The Interplay between Fe3O4 Superparamagnetic Nanoparticles,Sodium Butyrate,and Folic Acid for Intracellular Transport

Combined treatments which use nanoparticles and drugs could be a synergistic strategy for the treatment of a variety of cancers to overcome drug resistance, low efficacy, and high-dose-induced systemic toxicity. In this study, the effects on human colon adenocarcinoma cells of surface modified Fe₃O₄ magnetic nanoparticles (MNPs) in combination with sodium butyrate (NaBu), added as a free formulation, were examined demonstrating that the co-delivery produced a cytotoxic effect on malignant cells. Two different MNP coatings were investigated: a simple polyethylene glycol (PEG) layer and a mixed folic acid (FA) and PEG layer. Our results demonstrated that MNPs with FA (FA-PEG@MNPs) have a better cellular uptake than the ones without FA (PEG@MNPs), probably due to the presence of folate that acts as an activator of folate receptors (FRs) expression. However, in the presence of NaBu, the difference between the two types of MNPs was reduced. These similar behaviors for both MNPs likely occurred because of the differentiation induced by butyrate that increases the uptake of ferromagnetic nanoparticles. Moreover, we observed a strong decrease of cell viability in a NaBu dose-dependent manner. Taking into account these results, the cooperation of multifunctional MNPs with NaBu, taking into consideration the particular cancer-cell properties, can be a valuable tool for future cancer treatment.     

Preparation,physicochemical characterization,and AC induction heating properties of colloidal aggregates of ferrimagnetic cobalt ferrite nanoparticles coated with a bio-compatible polymer

AC induction heating properties of colloidal nano-aggregates of ferrimagnetic cobalt ferrite magnetic nanoparticles (MNPs) are reported in this study. Bio-compatible chitosan polymer-coated CoFe₂O₄ MNPs are synthesized using a co-precipitation method. Powder X-ray diffraction indicates the formation of mixed spinel structures for the uncoated (CP) and chitosan-coated (CP–CHN) MNPs, which is also supported by the cation distributions obtained from the Mössbauer spectra. The presence of chitosan coating on the surface of the CP-CHN MNPs is confirmed using X-ray photoelectron and Fourier transform infrared spectroscopy studies. Transmission electron microscopy shows primary particle sizes of ∼13 nm, which is larger than the superparamagnetic size limit of the CoFe₂O₄ MNPs. Hence, the CP and CP-CHN MNPs exhibit ferrimagnetic behaviour at room temperature with estimated saturation magnetization values of ∼77.4 emu/g and ∼74.4 emu/g, respectively. The average hydrodynamic diameter is found to be ∼90 ± 8 nm for an aqueous dispersion of the CP-CHN MNPs, which indicate the formation of colloidal nano-aggregates due to the ferrimagnetic interaction of the primary MNPs. The CP-CHN sample exhibits a significantly high AC induction heating efficiency of ∼267.2 ± 4.0 W/g_Fe, where the higher heating efficiency is attributed to the combination of hysteresis and relaxation-mediated magneto-thermal energy conversion, as confirmed using Stoner-Wohlfarth model-based dynamic hysteresis loop calculations. Further, the heating efficiency decreases with increasing sample concentration due to an increase in dipolar interaction, which is confirmed using semi-empirical calculations, where a lowering of the initial susceptibility is observed at higher concentrations. The higher AC induction heating efficiency, coupled with the demonstrated significant bio-compatibility during in vitro cytotoxicity studies, make the cobalt ferrite nano-aggregates potential candidates for magnetic hyperthermia.     

Syntheses of Fe-Ni Magnetic Nanoparticles and Their Applications on Biologic Labeling

In this study, we compared FeNi alloy magnetic nanoparticles (MNPs) prepared by either combustion or chemical precipitation methods. We found that the FeNi MNPs generated by combustion method have a rather high saturation magnetization Ms of～180 emu/g and a coercivity field Hc of near zero. However, the alloy nanoparticles are easily aggregated and are not well dispersive such that size distribution of the nanoparticle clusters is wide and clusters are rather big (around 50～700 nm). To prepare a better quality and well dispersed Fe-Ni MNPs, we also developed a thermal reflux chemical precipitation method to synthesize FeNi3 alloy MNPs. The precursor chemicals of Fe(acac)3 and Ni(acac)2 in a molecular ratio 1,2-hexadecandiol and tri-n-octylphosphine oxide (TOPO) were used as reducer and surfactant, respectively. The chemically precipitated FeNi3 MNPs are well dispersed and have well-controlled particle sizes around 10～20 nm with a very narrow size distribution (±1.2 nm). The highly monodispersive FeNi3 MNPs present good uniformity in particle shape and crystallinity on particle surfaces. The MNPs exhibit well soft magnetism with saturation magnetization of ～61 emu/g and Hc～0. The biomedically compatible FeNi MNPs which were coated with biocompatible polyethyleneimine (PEI) polymer were also synthesized. We demonstrated that the PEI coated FeNi MNPs can enter the mammalian cells in vitro and can be used as a magnetic resonance imagine (MRI) contrast agent. The results demonstrated that FeNi MNPs potentially could be applied in the biomedical field. The functionalized magnetic beads with biocompatible polymer coated on MNPs are also completed for biomedical applications.     

Synthesis of thermoresponsive copolymer/silica-coated magnetite nanoparticle composite and its application for heavy metal ion recovery

To enhance chemical stability and suppress of aggregation of magnetite nanoparticles (MNPs), which are used as a support for thermoresponsive copolymer immobilization, silica coating of the MNPs is applied via the electrooxidation method. Although the resulting silica coated-MNPs also formed aggregates, the size distribution of the aggregate shifted to smaller size range. Because of that, the surface area available for copolymer immobilization increased approximately 6.7 times at maximum as compared with that of the uncoated MNPs. It contributed to the increase of the amount of the immobilized copolymer on the silica-coated MNPs, which is approximately four times larger than that on the uncoated MNPs. Fe₃O₄ dissolution test confirmed enhancement of chemical stability of MNPs. The thermoresponsive copolymer immobilized on the silica-coated MNPs shows the ability to recycle Cu(II) ion from Cu(II) containing solution by changing temperature with significantly shorter time than those in other thermoresponsive adsorbents in gel form.     

The effect of two novel amino acid-coated magnetic nanoparticles on survival in vascular endothelial cells,bone marrow stromal cells,and macrophages

Magnetic nanoparticles (MNPs) have been popularly used in many fields. Recently, many kinds of MNPs are modified as new absorbents, which have attracted considerable attention and are promising to be applied in waste water. In our previous study, we synthesized two novel MNPs surface-coated with glycine or lysine, which could efficiently remove many anionic and cationic dyes under severe conditions. It should be considered that MNP residues in water may exert some side effects on human health. In the present study, we evaluated the potential nanotoxicity of MNPs in human endothelial cells, macrophages, and rat bone marrow stromal cells. The results showed that the two kinds of nanoparticles were consistently absorbed into the cell cytoplasm. The concentration of MNPs@Gly that could distinctly decrease survival was 15 μg/ml in human umbilical vascular endothelial cells (HUVECs) or bone marrow stromal cells (BMSCs) and 10 μg/ml in macrophages. While the concentration of MNPs@Lys that obviously reduced viability was 15 μg/ml in HUVECs or macrophages and 50 μg/ml in BMSCs. Furthermore, cell nucleus staining and cell integrity assay indicated that the nanoparticles induced cell apoptosis, but not necrosis even at a high concentration. Altogether, these data suggest that the amino acid-coated magnetic nanoparticles exert relatively high cytotoxicity. By contrast, lysine-coated magnetic nanoparticles are more secure than glycine-coated magnetic nanoparticles.     

Magnetic Nanoparticles as Intraocular Drug Delivery System to Target Retinal Pigmented Epithelium (RPE)

One of the most challenging efforts in drug delivery is the targeting of the eye. The eye structure and barriers render this organ poorly permeable to drugs. Quite recently the entrance of nanoscience in ocular drug delivery has improved the penetration and half-life of drugs, especially in the anterior eye chamber, while targeting the posterior chamber is still an open issue. The retina and the retinal pigment epithelium/choroid tissues, located in the posterior eye chamber, are responsible for the majority of blindness both in childhood and adulthood. In the present study, we used magnetic nanoparticles (MNPs) as a nanotool for ocular drug delivery that is capable of specific localization in the retinal pigmented epithelium (RPE) layer. We demonstrate that, following intraocular injection in Xenopus embryos, MNPs localize specifically in RPE where they are retained for several days. The specificity of the localization did not depend on particle size and surface properties of the MNPs used in this work. Moreover, through similar experiments in zebrafish, we demonstrated that the targeting of RPE by the nanoparticles is not specific for the Xenopus species.     

Exploiting Size-Dependent Drag and Magnetic Forces for Size-Specific Separation of Magnetic Nanoparticles

Realizing the full potential of magnetic nanoparticles (MNPs) in nanomedicine requires the optimization of their physical and chemical properties. Elucidation of the effects of these properties on clinical diagnostic or therapeutic properties, however, requires the synthesis or purification of homogenous samples, which has proved to be difficult. While initial simulations indicated that size-selective separation could be achieved by flowing magnetic nanoparticles through a magnetic field, subsequent in vitro experiments were unable to reproduce the predicted results. Magnetic field-flow fractionation, however, was found to be an effective method for the separation of polydisperse suspensions of iron oxide nanoparticles with diameters greater than 20 nm. While similar methods have been used to separate magnetic nanoparticles before, no previous work has been done with magnetic nanoparticles between 20 and 200 nm. Both transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis were used to confirm the size of the MNPs. Further development of this work could lead to MNPs with the narrow size distributions necessary for their in vitro and in vivo optimization.     

Preparation of novel porous calcium silicate scaffold loaded by celecoxib drug using freeze drying technique: Fabrication,characterization and simulation

In the current investigation, the microarchitecture of bio-nanocomposite scaffold, which is fabricated by natural synthetic diopside and composed of magnetite nanoparticles (MNPs), is considered. The MNPs are tested with various weight fractions (0, 5, 10, and 15 wt%) and are manufactured by the freeze-drying technique using sodium alginate as base matrix for the first time. Due to the limitation of the mechanical properties of calcium phosphates (CaPs) and bioactive glasses (BG), clinical usage of calcium silicate ceramics (CSC) are greatly affected. Therefore, CSCs are produced with the incorporation of metal oxides into the base binary xCaO-ySiO₂, as well as the substitution of calcium ions. Furthermore, mechanical and biological properties of CSCs are enhanced, which are a result of the ability to give out bioactive ions and their distinct compositions. After that, the porous bio-nanocomposite scaffolds are investigated for biological and mechanical properties corresponding to hardness versus elastic modulus, apatite formation versus biodegradation rate, wetting properties versus roughness and electrical conductivity of the sample. Then, the composition, microstructure, and physical characteristics are also examined using different techniques such as X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) which is equipped with energy-dispersive X-ray spectroscopy (EDX). The obtained outcomes show that addition of diopside bioceramic enhances the mechanical and physical properties of the samples. It is shown that the prepared porous bio-nanocomposite scaffolds, containing 10 wt% MNPs, represents a better agreement in serving as a bone graft for the cancer disease treatment and hyperthermia term. The results indicate that the specimen with 10 wt% MNPs in the bio-nanocomposite release the celecoxib drug easier, however, its has better porosity and mechanical behavior that make it suitable candidate for bone implantations.     

Fabrication of Poly（y-glutamic acid）-coated Fe3O4 Magnetic Nanoparticles and Their Application in Heavy Metal Removal

In this study, poly（y-glutamic acid）-coated Fe3O4 magnetic nanoparticles （y-PGA/Fe304 MNPs） were successfully fabricated using the co-precipitation method. Fe3O4 MNPs were also prepared for comparison. The av erage size and specific surface area results reveal that 7-PGA/Fe304 MNPs （52.4 nm, 88.41 m2.g-1） have smaller particle size and larger specific surface area_ than Fe3O4 MNPs （62.0 nm, 76.83 mLg-1）. The y-PGA/Fe3O4 MNPs     

Trihydrazinotriazine-grafting Fe3O4/SiO2 core-shell nanoparticles with expanded porous structure for organic reactions

This study focuses on the synthesis and characterization of a novel magnetic nanocomposite 2,4,6-trihydrazino-1,3,5-triazine (THDT)-functionalized with silica-coated iron oxide magnetic nanoparticles (MNPs). This nanocomposite has porous morphology decorated with the spherical MNPs. Through co-precipitation of iron salts, MNPs were obtained. The prepared THDT was placed on the chlorine surface-modified MNPs. The present environment-friendly nanocatalyst intensely accelerated the synthesis of highly functionalized tetrahydrobenzo[b]pyran derivatives as well as reduced the reaction times and increased yields of the products.     

Fabrication of a Chitosan‐Coated Magnetic Nanobiocatalyst for Starch Hydrolysis

The preparation of chitosan‐coated magnetic nanoparticles (MNPs) and covalent immobilization of α‐amylase for starch hydrolysis was investigated. Surface morphology, chemical composition, and structural characteristics of the MNPs were analyzed by scanning electron microscopy, energy dispersion spectrometry, and X‐ray diffractometry, respectively. Surface functional groups of MNPs, chitosan‐coated MNPs, and α‐amylase‐immobilized MNPs were characterized by Fourier transform infrared spectroscopy. Response surface methodology based on three levels was implemented to optimize three immobilization conditions and a regression model was developed. α‐Amylase‐immobilized MNPs provided better stability towards pH and temperature. The prepared thermostable nanobiocatalyst is well‐suited for industrial processes involving starch hydrolysis.     

Phosphocholine-Modified Magnetic Nanoparticles for Isolation of C-Reactive Protein from Human Serum

In the present study, we describe the easy isolation of C-reactive protein (CRP) from human serum using phosphocholine-modified magnetic nanoparticles (MNPs). A phosphocholine-based monomer, 3-(4)-vinylbenzyl-12-phosphorylcholine dodecanoate (VPC), was polymerized on methacrylate-coated MNPs and the resulting MNPs (VPC-MNPs) were assessed for their ability to isolate CRP from a human serum sample. CRP could be isolated from human serum with one adsorption step to VPC-MNPs within 1 h. The high purity of the isolated CRP fraction determined by SDS-PAGE indicates a good selectivity of VPC-MNPs for CRP binding. These results demonstrate the feasibility of using ligand-functionalized MNPs for rapid, easy, and efficient protein isolation.     

Size-regulated group separation of CoFe2O4 nanoparticles using centrifuge and their magnetic resonance contrast properties

Magnetic nanoparticle (MNP)-based magnetic resonance imaging (MRI) contrast agents (CAs) have been the subject of extensive research over recent decades. The particle size of MNPs varies widely and is known to influence their physicochemical and pharmacokinetic properties. There are two commonly used methods for synthesizing MNPs, organometallic and aqueous solution coprecipitation. The former has the advantage of being able to control the particle size more effectively; however, the resulting particles require a hydrophilic coating in order to be rendered water soluble. The MNPs produced using the latter method are intrinsically water soluble, but they have a relatively wide particle size distribution. Size-controlled water-soluble MNPs have great potential as MRI CAs and in cell sorting and labeling applications. In the present study, we synthesized CoFe₂O₄ MNPs using an aqueous solution coprecipitation method. The MNPs were subsequently separated into four groups depending on size, by the use of centrifugation at different speeds. The crystal shapes and size distributions of the particles in the four groups were measured and confirmed by transmission electron microscopy and dynamic light scattering. Using X-ray diffraction analysis, the MNPs were found to have an inverse spinel structure. Four MNP groups with well-selected semi-Gaussian-like diameter distributions were obtained, with measured T₂ relaxivities (r₂) at 4.7 T and room temperature in the range of 60 to 300 mM⁻¹s⁻¹, depending on the particle size. This size regulation method has great promise for applications that require homogeneous-sized MNPs made by an aqueous solution coprecipitation method. Any group of the CoFe₂O₄ MNPs could be used as initial base cores of MRI T₂ CAs, with almost unique T₂ relaxivity owing to size regulation. The methodology reported here opens up many possibilities for biosensing applications and disease diagnosis.

PACS

75.75.Fk, 78.67.Bf, 61.46.Df     

“双碳”下金属纳米颗粒优化暗发酵制氢策略及前景

氢能是“双碳”下推动化石能源低碳转型的重要方向,微生物暗发酵制氢是实现生物质绿氢转化的有效途径。其中,利用具有量子尺寸效应、比表面积大和电导率高的金属纳米颗粒（MNPs）优化暗发酵制氢技术是近年研究热点。综述和评论了国内外添加MNPs用于优化暗发酵制氢性能的作用机制、技术难点和制氢效果等,重点阐述并比较了铁、镍和锌基三类热门MNPs优化策略在提高产氢酶系活性、增强代谢产氢途径和优化微生物群落结构等方面的作用,展望了暗发酵制氢可深入MNPs优化氢化酶活性、拓宽生物质发酵底物以及产氢菌筛选和反应器设计、生物质发酵技术开发等研究方向和应用前景。     

Novel magnetic nanoparticles Fe3O4-immobilized domino Knoevenagel condensation,Michael addition,and cyclization catalyst

The synthesis, characterization and applications of guanidine supported on magnetic nanoparticles Fe₃O₄ (MNPs–Guanidine) as a novel magnetically separable base nanocatalyst are described. We have studied the application of this new catalyst for the Knoevenagel condensation reaction of aromatic aldehydes with malononitrile or cyanoacetate in PEG/water = 1:1 at room temperature. Also, the three-component and one-pot synthesis of 2-amino-4H-chromenes and 2-amino-4H-benzo[h]chromenes by condensing aldehydes, malononitrile and cyclic 1,3-dicarbonyl/α-naphthol, in the presence of catalytic amount of MNPs–Guanidine under same conditions is investigated. The supported catalyst could simply be separated and recovered from the reaction mixture with the assistance of an external magnet and reused several times.     

Synthesis and magneto-structural properties of chitosan coated ultrafine cobalt ferrite nanoparticles for magnetic fluid hyperthermia in viscous medium

AC induction heating mediated magnetic fluid hyperthermia of superparamagnetic nanoparticles (MNPs) is being widely explored for localized thermo-therapy of tumours. One of the primary hindrances for rapid adaptation of this technique is the loss of heating efficiency when the MNPs are placed within the viscous tissue medium, which necessitates undesired increase in MNP concentrations or exposure time during practical applications. With an objective to mitigate this, here we report the viscosity independent magnetic hyperthermia properties of biocompatible ultrafine (average size ～ 2.5 nm) chitosan-coated superparamagnetic CoFe₂O₄ MNPs synthesized using a low-cost co-precipitation technique. The presence of the chitosan coating is confirmed from Fourier transform infrared and X-ray photoelectron spectroscopy. The superparamagnetic nature of the synthesized MNPs at 300 K is confirmed from Mössbauer spectroscopy, isothermal and temperature dependent magnetization studies. Experimental findings indicate a higher field-induced heating efficiency for the chitosan-coated MNPs due to superior colloidal stability. The ultrafine size, combined with higher anisotropy energy density, results in viscosity independent Nèel relaxation-dominated magneto-thermal energy conversion for the CoFe₂O₄ MNPs. Experimental results reveal negligible loss of heating efficiency due to partial abrogation of Brownian relaxation when the chitosan-coated MNPs are immobilized in a tissue-equivalent agar medium, which is beneficial for practical applications. The heating efficiency of ～72.1 ± 2.8 W/g_Fe (at 33.1 kA/m and 126 kHz), obtained in the present study for the chitosan-coated MNPs, is higher than the previously documented values for ultrafine CoFe₂O₄ MNPs, which is useful for reducing the exposure time during practical applications. Further, the chitosan coating rendered the ultrafine CoFe₂O₄ MNPs bio-compatible against L929 cell line. The satisfactory magnetic fluid hyperthermia efficiency, negligible room temperature coercivity, retention of the field-induced heating efficiency in tissue-equivalent agar medium due to Nèel-dominated relaxation dynamics and superior biocompatibility, make the chitosan-coated ultrafine CoFe₂O₄ MNPs an attractive candidate for practical MFH applications.     

金属纳米颗粒辅助木质纤维素暗发酵生物制氢的研究进展

通过文献计量学分析表明暗发酵制氢是目前研究最热门的生物制氢方法,Fe、Ni、Co、Ag等金属纳米颗粒作为该领域研究热点可改善暗发酵制氢存在底物转化率与产氢效率均有待提高的难题。介绍了金属纳米颗粒的特点、生物相容性及其与酶、微生物细胞的作用机理,进一步从促进木质纤维素水解影响产氢、对水解酶的固定化影响产氢、提高氢化酶活性影响产氢、调控发酵微生物细胞代谢和促进细胞电子传递影响产氢、改善微生物群落结构影响多菌群协同产氢等几个方面对典型金属纳米颗粒辅助木质纤维素暗发酵产氢的研究现状进行综述,并对金属纳米颗粒应用于暗发酵产氢存在的难点及前景方向进行了展望。     

Establishment of a method to determine the magnetic particles in mouse tissues

This work is aimed to evaluate a method to detect the residual magnetic nanoparticles (MNPs) in animal tissues. Ferric ions released from MNPs through acidification with hydrochloric acid can be measured by complexation with potassium thiocyanate. MNPs in saline could be well detected by this chemical colorimetric method, whereas the detected sensitivity decreased significantly when MNPs were mixed with mouse tissue homogenates. In order to check the MNPs in animal tissues accurately, three improvements have been made. Firstly, proteinase K was used to digest the proteins that might bind with iron, and secondly, ferrosoferric oxide (Fe₃O₄) was collected by a magnetic field which could capture MNPs and leave the bio-iron in the supernatant. Finally, the collected MNPs were carbonized in the muffle furnace at 420°C before acidification to ruin the groups that might bind with ferric ions such as porphyrin. Using this method, MNPs in animal tissues could be well measured while avoiding the disturbance of endogenous iron and iron-binding groups.     

Morphological effect of oscillating magnetic nanoparticles in killing tumor cells

Forced oscillation of spherical and rod-shaped iron oxide magnetic nanoparticles (MNPs) via low-power and low-frequency alternating magnetic field (AMF) was firstly used to kill cancer cells in vitro. After being loaded by human cervical cancer cells line (HeLa) and then exposed to a 35-kHz AMF, MNPs mechanically damaged cell membranes and cytoplasm, decreasing the cell viability. It was found that the concentration and morphology of the MNPs significantly influenced the cell-killing efficiency of oscillating MNPs. In this preliminary study, when HeLa cells were pre-incubated with 100 μg/mL rod-shaped MNPs (rMNP, length of 200 ± 50 nm and diameter of 50 to 120 nm) for 20 h, MTT assay proved that the cell viability decreased by 30.9% after being exposed to AMF for 2 h, while the cell viability decreased by 11.7% if spherical MNPs (sMNP, diameter of 200 ± 50 nm) were used for investigation. Furthermore, the morphological effect of MNPs on cell viability was confirmed by trypan blue assay: 39.5% rMNP-loaded cells and 15.1% sMNP-loaded cells were stained after being exposed to AMF for 2 h. It was also interesting to find that killing tumor cells at either higher (500 μg/mL) or lower (20 μg/mL) concentration of MNPs was less efficient than that achieved at 100 μg/mL concentration. In conclusion, the relatively asymmetric morphological rod-shaped MNPs can kill cancer cells more effectively than spherical MNPs when being exposed to AMF by virtue of their mechanical oscillations.     

Intracellular Exposure Dose-Associated Susceptibility of Steatotic Hepatocytes to Metallic Nanoparticles

Non-alcoholic fatty liver disease (NAFLD), mainly characterized by the accumulation of excess fat in hepatocytes, is the most prevalent liver disorder afflicting ~25% of adults worldwide. In vivo studies have shown that adult rodents with NAFLD were more sensitive to metallic nanoparticles (MNPs) than healthy MNPs. However, due to the complex interactions between various cell types in a fatty liver, it has become a major challenge to reveal the toxic effects of MNPs to specific types of liver cells such as steatotic hepatocytes. In this study, we reported the susceptibility of steatotic hepatocytes in cytotoxicity and the induction of oxidative stress to direct exposures to MNPs with different components (silver, ZrO₂, and TiO₂ NPs) and sizes (20–30 nm and 125 nm) in an oleic acid (OA) -induced steatotic HepG2 (sHepG2) cell model. Furthermore, the inhibitory potential of MNPs against the process of fatty acid oxidation (FAO) were obvious in sHepG2 cells, even at extremely low doses of 2 or 4 μg/mL, which was not observed in non-steatotic HepG2 (nHepG2) cells. Further experiments on the differential cell uptake of MNPs in nHepG2 and sHepG2 cells demonstrated that the susceptibility of steatotic hepatocytes to MNP exposures was in association with the higher cellular accumulation of MNPs. Overall, our study demonstrated that it is necessary and urgent to take the intracellular exposure dose into consideration when assessing the potential toxicity of environmentally exposed MNPs.