Materials Letters ∎ (∎∎∎∎) ∎∎∎–∎∎∎

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4 Materials Letters
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journal homepage: www.elsevier.com/locate/matlet
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Biocompatibility and superparamagnetism in novel
13 silica/CaFe2O4 nanocomposite
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15 Q1 Lavanya Khanna 1, N.K. Verma
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17 Nano Research Lab, School of Physics and Materials Science, Thapar University, Patiala 147004, India

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20 art ic l e i nf o a b s t r a c t
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22 Article history: Q2 Calcium ferrite nanoparticles (CaFe2O4 NPs) have been successfully passivated by silica coating. The
Received 18 July 2013 particle size distribution was observed in the narrow range of 3–6 nm, without any agglomeration,
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Accepted 26 April 2014 suggesting the stabilization of high surface energy and magnetic interaction on the nanoparticles'
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surface, due to silica. Presence of amorphous silica resulted in less peak intensities of the orthorhombic
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Keywords: CaFe2O4 core. The characteristic IR bands of silica confirmed its coating thereby making the nanocom-
26 Silica–CaFe2O4 nanoparticles posite viable for bio-conjugation (active silanol groups in the silica layer covalently attach to bio-
27 Magnetic materials molecules). The nanocomposite exhibited superparamagnetic behavior (Ms ¼ 8.55 emu/g) and high
28 Nanocomposites biocompatibility below 500 mg/ml. The nanosized, superparamagnetic and highly biocompatible synthe-
29 Superparamagnetic
sized nanocomposite finds potential applications in biomedicinal hybrids.
Biocompatible
30 & 2014 Published by Elsevier B.V.
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1. Introduction is reported in this work. Their structural, morphological, magnetic
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properties and biocompatibility have been studied.
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Magnetic nanoparticles (MNPs) have attracted a great deal of These find applications in magnetic cell-separation of biological
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attention in biomedical field [1], but, there are some limitations entities, therapeutic drug delivery vehicles, contrast enhancement
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associated with them. Their surface inertness restricts the number agents in magnetic resonance imaging (MRI) and magnetic
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of molecules that can be bonded to the surface [2]. Also, their biosensing.
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tendency to agglomerate due to high surface energy and magnetic
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interactions lead to increase in size. This results in clogging of the
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capillaries and adsorption of plasma proteins leading to quick 2. Experiment
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clearance by the macrophages in the Reticuloendothelial system in
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the body, before reaching the target cells [2]. So, it is highly Silica coating on calcium ferrite nanoparticles was done by the
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desired that the synthesized nanomaterial be agglomeration-free modified Stober process [5]. Briefly, 150 ml ethanol, 3 ml water,
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and biocompatible. Surface stabilization of the MNPs serves both 5.1 ml ammonium hydroxide and 900 ml TEOS were added in
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these purposes. It is obtained by passivating the surface of MNPs a beaker and kept at 40 1C, while stirring. After 20 min, 6 ml
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with the most employed stabilizer i.e. silica. It screens the aqueous solution of calcium ferrite nanoparticles with the con-
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magnetic dipolar interaction between MNPs, thus preventing their centration of 4 mg/ml was added and the reaction was allowed to
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aggregation and favouring their dispersion in liquid media. It proceed for 5 h. Nanoparticles were magnetically separated,
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protects leaching of MNPs in an acidic environment, eliminates washed and dried to obtain silica coated calcium ferrite nano-
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protein adsorption and facilitates bio-conjugation [2,4,5]. particles.
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In addition, high inherent toxicity of transition metals (Ni, Co, The structure and morphology were studied by X-ray diffrac-
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Mn) in ferrites raises concern on their biocompatibility [3]. Ferrites tion (XRD; X’PERT PRO Panalytical, MRD ML) and Transmission
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of calcium are more biocompatible since calcium is inherently Electron Microscope (TEM; Hitachi (H-7500)), respectively. FT-IR
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non-toxic. A novel, agglomeration-free and highly biocompatible spectrum was recorded on Perkin Elmer-Model RZX. The magnetic
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nanocomposite composed of inherently non-toxic, superpara- property was determined by vibrating sample magnetometer
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magnetic calcium ferrite nanoparticles and biocompatible silica (VSM; Princeton Applied Research Model 151/155). The viability
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60 of synthesized nanocomposite on T cell lines (Jurkat cells) was
61 investigated by MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl-
62 E-mail address: lavanshya@yahoo.co.in (L. Khanna). tetrazolium bromide, a tetrazole) colorimetric assay at 5, 25, 50,
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Tel.: þ91 175 2393343; fax: þ91 175 2364498/2393002. 100, 250 and 500 mg/ml of nanoparticles (n ¼5). Cell viability was
64 http://dx.doi.org/10.1016/j.matlet.2014.04.168
65 0167-577X/& 2014 Published by Elsevier B.V.
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Please cite this article as: Khanna L, Verma NK. Biocompatibility and superparamagnetism in novel silica/CaFe2O4 nanocomposite.
Mater Lett (2014), http://dx.doi.org/10.1016/j.matlet.2014.04.168i
 2 L. Khanna, N.K. Verma / Materials Letters ∎ (∎∎∎∎) ∎∎∎–∎∎∎

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Fig. 1. (a) X-ray pattern (b) FT-IR spectrum and (c) M–H curve of the nanocomposite.
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41 calculated the following equation: Table 1
Description of the bands in FT-IR spectrum.
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I sample
43 Cell viability ð%Þ ¼  100 ð1Þ IR region or bands (cm  1) Descriptions
I control
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45 Isample is the mean absorbance of nanoparticles treated wells and 3410 v(O–H) stretching [4]
46 Icontrol is the mean absorbance of control wells without nanopar- 1635 v(O–H) scissor bending [8]
1450 v (Si–CH2) scissoring [7]
47 ticles treatment [1]. Statistical analysis was done by student's
1083 vas(Si–O–Si) stretching [2,8]
48 paired t-test. 876 v (Fe–O–H) bending [9]
49 857 v (Fe–O–H) bending [9]
50 712, 585, 466 v(Fe–O) stretching [10]
51 3. Results and discussion
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53 Fig. 1(a) shows the XRD pattern, where all the peaks are interacting superparamagnetic particles, demagnetizing effects
54 indexed and well-matched to the orthorhombic structure (JCPDS due to dipolar interactions reduce both the magnetic squareness
55 Card no. 74-2136) of CaFe2O4. Less intensity of CaFe2O4 core is (MR/MS ratio) and coercivity (HC) values [11]. Magnetic squareness
56 attributed to the silica coating which diminishes the peak inten- value ascribable to the above is 0.1, i.e more than 90% of magnet-
57 sities [6]. Also, the broad band near 2θ 15–201 corresponds to the ism is lost on removal of the magnetic field [12]. As, the obtained
58 amorphous silica surrounding CaFe2O4 NPs. Characteristic bands squareness value is 0.12, therefore, it can be comprehended that
59 corresponding to silica, in FT-IR spectrum (Fig. 1(b), Table 1), the nanocomposite exhibits superparamagnetism. Superparamag-
60 confirms the formation of silica coating on nanoparticles' surface. netic behavior of a particle is directly related to its magneto-
61 In the M–H curve, Fig. 1(c), the nanocomposite exhibits super- crystalline anisotropy [13]. It is further correlated to L–S coupling,
62 paramagnetic behavior, with magnetic saturation (Ms), remanent as the magneto-crystalline anisotropy originates from the coupling
63 magnetization (MR) and squareness value (MR/MS ratio), 8.55 emu/g, between electron spins and the angular momentum of the
64 1.05 emu/g and 0.12, respectively. Squareness value is instrumental electron orbital (L–S coupling) [13]. The observed magnetization
65 in determing superparamagnetic behavior. For non-interacting of the nanocomposite is due to non-collinear spin structure,
66 superparamagnetic particles, the squareness value is 0.5, in case of originating from the pinning of the surface spins and silica coating

Please cite this article as: Khanna L, Verma NK. Biocompatibility and superparamagnetism in novel silica/CaFe2O4 nanocomposite.
Mater Lett (2014), http://dx.doi.org/10.1016/j.matlet.2014.04.168i
 L. Khanna, N.K. Verma / Materials Letters ∎ (∎∎∎∎) ∎∎∎–∎∎∎ 3

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Fig. 2. TEM micrographs at (a, b) different magnifications, (c) enlarged view of the dotted box, and (d) histogram of the nanocomposite.
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42 at the interface of nanoparticles [14]. As the particle size decreases, also been calculated in order to understand the effect of synthe-
43 a large percentage of the atoms become surface atoms, resulting in sized nanocomposite on the cells. The enhanced cell viability is
44 pronounced surface and interface effects leading to increase in the attributed to good dispersibilty of silica coated CaFe2O4 NPs in the
45 ratio of dead layer (non-magnetic silica) to magnetic core [11], medium. This results in their relatively less aggregation, facilitates
46 thus diluting the magnetization value. their intracellular uptake and hampers cellular damage [17]. Silica
47 TEM micrographs in Fig. 2(a, b) reveal the spherical morphol- protects MNPs from acidic erosion. Correspondingly, the particle
48 ogy of the synthesized nanocomposite. Fig. 2(c) represents the stability translates into reduced cytotoxicity [18]. A significant cell
49 enlarged view of the dotted box in Fig. 2(b); where the core–shell viability has been observed below 500 mg/ml, and a fall occurs at
50 formation and the contrast of dark CaFe2O4 core and amorphous 500 mg/ml; this may be attributed to shortage of culture media
51 silica surrounding it, are well-observed. Formation of agglomeration- required for the growth of the cells while running the assay, rather
52 free nanocomposite is credited to the presence of silica shell. than any toxicity of the synthesized nanocomposite. On exposure,
53 It stabilizes the high surface energy and also forms a magnetically the nanocomposite first adhere to cells, then are internalized by
54 dead layer that inhibits the magnetic and inter-particle interactions endocytosis, finally get accummulated in digestive vacuoles [19].
55 of the nanocrystallites, thus preventing their agglomeration. Fig. 2 Therefore, at higher concentration, the overloading of particles
56 (d) shows the histogram, plotted by the analyze-it software. It reveals results in fatality of cells. The enhanced cellular viability of the
57 that the highest frequency (diameter) falls in the size range of nanocomposite as compared to reported materials [1,4,17] makes
58 3.5–4.0 nm. Relative standard deviation (RSD¼(Mean/S.D.  100)) is it a potential candidate for biomedicinal applications.
59 the parameter for determining narrow or wide distribution [16]. RSD
60 lower than 15% corresponds to a narrow distribution [15]. In the
61 present study, RSD has been calculated to be 14.5%; this being lower 4. Conclusions
62 than 15%, the obtained distribution is considered to be narrow.
63 Dose-dependent cytotoxicity of the nanocomposite tested on Biocompatibility and superparamagnetism in novel silica
64 T-cell lines (Jurkat cells) using MTT assay reveals their non-toxic coated CaFe2O4 nanocomposite have been reported. XRD pattern
65 behavior. Fig. 3 (a) shows the optical density (OD) values (standard reveals the orthorhombic structure of CaFe2O4, with a broad band
66 error (SE) bars). The statistical analysis using paired t-test [16] has near 2θ  15–201 pertaining to the presence of amorphous silica.

Please cite this article as: Khanna L, Verma NK. Biocompatibility and superparamagnetism in novel silica/CaFe2O4 nanocomposite.
Mater Lett (2014), http://dx.doi.org/10.1016/j.matlet.2014.04.168i
 4 L. Khanna, N.K. Verma / Materials Letters ∎ (∎∎∎∎) ∎∎∎–∎∎∎

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47 INSPIRE Fellowship to carry out this research work.
48

Please cite this article as: Khanna L, Verma NK. Biocompatibility and superparamagnetism in novel silica/CaFe2O4 nanocomposite.
Mater Lett (2014), http://dx.doi.org/10.1016/j.matlet.2014.04.168i