Applied Surface Science 501 (2020) 144267

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Applied Surface Science

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Full Length Article

Evaluation of physico-chemical properties and biocompatibility of new T

surface functionalized Fe3O4 clusters of nanoparticles
⁎
T. Radua, A. Petrana, , D. Olteanub, I. Baldeab, M. Potarac, R. Turcua
a
National Institute for Research and Development of Isotopic and Molecular Technologies, 67-103 Donat Str., 400293 Cluj-Napoca, Romania
b
Department of Physiology, Iuliu Hatieganu University of Medicine and Pharmacy, Clinicilor 1-3, 400012 Cluj-Napoca, Romania
c
Nanobiophotonics and Laser Microspectroscopy Center, Interdisciplinary Research Institute in Bio-Nano-Sciences, Babes-Bolyai University, T. Laurian Str. 42, 400271
Cluj-Napoca, Romania

A R T I C LE I N FO A B S T R A C T

Keywords: In this research, magnetic nanoclusters of Fe3O4 (MNC) were synthesized by solvothermal method using dif-
Magnetite nanoclusters ferent organic acids as surfactants such malic acid, aspartic acid and sodium tartrate in order to obtain new
Solvothermal nanostructured materials with applications in medicine. The interaction process of these acids on the surface of
Malic acid magnetic nanoclusters is important in understanding the change induced on the surface properties of the ob-
Aspartic acid
tained MNC. Structural analysis and physico-chemical characterization of the new synthesized materials were
Sodium tartrate
Red blood cells
performed by means o X-ray photoelectron spectroscopy (XPS), transmission electron microscopy, zeta potential
and magnetic measurements to provide their spherical shape, stability and specific functional groups and the
influence of the surfactants on their magnetic properties. XPS core level and valence band photoemission spectra
for all investigated samples is discussed in terms of changes induced in the electronic structure linked to var-
iation of the Fe2+ cations at the samples surface. This is of fundamental importance to better understand the
electronic structure and magnetism of the obtained MNC in order to tailor their electronic properties by surface
engineering for specific biomedical applications. The biological effects of MNC in rat blood were studied by
hemolysis and erythrocyte antioxidant systems (superoxide dismutase and catalase) showing no negative effect
on the cells.

1. Introduction with other molecules needed in certain applications e.g., targeted

Among biomedical applications of iron oxide nanoparticles, drug
delivery systems, medical diagnosis, hyperthermia are those based
primarily on magnetic properties of the single domain nanoparticles
magnetism [1–3]. Moreover, the control of the synthetic conditions and
functionalization of the magnetic nanoparticle surface is crucial be-
cause it dictates their physico-chemical properties, colloidal stability
and biological behaviour. Previous study shows that when solvothermal
method is used to produce MNCs the obtained clustersare in the range
of 250–400 nm, mainly as agglomerate of smaller particles of 9–30 nm
sizes, due to their high surface energy [4]. The crystal size of magnetic
nanoparticles and the way of packing in the crystal lattice influence the
superparamagnetic behaviours and magnetization of the obtained par-
ticles, important demands for biomedical applications [5].
For bio-medical applications the surface shell of the MNC it is re-
quired to be nontoxic, stable and biocompatible. It is also necessary to
supply specific functional groups for the further bonding or interaction
therapy. Therefore, clusters of Fe3O4 synthesizedby solvothermal
method, stabilized with biocompatible surfactants such sodium tartrate,
aspartic acid or malic acid may be good candidates to fulfil the above
mentioned criteria. Until now, none of these acids have been studied as
surfactantsfor Fe3O4-MNC synthesis by solvothermal method and no
study concerning their interaction and effect with human blood were
performed.
It has been reported recently that using EDTA as stabilizer, the
obtained MNC are biocompatible and show affinity for protein inter-
action [6]. Unlike EDTA, the surfactants proposed here have extra
functionalities such hydroxyl and amino which may influence their
physico-chemical properties or protein adsorption, biologic response
and cytotoxicity of MNC.
Various bio-distribution studies report different types of nano-
particles (NPs) being localized in blood, spleen, liver or kidney [7].
Once in the body they interact with the red blood cells (RBC). Therefore
a preliminary study is needed to understand how NPs affect the RBC in

⁎
Corresponding author.
E-mail address: anca.petran@itim-cj.ro (A. Petran).

https://doi.org/10.1016/j.apsusc.2019.144267
Received 2 July 2019; Received in revised form 30 September 2019; Accepted 1 October 2019
Available online 11 October 2019
0169-4332/ © 2019 Elsevier B.V. All rights reserved.
T. Radu, et al.
the perspective of their applications in medicine.
Though previous studies established already the effect of particles
dimension and hydrophilicity on the blood clearance, the results re-
ported on the effect of particles surface charge are contradictory, being
directly connected to the type of particles, density of charge, and the
nature of the functional groups on their surface [8–11].
Taking into account the above mentioned issue we study the mor-
phology, elemental composition, electronic structure, chemical surface
and magnetic properties of the obtained materials by transmission
electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS),
valence band XPS (VB-XPS) and magnetization measurements. The
surfactants used here have also been studied for their potential to tailor
the surface electronic structure of the obtained MNC by surface en-
gineering. Therefore we address our study also to the investigation of
the surface electronic structure modifications and its effect on the
magnetic properties.
In order to have a comprehensive picture of their surface chemical
nature and surface charge effects, zeta potential measurements as a
function of pH was measured with a special emphasis in the pH region
with values close to those of human body fluids. The effect on the rat
blood cells, including haemolysis and the erythrocyte antioxidant sys-
tems (superoxide dismutase and catalase) was also investigated.
2. Experimental
2.1. Materials and synthesis method
MNC synthesis: FeCl3·6H2O (7.3 mmol, 2 g) and NaAc (36 mmol,
3 g) were suspended in a mixed solvent of DEG and EG (50:50% from
40 ml total volume) to form a concentrated uniformed dispersed mix-
ture under mechanical stirring at 60 °C for 3 h. The acidsmalic, aspartic
andsodium tartrate, (1.8 mmol corresponding to 0.42 g sodium tartrate,
0.246 g malic acid and 0.242 g aspartic acid) were separately suspended
in 20 ml mixture of DEG and EG (50:50% from 20 ml of the total vo-
lume) at room temperature for 3 h. After that the two suspensions were
mixed together in a Teflon-lined stainless-steel autoclave (100 ml vo-
lume) and then sealed and heated at 200 °C. After 14 h reaction time,
the autoclave was cooled to room temperature. The dark suspension
was washed several times with methanol: water mixture and distilled
water. The magneticclusters werekeptin10 ml water, asasuspension,
andpartofthe samples were dried and used for different analyses. For a
clearer description of the obtained results we will introduce here the
following notations: MNC-T, MNC-A and MNC-M for the MNC stabilized
with sodium tartrate, aspartic acid and malic acid respectively.
Red Blood Cells (RBCs) collection and handing. Heparin-stabi-
lized Wistar rat RBCs were freshly collected. Briefly, a 2 ml sample of
whole blood was added to 4 ml of phosphate-buffered saline (PBS), and
the RBCs were isolated from serum by centrifugation at 1500 rmp for
10 min. The RBCs were further washed twice with PBS solution.
Following the last wash, the RBCs were diluted to 4 ml with PBS solu-
tion. Then, the diluted RBC suspension was added on MNC suspension
in PBS to make the final MNCs concentration 10 μg/mL. Deionized
water and PBS were added to RBC suspensions as the positive and ne-
gative controls, respectively. All samples were prepared in triplicate
and the suspension was briefly vortexes before leaving at static condi-
tions at 24 °C for 2 h. Finally, the mixtures were centrifuged at
1500 rmp for 10 min, and the RBCs were isolated and washed twice
with PBS [12].
2.2. Haemolysis assay
RBCs were incubated with MNC (10 μg/mL) for 2 h and centrifuged
to isolate the RBC as described above. The absorbance values of the
supernatant at 570 nm were measured.
2
Applied Surface Science 501 (2020) 144267
2.3. Antioxidant enzymes activity in the RBS assay
The diluted RBC suspension was mixed with MNCs at a concentra-
tion of 10 μg/mL in PBS solution and incubated at 24 °C for 2 h. Then,
the cells were lysed and the supernatant was collected for the mea-
surement of SOD activity.
The protein levels in RBCs lysates were measured with the Bradford
method [13].
Superoxide dismutase (SOD) activity was determined using SOD
Assay Kit-WST(Sigma) as indicated by the producer. The absorbance
was read at 450 nm using an ELISA plate reader (Tecan, Männedorf,
Switzerland); the % inhibition rate was calculated.
Catalase activity (CAT) was assayed according to Pippenger
method16. Catalase activity in homogenates was measured in a reac-
tion mixture containing 10 mM hydrogen peroxide in 50 mM kalium
phosphate buffer, pH7.4. The reduction in absorbance at 240 nm was
recorded for 3 min. The enzyme quantity which produced an 0.43 re-
duction in absorbance at 250 per minute in this system was defined as
one unit of catalase activity. The activity was expressed as units/mg
protein [14].
2.4. Characterization methods
Transmission electron microscopy (TEM) was performed by a
Hitachi SU 8230 equipped with energy dispersive X-ray analysis (EDX)
in order to determine the morphology and elemental composition of the
prepared samples. The electron energy used was 30 keV. In order to
amplify the secondary electron signal, the samples prepared in the form
of powders were metalized with an Au thin layer of 10 nm in an au-
tomatic Sputter Coater, in argon atmosphere.
The surface chemical composition of the samples was investigated
by X-ray Photoelectron Spectroscopy (XPS) using a spectrometer SPECS
equipped with an Al/Mg dual-anode X-ray source, a PHOIBOS 150 2D
CCD hemispherical energy analyzer, and a multichanneltron detector
with vacuum maintained at 1 × 10−9 Torr. The Al Kα X-ray source
(1486.6 eV) was operated at 200 W. The XPS survey spectra were re-
corded at 30 eV pass energy and 0.5 eV/step. The high-resolution
spectra for the individual elements (Fe, C, O, N, VB) were recorded by
accumulating 10 scans at 30 eV pass energy and 0.1 eV/step. Data
analysis and deconvolutions was performed using CasaXPS software
with a Gaussian-Lorentzian product function and a nonlinear Shirley
background subtraction. Peak shifts due to any apparent charging were
normalized with the C 1s peak set to 284.8 eV.
Magnetic measurements on powder samples at room temperature
was performed using a Vibrating Sample Magnetometer Cryogenics.
Caracterization of the MNC colloidal suspension was performed by
measuring the zeta potential as a function of the suspension pH at 25 °C
by laser Doppler micro-electrophoresis technique using a Malvern
Zetasizer Nano ZS-90 instrument. The pH range was 2-11 adjuted by
using HCl and NaOH. The equipment contains a He–Ne laser operating
at a wavelength of 633 nm and an avalanche photodiode detector. To
ensure the reproducibility of our results the measurements were per-
formed in triplicate and the mean value was reported.
3. Results
3.1. Samples preparation and physico-chemical caracterization
The scheme of reaction is shown in Fig. 1. For the prepared samples
all the reactions parameters were identical, only the surfactant differs
from sample to sample, as described in the experimental section.
To investigate the effects of organic acids on the shape and sizes of
the obtained MNC, TEM analyzes were performed. As can be seen from
the obtained TEM images and histograms (Fig. 2), the MNC synthesized
by solvothermal method show regular spherical shapes with sizes
within the ranges: 40–120 nm for MNC-T, 70–280 nm for MNC-A and
T. Radu, et al. Applied Surface Science 501 (2020) 144267

Fig. 1. Synthesisof Fe3O4 magnetic nanoclustersby solvothermal method.

Fe2p MNC-A

3+
I (a. u.) Fe
2+ 3+
Fetet Feoct
3+
Fe
2+
satellites Feoct

712 710 708

MNC-T
MNC-A
MNC-M (b)
(a)
740 720 700 740 720 700
BE (eV) BE (eV)
Fig. 3. (a) Normalized XPS Fe2p high resolution spectra obtained for the
analysedMNC; (b) Deconvoluted Fe2p XPS spectra obtained after Shirley
background subtraction.

investigated samples atoms of Fe, C and O were identified in different

amounts while for the MNC-A a small amount of nitrogen was also
detected as expected from the aspartic acid molecule. Deconvolution of
Fig. 2. Left: TEM images for the MNC samples; scale bar 300 nm. Right: MNC
the Fe2p XPS high resolution spectra shown in Fig. 3b provides the
size histogram with a dashed black line representing the size distribution fit to a
characteristic components for inverse spinel structure of magnetite. The
normal function used to determine the distribution mean-size.
Fe2+ and Fe3+ specific lines for orthorhombic and tetrahedral config-
uration are observed at 709.4 and 710.5 eV respectively while Fe3+
70–190 nm for MNC-M. The size distribution for each sample was fitted tetrahedral species is observed at 713 eV [16,17]. Two satellites peaks
with a normal function used in generally to describe granular random are also identified in the deconvolution of the spectra at 716 and
systems. The average size of MNC-Tis ~ 86 nm which is smaller as 718.5 eV which can be attributed to shake up or charge transfer process
compared with MNC-Mand MNC-A for which the obtained mean size is [18]. The obtained Fe2p XPS spectrum for MNC-T and MNC-M (not
123 and 172 nm respectively. shown) is very similar with the MNC-A, which proves that the samples
XPS survey spectra were analyzed in order to determine the ele- core is magnetite. As shown in inset of Fig. 2, a small shift to lower
mental composition of the samples surface. As shown in Table 1 for the binding energy, was observed for MNC-A relative to the MNC-T and
MNC-M. Similar behaviour was observed by other studies previous re-
Table1 ported [19–21] and was correlated with a small decrease in the Fe3+/
Elemental composition at the surface of MNCs determined from survey spectra. Fe2+ ratio in the sample. The enhancement in the Fe2+ component of
the Fe 2p spectra is visible also in the region of the satellite peaks which
sample Elemental concentration (at %)
shows an increased intensity of Fe2+ satellite in the Fe2p spectra [22]
Fe O C N From the deconvolution of the Fe2p XPS spectra the Fe3+/Fe2+
ratio for MNC-A was 1.9 while for MNC-T and MNC-M the values ob-
MNC-A 40 46.2 12.5 1.2
tained were 2.1 and 2.2 respectively.
MNC-M 36.4 54.7 8.8 –
MNC-T 31.9 53.6 14.5 – High resolution XPS spectra obtained for C1s for the three analyzed
samples is shown in Fig. 4. As one can see, the spectra have similar

3
T. Radu, et al. Applied Surface Science 501 (2020) 144267

MNC-A MNC-T MNC-M

C-O/C-N
C-C C-O
C-O C-C
I (a. u.) COO
- C-C COO
-

-
COO

(a) (b) (c)

300 290 280 300 290 280 300 290 280

BE (eV) BE (eV) BE (eV)
Fig. 4. C1s high resolution XPS spectra each deconvoluted into three components characteristic for the chemical bonding present at the surface.

shape, only the contribution of the identified components varies slightly
according with the ration of the chemical bonds present at the surface:
CeC at 284.5 eV, CeO at 285.6 eV and COO− at 288.2 eV respectively.
The data obtained from the deconvolution of C1s XPS spectra are
summarized in Table 2.
The identified chemical bonds in the C1s spectra are characteristic
for the type of organic acid of the prepared samples: malate, aspartate
and tartrate.
The O1s spectra (see SI Fig S1) confirms the presence of lattice O,
CeO and COO− at the sample surface [23,24].
The VB spectra of the obtained samples are shown in Fig. 5. The
obtained spectra are referred to the Fermi level (EF). The FWHM of the
VB-XPS spectra slightly varies from 5.8 eV for MNC-T to 6 eV for MNC-A
and 6.1 eV for MNC-M respectively (see Fig. 5). The clusters mean sizes
as shown in TEM images (Fig. 2) increases from 82.6 nm for MNC-T to
172 nm for MNC-A. That is why small differences are observed in the
VB spectra FWHM, knowing that variations in the width of spectrum
are correlated with the variation of coordination induced by particles
size modifications. Regarding the position in binding energy of the
VB-XPS MNC-M
MNC-T
MNC-A
EF = 0
obtained VB and XPS spectra, it was shown previously that in the case
of very small particle size the inner core electrons are affected more
strongly by the quantum confinement while the valence electrons are
strongly affected by the coordination variations [15]. For the MNCs
studied here a small shift was observed to higher binding energy for the
inner core spectra of the MNC-T and MNC-M as compared to MNC-A
(Fig. 4a). This observation can be related to smaller particles size in
these samples as compared to the MNC-A.
Moreover, the obtained VB-XPS spectra (Fig. 5) have similar shapes
with a well defined maximum at 6 eV attributed to contributions mainly
from Fe octahedral and O atoms, and a weak shoulder at about 3 eV due
to the Fe tetrahedral and octahedral sites. The only difference observed
is close to EF = 0 where the total density of states increases for the
MNC-A as compared to MNC-M and MNC-T. This result may be linked
with earlier reported theoretical and experimental results [25] which
prove that surface functionalization of the magnetite induce change in
its electronic properties due to an enrichment of the Fe2+ cation at the
sample surface. For MNC-A, XPS results show indeed a higher Fe con-
tent as compared to the other two MNC (see Table 2) which may be
linked with a higher content of Fe2+. The shoulder observed at 709 eV
in Fig. 3a characteristic for Fe2+ is more pronounced for MNC-A, which
is consistent with an increase in the Fe2+ content. However, an en-
richment of Fe2+ cations would be reflected to an enhanced magneti-
zation for this particular sample. Therefore in order to confirm the
observed findings magnetization measurements were performed. The
results obtained are shown in Fig. 6.
All the analysed MNCs show high saturation magnetization (Ms)

with values from 68 to 71 emu/g (Fig. 6a). Similar values were obtained
I (a. u.)

by other research groups for MNCs synthesized by solvothermal method

EF = 0

using different organic acids as surfactants [27,29]. The Ms for MNC-A

sample represents 83% from the bulk magnetite value while for the
MNC-T and MNC-M the value obtained for the saturation magnetization
is 74% from that expected for the bulk. This observation confirms
theoretical predictions for the Ms behaviour of clusters of dense parti-
cles where, even in a simplified model, the particle radius R, the cluster

Table 2
4 2 0 -2 The ratio of the identified chemical bonds as determined from the deconvolu-
tion of the C1s XPS spectra.
10 5 0 sample CeC (284.4 eV) CeO (285.6 eV) COO− (288.2 eV)
BE (eV) MNC-T 32% 36.3% 31.7%
MNC-A 36.1% 32.5% 31.2%
Fig. 5. Normalized VB-XPS spectra of MNCs. Inset: The VB-XPS spectrum of
MNC-M 30% 36.2% 33.6%
magnetite in the region close to EF.

4
T. Radu, et al. Applied Surface Science 501 (2020) 144267

Ms for bulk magnetite

80 2.0 2.0
MNC-T 3
MNC-A
40 MNC-M
1.5 1.5
M (emu/ g)
M (emu/g)

2+
Bc (mT)

Fe /Fe
0

3+
0
1.0 1.0
-3
-40 -4 0 4
Bc (mT)
0.5 0.5
(a) (b)
-80
-4 0 4 80 120 160
B (T ) D (nm)
Fig. 6. (a) Saturation magnetization measurements performed for MNCs.
Horizontal dashed line represents the Ms reported value for bulk magnetite
92 emu/g [26]. Inset: low field region showing Bc values. (b) Left: Variation of
Bc as a function of particle diameter; Right: Variation of Fe3+/Fe2+ as a
function of particle diameter. Fig. 7. The influence of stabilizer used for each type of sample on the ς po-
tential.

radius and the total number of the nanoparticles in the cluster Np have
to be taken into account [28]. MNC-T, MNC-M and MNC-A respectively. For in vivo studies the de-
Moreover, lower concentration of C-O bonds and higher Fe content sired pH is in the range 6–8, similar with the human body fluids (with
is present at the surface of MNC-A sample as shown by XPS. Therefore, the exception of the stomach pH). In this range our clusters presents no
the observed decrease of Ms for MNC-M and MNC-T can also be related charge fluctuations of the ς potential values. These values suggest their
with surface effects. It is known that near the surface the magnetization colloidal stability making them proper for different bio-applications.
is lower than in the interior, therefore the surface effects become more
important with reduction of particle size. 3.2. The blood biological effects of MNC
With increasing the diameter of the MNC a decreasing tendency of
Fe3+/Fe2+ ratio is observed (see Fig. 6b) as determined by XPS. Thus, 3.2.1. Haemolysis
as Fe2+ is taken into account for the resultant magnetic moment in Previous research results on interaction between human RBC and
Fe3O4, the observed decrease in Ms with decrease in Fe2+ (Fig. 6a) is nanoparticles of various materials[26], diameter ≤0.2 μm, with dif-
expected. The coercivity, Bc, of the samples is presented in the inset of ferent surface charge showed that negatively and neutral particles were
Fig. 6a which shows the low field region of the magnetization mea- found within the blood cell while positive surface charged particles
surements. The results prove that all the obtained samples are super- where only attached to the surface cell. To assess the impact of MNC on
paramagnetic with low Bc: 2.1 mT for MNC-M, 0.8 mT for MNC-A and erythrocytes, a haemolysis test was performed by measuring the hae-
0.5 mT for MNC-T. moglobin release after exposure to the different MNC at a concentration
It is worth to mention that the superparamagnetic behaviour of the of 10 μg/mL (Fig. 8). For this study we used distilled water as positive
obtained samples is favourite for biological applications, since their net control and PBS as negative control. There is no significant statistical
magnetization at room temperature is zero and therefore do not ag- difference between the use of PBS (negative control) and the exposure
glomerate. of the RBCs to the MNC, but a tendency towardsmore haemolysis is seen
However, the Bc values are very sensitive to particles size variations for the MNC-A.
as shown in Fig. 6b. The values of Bc increase with increasing size of
MNC similar to the observed Ms behaviour. 3.2.2. Antioxidant enzymes
The stability and charge of the three MNCs was studied by zeta We found no statistically significant difference between the levels of
potential varying the pH from 2 to 11, (Fig. 7). The general tendency of SOD (Fig. 9) and CAT (Fig. 10) of the RBCs exposed to the 3 types of
zeta potential is similar for all three samples, positive values at acidic
pH and negative values starting from pH ~ 6; at pH = 2 the obtained
values are positive for all MNCs: 46.4 mV (MNC-T), 40.9 mV (MNC-A)
and 37 mV (MNC-M). For increasing pH value to 5, the zeta potential
value for the sample MNC-M remains positive, about 15 mV while for
other two samples the zeta potential has negative values of about
−4 mV (MNC-T) and −8 mV (MNC-A). PH = 6 gives no significant
changes for MNCs, −4 and −10 for the MNC-M and MNC-T while for
the MNC-A sample the zeta potential has positive value, 5 mV. For
pH = 7 a decrease of the zeta potential to −1.6 mV is observed for
MNC-A while for MNC-M and MNC-T, the zeta potential has very small
variations, −10 and −13 mV respectively. For the pH values 8 and 9
the zeta potential remains below −10 mV and particularly for the
samples MNC-T and MNC-M the smallest values for zeta potential, −1
and 0 respectively was observed. A strong decrease of zeta potential to
Fig. 8. Hemolysis of MNC-T, MNC-A and MNC-M samples. Distilled water and
negative values was observed for pH = 11, −48, −50 and −54 mV for
PBS were used as positive and negative control respectively.

5
T. Radu, et al.
Fig. 9. CAT for MNC-T, MNC-A and MNC-M relative to the reference PBS ex-
pressed in expressed as units/mg protein.
Fig. 10. SOD level after interaction with MNCs.
MNCs at a concentration of 10 μg/mL as compared to PBS after 2 h of
exposure.
Erythrocytes act against oxidative damage, equipped with an anti-
oxidant system that contains enzymes: SOD, glutathione peroxidase,
glutathione reductase and CAT [28]. Previous studies haverevealed that
iron NPs determine dose-dependent oxidative stress and reducecell
viability, as theyproduce free radical species which reduce the levels of
cellularantioxidants such as SOD and CAT that lead to oxidative da-
mage [30,31]. When erythrocytes are facing oxidative stress, RBCs’
antioxidant enzymes cannot protect them against oxidative damage of
free radicals, and RBC lesions can occur due to oxidative damage. MNC
used in this study don’t influence significantly the levels of SOD and
CAT which reveals the compatibility of these MNCs with the red blood
cellsrespectively their friendly interaction with human body fluids.
4. Conclusions
New MNC stabilized with biocompatible organic acids (sodium
tartrate, aspartic acid and malic acid) were synthesized by solvothermal
method using same reaction parameters. Uniform superparamagnetic
clusters with the average size of 86, 123 and 172 nm depending on the
shell type and high magnetization values between 68 and 71 emu/g
were obtained. As shown by XPS, the chemical bonds characteristic for
each surfactant are present at the MNC surface. VB-XPS results de-
monstrated that by using the surfactants proposed here, the electronic
structure at the MNC surface may be tuned so that variation of Ms is
obtained. ς potential shows negative values with no fluctuations, for all
three types of MNCs in pH range 6 to 8; negative charge is attributed to
the carboxylic moieties of the surfactants. The biocompatibility of the
MNCs was proved by the interaction with rat RBCs using haemolysis
and erythrocyte antioxidant systemstests which shows no negative ef-
fect between clusters and RBCs. These findings together with the phy-
sico-chemical properties of the obtained MNC (saturation magnetiza-
tion, dimensions, colloidal stability) make them proper for biomedical
6
Applied Surface Science 501 (2020) 144267
applications.
Acknowledgments
Financial support from the National Authority for Scientific
Research and Innovation - ANCSI, Core Programme, Project No 36N/
2019 PN19-35 02 03 is gratefully acknowledged. The authors ac-
knowledge Dr. Cristian Leostean for conducting XPS and magnetic
measurements. This work was partially supported through the infra-
structure obtained in the project Research Center and Advanced
Technologies for Alternative Energies - CETATEA - 623/11.03.2014.
Declaration of Competing Interest
There are no conflicts to declare.
Appendix A. Supplementary material
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.apsusc.2019.144267.
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