bio-magnetically targeted therapy

Amino-functionalized superparamagnetic iron oxide nanoparticles (SPION) were synthesized by coprecipitation method. The particles were characterized by X-ray diffraction (XRD), vibrating sample magnetometer (VSM), scanning electron micrographs (SEM), transmission electron micrographs (TEM) and atomic force micrographs (AFM). The size of the modified particles varied in the range 10–15 nm and did not change significantly after modification. Hepama-1, an excellent humanized monoclonal antibody directed against liver cancer, was conjugated to the SPION to prepare immuno-magnetic nanoparticles (IMN). A direct labeling method was employed to radiolabel IMN with rhenium-188. The radiolabeling efficiency was about 90% with good in vitro stability. ¹⁸⁸Re labeled IMN could markedly kill SMMC-7721 liver cancer cells. Such SPION might be very useful for bio-magnetically targeted radiotherapy in liver cancer treatment.     

In situ synthesis and magnetic studies of iron oxide nanoparticles in calcium-alginate matrix for biomedical applications

In this work we applied a new route to synthesize magnetic iron oxide nanoparticles into alginate polymer for future application as drug delivery system activated by magnetic external stimuli. Calcium-alginate was used to encapsulate iron oxide nanoparticles, and as scaffold for particle nucleation and its influence on particles size and magnetic properties were studied. The iron oxide mean sizes were between 4.3 and 9.5 nm. Iron is dispersed throughout the polymer matrix mainly as iron oxide particles, and a small fraction as iron (III) occupying calcium sites in the polymer network. The temperature dependence of the Mössbauer spectra is typical of superparamagnetic particles in agreement with the magnetic susceptibility data.     

Uptake and biocompatibility of functionalized poly(vinylalcohol) coated superparamagnetic maghemite nanoparticles by synoviocytes in vitro

Superparamagnetic iron oxide nanoparticles (SPION) were coated with either Polyvinyl alcohol (PVA) or Vinyl alcohol/vinyl amine copolymer and further functionalized with the fluorochromes Cy3.5 or Texas Red. A colloidally stable suspension of nanoparticles was incubated on sheep synovial cells in vitro for 3, 24, 72, and 120 hours. Nanoparticle internalization into synoviocytes as well as biocompatibility was visualized using light, fluorescence and confocal microscopy and fluorochrome labeled cells were quantified by flow cytometry. Data were analyzed by ANOVA factorial tests. Amino-PVA-SPION alone was detectable in cytoplasmic endosome-like structures after 3 hours of incubation but resulted in early cell death after 24 hours. Although amino-PVA-Cy3.5-SPION and PVA-TexasRed-SPION were taken up more slowly and less intensely, both labeled more than 80% of the cells in culture, but did not significantly change cell morphology or vitality at any time of evaluation in comparison to control cells. Results indicate that functionalized amino PVA-coated SPION are biocompatible, were successfully internalized by synoviocytes and hold promise for future biomedical applications utilizing magnetic drug targeting in joint disease.     

Gene expression in synovial membrane cells after intraarticular delivery of plasmid-linked superparamagnetic iron oxide particles--a preliminary study in sheep

This study evaluated in vivo gene delivery and subsequent gene expression within cells of the synovium in the presence of static and pulsating magnetic field application following intraarticular injection of superparamagnetic iron oxide nanoparticles linked to plasmids containing reporter genes encoding for fluorescent proteins. Plasmids encoding genes for either green fluorescent protein or red fluorescent protein were bound to superparamagnetic nanoparticles coated with polyethyleneimine. Larger (200-250 nm) and smaller (50 nm) nanoparticles were compared to evaluate the effects of size on transfection efficiency as well as any associated intraarticular reaction. Comparisons between groups were evaluated at 24, 72, and 120 h time periods. Inflammatory response was mild to moderate for all injected particles, but was present in the majority of synovial membrane samples evaluated. Larger particles tended to be associated with more inflammation than smaller ones. Nevertheless, intraarticular application of both experimental and control nanoparticles were well tolerated clinically. Gene expression as determined by observation of either green or red intracellular fluorescence was difficult to assess by both epifluorescent light, and confocal microscopy. An insufficient concentration of nanoparticles in relation to joint volume likely resulted in a limited number of samples with positive evidence of iron staining and with suspected positive evidence of cells expressing fluorescent proteins. Our results indicate that intraarticular administration of functionalized superparamagnetic iron oxide nanoparticles resulted in a mild to moderate synovitis and there was in conclusive evidence of gene expression. Further research is warranted to determine the best and most effective reporter assay for assessment of the in vivo gene delivery into the joints. In addition, the best suited concentration and size of nanoparticles, which will optimize gene delivery and expression, while minimizing intraarticular inflammation, needs to be determined.     

Magnetomicelles: composite nanostructures from magnetic nanoparticles and cross-linked amphiphilic block copolymers

We report the synthesis, characterization, and covalent surface chemistry of "magnetomicelles", cross-linked, amphiphilic block-copolymer micelles that encapsulate superparamagnetic iron oxide nanoparticles. Because these composite nanostructures assemble spontaneously from solution by simultaneous desolvation of nanoparticle and amphiphilic poly(styrene(250)-block-acrylic acid(13)) components, explicit surface functionalization of the particles is not required, and the encapsulation method was applied to different magnetic nanoparticle sizes and compositions. TEM images of the magnetomicelles illustrated that the number of encapsulated particles could be dictated rationally by synthetic conditions. The magnetic properties of the particles were characterized by SQUID magnetometry and followed the general Langevin magnetic model for superparamagnetic materials. The micellar shells of these particles were functionalized using covalent chemistry that would not ordinarily be possible on the magnetic particle surface. As a result, this noncovalent approach provides a new route to technological applications of hydrophobic magnetic nanomaterials that lack appropriate conjugate surface chemistry.     

Preparation and evaluation of magnetic nanoparticles for cell labeling

Cell labeling and tracking are becoming increasingly important in the fields of stem cell transplantation. To track the migration and distribution of the implanted cells is critical for understanding the beneficial effects of stem cell therapy. The aim of this study is to synthesize new superparamagnetic nanoparticles and investigate the feasibility of magnetic labeling of bone marrow mesenchymal stem cells (MSCs). Monodisperse hydrophobic magnetite (Fe3O4) nanoparticles were prepared through high temperature decomposition of Fe(acac)3 and a long-chain alcohol. The nanoparticles were further modified with a bipolar surfactant, 2,3-dimercaptosuccinic acid (DMSA) and then transformed into water-soluble iron oxide nanoparticles (WION). The magnetic particles showed uniform size (10 nm), high efficiency and stability in MSCs labeling. The labeled cells were cultured until passage 8, there is no reduction in magnetic tropism and the percentage of labeled cells. The results of MTT proliferation assay and flow cytometry analysis show that the WION are biocompatible. The labeling process does not cause cell death and apoptosis, and has no side effect on growth capacity of the cells. In conclusion, the successful and stable labeling of MSCs and the efficient magnetic tropism indicate that this WION can be used for tracking of MSCs in future MSCs-based stem cell therapy.     

SPION nanoparticles as an efficient probe and carrier of DNA to umbilical cord blood-derived mesenchymal stem cells

Superparamagnetic iron oxide nanoparticles (SPION) were used to transfer gene into umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) in this study. This novel transfection method using SPION is safe and effective to UCB-MSCs, and would be a tool for genetic optimization with a significant potential for cell tracing.     

Sorafenib delivery nanoplatform based on superparamagnetic iron oxide nanoparticles magnetically targets hepatocellular carcinoma

Currently,sorafenib is the only systemic therapy capable of increasing overall survival of hepatocellular carcinoma patients.Unfortunately,its side effects,particularly its overall toxicity,limit the therapeutic response that can be achieved.Superparamagnetic iron oxide nanoparticles (SPIONs) are very attractive for drug delivery because they can be targeted to specific sites in the body through application of a magnetic field,thus improving intratumoral accumulation and reducing adverse effects.Here,nanoformulations based on polyethylene glycol modified phospholipid micelles,loaded with both SPIONs and sorafenib,were successfully prepared and thoroughly investigated by complementary techniques.This nanovector system provided effective drug delivery,had an average hydrodynamic diameter of about 125 nm,had good stability in aqueous medium,and allowed controlled drug loading.Magnetic analysis allowed accurate determination of the amount of SPIONs embedded in each micelle.An in vitro system was designed to test whether the SPION micelles can be efficiently held using a magnetic field under typical flow conditions found in the human liver.Human hepatocellular carcinoma (HepG2) cells were selected as an in vitro system to evaluate tumor cell targeting efficacy of the superparamagnetic micelles loaded with sorafenib.These experiments demonstrated that this delivery platform is able to enhance sorafenib's antitumor effectiveness by magnetic targeting.The magnetic nanovectors described here represent promising candidates for targeting specific hepatic tumor sites,where selective release of sorafenib can improve its efficacy and safety profile.     

Preparation of Water-Dispersible and Biocompatible Iron Oxide Nanoparticles for MRI Agent

Highly monodisperse superparamagnetic iron oxide nanoparticles (SPIONs, 7.5 nm gamma- F₂O₃) were synthesized by thermal decomposition of iron pentacarbonyl and consecutive aeration in organic medium. By treating with a small amount of iron pentacarbonyl, Fe-rich surface has been formed on SPION. Water-dispersible SPIONs (SPION-MPA) were prepared by Fe-S covalent conjugation between Fe-rich SPION and mercaptopropionic acid (MPA) and then, transformed to SPION-MPA-dextran composite by physical adsorption of biocompatible polymer dextran. The hydrodynamic diameter of SPION-MPA-dextran was in the range of 225~237 nm in water. MR contrast and spin-spin relaxation intensity of our SPION-MPA-dextran were similar to those of the commercial products, Ferridex and Resovist.     

Characterization of the biocompatible magnetic colloid on the basis of Fe3O4 nanoparticles coated with dextran, used as contrast agent in magnetic resonance imaging

The magnetic resonance imaging contrast agent, the so-called Endorem colloidal suspension on the basis of superparamagnetic iron oxide nanoparticles (mean diameter of 5.5 nm) coated with dextran, were characterized on the basis of several measurement techniques to determine the parameters of their most important physical and chemical properties. It is assumed that each nanoparticle is consisted of Fe3O4 monodomain and it was observed that its oxidation to gamma-Fe2O3 occurs at 253.1 degrees C. The M?ssbauer spectroscopy have shown a superparamagnetic behavior of the magnetic nanoparticles. The Magnetic Resonance results show an increase of the relaxation times T1, T2, and T2* with decreasing concentration of iron oxide nanoparticles. The relaxation effects of SPIONs contrast agents are influenced by their local concentration as well as the applied field strength and the environment in which these agents interact with surrounding protons. The proton relaxation rates presented a linear behavior with concentration. The measured values of thermo-optic coefficient dn/dT, thermal conductivity kappa, optical birefringence delta n0, nonlinear refractive index n2, nonlinear absorption beta' and third-order nonlinear susceptibility |chi(3)| are also reported.