Facile fabrication route for binary transition-metal oxides Janus ...

Facile

fabrication

route

for

binary

transition-metal oxides Janus nanoparticles for cancer theranostic applications M. Zubair Iqbal1, Wenzhi Ren1, Madiha Saeed1, Tianxiang Chen1, Xuehua Ma1, Xu Yu1, Jichao Zhang2, Lili Zhang2, Aiguo Li2, Aiguo Wu1* 1

Key Laboratory of Magnetic Materials and Devices, CAS & Key Laboratory of Additive Manufacturing Materials of Zhejiang Province, & Division of Functional Materials and Nano devices, Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, No. 1219 ZhongGuan West Road, 315201, Ningbo, China 2 Shanghai Synchrotron Radiation Facility, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201204, P. R. China

A unique solution based method is employed to fabricate Mn3O4-TiO2/ZnO/Fe3O4 multifunctional binary transition-metal oxide based Janus nanoparticles (JNPs) using the concept of epitaxial growth and lattice mismatch among synthesized materials. These multifunctional Mn3O4-TiO2 Janus nanoparticles enhance T1 MRI contrast agent in heart, liver and kidneys with excellent tumor ablation ability under the photodynamic therapy function.

Nano Research DOI (automatically inserted by the publisher) Research Article

Facile fabrication route for binary transition-metal oxides Janus nanoparticles for cancer theranostic applications 1

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M. Zubair Iqbal , Wenzhi Ren , Madiha Saeed , Tianxiang Chen , Xuehua Ma , Xu Yu , Jichao Zhang , Lili 2 2 1 Zhang , Aiguo Li , Aiguo Wu () 1

Key Laboratory of Magnetic Materials and Devices, CAS & Key Laboratory of Additive Manufacturing Materials of Zhejiang Province, & Division of Functional Materials and Nano devices, Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, No. 1219 ZhongGuan West Road, 315201, Ningbo, China 2 Shanghai Synchrotron Radiation Facility, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201204, P. R. China

Received: day month year

ABSTRACT

Revised: day month year

Janus nanoparticles with multiple configurations of molecular imaging, targeting and therapeutic functions in cancers made them very attractive for biomedical applications. However, smart strategies for the controlled synthesis in liquid phase and explore the appropriate applications of Janus nanoparticles remain a challenge. In current investigation, a unique solution based method was employed to fabricate Mn3O4-TiO2/ZnO/Fe3O4 multifunctional binary transition-metal oxide based Janus nanoparticles (JNPs) using the concept of epitaxial growth and lattice mismatch among synthesized materials. Transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) results have demonstrated that the prepared materials are embedded in the form of dimmers with good dispersion and homogeneous growth in non-polar solvent. Pluronic® F-127 coated Mn3O4-TiO2 JNPs were employed as contrast agent in T1- weighted magnetic resonance imaging (MRI) and photodynamic therapy (PDT) for cancers in vitro and in vivo. In vivo T1-weighted MR imaging of heart, liver, and kidney in mice after intravenous injection of nanoparticles further verified the high sensitivity and biocompatibility of as-synthesized Mn3O4-TiO2 JNPs. Synchrotron X-ray fluorescence microscopy (SXRF) microscopy mapping results showed the stability of nanocomposites and strength of penetrating inside cytoplasm and around nucleus. Inorganic photosensitizers TiO2 demonstrated potential PDT tumor ablation performance in vitro and in vivo at very low intensity of UV (5.6 mW/cm2) because of their ultra-small size and photodegradable stability. These results reveal that multifunctional Mn3O4-TiO2 Janus nanoparticles enhance T1 MRI contrast agent with excellent ability of photodynamic therapy function which might be a novel candidate for cancer theranostic in future.

Accepted: day month year (automatically inserted by the publisher) © Tsinghua University Press and Springer-Verlag Berlin Heidelberg 2014

KEYWORDS Janus Nanostructures, Manganese Oxide, Magnetic Resonance Imaging, Photodynamic Therapy, Breast Cancer

Address correspondence to Aiguo Wu, [email protected]

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synthesis.

1 Introduction Excellent control on the heterogeneous nucleation of unfamiliar materials with anisotropic products in solution is generally a challenging assignment. The successful heterogeneous nucleation of alien metals in a single system shaped them into attractive configuration named Janus structure. Therefore, Janus nanoparticles are non-centrosymmetric nanoparticles with two distinct faces and exhibit multifunctional properties which cannot be observed in the individual component. These synergetic/collective characteristics result form a combined chemical, magnetic, optical and electronic communication at the solid-state heterojunction of JNPs [1, 2]. At nanoscale, these interfacial characteristics are not restricted only to compartmentalized areas, but these may spread throughout the particle by crystal growth of constituent domains resulting enhancement in functionality. These fascinating features of Janus colloidal nanomaterials or two-colored Janus particles make them promising candidates for a range of applications including heterogeneous catalysis [3, 4], sensing devices [5], solar energy conversion into electrical current [6], imaging probes [7, 8], active targeting [9] and flexible displays [10]. Facile synthesis of Janus particles in liquid solution e.g., hydrophobic and hydrophilic interactions with anisotropic shape remain a big challenge and many researchers around the world are deeply interested in developing the facile fabrication techniques to prepare Janus colloidal nanostructures in a controlled manner [11-13]. Some important examples of these distinct Janus nanostructures are silica-PS heterodimer, gold-silica heterodimers, CdS-FePt [14], Au-Fe3O4 [15], Fe3O4/silica/polystyrene [16], PI-b-PS and PI-b-P2VP Janus nanoparticle [17], Au-TiO2, Au-Cu2S [18], and CdSe/CdS/ZnS−Au@hollow SiO2 yolk/shell nanospheres [19]. Mostly JNPs consist of metal-metal, metal-semiconductor, metal-polymer and polymer-semiconductor materials [14, 15, 20-24]. Nevertheless, only a handful of investigations are available on the transition-metal oxide-oxide anisotropic structure due to the complexity of

“Seeded growth” technique has been widely exploited as an effective tool for the fabrication of metal-inorganic heterostructures in which metal NPs assist as principal seeds or catalysis and accommodate the secondary particles onto it [25, 26]. Further, self-assembly contributes an important function in the formation of Janus colloidal nanoparticles and can be feasible both in bulk and solution based synthesis. This whole process is based on a complicated interplay of several factors such as entropy, enthalpy and the dynamics of self-assembly [20-22, 27, 28]. It is still unclear why spatial arrangements or junctions develop between large lattice mismatch materials during self-assembling process and requiring ingenious strategies. In both cases, lattice (matched or not) may undergo from an extensive distribution of geometries, making it very complicated to control [29]. The epitaxially joint which shaped the metal and oxide material into dumbbell-like morphology is due to the difference in the lattice parameters and their orientation depends upon how they share a coherent interface. Apart from lattice mismatch phenomena, synthesis solvents contribute a significant character in controlling the nucleation positions (reducing the electron density and hindering) on the metal seeds [15]. There are a number of researches present in literature on the preparation methods of JNPs [23, 30-33]. However, the implementations of such techniques involve the use of sophisticated equipment, complicated procedures, and rigorous experimental conditions. Furthermore, most of them are template assisted methods, feasible for larger or submicron Janus colloidal structures, but unsuitable to be used in biomedical imaging applications. Therefore, an effective and simple process with high rate of reproducibility is essential to synthesize such particles for practical interests. In present research, we demonstrate a step forward towards topologically controlled synthesis of Janus heterostructures / hetero-dimers in solution, completely made of binary transition-metal oxides (BTMOs). Also, we report a heteroepitaxial growth

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mechanism between as-synthesized BTMOs which leads to the formation of nano-heterodimers using lattice mismatch process and rarely reported before. We employ a simple and convenient two-step solvothermal fabrication route to prepare novel Janus nanostructure based on titanium dioxide (TiO2), zinc oxide (ZnO), iron oxide (Fe3O4), and manganese oxide (Mn3O4), technologically promising materials which have attained a significant attention owing to their potential applications in numerous fields such as dye-sensitive solar cells, photocatalysis, photochromic devices, gas sensing, energy storage and biomedical applications.

2 Materials and Methods 2.1 Synthesis of binary transition-metal oxides Janus nanoparticles The ultra-small hydrophobic TiO2 nanoparticles were fabricated using previous reported method by titanium (IV) butoxide [34]. Briefly, 1.7 g of titanium (IV) butoxide dissolved in the mixture of 7 mL oleic acid, 5.5 mL oleylamine and 4 mL ethanol during stirrer at room temperature for 20 min. The obtained solution put in a 40 mL Teflon tube. Then, the Teflon tube placed into a 100 mL Teflon-lined stainless steel autoclave containing 3:1 ratio of ethanol to water. Finally, the autoclave put in oven at 180 °C for 18 h. After completing the reaction, the resulting white precipitate was collected and washed several times with absolute ethanol. At last, the as-synthesized TiO2 NPs were dispersed in 10 mL non-polar solvent such as hexane or toluene. To synthesize novel Mn3O4-TiO2 Janus nanoparticles, 0.13 g of Mn(acac)3 were dissolved in 5 mL of octadecane and 13 mL of n-octyl alcohol and stirrer for 30 min. Subsequently, 3.5 mg/mL of TiO2 nanoparticles dispersed in 8 mL n-hexane and then above solution was added. The obtained mixture continued to be stirrer for 3.5 h at room temperature and then increase the temperature to 80 °C for 30 min to evaporate the n-hexane. Following by, the obtained final solution poured in a 40 mL Teflon-lined stainless steel autoclave and placed in the heating

oven at 230 °C for 3 h. The same experiment was performed by replacing TiO2 nanoparticles with oleic acid coated ZnO and Fe3O4 by keeping all other parameters unchanged. Finally, after completion of reaction, the resulting precipitate was collected, and washed several times with the ethanol. The yield of the Mn3O4-TiO2 was approximately Mn: 3.8 mg/mL and Ti: 3.1 mg/mL observed by ICP-MS and dispersed in 7 mL cyclohexane for further analysis. Finally, FDA approved triblock copolymer, Pluronic® F-127 [poly (ethylene oxide)-poly (propylene oxide)-poly (ethylene oxide) PEO-PPO-PEO] was used to transfer the nanoparticles phase from organic to aqueous solution. In phase transfer process, 700 mg of PF-127 was dissolved into 70 mL CHCl 3 and stirred for 20 min to obtain transparent solution. Then, 1 mL of oleic acid coated Mn3O4-TiO2 added gradually into above solution and stirred for 4 h. After that, 5 mL water added into the solution and chloroform was removed slowly using a rotary evaporator at 40 °C. NPs were washed with ethanol using rotary evaporator and then water solvable (7 mL) nanoparticles were used for further examinations. The summarize form of the experimental procedure and growth mechanism can be shown in schematic diagram. 2.2 Characterizations Powder X-ray diffraction (XRD) pattern of the prepared material was recorded on a D8 Focus XRD diffractometer (Bruker) with Cu Kα radiation (λ=0.154 nm). Transmission electron microscopy (TEM) images, high-resolution TEM (HRTEM) images and energy dispersive analysis (EDS) of the prepared nanocrystals were attained on JEOL-2100 electron microscope. High-angle annular dark-field (HAADF)-scanning transmission electron microscopy (STEM) coupled with EDS elemental mapping was performed on Tecnai G2 20 S-TWIN. Dynamic light scattering (DLS) was performed on Zetasizer Nanoseries (Nano-ZS, Malvern Instruments). Determination of the manganese and titanium contents in a sample were measured by inductively coupled plasma mass spectrometry (ICP-MS) with an Optima 2100 instrument from Perkin Elmer.

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2.3 MRI Relaxation Properties of JNPs@PF-127 The longitudinal and transverse relaxation time (T1 and T2) and relaxivities (r1 and r2) of JNPs were measured on a 0.47 T MRI instrument. PF-127 coated Mn3O4-TiO2 nanoparticles were dissolved in water with variable concentrations of Mn (0.13, 0.39, 0.65, 0.91 and 1.17 mM) to measure the relaxation times and T1 imaging. Here, we used the Mn concentration and neglect the Ti factor in the sample because Mn is the main source of MR signal and imaging. Longitudinal (r1) and transverse (r2) relaxivities were calculated from inverse of relaxation times (1/T1 and 1/T2) and plotted against different concentrations of Mn. T1-weighted MR images were performed running a standard spin echo (SE) sequence with TR = 300 ms and TE = 18 ms on a 0.55 T MR imager (MeoMR60, Shanghai Niumag Corporation).

avoid protein degradation. After the incubation time, centrifugation was performed at 10000 rmp for 10 min on the NP-protein corona complex to separate from the supernatant. The above solution was washed twice with phosphate-buffered saline (PBS) and finally also spread in PBS solution. Serum-contained, serum-free JNPs, supernatant and precipitate were characterized by UV-Vis spectrophotometry. The absorption spectra were executed from 200 to 900 nm using a T10CS instrument (Beijing Purkinje General Instrument Co., Ltd, China). The pure JNPs nanoparticles were incubated with high and low concentration of FBS and also characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to observe the NPs-protein conjugate [35, 36].

2.4 In vitro Cytotoxicity

2.6 In vivo MR-Imaging

The cytotoxicity of the Mn3O4-TiO2 JNPs was evaluated using the colorimetric methyl thiazolyl tetrazolium (MTT) assay in MCF-7 cells. Briefly, MCF-7 cells were cultured in a 96-well plate and incubated for 24 h in Dulbecco’s Modified Eagle Medium (DMEM, GIBCO, high glucose, C11995) supplemented with 10 % fetal bovine serum (FBS, Thermo, SH3007003). Then, JNPs (Mn: 554 µg/mL and Ti: 443 μg/mL) were added into each well at different Mn concentrations (0, 20, 40, 60, 80, 100 and 120 ug/mL, diluted in DMEM) for 24h at 37 °C under 5% CO2, where as Ti concentration was (0, 16, 32, 48, 64, 80 and 96 ug/mL). Later, 10 μL of MTT solution (5 mg/mL) was added to each well and was incubated for another four hours at 37 °C under 5% CO2 for formazan product formation. Then, the culture medium was carefully detached and 100 μL of DMSO solution was added to dissolve the MTT formazan crystals. Finally, the cell viability was calculated by measuring the absorption of each well using a micro-plate reader (iMark 168-1130, Bio-rad, USA) at a wavelength of 550 nm.

In the mice experiments, the animal care and handing procedures were in agreement with the guidelines of the Regional Ethics Committee for Animal Experiments at Ningbo University [Permit No# SYXK (Zhe 2013-0191)]. In vivo MRI experiments were carried out on nude mice (weight, 15 g) with intravenous injection of 150 µL aqueous solutions of JNPs (Mn: 554 µg/mL and Ti: 443 μg/mL). T1-weighted MR Contrast-enhanced images of mice heart, liver and kidneys were measured on a 0.55 T MRI instrument (MeoMR60, Shanghai Niumag Corporation) at 32 °C. The imaging parameters were customized as follows: TR= 300 ms, TE= 20 ms, bandwidth= 23.318 kHz, slice Width= 3.0 mm, slice Gap= 0.5 mm, FOVRead= 100 mm and FOVPhase= 100 mm.

2.5 NPs-protein corona complexes 1 mL volume of PF-127 coated Mn3O4-TiO2 nanoparticles (Mn: 554 μg/mL and Ti: 443 μg/mL) were incubated with 80 % of fetal bovine serum (FBS) for one hour on ice to slowdown the reaction and to

2.7 In vivo tumor MR-Imaging Two female BALB/c nude mice subcutaneously injected with 5 × 106 MCF-7 cells/mouse on the right backside. After 3 weeks of post-injection, the tumor nodule was reached a volume of about 270 mm3. Subsequently, the mouse was anesthetized and then 150 uL (Mn: 554 µg/mL and Ti: 443 μg/mL) of multifunctional PF-127 coated Mn3O4-TiO2 nanoparticles in PBS solution were delivered at tumor site. The T1 MR-imaging of the tumor was performed using TR= 300 ms, TE= 20 ms, bandwidth=


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23.318 kHz, slice Width= 3.0 mm, slice Gap= 0.5 mm, FOVRead= 100 mm and FOVPhase= 100 mm. Two dimensional spin-echo T1-weighted MR-images were attained before and 10, 20, 30, 60 and 120 min post injection of the JNPs. 2.8 Synchrotron X-ray fluorescence microscopy (SXRF) mapping The cells were grown on sterile Malay film. As-prepared JNPs (Ti: 335 µg/mL and Mn: 419 µg/mL) was incubated with MCF-7 cell 2 h, then washed three times with PBS and were fixed with 4 % formaldehyde solution for 30 min and then again washed with deionized water. SXRF maps of cells were conducted at hard X-rays BL15U beam line at Shanghai Synchrotron Radiation Facility (Shanghai, China). Energy of X-ray was 10 keV, and beam spot was 0.5 × 0.5 um. Scan time was 1 second at each step.

Elemental maps of Ti, Mn, Fe, Cu, Cl, S, Ca, and Zn in cells were attained as described previously [37]. 2.9 In Vitro Photodynamic Therapy The colorimetric methyl thiazolyl tetrazolium (MTT) assay was executed to assess the UV irradiation effect of the PF-127 coated Mn3O4-TiO2 NPs on cell viability. MCF-7 cells were seeded into a 96-well plate and cultured for 24 h in Dulbecco’s Modified Eagle Medium (DMEM, high glucose, GIBCO, C11995) supplemented with 10% fetal bovine serum (FBS, Thermo, SH3007003). Subsequently, the cells were incubated with PF-127 coated Mn3O4-TiO2 JNPs with different Ti concentrations (0, 40, 80, and 120 μg/mL, diluted in DMEM) for 24h at 37 °C under 5% CO2, where as Mn concentration was (0, 50, 100 and 150 μg/mL).

Scheme: Schematic illustration of the growth mechanism accompanied by TEM results and coating process with Pluronic® F-127 of Mn3O4-TiO2 transition-metal oxides Janus nanostructure. The principle of use in MRI and PDT is also depicted.


Research

When they were incubated for 4 h, replaced the

respectively. The behavioral changes of the mice

culture media by new media, UV irradiation with

were observed for fifteen days and then mice were

power intensity 5.6mW/cm was carried out, and

sacrificed. The major organs of the mice including

the irradiation time was 0 min, 10 min, and 30 min.

liver,

Then, the cells were incubated another 24 hours.

conserved in a 10% formalin solution, stained with

Then, MTT solution (10 μL, 5 mg/mL) was injected

Hematoxylin and Eosin (H&E) for histopathological

to each well, and was incubated for another 4 h

bioanalysis to assess the toxicity of Mn3O4-TiO2

before the addition of 100 μL DMSO (Amresco,

JNPs by an optical microscope (Leica, DMI3000).

2

heart,

lung,

spleen,

and

kidney

were

code 0231) for dissolution of the precipitation. Finally, the absorption of each well was executed using micro-plate reader (iMark 168-1130, Bio-rad, USA) at a wavelength of 550 nm.

The

2.10 In vivo Photodynamic ablation of MCF-7 For in vivo PDT, female Balb/c nude mice (18-20 g, 4-6 weeks old) were used in this experiment. Firstly, the tumors were originated by injection of MCF-7 cells (1×107/100 μL) onto the right back side of each female Balb/c mouse. After growing for 1 the

MCF-7

tumor-bearing

mice

fabrication

of

heterostructure

micro/nano-particles that deviate importantly from

tumors

week,

3 Results and Discussion

were

randomly divided into five groups (n = 4, each group) and were treated by intratumoral injection with 150 μL PBS (the control group), PBS combined with UV, Mn3O4-TiO2 JNPs, and JNPs combined with UV irradiation. Moreover, the JNPs-UV further divided into two groups; (1) single treatment named JNPs+UV(1), and twice treatment named JNPs+UV(2). In the JNPs+ UV(2), the tumor site was irradiated once more time by UV after 11 days of first treatment. The injected amount of JNPs was 150 μL *dose concentration is according to the ICP-MS analysis (Ti: 443 μg/mL and Mn: 554 µg/mL)]. The photodynamic treatment was carried out after 20 min of intratumoral administration of JNPs. The power density was 5.6 mW/cm2 and the irradiation time was 30 min. 2.11 In vivo biocompatibility For in vivo toxicity/side effect of prepared JNPs were observed by histopathological bioanalysis on three mice. 150 μL of saline as a control injected to one mouse and 150 µL (Mn: 554 µg/mL and Ti: 443 μg/mL) JNPs intravenously injected to other mice

centrosymmetric core/shell nature morphology may presents a various set of building blocks for self-assembly of fascinating architectures. The formation of hybrid nano-crystals from identical lattice constants is common [29]. However, in divergent lattice or mismatch lattice systems, it is uncertain why strained interfaces should appear. Lattice (matched or not) may undergo from an extensive distribution of geometries which is very complicated to control. The Mn3O4-TiO2/ZnO/Fe3O4 Janus nanostructures used in this work were synthesized using the concept of heteroepitaxial growth of one particle on the surface of core particle. In this facile protocol, the starting transition-metal oxides (TiO2, ZnO, and Fe3O4) dissolved in non-polar

solvent

dispersion

and

to

then

obtain Mn(acac)3

homogeneous dissolved

in

nonionic surfactants introduced which leads to the development of an unstructured shell around the hydrophobic nanoparticles. Later, this amorphous structure is transformed to a Mn3O4 shell by successive addition of Mn3+ ions. Moreover, reaction time and temperature are two essential parameters in the configuration of the hybrid nanostructure. Therefore, increase in the reaction time and lattice mismatch between Mn3O4 and binary oxides (TiO2, ZnO, and Fe3O4), the shell started ruptures. Further,

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in a de-wetting phenomenon, all of the shell

and 0.248 nm are consistent with the interplanar

nucleates and anneal at one particular position in

spacing planes of (101) in anatase TiO2 and (211) in

the surface of binary oxides nanocrystals.

hausmannite Mn3O4 structure. The small particle

The sketch of the fabrication mechanism of the

about 5 nm is referred to Mn3O4 and a large particle

different stages can be seen in schematic illustration.

is about 10 nm belongs to TiO2. The average size of

Fig. 1 (a, b) shows the low magnification TEM and

the Mn3O4-TiO2 hybrid structure is about 15 ± 1nm.

high magnification HRTEM images of JNPs images

HAADF-STEM

and revealed that Mn3O4-TiO2 nanocomposite are in

mapping

the form of Janus structure with good dispersion

elemental/compositional distribution for individual

and homogeneous growth in non-polar solvent. A

Janus nanoparticles as shown in Fig. 1(d-g). The

high resolution HRTEM image [shown in Fig. 1 (c)]

analysis demonstrated that Mn, Ti and O elements

confirmed that these two materials are embedded

are present in the single nano-crystal which further

with each other in the form of dimmer. The HRTEM

confirmed that Janus structure composed of only

image showed the lattice spacing values of 0.356 nm

two materials Mn3O4 and TiO2.

was

coupled used

with to

EDS

elemental

investigate

the

Figure 1. TEM micrographs; (a) low magnification TEM, (b) high magnification and (c) HRTEM images of Mn3O4-TiO2 Janus nanostructures. (d-g) HAADF-STEM images and corresponding EDS elemental mapping of individual JNP. (h) Powder X-ray diffraction (XRD) of TiO2, Mn3O4 and Mn3O4-TiO2 Janus nanoparticles and (i) EDS spectra of Mn3O4-TiO2. (g) Cell viability of


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Pluronic® F-127 coated Mn3O4-TiO2 Janus nanoparticles with different concentration.

EDS elemental mapping pictures prove that oxygen

as-synthesized Mn3O4-TiO2 Janus nanostructures.

is well distributed, big particle represents titanium

Particularly, two prominent peaks (apart from some

and dimmer part manganese shell is nucleating at

smallest) are present at 2for plane Mn3O4

one position to form a Janus shape. In addition, this simple method is useful to synthesize Mn3O4-ZnO

(004), TiO2 (004) and 2 for plane Mn3O4 (312), TiO2 (105) show that the nature of the

and

hybrid/Janus

as-prepared material is heterostructure. Sharp

structure as shown in supporting Fig. S1 (a-d) and

diffraction peaks suggest the excellent crystallinity

supposed that also applicable on other kind of

of the synthesized material which also observed in

oxide materials. We selected Mn3O4-TiO2 Janus

HRTEM image. The small deviation is observed in

nanoparticles for further investigations and to

the intensity due to the oriented growth caused by

observe their properties in MR imaging and

the dual structural properties. In addition, no

photodynamic

in vivo.

impurity peak is detected in synthesized material.

Furthermore, FDA approved triblock polymer,

Moreover, the phase purity of as synthesized binary

Pluronic® F-127 was used for coating Janus

transition oxide Janus nanoparticles was examined

nanoparticles to enhance the compatibility and

using EDS. In EDS results, Fig. 1(i) and Fig. S3,

phase transfer to aqueous media. Fig. S2 (a)

show the EDS analysis of the final products which

manifests the JNPs@PF-127 TEM images. The

contained only Mn, Ti, Zn, Fe, O, C and Cu

coating results showed that successful modification

elements, in which the elements of Mn, Ti, Zn, Fe

of the JNPs using thin layer and stable aqueous

and

dispersion.

the

Mn3O4-TiO2/ZnO/Fe3O4 Janus nanostructure, while

hydrodynamic size analysis of the coated JNPs, one

C and Cu are from a copper grid (TEM sample

about at 27 nm and other at 105 nm as shown in Fig.

holder) with carbon film. Therefore, the XRD and

S2 (b). The difference between dynamic light

HAADF-STEM

scattering and micrographs results is because of soft

demonstrated the purity of the as-synthesized

polymer aggregation in DLS, which could not be

material.

seen in TEM pictures.

For biomedical applications, materials are probably

Fig. 1 (h) shows typical XRD pattern of TiO2, Mn3O4

to have an intrinsically low toxicity in aqueous and

and

their

Mn3O4-Fe3O4

with

therapy

Two

peaks

Mn3O4-TiO2

Janus

similar

in vitro

are

and

observed

nanoparticles.

in

Mn3O4



O

are

from

the

coupled

cytotoxicity

with

assessment

EDS

is

binary

results

compulsory.

nanoparticles are synthesized with the similar

Therefore,

experimental condition as Mn3O4-TiO2 Janus hybrid

Mn3O4-TiO2@PF-127

nanostructures without TiO2 as described in the

examined using MTT assays. Fig. 1(j) illustrates the

experiment procedure. All the diffraction spectra

cytotoxicity of MCF-7 breast cancer cells incubated

for TiO2 and Mn3O4 in Fig. 1(e) are well matched to

with various concentrations of JNPs for 24h. As seen

the

tetragonal

anatase

structure

with

the

prepared

biocompatibility Janus

nanoparticles

of was

lattice

in viability analysis, the JNPs showed no toxicity

parameters of a = b = 3.785 Ǻ, c = 9.514 Ǻ and α = β =

effect even at high concentration. The result

γ = 90o (JCPDS file No.21-1272), and tetragonal

suggested

hausmannite structure with the lattice constant of a

biocompatibility and have potential for further in

= b = 5.762 Ǻ, c = 9.47 Ǻ and α = β = γ = 90o (JCPDS

vivo investigations.

file No.24-0734), respectively. All the diffraction

Furthermore, before proceed towards in vivo studies,

peaks of TiO2 and Mn3O4 are observed in

adsorption of proteins and biological fate on the


that

Mn3O4-TiO2@PF-127

has

good

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surface of as-prepared JNPs were examined using

hour.

serum free and fetal bovine serum (FBS) containing

detached

medium. When nanoparticles are exposed to

centrifugation process and then nanoparticles were

biological environment via different routes, they

washed two times with PBS to remove unattached

uptake some natural fluid (blood plasma) from

proteins. Fig. 2(c) demonstrates the SDS-PAGE

biological atmosphere named “protein corona”

coomassie stained gel lanes including commercially

which has considerable influence in biological

available ProteinRuler® IV (30-200 kDa), supernate -

applications

and

precipitant at high concentration of FBS incubated

and

with fabricated nanoparticles, JNPs incubated with

SDS-PAGE were employed to observe nude and the

80%, 60%, 40%, 20% FBS, and JNPs free 80% FBS

corona conjugates NPs. PF-127 coated Mn3O4-TiO2

diluted

JNPs show a weak absorption peak at λ = 292 and

disconnected from medium according to their

strong intensity at λ = 302 nm as illustrated in Fig.

molecular weight. The faint / feeble bands are

2(a). Absorption spectra of serum which is diluted

detected at 80 kDa, 60 kDa and 30 kDa related to

with phosphate-buffered saline (PBS) also shows

hard corona and these bands are neglect able at low

weak absorption peaks at λ = 410 and strong weak

concentration of FBS as shown in Fig. 2(c). Further,

intensity at λ = 291 nm as illustrated in Fig. 2(b). In

the zeta-potential was measured to observe the

addition, when the JNPs incubated with FBS, the

change in surface charges. The zeta potential of

absorption performance of serum contained JNPs is

Mn3O4-TiO2 JNPs

similar with serum free nanoparticles. However,

JNPs@PF-127 in aqueous media is observed 0.7 mV

after

nd

and -11 mV respectively. Whereas, the zeta potential

supernate showed absorption at λ = 292. Therefore,

of JNPs-FBS conjugate in aqueous media is obtained

to overcome this absorption confusion, sodium

about -7.86 mV and decreased from -11 mV

dodecyl sulfate-polyacrylamide gel electrophoresis

showing that a small number of positively charged

(SDS-PAGE)

such

nanomedicine.

the

as

UV-Vis

centrifugation

experiment

nanotoxicology spectrophotometry

process,

1

st

and

2

Soft

coronas

from

with

the

PBS.

(unbound

proteins)

were

prepared

JNPs

high

The

in

by

proteins/serum

non-polar

solvent

was

and

was carried out for

protein from plasma serum absorbed by negatively

profound investigation of NPs-protein corona

charged NPs due to the electrostatic interaction

complex. In this experiment, as-prepared JNPs were

between protein and JNPs.

incubated with different concentration of FBS for an

Figure 2. (a) UV-Vis absorbance spectra of naked JNPs@PF-127, (b) FBS, NPs-FBS conjugation, naked and NPs protein coronas conjugate and (c) SDS-PAGE gel of the protein corona of JNPs after incubation with fetal bovine serum (FBS) medium for an hour www.theNanoResearch.com∣www.Springer.com/journal/12274 | Nano

Research

Same

phenomena,

neutralization

effiect

has

sensitivity

compared

with

nuclear

imaging.

observed in both negatively and positively charged

However, it was observed that the development of

NPs [35, 38]. The SDS-PAGE, zeta-potential and

contrast agents has facilitated MRI to become the

UV-Vis results showed that a few positively

most sensitive imaging technique not only in

charged proteins connect onto the surface of

clinical diagnosis but also a useful instrument for

JNPs@PF-127 from FBS medium to form a hard

biomedical research [39]. Contrast agents enhance

corona without disturbing the external biological

sensitivity

surroundings which may demonstrate

shortening of proton relaxation time. Therefore, MR

to

be

and/or

specificity

owing

to

the

excellent properties for in vivo applications.

signal intensity bases on the local value of

In order to measure the potential application of

transverse (1/T2) or longitudinal (1/T1) relaxation

as-synthesized material as a MR contrast agent,

rate of water protons.

magnetic relaxation properties were investigated. In

Unfortunately,

the beginning, MRI was developed to present

gadolinium-based T1 MR contrast agents are

molecular

associated with nephrogenic systemic fibrosis (NSF).

imaging

noninvasively.

The

major

the

commercially

existing

problem of MRI in definitive diagnoses is its lower

Figure 3. (a, b) T1, T2 relaxivities plot and (c-e) T1-weighted MR images of the Mn3O4-TiO2@PF-127 Janus nanostructure obtained using aqueous suspensions at various Mn concentrations.

In addition, these Gd- chelates are small molecules

the Mn3O4 (5 nm) nanoparticles. Ultra small

showing low sensitivity and short circulation time

nanoparticles

inside the body which hinders high-resolution

because of atoms/molecules are positioned on their

imaging

oxide

surface. Therefore, Mn3+ ions on the surface of

nanoparticles have attained significant attraction as

nanoparticles increase the magnetic moment which

promising alternative T1 MRI contrast agents

leads to the enhancement of longitudinal contrast

because of reduced toxicity than lanthanide as Mn

effect [41, 42]. The positive MRI signal intensity can

involved in a number of physiologic processes

also be distinguished in color images (Fig. 3e).

within the body, and posses long blood circulation

Based on the above mentioned MR relaxivities and

time. However, there is still a challenge to enhance

imaging results, as-prepared thin layer coated

the relaxivity of Mn-based contrast agents.

Mn3O4–TiO2 Janus nanoparticles have excellent

The magnetic relaxivities of JNPs in aqueous

T1-weighted MRI performance.

suspensions were executed using a 0.47 T scanner at

The ideal MR contrast agents should significantly

room temperature with various concentrations. The

enhance image contrast and offer a sufficient time

obtained quantitative data related to contrast

window for MR examination. In vivo animal

properties of the prepared Janus nanoparticles, the

experiment was executed at 0.55 T MR field to

relaxivities values r1 and r2 are shown in Fig. 3 (a, b).

investigate the circulation and biological response

The r1 value is 2.95 s-1mM-1 and r2 is 12.7 s-1mM-1,

of the as-prepared Janus nanoparticles inside mice

giving r2/r1 ratio of 4.3, suggesting that the prepared

body. The aqueous dispersed Mn3O4-TiO2 Janus

PF-127 coated Mn3O4-TiO2 Janus nanoparticles are

nanoparticles (150 ul of Mn: 554 µg/mL and Ti: 443

an excellent T1-weighted contrast agent. MR

μg/mL) were injected into a mouse via tail vein and

imaging was performed to further confirm the

a series of T1-weighted MR images were obtained

acquired quantitative data. T1-weighted MR signals

with respect to time using 0.55 T field. After

enhancements were measured at MRI 0.55 T

intravenous

analyzing

[40].

In

contrast,

nanoaprticles

the mice body. We observed a great contrast

Mn3O4–TiO2 Janus nanoparticles in comparison of

enhancement in the heart, liver and kidneys after

water. The MR imaging results revealed that

intravenous injection (Fig. 4). T1-weighted MR

imaging signal intensity is increasing with the

images of the major organs were obtained before

increment in Mn concentration. The MRI signal of

and after intravenous injection. The positive signal

JNPs appeared brighter and brighter as compared

enhancement is observed throughout the mice body.

to the MR image of water. The coating of thin layer

Especially, in the heart which suddenly increased

triblock polymer with hydrophilic surface played

after 5 min of injection and slightly decreased over

an effective expression in enhancement of positive

time and saturated due to blood circulation system

signal intensity. Water molecules are attached

as shown in Fig. 4 (a, b). Subsequently, the

directly to the intricated paramagnetic ion and

nanoparticles showed T1 image enhancement in the

generate the strongest effect on the overall relaxivity.

liver area and increased contrast as the time passes.

The

of

The highest T1 MR contrast is obtained at 60 min

as-synthesized JNPs is better than many reported

after injection and decreased slowly over time. The

manganese

in

significant and obvious difference between control

supporting information Table S1. The higher r1

and enhanced MRI contrast with as-synthesized

relaxivity at 0.47 T is attributed to ultra small size of

JNPs can be distinguished in pseudo colored

calculated based

of

longitudinal nanomaterials

shows

Janus

directly reached to the heart and then spread into

images

3(c-e)

the

properties

the

MR

Fig.

injection,

outstanding

coated

T1-weighted

instrument.

manganese

present

PF-127

relaxivity as

listed

12

Nano Res.

pictures [Fig. 4(b, d)] pre and post intravenous injection.

Figure 4. In vivo MR imaging. (a) T1-weighted MR images of a mice heart pre-injection and 5min, 10 min, 40 min, 100 min and 140 min after intravenous injection. (c) T1-weight MR images of mice liver pre and 5 min, 20 min, 30 min, 40 min and 50 min post intravenous injection. (e) T1-weighted MR images of mice left and right kidneys before and 5min, 30 min, and 65 min post injection.


Nano Res.

13

(b, d, f) The pseudo color pictures consistent to (a), (c) and (e) respectively

Figure 5. In vivo signal intensity of MR imaging in heart, liver and kidney with respect to time

Figure 6. T1- MR-imaging of MCF-7 tumor bearing mice before injection and post intratumoral injection of 150 µL (Mn: 554 µg/mL and Ti: 443 μg/mL) aqueous solution containing JNPs.

Further, the accumulation of Mn3O4-TiO2@PF-127

maintained for about 2 hours. Hence, as-prepared

Janus nanoparticles is highest in the kidneys. Fig. 4

heterojunction particles seem very appropriate to

(e, f) shows the obvious contrast enhancement in

detect the heart and kidney related problems.

the kidneys which is started since injection and

Moreover, the T1 MRI signal intensity of heart, liver


Research

14

Nano Res.

and kidneys with respect to time are given in Fig. 5.

can be seen in Fig. 6(c). These results demonstrated

The signal intensity gradually enhances in the liver

that as-synthesized JNPs@PF-127 have excellent

after decrease the signal in heart. Later, very stable

imaging

signal observe in left and right kidneys form 30 min

T1-weighted MRI contrast agent and have great

to 100 min and then started decline. The results

potential for clinical applicability.

clearly

metabolic

Synchrotron X-ray fluorescence microscopy (SXRF)

response of fabricated novel nanoparticles. Based

is a new promising micro analytical technique

on the above investigated results, Pluronic® F-127

which is fully compatible for biological samples and

coated Mn3O4-TiO2 Janus nanoparticles are the

excellent for direct visual distribution of metal

excellent whole body imaging candidates for

oxide NPs [43]. Here, we also used SXRF

T1-weight

microscopy to observe the direct distribution of as

demonstrated

contrast

the

physical

agent

with

good

enhancement

ability

in

tumor

as

biocompatibility.

prepared JNPs by mapping of the individual MCF-7

Further, we performed in vivo MRI experiments

cancer cells. A hard X-ray beam of 10 keV energy

with MCF-7 tumor-bearing mice to investigate the

was utilized at Shanghai Synchrotron Radiation

as-prepared JNPs effect on tumor imaging. T1-MR

Facility to excite the required elemental S, Ti and

images

after

Mn in cancer cells, which have X-ray emission lines

intratumoral delivery of JNPs using 0.55 T MRI

in 10 keV. SXRF map images are taken from the

instrument. Excellent positive signal improvement

normal cell as control and JNPs incubated cell. The

is observed after 20 min of post-injection which

intracellular distribution results show that there is

made tumor obvious as compared to pre-injection

no signal observed against Mn and Ti elements in

[Fig. 6 (a, b)]. After two hours, the brightness still

the control cell.

were

carried

out

before

and

Figure 7. X-ray fluorescence maps of MCF-7 cells incubated with JNPs. Cells were incubated with JNPs at equivalent Ti concentration (335 µg/mL) and Mn concentration (419 µg/mL) for 2 h. Scanning was executed using 10 keV incident energy with the scale bar as 20 mm. Beam spot was 0.5×0.5 mm and scan time was 1 second at each step. Blue fluorescence indicated biogenic elemental sulfur in cells. Titanium of TiO2 was showed as red and Mn of Mn3O4 was indicated as green. | www.editorialmanager.com/nare/default.asp

Whereas, cell incubated with JNPs shows the strong

observed of UV light combined with Mn3O4–TiO2

Mn (green) and Ti (red) signal and are mainly

Janus nanoparticles at low concentration and the

distributed in the cytoplasm and some penetrated

viability is about 72%. However, at 120 µg/mL of

around the nuclei of MCF-7 cell (Fig. 7), suggesting

Mn3O4–TiO2 Janus nanoparticles and UV light

accumulation

Janus

irradiated for 30 min, the cell viability decreased

nanoparticles by cancer cells within 2 h incubation

obviously to 50% at very low intensity of UV (5.6

period. The biogenic sulfur (blue) is native to cells

mW/cm2). This cell ablation can be improved using

and its outline matches outline of the whole cell.

direct contact of UV light with the cancer cells by

The overlay images again proved that the both

removing the cover of the cell culture plate.

particles are embedded with each other in the form

Furthermore, the appropriate concentration of

of Janus structure. SXRF results analysis suggested

Mn3O4–TiO2 Janus nanoparticles, the irradiation

that

time of UV light and intensity power were very

these

and

internalization

particles

may

of

have

potential

photodynamic effect in vitro.

essential for the PDT performance. Therefore, based on the above mentioned results as prepared Mn3O4–TiO2 Janus nanostructured material showed the ability to generate

O2 which leads to cell

1

ablation under UV irradiation in vitro. After the good biocompatibility and in vitro positive PDT behavior of JNPs, we further investigated the efficacy of as-prepared Janus nanoparticles as PDT agents in vivo. MCF-7 tumor-bearing nude female mice

divided

into

treated/operated methods. performed

five

intratumoraly

Photodynamic after

groups

20

with

therapy min

(n=4)

of

and

different was

also

intratumoral

administration of JNPs, as they acquired same time Figure 8. Cell viability of MCF-7 cells incubated with various

to spread in the tumor area as presented in Fig. 6(b).

concentrations of Mn3O4–TiO2 Janus nanoparticles and with

Tumor site was exposed to the very low power UV

and without UV light irradiation

light and other body covered with foil paper to avoid UV light effects on mice skin. After various

Furthermore, photodynamic effect was observed

treatments of five groups, such as control, only NPs,

under UV light irradiation with diffierent time, and

only UV irradiation and NPs plus UV irradiation

then the cell viability of MCF-7 cells incubated with

[NPs+UV (1) and NPs+UV(2)], the result of each

Mn3O4–TiO2

was

group at 21 days is illustrated in Fig. 9. MCF-7

Janus

nanocomposites

investigated using MTT assay method.

Fig. 8

tumor bearing mice with various treatments is

shows the cell viability results with and without

shown in Fig. 9(a, b). Significance change is

JNPs at 0, 10, and 30 min under UV light irradiation.

observed between control group and as-synthesized

It is clear from the MTT analysis that UV light

JNPs combined with UV irradiation. It is apparent

combined with JNPs has significant influence

that the tumor size and weight is very large without

against the growth of cells. In the absence of the

any treatment and injection. In addition, when JNPs

JNPs, the viability of cells is similar before and after

or UV used as separately an operating source, there

UV irradiation process. Any prominent effect is not

is still no obvious alteration observed.

16

Nano Res.

Figure 9. (a) Representative digital snapshots of the mice taken after various intratumoral treatments: control, only UV irradiation, JNPs and JNPs combined with UV irradiation treated once and twice. (b) Photographs of tumor tissue and weight obtained from each group at 21 days. (c) The relative tumor volume and (d) bodyweight of MCF-7 tumor bearing mice in different groups after treatment. However, combined JNPs with UV irradiation

treated with NPs plus UV indicates that the UV

showed promising tumor ablation effect and almost

irradiation and JNPs together played a fundamental

disappear after twice therapy treatment which can

role in the therapy result. It was also reported that

be seen in Fig. 9(a, b). The main difference between

manganese dioxide NPs could have the ability to

NPs+UV(1) and NPs+UV(2) is that the tumor site

raise tumor oxygen level by interacting with

was irradiated once again 30 min by UV after 11

endogenous H2O2 within hypoxic tumor site which

days of first treatment in NPs+UV(2) group. The

also support tumor ablation [44, 45]. Moreover, the

remarkable difference between control and tumor

tumor sizes of the control, nanoparticles and UV


Figure 10. Representative H&E stained histopathological bioanalysis images of most important organs of the healthy mice (control) and after 15-day post intravenous injection of Mn3O4-TiO2 Janus nanoparticles

group are increased notably, while the sizes of NPs-UV-1 (treated once) and NPs-UV-2 (treated twice) showed the excellent results of treatment almost stop to increase and even decrease as illustrated in Fig. 9 (b, c). None of the mice died and no abnormality in body weight, drinking, eating, or other

daily

activity

was

observed

during

photodynamic therapy operation (Fig. 9d). Finally,

we

also

histopathological

carried

out

bioanalysis

to

H&E

stained

evaluate

the

potential side effects (toxicity) of the as-prepared JNPs on the major organs of mice in vivo. The mice were sacrificed after fifteen days of post-injection of saline (control) and JNPs. The major organs such as the heart, liver, spleen, lung and kidney were collected and observed by histological analysis of tissues. Fig. 10 demonstrates that there are no distinct histopathologic alterations like injure, and damage found in the tissues / lesions such as necrosis, pulmonary or inflammatory fibrosis as compared

to

control.

The

histopathological

bioanalysis

strong

of

evidence

biocompatibility

of

safe

H&E further

use

Pluronic®

Mn3O4-TiO2 Janus nanoparticles.

and F-127

stained provided excellent coated

4 Conclusions In summary, a simple hydothermal/solvothermal method used to synthesize multifunctional binary transition-metal oxide Mn3O4-TiO2/ZnO/Fe3O4 Janus shaped nanoparticles. The epitaxial growth of secondary particle on the surface of core particle resulted in heterodimer structures with excellent dispersion and homogeneous growth in non-polar solvent. All the mixed diffraction peaks of TiO2 and Mn3O4 demonstrate hybrid nature of as-synthesized Mn3O4-TiO2 Janus nanostructures. HAADF-STEM coupled with EDS elemental mapping further confirm that JNPs are embedded with each other in the form of dimmers. The average size of the JNPs was about 15 ± 1 nm, whereas the average size of Pluronic® F-127 coated JNPs was about 100 nm because of soft polymer aggregation. Further, the PF-127 coated Mn3O4-TiO2 JNPs exhibited a good r1 relaxivity of 2.95 mM-1s-1 and low r2/r1 ratio of 4.3 suggesting

that

the

prepared

PF-127

coated

Mn3O4-TiO2 JNPs are an excellent T1-weighted contrast agent. In vivo T1-weighted magnetic resonance imaging of the heart, liver, and kidney in mice after intravenous injection of JNPs further

18

Nano Res.

confirms the biocompatibility and high sensitivity

Facility at Line BL15U (No. h15sr0021) used for

of

X-ray fluorescence imaging.

as-synthesized

JNPs.

Additionally,

SXRF

microscopy mapping results showed the stability of nanocomposites and strength of penetrating inside

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Nanotoxicology: An Emerging Discipline Evolving from Studies

of

Ultrafine

Particles.

Environmental

Health

Perspectives 2005, 113, 823-839. [42] Xing, R.; Zhang, F.; Xie, J.; Aronova, M.; Zhang, G.; Guo, N.; Huang, X.; Sun, X.; Liu, G.; Bryant, L. H.; Bhirde, A.; Liang, A.; Hou, Y.; Leapman, R. D.; Sun, S.; Chen, X. Polyaspartic acid coated manganese oxide nanoparticles for efficient liver MRI. Nanoscale 2011, 3, 4943-4945. [43] Paunesku, T.; Wanzer, M. B.; Kirillova, E. N.; Muksinova, K. N.; Revina, V. S.; Romanov, S. A.; Lyubchansky, E. R.; | www.editorialmanager.com/nare/default.asp

Nanoparticles

Response

by

Modulate

Priming

Solid

Tumor

Tumor-Associated

Nano Res.

Electronic Supplementary Material

Facile fabrication route for binary transition-metal oxides Janus nanoparticles for cancer theranostic applications 1

1

1

1

1

1

2

M. Zubair Iqbal , Wenzhi Ren , Madiha Saeed , Tianxiang Chen , Xuehua Ma , Xu Yu , Jichao Zhang , Lili 2 2 1 Zhang , Aiguo Li , Aiguo Wu () 1

Key Laboratory of Magnetic Materials and Devices, CAS & Key Laboratory of Additive Manufacturing Materials of Zhejiang Province, & Division of Functional Materials and Nano devices, Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, No. 1219 ZhongGuan West Road, 315201, Ningbo, China 2 Shanghai Synchrotron Radiation Facility, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201204, P. R. China Supporting information to DOI 10.1007/s12274-****-****-* (automatically inserted by the publisher)

Figure S1. TEM images of the heterojunction colloidal nanoparticles, low and high magnification of (a, b) Mn3O4-ZnO and (c, d) Mn3O4-Fe3O4 Janus nanoparticles Address correspondence to M. Zubair Iqbal, [email protected] and Aiguo Wu, [email protected] www.theNanoResearch.com∣www.Springer.com/journal/12274 | Nano

Research

Nano Res.

Figure S2. (a) TEM image and (b) size distribution analysis of the PF-127 coated Mn3O4-TiO2 Janus nanoparticles

Figure S3.EDS spectra of (a) Mn3O4-ZnO and (b) Mn3O4-Fe3O4 Janus nanoparticles

Figure S4.Cell viabilities of (a) Fe3O4 nanoparticles and (b) TiO2 Nanoparticles with different concentrations


Nano Res.

Table 1. Relaxivity Data of Manganese Oxide-Based Nanoparticles Name

Core Material

r1 (mM-1S-1)

B0 (T)

Ref.

Mn3O4-TiO2 JNPs

Mn3O4

2.95

0.47

This Work

HMONs

MnO

2.58

7

[1]

MnO

MnO

0.12

3

[2]

HMnO@mSiO2

MnO

1.72

1.5

[3]

Mn3O4 nanocubes

Mn3O4

1.08

3

[4]

Mn3O4 nanospheres

Mn3O4

1.31

3

Mn3O4 nanoplates

Mn3O4

2.06

3

Mn-MSNs

MnO(Mn3O4)/SiO2

2.28

3

[5]

Spherical Mn3O4

Mn3O4

2.38

1.5

[6]

HSA-MONP

MnO

1.97

7

[7]

HMONs

Mn3O4

1.42

3

[8]

Mn3O4@SiO2

Mn3O4

0.47

3

[9]

Fe3O4/MnO hybrid

Fe3O4/MnO

1.4

3

[10]

Nanoparticle

annotations:

JNPs-Janus

nanoparticles,

HMONs-hollow

manganese

oxide

nanoparticles,

HMnO@mSiO2-mesoporous silica-coated hollow MnO nanoparticles, HAS-MNOP-human serum albumin-coated manganese oxide nanoparticles, References [1] Hsu, B. Y. W.; Ng, M.; Zhang, Y.; Wong, S. Y.; Bhakoo, K.; Li, X.; Wang, J. A Hybrid Silica Nanoreactor Framework for Encapsulation of Hollow Manganese Oxide Nanoparticles of Superior T1Magnetic Resonance Relaxivity. Advanced Functional Materials 2015, 25, 5269-5276. [2] Gilad, A. A.; Walczak, P.; McMahon, M. T.; Na, H. B.; Lee, J. H.; An, K.; Hyeon, T.; van Zijl, P. C. M.; Bulte, J. W. M. MR tracking of transplanted cells with “positive contrast” using manganese oxide nanoparticles. Magnetic Resonance in Medicine 2008, 60, 1-7. [3] Kim, T.; Momin, E.; Choi, J.; Yuan, K.; Zaidi, H.; Kim, J.; Park, M.; Lee, N.; McMahon, M. T.; Quinones-Hinojosa, A.; Bulte, J. W.; Hyeon, T.; Gilad, A. A. Mesoporous silica-coated hollow manganese oxide nanoparticles as positive T1 contrast agents for labeling and MRI tracking of adipose-derived mesenchymal stem cells. Journal of the American Chemical Society 2011, 133, 2955-2961. [4] Huang, C.-C.; Khu, N.-H.; Yeh, C.-S. The characteristics of sub 10 nm manganese oxide T1 contrast agents of different nanostructured morphologies. Biomaterials 2010, 31, 4073-4078. [5] Chen, Y.; Chen, H.; Zhang, S.; Chen, F.; Sun, S.; He, Q.; Ma, M.; Wang, X.; Wu, H.; Zhang, L.; Zhang, L.; Shi, J. Structure-property relationships in manganese oxide - mesoporous silica nanoparticles used for T1-weighted MRI and simultaneous anti-cancer drug delivery. Biomaterials 2012, 33, 2388-2398. [6] An, K.; Park, M.; Yu, J. H.; Na, H. B.; Lee, N.; Park, J.; Choi, S. H.; Song, I. C.; Moon, W. K.; Hyeon, T. Synthesis of Uniformly Sized Manganese Oxide Nanocrystals with Various Sizes and Shapes and Characterization of Their T1 Magnetic Resonance Relaxivity. European Journal of Inorganic Chemistry 2012, 2012, 2148-2155. www.theNanoResearch.com∣www.Springer.com/journal/12274 | Nano

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[7] Schladt, T. D.; Schneider, K.; Shukoor, M. I.; Natalio, F.; Bauer, H.; Tahir, M. N.; Weber, S.; Schreiber, L. M.; Schroder, H. C.; Muller, W. E. G.; Tremel, W. Highly soluble multifunctional MnO nanoparticles for simultaneous optical and MRI imaging and cancer treatment using photodynamic therapy. Journal of Materials Chemistry 2010, 20, 8297-8304. [8] Shin, J.; Anisur, R. M.; Ko, M. K.; Im, G. H.; Lee, J. H.; Lee, I. S. Hollow Manganese Oxide Nanoparticles as Multifunctional Agents for Magnetic Resonance Imaging and Drug Delivery. Angewandte Chemie International Edition 2009, 48, 321-324. [9] Yang, H.; Zhuang, Y.; Hu, H.; Du, X.; Zhang, C.; Shi, X.; Wu, H.; Yang, S. Silica-Coated Manganese Oxide Nanoparticles as a Platform for Targeted Magnetic Resonance and Fluorescence Imaging of Cancer Cells. Advanced Functional Materials 2010, 20, 1733-1741. [10] Im, G. H.; Kim, S. M.; Lee, D. G.; Lee, W. J.; Lee, J. H.; Lee, I. S. Fe(3)O(4)/MnO hybrid nanocrystals as a dual contrast agent for both T(1)- and T(2)-weighted liver MRI. Biomaterials 2013, 34, 2069-2076.
