Superparamagnetic Iron Oxide Nanoparticles: Cytotoxicity, Metabolism, and Cellular Behavior in Biomedicine Applications.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely investigated and applied in the field of biomedicine due to their excellent superparamagnetic properties and reliable traceability. However, with the optimization of core composition, shell types and transfection agents, the cytotoxicity and metabolism of different SPIONs have great differences, and the labeled cells also show different cellular behaviors. Therefore, a holistic review of the construction and application of SPIONs is desired. This review focuses the advances of SPIONs in the field of biomedicine in recent years. After summarizing the toxicity of different SPIONs, the uptake, distribution and metabolism of SPIONs in vitro were discussed. Then, the regulation of labeled-cells behavior is outlined. Furthermore, the major challenges in the optimization process of SPIONs and insights on its future developments are proposed.

Variation in the concentration and regional distribution of magnetic nanoparticles in human brains, with and without Alzheimer's disease, from the UK.

The presence of magnetic nanoparticles (MNPs) in the human brain was attributed until recently to endogenous formation; associated with a putative navigational sense, or with pathological mishandling of brain iron within senile plaques. Conversely, an exogenous, high-temperature source of brain MNPs has been newly identified, based on their variable sizes/concentrations, rounded shapes/surface crystallites, and co-association with non-physiological metals (e.g., platinum, cobalt). Here, we examined the concentration and regional distribution of brain magnetite/maghemite, by magnetic remanence measurements of 147 samples of fresh/frozen tissues, from Alzheimer's disease (AD) and pathologically-unremarkable brains (80-98 years at death) from the Manchester Brain Bank (MBB), UK. The magnetite/maghemite concentrations varied between individual cases, and different brain regions, with no significant difference between the AD and non-AD cases. Similarly, all the elderly MBB brains contain varying concentrations of non-physiological metals (e.g. lead, cerium), suggesting universal incursion of environmentally-sourced particles, likely across the geriatric blood-brain barrier (BBB). Cerebellar Manchester samples contained significantly lower (~ 9×) ferrimagnetic content compared with those from a young (29 years ave.), neurologically-damaged Mexico City cohort. Investigation of younger, variably-exposed cohorts, prior to loss of BBB integrity, seems essential to understand early brain impacts of exposure to exogenous magnetite/maghemite and other metal-rich pollution particles.

Patterns of use, effectiveness and safety of gadolinium contrast agents: a European prospective cross-sectional multicentre observational study.

BACKGROUND: The EU gadolinium-based contrast agents (GBCA) market has changed in recent years due to the European Medicines Agency decision to suspend the marketing authorisation of linear GBCA and the marketing authorisation of new generic macrocyclic GBCA. The study aims to understand the patterns of (GBCA) use, and to study the effectiveness and safety of GBCA in routine practice across Europe. METHODS: Prospective, cross-sectional, multicentre, observational study in patients undergoing contrast-enhanced magnetic resonance. Reported usage patterns included indication, referral and examination details. Assessment of effectiveness included changes in radiological diagnosis, diagnostic confidence and image quality. Safety data were collected by spontaneous patient adverse event (AE) reporting. RESULTS: 2118 patients were included from 8 centres across 5 European countries between December 2018 and November 2019. Clariscan, Dotarem (gadoteric acid), Gadovist (gadobutrol) and ProHance (gadoteridol) were utilised in 1513 (71.4%), 356 (16.8%), 237 (11.2%) and 12 (0.6%) patients, respectively. Most were performed in CNS-related indications (46.2%). Mean GBCA doses were 0.10 mmol/kg body weight, except for Gadovist (mean 0.12 mmol/kg). GBCA use increased confidence in diagnosis in 96.2% of examinations and resulted in a change in radiological diagnosis in 73.9% of patients. Image quality was considered excellent or good in 96.1% of patients and across all GBCA. Four patients reported AEs (0.19%), with only 1 (0.05%) considered serious. CONCLUSIONS: This European study confirmed that GBCAs are used appropriately in Europe for a wide range of indications. The study demonstrated a significant increase in diagnostic confidence after GBCA use and confirmed the good safety profile of GBCAs, with comparable results for all agents used.

Magnetic-core-based silibinin nanopolymeric carriers for the treatment of renal cell cancer.

AIMS: Silibinin offers potential anticancer effect with less aqueous solubility and high permeability. The present study aimed to develop biocompatible magnetic-core-based nanopolymeric carriers of poly (D, l-lactide-co-glycolic) acid (PLGA) encapsulated silibinin for the sustained release action on renal cancerous cell. MAIN METHODS: The synthesized iron oxide nanoparticles were prepared by precipitation method via encapsulation of silibinin in PLGA network using double emulsion method. The nanoparticle formulations were characterized for morphological, physicochemical properties (HRTEM, FTIR, Raman Spectroscopy and VSM), in vitro drug release and cytotoxicity study on kidney cancer cells (A-498). The safety of magnetic-core-based silibinin nanopolymeric carriers was conducted by i.v. administration at a dose of 50 mg/kg in mice. KEY FINDINGS: The mean particle size, zeta potential and % encapsulation efficiency of magnetic-core-based silibinin nanopolymeric carriers were found to be 285.9 ± 0.28 nm, -14.71 ± 0.15 mV and 84.76 ± 1.29%, respectively. The saturation magnetization of magnetic core and optimized nanoparticles were reported as 36.35 emu/g and 12.78 emu/g, respectively. HRTEM analyses revealed the spherical shapes of the particles with uniform size distribution. The in vitro release profile of silibinin from the nanoparticles exhibited a sustained delivery for 15 days and displayed better cytotoxicity against human kidney cancer cells (A-498) than silibinin. In vivo study showed the safety of magnetic-core-based silibinin nanopolymeric carriers in mice. SIGNIFICANCE: The magnetic-core-based silibinin nanopolymeric carriers will act as a potential carrier for targeted transportation of actives in cancer therapy.

Brave new world revisited: Focus on nanomedicine.

Nanomedicine is at a crossroads: with relatively few success stories in terms of clinical translation despite more and more research on increasingly sophisticated nanomaterials, it is important to consider whether we are on the right track. Indeed, it is crucial that we address the fact that while considerable efforts are being made to overcome barriers to translation from the bench to the clinic, scientists are still struggling to decipher fundamental aspects of nanomaterial interactions with biological systems. We believe that a key to the successful adoption of nanomedicines in oncology and beyond lies in a deeper understanding of underlying biological processes and in decoding interactions between engineered nanomaterials and biological systems. Here we provide an overview of progress in nanomedicine during the past 5 years.

Promising insights into the kosmotropic effect of magnetic nanoparticles on proteins: The pivotal role of protein corona formation.

Increasing concerns about biosafety of nanoparticles (NPs) has raised the need for detailed knowledge of NP interactions with biological molecules especially proteins. Herein, the concentration-dependent effect of magnetic NPs (MNPs) on bovine serum albumin and hen egg white lysozyme was explored. The X-ray diffraction patterns, zeta potential, and dynamic light scattering measurements together with scanning electron microscopy images were employed to characterize MNPs synthesized through coprecipitation method. Then, we studied the behavior of two model proteins with different surface charges and structural properties on interaction with Fe3 O4 . A thorough investigation of protein-MNP interaction by the help of intrinsic fluorescence at different experimental conditions revealed that affinity of proteins for MNPs is strongly affected by the similarity of protein and MNP surface charges. MNPs exerted structure-making kosmotropic effect on both proteins under a concentration threshold; however, binding strength was found to determine the extent of stabilizing effect as well as magnitude of the concentration threshold. Circular dichroism spectra showed that proteins with less resistance to conformational deformations are more prone to secondary structure changes upon adsorption on MNPs. By screening thermal aggregation of proteins in the presence of Fe3 O4 , it was also found that like chemical stability, thermal stability is influenced to a higher extent in more strongly bound proteins. Overall, this report not only provides an integrated picture of protein-MNP interaction but also sheds light on the molecular mechanism underling this process.

Hemolysis tendency of anticancer nanoparticles changes with type of blood group antigen: An insight into blood nanoparticle interactions.

Different blood groups of ABO system have specific antigen which bestows them with different biochemical properties and hence they can show different hemolytic activity. In this report, hemolytic activity of thiol-functionalized Fe3O4-Au nanoparticles were studied in presence and absence of doxorubicin and the effect of various thiol coatings were correlated towards their hemolysis tendency. The nanoparticles were functionalized with four different amino thiols, cysteamine (CEA), cystamine (CA), cysteine (Cys) and cystine (Cyt) to form Fe3O4-Au CEA, Fe3O4-Au CA, Fe3O4-Au Cys and Fe3O4-Au Cyt nanoparticles which were loaded with anticancer drug, doxorubicin. The functionalization was characterized using ATR-FTIR, HR-TEM, XPS and other spectroscopic methods. Maximum drug encapsulation efficiency of 83% was observed with Fe3O4-Au CA nanoparticles. In-vitro experiments were performed on HeLa cells to check the cellular uptake and cytotoxicity using MTT assay. Hemolytic activity was then analyzed with all the blood groups (positive and negative). The amino acid functionalized, Fe3O4-Au Cys and Fe3O4-Au Cyt nanoparticles, shows lesser hemolysis compared to amino thiol functionalized Fe3O4-Au CEA, and Fe3O4-Au CA nanoparticles. In positive blood groups, the Fe3O4-Au CA nanoparticles shows the highest rate of hemolysis followed by Fe3O4-Au CEA, while the lowest hemolysis rate was observed for Fe3O4-Au Cyt nanoparticles. For negative blood groups, the thiol coated nanoparticles show more abrupt hemolysis rate depending upon the type of antigen.

Biocompatibility assessment of Fe3O4 nanoparticles using Saccharomyces cerevisiae as a model organism.

Using Saccharomyces cerevisiae as an experimental model, the potential toxicological effects of Fe3O4 nanoparticles (Fe3O4-NPs) were investigated following exposure to 0-600 mg/L for 24 h. Results revealed that cell proliferation was significantly inhibited by Fe3O4-NPs with an IC50 value of 326.66 mg/L. Mortality showed a concentration-dependent increase, and the highest concentration in this study (600 mg/L) resulted in 22.30% mortality. In addition, Effects on proliferation and mortality were accounted for Fe3O4-NPs rather than iron ion released from Fe3O4-NPs. Scanning and transmission electron microscope observation showed that Fe3O4-NPs extensively attached on the cell surfaces, causing cells to deform and shrink. Moreover, Fe3O4-NPs could be internalized in S. cerevisiae cells via endocytosis and then be distributed in cytoplasm and vesicles. The data of uptake kinetics demonstrated that the maximal accumulation (4.898 mg/g) was reached at 15 h. Besides, percentage of late apoptosis/necrosis was observably increased (p < 0.01) at 600 mg/L (15.80%), and the expression levels of apoptosis-related genes (SOD, Yca1 and Nuc1) were dramatically increased following exposure to Fe3O4-NPs for 24 h. As expected, mitochondrial transmembrane potential was significantly decreased (p < 0.01) at 50-600 mg/L, and biomarkers of oxidative stress (ROS, CAT and SOD) were also markedly changed following exposure. Altogether, the combined results so far indicated Fe3O4-NPs could induce S. cerevisiae cell apoptosis that mediated by mitochondrial impairment and oxidative stress.

Targeting of temozolomide using magnetic nanobeads: an in vitro study

Temozolomide, a chemotherapeutic drug that is often administered for the treatment of brain cancer has severe side effects and a poor aqueous solubility. In order to decrease the detrimental effect of the drug over healthy cells, a novel drug delivery vehicle was developed where the therapeutic drug was encapsulated within the hydrophobic cavities of b-CD modified magnetite nanoparticles, which are embedded in chitosan nanobeads prepared by salt addition. In-vitro studies have shown that the magnetic properties of the novel delivery vehicle are adequate for targeted drug delivery applications under an external magnetic field. Additionally, an increase in the amount of chitosan was shown to exhibit a strong shielding effect over the magnetic properties of the delivery vehicle, which lead to deterioration of the amount of captured drug at the targeted area, suggesting a delicate balance between the amounts of constituents composing the drug delivery vehicle.

Biochemical functionality of magnetic particles as nanosensors: how far away are we to implement them into clinical practice?

Magnetic nanosensors have become attractive instruments for the diagnosis and treatment of different diseases. They represent an efficient carrier system in drug delivery or in transporting contrast agents. For such purposes, magnetic nanosensors are used in vivo (intracorporeal application). To remove specific compounds from blood, magnetic nanosensors act as elimination system, which represents an extracorporeal approach. This review discusses principles, advantages and risks on recent advances in the field of magnetic nanosensors. First, synthesis methods for magnetic nanosensors and possibilities for enhancement of biocompatibility with different coating materials are addressed. Then, attention is devoted to clinical applications, in which nanosensors are or may be used as carrier- and elimination systems in the near future. Finally, risk considerations and possible effects of nanomaterials are discussed when working towards clinical applications with magnetic nanosensors.

Systemic exposure to a single dose of ferucarbotran aggravates neuroinflammation in a murine model of experimental autoimmune encephalomyelitis.

BACKGROUND: Medicinal preparations of iron oxide nanoparticles (IONPs) have been used as MRI contrast agents for the diagnosis of hepatic tumors and the assessment of neuroinflammation and blood-brain barrier integrity. However, it remains mostly unclear whether exposure to IONPs affects neuroinflammation under disease conditions. The present study aims to investigate the impact of IONPs on autoimmune-mediated neuroinflammation using a murine model of experimental autoimmune encephalomyelitis (EAE) that mimics human multiple sclerosis. METHODS: Mice were either left untreated or immunized with myelin oligodendrocyte glyco-protein on day 0 followed by two injections of pertussis toxin for EAE induction. The EAE mice were intravenously administered with a single dose of the carboxydextran-coated IONPs, ferucarbotran (20 mg Fe/kg) and/or saline (as vehicle) on day 18. Symptoms of EAE were daily monitored until the mice were killed on day 30. Tissue sections of the brain and spinal cord were prepared for histopathological examinations. Iron deposition, neuron demyelination and inflammatory cell infiltration were examined using histochemical staining. The infiltration of microglial and T cells, and cytokine expression were examined by immunohistochemical staining and/or reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Iron deposition was detected in both the brain and spinal cord of EAE mice 3 days post-ferucarbotran treatment. The clinical and pathological scores of EAE, percentage of myelin loss and infiltration of inflammatory cells into the spinal cord were significantly deteriorated in EAE mice treated with ferucarbotran. Furthermore, ferucarbotran treatment increased the number of CD3+, Iba-1+, IL-6+, Iba-1+TNF-α+ and CD3+IFN-γ+ cells in the spinal cord of EAE mice. CONCLUSION: A single exposure to ferucarbotran exacerbated neuroinflammation and disease severity of EAE, which might be attributed to the enhanced activation of microglia and T cells. These results demonstrated that the pro-inflammatory effect of ferucarbotran on the central nervous system is closely associated with the deterioration of autoimmunity.

Uptake and translocation of magnetite (Fe3O4) nanoparticles and its impact on photosynthetic genes in barley (Hordeum vulgare L.).

This study investigates the fate and impact of iron oxide or magnetite (Fe3O4, ∼13 nm in size) nanoparticles (NPs) in barley (Hordeum vulgare L.), a common crop cultivated around the world. Barley seedlings were grown in hydroponic culture for three weeks to include NPs (125, 250, 500, and 1000 mg/L). Transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) techniques were used to assess their uptake and translocation. Photosynthesis marker genes were quantified by RT-qPCR. Results revealed that increasing doses of Fe3O4 NPs were gradually enhanced the plant growth up to 500 mg/L, which promoted the fresh weight (FW) respectively ∼19% and ∼88% for leaf and root tissues than the ones for control. No phytotoxic effect was recorded even at high NPs doses. NPs inclusion increased some phenological parameters such as chlorophyll, total soluble protein, number of chloroplasts, and dry weight. High NPs doses dramatically reduced the catalase activity and hydrogen peroxide content, suggesting a possible function of NPs as nanozyme in vivo. TEM observations showed that Fe3O4 NPs penetrated and internalized in the root cells. In leaves, they were mostly existed at the surrounding cell wall, suggesting their translocation from root to shoot without cellular penetration. Further analysis by using VSM confirmed the existence of Fe3O4 NPs in leaves which result in dramatic alterations of the photosystem genes (PetA, psaA, BCA and psbA). In conclusion, barley plants uptake and translocate Fe3O4 NPs, which promoted the plant growth probably due to the promoted gene expression and efficient photosynthetic activity.

Toxic effects of magnetic nanoparticles on normal cells and organs.

Magnetic nanoparticles (MNPs) are promising candidates for drug delivery and treatment of various disorders. Toxicity evaluation is a critical point in the development of nanoformulations and therefore, draws considerable attention. Formulations involving individual or combinatorial nanoparticle suspensions might be used for targeted delivery and treatment. This might be a evaluated further for safety related issues considering future medications based on MNPs. Nanoparticle distribution in the body is dependent on its surface characteristics. Size, dose and routes of nanoparticle entry have to be taken into consideration for future assays.

Study of biodistribution and systemic toxicity of glucose functionalized SPIO/DOX micelles.

The present study examined the cytotoxicity and magnetic resonance imaging (MRI) distribution of cancer-targeted, MRI-visible polymeric micelles that encapsulate doxorubicin (DOX) and superparamagnetic iron oxide (SPIO) and are conjugated with glucose as a targeting ligand. In this study, the micelles were investigated the clinical potential of glucose-micelles, in vitro cytotoxicity assays of nonencapsulating or SPIO-and-DOX-coencapsulating micelles were performed on L929 mouse fibroblasts, and we found that glucose-micelles did not exert in vitro cytotoxic effects. Next, in vitro MRI detectability of glucose SPIO micelles was evaluated at the loaded SPIO content of 2.5% and 50%, and it was found that glucose-micelles can increase MRI relaxivity (r2*) at high SPIO loading. Furthermore, 50% SPIO micelles persisted in the blood circulation for up to 5 days (slow liver clearance) as determined by in vivo MRI. For in vivo toxicity evaluation, 50% SPIO/DOX micelles at a dose up to 18 (mg DOX)/(kg body weight) showed no impact on animal health according to clinical chemistry and clinical hematology laboratory testing. Altogether, these results indicate that glucose-micelles can serve as an effective and safe drug delivery system.

Dose-, treatment- and time-dependent toxicity of superparamagnetic iron oxide nanoparticles on primary rat hepatocytes.

AIM: As a first study in literature, to investigate concentration-dependent (0-400 µg/ml) and exposure-dependent (single dosing vs cumulative dosing) effects of superparamagnetic iron oxide nanoparticles (d = 10 nm) on primary rat hepatocytes in a time-dependent manner. MATERIALS & METHODS: Sandwich-cultured hepatocyte model was used to evaluate viability, hepatocyte specific functions and reactive oxygen species level. RESULTS: In terms of all parameters, generally statistically more significant effects were observed in a concentration- and time-dependent manner. In terms of hepatocyte-specific functions, cumulative dosing caused significantly (p < 0.05) more deleterious effects at 48th hour. CONCLUSION: A combination of various biomarkers should be employed for the evaluation of the effect of superparamagnetic iron oxide nanoparticles on liver, and each biomarker should be analyzed in a time- and exposure-dependent manner.

Colloidal stability of superparamagnetic iron oxide nanoparticles in the central nervous system: a review.

Superparamagnetic iron oxide nanoparticles (SPIONs) consist of nanosized metallic-based particles with unique magnetic properties. Their potential in both diagnostic and therapeutic applications in the CNS is at the source of an expanding body of the literature in recent years. Colloidal stability of nanoparticles represents their ability to resist aggregation and is a central aspect for the use of SPION in biological environment such as the CNS. This review gives a comprehensive update of the recent developments and knowledge on the determinants of colloidal stability of SPIONs in the CNS. Factors leading to aggregate formation and the repercussions of colloidal instability of SPION are reviewed in detail pertaining to their use in the CNS.

Biological responses to core-shell-structured Fe3O4@SiO2-NH2 nanoparticles in rats by a nuclear magnetic resonance-based metabonomic strategy.

BACKGROUND: Core-shell-structured nanoparticles (NPs) have attracted much scientific attention due to their promising potential in biomedical fields in recent years. However, their underlying mechanisms of action and potential adverse effects following administration remain unknown. METHODS: In the present study, a 1H nuclear magnetic resonance-based metabonomic strategy was applied to investigate the metabolic consequences in rats following the intravenous administration of parent NPs of core-shell-structured nanoparticles, Fe3O4@SiO2-NH2 (Fe@Si) NPs. RESULTS: Alterations reflected in plasma and urinary metabonomes indicated that Fe@Si NPs induced metabolic perturbation in choline, ketone-body, and amino-acid metabolism besides the common metabolic disorders in tricarboxylic acid cycle, lipids, and glycogen metabolism often induced by the exogenous agents. Additionally, intestinal flora metabolism and the urea cycle were also influenced by Fe@Si NP exposure. Time-dependent biological effects revealed obvious metabolic regression, dose-dependent biological effects implied different biochemical mechanisms between low- and high-dose Fe@Si NPs, and size-dependent biological effects provided potential windows for size optimization. CONCLUSION: Nuclear magnetic resonance-based metabonomic analysis helps in understanding the biological mechanisms of Fe@Si NPs, provides an identifiable ground for the selection of view windows, and further serves the clinical translation of Fe@Si NP-derived and -modified bioprobes or bioagents.

Preparation of an efficient and safe polymeric-magnetic nanoparticle delivery system for sorafenib in hepatocellular carcinoma.

AIMS: Superparamagnetic iron oxide nanoparticles (SPIONs), as drug delivery vehicles, offer to eliminate the concerns associated with hydrophobic anti-cancer agents. The current study was intended to fabricate a SPION based delivery system for sorafenib that can simultaneously enable targeted delivery of sorafenib and expand its therapeutic index against hepatocellular carcinoma (HCC). MAIN METHODS: Co-precipitation and physical entrapment methods were employed for the synthesis of sorafenib loaded PVA coated SPIONs. Physicochemical characterizations were done using TEM, XRD, FTIR, Raman spectra and VSM measurements. The superior activity of nanoconjugate was demonstrated by AO/EB staining, FACS, immunofluorescence and Western blot. The safety of the sorafenib conjugated nanoparticles were verified in Wistar rats. KEY FINDINGS: The synthesized nanoparticles were in the size range of 5-15 nm. The adsorption of PVA to the SPIONs and the conjugation of sorafenib to the nanocarrier were confirmed by XRD, FTIR and Raman spectra analyses. VSM study ascertained the superparamagnetic nature of the nanoconjugate. Cellular uptake studies suggested its efficient entrapment in HepG2 cells. MTT assay showed that the cytotoxicity of sorafenib loaded PVA/SPIONs was comparable or higher than free sorafenib. The activation of apoptosis and autophagy pathways in HepG2 by the nanoconjugate was evidenced. Acute toxicity testing in Wistar rats supported the safe administration of the nanoconjugate and established its localization in animal tissues by Perl's Prussian Blue reaction. SIGNIFICANCE: The novel combination of sorafenib with PVA/SPIONs showed better anticancer efficiency than free sorafenib demonstrative of its potential in cancer chemotherapy.

Cellular and Molecular Toxicity of Iron Oxide Nanoparticles.

Iron oxide nanoparticles (ION) have attracted much attention because of their particular physico-chemical properties, including superparamagnetism. These features make them suitable for many purposes and several interesting biomedical applications, such as to increase contrast in magnetic resonance imaging (MRI), as drug delivery systems and as hyperthermia agents. However, they have also shown to be easily accumulated in diverse tissues and induce toxicity at different levels. This chapter reviews the different cellular and molecular effects induced by ION reported from in vitro studies with human and non-human cell lines. Those effects are mainly dependent on ION type and concentration, time of exposure, presence and nature of coating, and cell type evaluated. They include decreases in viability, plasmatic membrane disruption, oxidative damage, mitochondrial alterations, cell cycle impairments, cytoskeleton disruption, cell death, and alterations in cell motility, and in cell integrity. Despite these negative effects, the numerous advantages of ION together with their promising applications in biomedicine, make it necessary to clearly define their toxicity in order to discard potential health risks and to reach optimal benefits of their use.

Comparative study of nano and bulk Fe3O4 induced oxidative stress in Wistar rats.

CONTEXT: Magnetic nanomaterials (Fe3O4 NMs) have become novel tools with multiple biological and medical applications because of their biocompatibility. However, adverse health effects of these NMs are of great interest to learn. OBJECTIVE: This study was designed to assess the size and dose-dependent effects of Fe3O4 NMs and its bulk on oxidative stress biomarkers after post-subacute treatment in female Wistar rats. METHODS: Rats were daily administered with 30, 300 and 1000 mg/kg b.w. doses for 28 d of Fe3O4 NMs and its bulk for biodistribution and histopathological studies. RESULTS: Fe3O4 NMs treatment caused significant increase in lipid peroxidation levels of treated rats. It was also observed that the NM treatment elicited significant changes in enzyme activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase  and glutathione-S-transferase in treated rat organs with major reduction in glutathione content. Metal content analysis revealed that tissue deposition of NM in the organs was higher when compared to bulk and caused histological changes in liver. CONCLUSION: This study demonstrated that for same dose, NM showed higher bioaccumulation, oxidative stress and tissue damage than its bulk. The difference in toxic effect of Fe3O4 nano and bulk could be related to their altered physicochemical properties.