NANO

A facile synthesis, characterization of spinel structured nickel chromite (NiCr2O4)

nanoparticles and their application in antibacterial and anticancer activities
--Manuscript Draft--

Manuscript Number: WSPC-NANO-D-23-00348

Full Title: A facile synthesis, characterization of spinel structured nickel chromite (NiCr2O4)
nanoparticles and their application in antibacterial and anticancer activities

Article Type: Research Paper

Section/Category: Chunli Bai (Nanomaterials and Nanostructures; Nanomedicine and Nanobiology)

Keywords: Sol-gel synthesis, NiCr2O4, Bioactivity, Anticancer activity

Corresponding Author: Tasaduk Ahmad Wani

Annamalai University
INDIA

Corresponding Author Secondary

Information:

Corresponding Author's Institution: Annamalai University

Corresponding Author's Secondary

Institution:

First Author: Ajith A

First Author Secondary Information:

Order of Authors: Ajith A

Tasaduk Ahmad Wani

Suresh G

Saravanan N

Rachid Masrour

Order of Authors Secondary Information:

Abstract: Herein, a sol-gel technique has been used to prepare spinel NiCr2O4 nanoparticles
(NCNPs) at 700°C to investigate their antibacterial and anticancer activities.
Thermogravimetric thermal analysis, X-ray diffraction, scanning electron microscopy,
energy dispersive spectroscopy, transmission electron microscopy, Fourier transform
infrared spectroscopy, X-ray photoelectron spectroscopy, ultraviolet diffuse reflectance
spectroscopy, and photoluminescence were used to characterize NCNPs. TG-DTA
confirmed the thermal durability of NCNPs above 700°C. XRD indicated cubic-shaped
NCNPs having a 38 nm crystallite size. FT-IR spectra confirmed the spinel lattice with
distinctive tetrahedral and octahedral stretching bands owing to metal-oxides below
700 cm-1. FESEM and TEM images revealed almost cubic-shaped NPs. The optical
bandgap as per UV-DRS spectra was 2.7 eV. The PL showed emission bands in the
visible and UV regions. The surface-phase-pure elemental and electronic arrangement
of the nanosamples were verified by X-ray photon spectroscopy. The in-vitro
antibacterial and anticancer applications of NCNPs were investigated. Among the
microorganisms, S. aureus exhibited the highest level of sensitivity (9 mm), while E.
coli had the lowest (5 mm) against NCNPs. The MTT test revealed the high cytotoxicity
of NCNPs against HeLa and MCF-7 cells, thus opening the way for NCNPs to be
considered as prospective candidates for bactericidal and anticarcinogenic agents.

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A facile synthesis, characterization of spinel structured nickel chromite (NiCr2O4)

nanoparticles and their application in antibacterial and anticancer activities

Ajith A1, Tasaduk Ahmad Wani2, Suresh G3, Saravanan N4, Rachid Masrour5

1,2,3 Department of Physics, Annamalai University, Annamalai Nagar, Tamil-Nadu,

India

4 Division of Biochemistry, Rani Meyyammai College of Nursing, Annamalai

University, Annamalai Nagar, Tamil-Nadu, India

5 Laboratory of Solid Physics, Faculty of Sciences Dhar El Mahraz, Sidi Mohamed

Ben Abdellah University, Box 1796, Fez, Morocco

Abstract: Herein, a sol-gel technique has been used to prepare spinel NiCr2O4 nanoparticles
(NCNPs) at 700°C to investigate their antibacterial and anticancer activities.
Thermogravimetric thermal analysis, X-ray diffraction, scanning electron microscopy, energy
dispersive spectroscopy, transmission electron microscopy, Fourier transform infrared
spectroscopy, X-ray photoelectron spectroscopy, ultraviolet diffuse reflectance spectroscopy,
and photoluminescence were used to characterize NCNPs. TG-DTA confirmed the thermal
durability of NCNPs above 700°C. XRD indicated cubic-shaped NCNPs having a 38 nm
crystallite size. FT-IR spectra confirmed the spinel lattice with distinctive tetrahedral and
octahedral stretching bands owing to metal-oxides below 700 cm-1. FESEM and TEM images
revealed almost cubic-shaped NPs. The optical bandgap as per UV-DRS spectra was 2.7 eV.
The PL showed emission bands in the visible and UV regions. The surface-phase-pure
elemental and electronic arrangement of the nanosamples were verified by X-ray photon
spectroscopy. The in-vitro antibacterial and anticancer applications of NCNPs
were investigated. Among the microorganisms, S. aureus exhibited the highest level of
sensitivity (9 mm), while E. coli had the lowest (5 mm) against NCNPs. The MTT test
revealed the high cytotoxicity of NCNPs against HeLa and MCF-7 cells, thus opening the
way for NCNPs to be considered as prospective candidates for bactericidal and
anticarcinogenic agents.

Keywords: Sol-gel synthesis, NiCr2O4, Bioactivity, Anticancer activity.

1. Introduction
Owing to their significant visible-light absorption chromium based spinel oxide compounds
with general formula MCr2O4 (M = Co, Zn, Ni, Mn etc.) have received a great deal of
attention in recent research field as a significant class of inorganic functional materials
because of their high melting point, narrow gap energy, stability, superior catalytic
characteristics, etc. [1]. Mined as an ore (FeO.Cr2O3), chromite mostly contains the base
metal chromium along with several transition elements. It mainly appears in metamorphic
rocks such as serpentinites and stratified ultramafic intrusive stones. Early igneous
differentiation also produced chromite ore deposits. [2].One particular spinel chromite,
Nickel Chromite (NiCr2O4) abbreviated as NC, crystallizes with the normal spinel structure,
as do many other A2+B23+O4 compounds. These have tetragonal and cubic symmetry in the
room-temperature phases, respectively. Because of their many applications in sensors,
semiconductive substances, magnetic properties of materials, catalyzing substances,
temperature-sensitive substances, as well as excellent industrial catalysts spinel-type oxides
are more desirable right now [3]. NiCr2O4, a P-type semiconductor material has been
recognized as an excellent photocatalyst [4] and are typically employed as catalytic materials.
[5]. These have numerous uses in various industries, including the paper, leather, and textile
ones, as well as being employed as indicators in analytical laboratories [6]. Large quantities
of dye material comprising colored dangerous compounds can combine with wastewater
during the manufacturing process of textile, paper, and chemical laboratories and then be
released into the natural resources of aquatic systems. [7]. Considering the rising
environmental and energy crises there is a massive demand or photocatalysts that can
effectively eliminate these toxicants from the biosystem. Thanks to appealing properties and
performance of the heterogenous photocatalysts for tackeling the above mentioned crises [8].
As a general rule, during photocatalytic solar energy is utilized for photodegradation of the
toxicants.

Metal oxide nanoparticles have been also utilized in the medical and pharmaceutical
industries recently [9]. Therefore, it is believed that nanomedicine is a cutting-edge contender
for effectively resolving these worldwide issues. Due to its unique and inspiring
physicochemical properties, such as its large surface area, ease of transformation, effective
magnetic, anti-carcinoma and bactericidal properties, it is one of the adaptable nanomaterials
that are frequently used in biomedical applications [10]. Owing to their high surface activity
and wide surface to volume ratio, NCNPs have notable cytotoxic and bactericidal properties
[10].
It is worth to note that cancer nanotechnology has opened up a new frontier of
multidisciplinary study spanning chemistry, medicine, engineering, and biology, with a focus
on big advances in cancer detection, diagnosis, and therapy [11]. When the NP diameter of a
spherical particle decreases, the surface size rises according to diameter squared leading in an
increase in surface activity relative to bulk materials with greater dimensions, which can
improve material biocompatibility [12]. NPs can exhibit cytotoxicity in carcinogenic cells by
changing their shape and decreasing viability while also producing oxidative stress [13].

Various chemical and physical techniques for the creation of metal nanoparticles have been
documented. Nanomaterial synthesis techniques including co-precipitation [14], thermal
decomposition [15], reflux condensation [16], sol-gel auto combustion [17], thermolysis [18],
Precipitation [19], spray pyrolysis [20], combustion [21], solution casting [22], microwave
[23], and solvothermal routines [24]. Among all of them we used sol-gel autocombustion
synthesis technique which is recognized as a simplest, cheap, reliable, reproducible,
controllable and low temperature method [25].

Our purpose is to synthesize NiCr2O4 nanoparticles (NCNPs) using a simple sol-gel auto-
combustion method and characterizing them by TG-DTA, XRD, FE-SEM, EDS, HR-TEM,
SAED, FTIR , XPS, UV-DRS and PL spectroscopy. Finally its antibacterial and cytotoxic
activity has been evaluated. The novelty of the present study lies in the fact that we have
reported of NiCr2O4 NPs and its antibacterial and anticancer activities in detail. There are
very few reports on it. Antibacterial activity of NCNPs was studied against Gram-positive (E.
coli and B. subtilis) and Gram-negative bacterias (S. aureus and B. aureus) at varied
concentrations by disc diffusion method whereas its cytotoxic action was evaluated by MTT
test against carcinogenic HeLA and MCF-7 cancerous cell lines.

2. Methodology

2.1. Materials

The Cr (NO3)3.9H2O; 99%, Ni (NO3)2.6H2O; 99.98% and C6H8O7; 99% (Sigma Aldrich)
were used as precursors. No additional purification was performed before using any of the
compounds.

2.2.Preparation of NiCr2O4 Nanocomposite

NCNPs were produced utilizing citric-acid assisted sol-gel auto-combustion technique. The
necessary precursors, including citric acid [(C6H8O7)], nickel nitrate [(NO3)2.6H2O; 99.98%],
and chromium nitrate [(NO3)3.9H2O; 99%], were accurately dissolved in distilled water while
maintaining nitrate-citrate ratio as 1: 2. To create a homogenous solution, all the precursors
were individually dissolved in 100 mL of distilled water. The prepared solutions were heated
on a magnetic plate at 80°C while being swirled magnetically. The stirring was ceased once
the hot liquids had transformed into a gel after two hours. The temperatures of the samples
were raised to 250°C, after the formation of the viscous gel. The gel eventually instantly
burned into a loose powder at this temperature after some time. Using an agate mortar and
pestle, a homogeneous dried brown-black xerogel was produced, which was then pulverized
into an ultrafine powder. In a muffle furnace, the powder was calcined at 700°C for two
hours. The sample was named as NCNPs. The procedure for the synthesis of NCNPs is
schematically shown in Figure 1.

NH4OH

Sol Formation

+
Heating cum stirring at 80 oC
Vigorous Stirring
Evaporation
B

Nitrates Citric Acid

Precursor Nitrates

Stop Stirring
Anealing

Chromite Nanopowder

Autocombustion at 250 oC Viscous Gel

A
Figure 1. Procedure for synthesis of NCNPs [27] .

2.3. Antibacterial activity

By using the Kirby Bauer diffusion method, bactericidal action of NCNPs was examined
against Gram-positive Staphylococcus aureus and Bacillus subtilis and Gram-negative
Escherichia coli and Salmonella sp. Firstly, 15-20 mL of sterile Muller Hinton Agar (MHA)
was added to sterile Petri plates. Then, inoculums from cultures that had been synthesizing
for 24 hours were applied to the MHA media's surface and let to dry for 10 minutes. The
produced NiCr2O4 NPs were impregnated onto sterile discs (size 6 mm) at concentrations of
50 g/mL. In order to allow for compound diffusion, the loaded discs were kept on top of the
MHA plates containing bacterial culture seeds and the necessary controls for 30 min. at room
temperature. A positive control for microorganisms was utilized, which was ampicillin (5
g/disk). The sterile disc served as a negative control and was treated with dimethyl-sulfoxide
(DMSO). After that, each disc was incubated for 24 hours at 37 °C. Following incubation, the
strong inhibitory zone around the discs on the agar surface was measured in millimeters to
determine the bacterial susceptibility. This outcome was analyzed and contrasted with that of
the antibiotic ciprofloxacin.

The bacterias were procured from Sesaam Biological Lab Services, No. 24, 2nd Avenue,
Ashok Nagar, Chennai. The NCIM numbers of the bacteria are: E.Coli (5662); Salmonella
sp. (2501); Staphylococcus aureus (2079); Bacillus subtilis (2097).

2.4. Anticancer activity

2.4.1. Cell Line and Culture

The Henrietta Lacks (HeLa) and Michigan Cancer Foundation (MCF-7) cell lines were
donated by the King Institute in Guindy, Chennai. The cells have been maintained at 37 °C in
a humidified environment of 50 g/ml CO2, a Minimal Essential Medium (MEM)
supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin
(100 g/ml), and a Minimum Essential Medium without these additives.

Reagents

MEM was provided by Hi Media Laboratories, and FBS by Cistron Laboratories. MTT
(methyl thiazolyl diphenyl-tetrazolium bromide) and dimethyl sulfoxide (DMSO) including
all other chemicals and reagents were purchased from Sigma-Aldrich, Mumbai.

2.4.2. In vitro Assay for Anticancer Activity (MTT Assay)

24-well plates with (1×105/well) were seeded and incubated at 37 °C with 5% CO2. When the
cell attained confluence, the sample was added, and it was cultured for 24 hours. After
incubation, the sample was removed from the well and rinsed in MEM without serum or
phosphate-buffered saline (pH 7.4). Each well received 100 µl (5 mg/ml) of 0.5% 3-(4,5-
dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT), which was then incubated
for 4 hours. Each well received 1 milliliter of DMSO after the incubation time. The
absorbance at 570 nm was determined using a UV spectrophotometer with DMSO as a blank.
The concentration required for 50% inhibition (IC50) was measured and visually
approximated. The cell viability (% ) was calculated using the formula shown below [26]

A
Viability (%) = A t ∗ 100 (2)
C

where At is the cell absorbance treated with the NCNPs and AC is the cell absorbance treated
with the culture medium only.

2.5. Characterization

The synthesized powdered samples of NCNPs were characterized by TG/DTA, XRD, SEM-
EDS, TEM-SAED, FT-IR, XPS, UV-visible DRS and PL spectroscopy. The thermal
behavior of the NCNPs was studied by using NETZSCH - STA - 449F 3 – JUPITER thermal
analyzer. The XRD analyses of the samples was conducted by PANalytical X’pert PRO
instrument with CuKα radiation (α = 1.541 Å ). FTIR spectra was recorded by Perkin Elmer
Model No. Spectrum Two Serial No. 105627 FT-IR spectrometer. FESEM, CARI ZEISS -
SIGMA 300 coupled with EDS was used for morphological studies of the samples. FEI
Tecnai G2 20 S-TWN HRTEM coupled with SAED was employed to analyze the internal
microstructure and d-spacing of the samples. The luminescence behaviour of the prepared
samples was studied using the SCHIMADZU FLUOROLOG – FL3 -11 spectrophotometers.
X-ray photoelectron spectra (XPS, ULVAC-PHI, Model: PHI5000) were recorded to analyse
the elemental composition of the nanosamples and binding states of the elements. The optical
studies of the samples were recorded using UV-DRS- SCHIMADZU 3600.

3. Results and Discussion

3.1. TG – DTA Investigations

Figure 2 depicts the TG/DTA curves of the initially produced samples. The TG curve
demonstrates that the as-prepared sample undergoes two significant stages of breakdown
before becoming NiCr2O4 powders. With a total weight loss of 4.8%, the first stage of weight
loss is from 34 °C to 310 °C. From 310 °C to 680 °C, or a total weight loss of 13.9%, is the
second stage of weight loss. The dehydration and breakdown of the organic component can
be attributed to the first stage of weight loss, while the reaction is mostly responsible for the
second stage. No further weight loss is seen above 700 °C, indicating the creation of a stable
oxide and demonstrating the thermal stability of spinel-type NiCr2O4 nanocrystalline at
higher temperatures.
 Figure 2.Thermal analysis curve of the as-prepared sample of NCNPs.

It is evident from the DTA curve that there are two endothermic peaks at roughly 321 °C and
412 °C. The evaporation of organic substances is what causes the peak to occur at about 321
°C. The breakdown of the sample as prepared is what caused the peak to appear at roughly
412 °C. NiCr2O4 is the end product, and no thermal breakdown can be seen at temperatures
above 700 °C.

3.2. XRD Analysis

XRD pattern of NCNPs in the 2𝜃 range of 10o to 80o is depicted in Figure 3. At 2θ values of
18.47o, 30.41o, 35.78o, 37.24o, 43.27o, 54.05o, 57.66o, 63.27o, and 74.85o, the intense and
narrow peaks for the sample notably predicts the crystalline sample. It was confirmed that the
cubic spinel crystal structure with the space group Fd-3 m formed when the evaluated hkl
values (111), (220), (311), (222), (400), (422), (511), (440), and (533) corresponding to the
above-mentioned 2𝜃 were perfectly matched with JCPDS file no. 88-0108. Furthermore, it
can be noticed that NCNPs diffraction pattern lacks any impurity peaks. The crystallite size
was computed typically by relation 3 [27].

kλ
D = βcosθ (3)

Where k is a constant, θ is the diffraction angle, λ is the wavelength, and β is the full width at
half maximum. The lattice parameter was estimated using the formula below [27 ].

𝑎 = d×√ℎ2 + 𝑘 2 + 𝑙 2 (4)

Where hkl are the miller indices.

The average crystallite size was determined as 38 nm. The lattice parameter of NCNPs was
8.338Å which is in line with previous reports[28].

The above formula V = 𝑎3 was used to compute the volume of a single cell. Formulas (5) and
(6) were used to calculate microstrain and dislocation density [29].

β
ε = 4cosθ (5)

1
δ = D2 (6)

where the diffraction angle is θ and the full width at half maximum (FWHM) of the line is β.
Depending on the material lattice parameter, the volume of unit cell values was 579.67 Å3.
The microstrain and dislocation density of the samples were calculated as 3.7181 × 10-3 line-
2
m-4 and 0.6925× 10-3 nm-1 respectively. Table 1 tabulates all the structural parameters of
NCNPs that were 0discussed and
2 4
determined above 6using XRD.
8 10
These results are quite
consistent with previous reports [30].
10

(311)
1200 NCNPs
8

1000
Intensity (a.u)

800 (400)

(220) 4
(440)
(222)
600
(511)
(111) 2
(422)
400 (533)
0
10 20 30 40 50 60 70 80
200
2-theta (degrees)

Figure 3: XRD spectra of NCNPs.

Table 1: Structural properties: Average crystalline (D), Lattice constant, (𝑎), Cell Volume
(V), Average Micro strain (Ԑ × 10−3 ), & Dislocation density (δ × 10−3 ) of NC NPs

Sample D(nm) 𝑎(Å) V(Å3) (Ԑ × 10−3 )line-2m-4 (δ × 10−3) nm-2

NCNPs 38 8.338 579.67 3.7181 0.6925

3.3. FTIR spectroscopy

Figure 4 shows the FTIR spectra of NCNPs using KBr pellets. Using KBr pellet as a
reference, the spectral plots displayed transmittance versus wavenumber. The A and B sites
and the ionic distribution of the cations play major roles in determining the characteristics of
0 2 4 6 8 10

spinel chromites.
100

90 10

80
8
Transmittance (%)

70

60 6

50
n 3= 3488 cm
-1
4
40
n 2= 470 cm
-1

30
2
n 1= 628 cm
-1
20
4000 3500 3000 2500 2000 1500 1000 500
0
-1
Wavenumber (cm )

Figure 4: FT-IR curve of NCNPs

The spinel configuration of the NCNPs may be confirmed by FTIR spectra, which consist of
two signature bands υ1 (high frequency) and υ2 (low frequency) at 470 cm-1 and 628 cm-1
respectively. υ1 is related to the stretching vibrations of tetrahedral A-site while as υ2
corresponds to the stretching vibrations of octahedral B-site. Both of the bands represent the
signature bands of the spinel structures with two sub-lattices, namely tetrahedral and
octahedral sites [27, 31]. A third significant absorption band around 3488 cm-1 can be
attributed to O-H stretching vibration [27]. Table 2 enlists the FTIR band locations that were
observed.

Table 2. FTIR bands of synthesized NiCr2O4 NPs.

Sample υ1 (𝑐𝑚−1 ) υ2 (𝑐𝑚−1 ) υ3 (𝑐𝑚−1 )

NCNPs 628 470 3488

3.4. FE-SEM with EDS study

The morphological study of NCNPs has been studied by FE-SEM. The FESEM image of
NCNPs is shown in Figure 5a. FESEM images prove the cubic-shaped particles which are
evenly distributed, nanosized, agglomerated.

Figure 5. (a, b): FESEM image and (c) EDS spectra of NiCr2O4 NPs.

The sample was also examined using the EDS technology as shown in Figure 5b. Ni, Cr, and
O were found to have atomic percentage compositions of 14.77%, 15.53%, and 69.70%,
respectively without any phase impurity (Table 3).

Table 3. Elemental composition of NCNPs via EDS spectra

Element line Mass % Atom %

OK 39.98 69.70

Cr K 28.94 15.53

Ni K 31.08 14.77

Total 100.0 100.0

3.5.TEM Analysis
To learn more about the sample's microstructure and crystallinity, HR-TEM with SAED
analysis was employed. HRTEM image with SAED pattern of NCNPs is depicted in Figure 6
(a,b). The d-spacing values as shown in Table 4, were also calculated from SAED pattern by
employing ImageJ software. The figure shows that the samples contain nearly cubic-shaped
particles with agglomeration. For better estimation of particle size of the samples, the particle
size of NCNPs was estimated to be 46 nm respectively.

Figure 6. (a) HR-TEM image and (b) SAED pattern NCNPs

Table 4: d-spacing values and corresponding hkl values of NZNPs

1/2r(nm-1) 1/r(nm-1) r (nm) d-spacing (nm) hkl values

1.031031 2.062062 0.484951 4.849515 111
1.691211 3.382422 0.295646 2.956461 220
1.989553 3.979106 0.251313 2.513127 311
2.092573 4.185146 0.23894 2.389403 222
2.507728 5.015456 0.199384 1.993837 400

Moreover, both spots and rings may be seen in the SAED (selected area electron diffraction)
pattern of the samples (Figure 6b) proving that NCNPs are crystalline in nature as predicted
by XRD data.
 3.6. UV-Vis- DRS
The optical behavior of NCNPs was examined by UV-DRS (Diffused Reflectance
Spectroscopy) spectra and is shown in Figure 7a. As anticipated, the reflectance edge for
synthesized NPs was 268 nm. The band gap of NCNPs was determined by using the
Kubelka-Munk function F[R2] verses energy and extrapolating the linear part of K-M plot to
F[R2] to zero as shown, yeiding direct bandgaps energy of the NPs [32].

[1−R]2
F[R] = (7)
2R

Where R = 0 Relative
2 4
diffusion 6 8
reflectance 10 0 2 4 6 8 10

10
24 (a) (b)

10
8
22
Reflectance %

8
[F[R]*hn]2

6
20

6
18 4

4
16 2

2 Eg = 2.7 eV
14 0
200 400 600 800 1000 1 2 3 4 5 6

l (nm) 0 Energy (eV)

Figure 7. (a) Diffuse reflectance spectra and (b) K – M plot of NCNPs

The graph plotted between hν and [F [R]. hν]2 , as shown in Figure 7b, shows the energy gap
value of 2.7 eV corresponding NCNPs.

The energy gap values were also calculated by using the following energy equation.

hc 1240
E = hν = = (8)
λ λ

Where h = Planck’s constant = 6.62176 × 10−34 Js,

c = Speed of light = 3 × 108 m/s

λ = Absorption edge or reflectance edge

From the spectra it is clear that the bandgap of NCNPs is under 3 eV due to the enhancement
in crystalline nature of the sample at higher temperature. Same outcomes were reported by
Othman et al. [33] and Prakash et al., [34].
Herein due to its appropraite bandgap (< 3 eV) and better surface and microstructure, NCNPs
sample is the best candidate for bioactivity applications and for further investigation.

3.7. Photoluminescence study

The luminescence property of NCNPs was examined using the PL technique as shown in
Figure 8. The excitation wavelength used to record the spectra was 350 nm. The band gap of
samples is about 2.7 eV, but the employed excitation energy is 3.0 eV. Thus, an electron in
the valence band can be directly excited to move to the conduction band. It was also possible
to excite an electron that was deep inside the band gap [35]. These spectra show two distinct
emission bands, one in the visible spectrum at 701 nm and the other in the UV region at 350
nm. It was indicated what the preceding researches had found [36]. Put out the idea that the
PL spectrum of both the samples usually consists of two emission bands: one in the visible
spectrum (around 4000 nm to 700
2 nm) and4 the other6in the UV8 spectrum
10(roughly 350 nm to

400 nm). 10
140000

120000
8

100000

80000 6
Intensity (a.u)

60000
4

40000

20000 2

0
0

300 350 400 450 500 550 600 650 700 750 800
l (nm)

Figure 8. PL spectra of synthesized NCNPs

The near-band edge (NBE) emission peak of NCNPs at 350 nm is the result of free
exciton radiative recombination [37]. At 2.7 eV below the conductive band, the
recombination of electrons with firmly locked holes will produce a red band at 701 nm [37].

3.8. XPS Analysis

 The elemental makeup of the NCNPs was confirmed by XPS analysis. The survey XPS
spectrum, as depicted in Figure 9a, revealed various peaks corresponding to O, Ni, and Cr
elements, indicating their presence in the sample. From instrument reference the
characteristic peak was used at 285.6 eV and Carbon is used to calibrate the positions of all
the elements [38]. As seen in Figure 9b, Ni 2p3/2 and its satellite are responsible for the peaks
at 856.4 eV and 861.7 eV, whereas Ni 2p1/2 and its satellite are responsible for the peaks at
872.7 eV and 879.6 eV [39]. The Cr3+ in the NC structure is represented by two peaks in the
Cr2p XPS spectrum [Figure 9c] at 576.4 eV (Cr 2p3/2) and 585.8 eV (Cr2p) [40]. The XPS
O1s peak in Figure 9d is divided into two peaks. Lattice oxygen of NPs is well supported by
the first peak
0
at 530.2
2
eV,
4
while
6
the 8surface10 hydroxyl groups are responsible for the second
2 4 6 8

peak at 554 eV [41]. All these findings are fully in line with already reported literature.
700000
10
(a) O 1s (b) Ni 2p Original Peak
Ni 2p

600000 Fit Peak 1 Ni 2p3/2

Fit Peak 2 8
Fit Peak 3
8 Ni 2p1/2
Cr 2p

500000 Fit Peak 4

Fit Peak 5
Fit Peak 6
400000
Intensity (a.u)

Cumulative Fit
Intensity (a.u)

6
300000 6

200000 4
Ni 2p

100000
2 4
0 0 2 4 6 8 10
0 2 4 6 8 10

-100000
0
1400 1200 1000 800 600 400 200 0 885 880 875 870 865 860 855 850 845
Binding Energy (eV) 10 Binding Energy (eV)
(c) Cr 2p3/2 (d) 10
O 1s Lattice O
70000 Cr 2p
8

60000 Cr 2p1/2 8
Intensity (a.u)

6
Intensity (a.u)

50000
6

40000 4

4
30000
2

20000 2

0
10000
590 585 580 575 570 544 542 540 538 536 534 532 530 528 526 0

Binding Energy (eV) Binding Energy (eV)

Figure 10. (a) XPS spectra of NCNPs, (b) Ni 2p, (c) Cr2p & (d) O 1s, in the NCNPs

3.9.Antibacterial Activity

Bactericidal activity of NCNPs against selected strains was investigated via disc diffusion
testing method as depicted in Figure 10. The produced NCNPs were tested against several
bacterial strains for bactericidal activity, and the zone of inhibition (ZOI) was noted.
Ampicillin and the synthesized NPs were used to observe the antibacterial efficacy, as shown
in Table 5. The concentration of the NCNPs and the bacterial strains affected the bactericidal
performance profoundly. It was also noted that the increased ZOI acquired with increasing
NCNPs concentrations used in antibacterial strains. Minimum and maximum activity against
NCNPs was 5 and 9 mm, respectively. Staphylococcus aureus was protected with maximal
sensitivity against synthesized NCNPs, while E. coli had the lowest sensitivity amoung the
microorganisms. NCNPs prepared showed an effective and good antibacterial activity against
Salmonella sp., E. coli, S. aureus, and B. subtilis. Additionally, antimicrobial activity of
NCNPs may boost a number of industries, particularly the medical sector. An appealing
component of nanobiotechnology is figuring out how to work to kill germs.

Figure 10 Antibacterial activity of synthesized NCNPs against Gram-negative (a and b) and

Gram-positive (c and d) bacteria
Table 5. ZOI (mm) of synthesized NiCr2O4 NPs against selected pathogenic
strains
Test pathogens ZOI (mm) of NiCr2O4 NPs (ug/ml) Control
25 50 75 (ampicillin)

E. coli 5 6 8 10
Salmonella sp. 7 8 8 11
Staphylococcus aureus 7 8 9 13
Bacillus subtilis 7 7 8 13

Mechanism of antibacterial activity

The principal bacterial-lysis mechanism may be the action of Reactive Oxygenated Species
(ROS) generated in the presence of NCNPs [42]. The formation of hydroxyl radicals and
superoxide ions shows that the presence of NCNPs assists in the formation of oxygen species.
This results in lipid peroxidation, DNA smash up, and protein oxidation, all of which are
capable of killing bacteria without causing much harm to nonbacterial cells. Teichoic acid in
the peptidoglycan layer and lipoteichoic acid in the membrane create negative charges on the
cell surface [43]. Positive charges from NCNPs are drawn to the cell surface as a result of
electrostatic interactions, and the difference in electrostatic gradient causes cell wall damage
[44]. As demonstrated in Figure 11, the cell wall membrane (negative charge) and Ni2+
(positive charge) are mutually attracted and decrease DNA replication ability; protein
denaturation leads in bacterial death. Several studies have demonstrated photogeneration of
reactive oxygen species (ROS) on the surface of metal-oxide NPs, as suggested by Karthik et
al. [45].

Gram-negatives are more prone to NCNPs as their cellular wall is thinner than that of Gram-
positive bacteria [46]. Because small particles have more electrostatic interactions, this
process is linked to sample size [47]. Thus, the inhibitory action of NCNPs is subjective to
numerous factors like concentration, size, and duration of contact.
 ROS = Reactive Oxygen Species
ZnCr2O4 NPs NCs = Nanocomposites

ROS

ROS GENERATION × A

B C D E

CELL DEATH
F G

A. Electron Transport Interruption E. Enzyme Disruption

B. DNA Replication Inhibition F. Physical Cell attachment of NCs
C. Ribosome Dysfunction G. Cell Membrene Disruption
D. Protein Demage

Figure 11. Possible antibacterial activity mechanism of NiCr2O4 NPs

3.10. Anticancer Activity

In this study, HeLa and MCF-7 cell lines were used to test the in vitro cytotoxic effect of
nikel chromite NPs. The cytotoxic impact of the synthesized NCNPs at various doses was
investigated. Here, regular cell lines were employed as the standard. The acquired results are
displayed in Tables 6 and 7. The fact that the cytotoxicity is dose-dependent is highlighted by
a close examination of these results. Maximum cell viability was attained at 1000 g/ml and
minimal viability was at 7.8 g/ml, despite the fact that the synthesized nanosamples were
successful in killing all of the selected cancer cell lines.

Table 6 S.No. Concentration(µg/ml) Cell viability(%)

The synthetic NC NPs has
anticancer effects on 1 1000 16.17
HeLa cell lines. 2 500 32.35
3 250 42.64
4 125 54.41
5 62.5 63.23
6 31.2 72.05
7 15.6 76.47
8 7.8 80.88
9 Cell control 100

Table 7 S.No. Concentration(µg/ml) Cell viability(%)

Effect of synthetic NC
1 1000 14.03
NPs on MCF-7 cell lines
2 500 24.56
that inhibits cancer.
3 250 40.35
4 125 56.14
5 62.5 71.92
6 31.2 80.72
7 15.6 89.47
8 7.8 92.98
9 Cell control 100
For the HeLa and MCF-7 cell lines, the IC50 was calculated and discovered to be 170.04
μg/ml and 174.96 μg/ml, respectively (Figures 12 and 13). The morphological study was also
carried out using light microscopy to comprehend the method of carcinogenic cell death
which has been included in supplemantry data.
 Figure 12 : Anticancer activity of NCNPs synthesized against HeLa cell lines.

Figure 13: Anticancer activityof NCNPs synthesized against MCF-7 cell line.

Same results were reported by Islam et al. [48] and Donga et al., and Chung et al. for ZnO
NPs. [49]. the mechanism involved in the inhibition of cancer cell lines by NCNPs leads to
DNA damage, supporting the fact that increase in the concentration of nanoparticles
improves the degree of apoptosis [50], a programmed cell death process in HeLa and MPF-7
cancer cell lines. These results are supported by previous reports [51, 52]. Furthermore the
size/shape of NCNPs can directly contact the cell surfaces and initiate cytotoxicity [53].
Thus, the current research suggests that NCNPs might be employed in some forms of anti-
cancer therapy.
Anticancer activity mechanism

The mechanism of anticancer action of NCNPs, like the mechanism of antibacterial activity,
is complex and poorly explored. The NCNPs may be responsible for the generation of ROS
due to intra-cellular Ni2+ ions resulting in the malfunction of cellular redox machinery [54].
When Ni-chromite NPs come into contact with cell walls, they produce membrane
deformation and rupture, ROS generation on the surface of the NCNPs and particle
dissolution, and the subsequent release of free Ni2+ ions, which causes ROS development
within the cells. One of the possible reasons of NCNPs apoptosis is ROS generation and
oxidative stress [55]. Due to DNA damage and activation of the caspase cascade of apoptosis
and mitochondrial dysfunction, mitochondria may be the most effective intracellular
producers of ROS [56]. By disrupting cellular redox equilibrium, peroxidation of lipids in cell
membranes, and a cascade of antioxidant responses, reactive oxygen species in mitochondria
can influence seriously apoptosis and mitochondrial respiration [56].

4. Conclusion

Chromium based spinel NCNPs were successfully synthesized at a reasonable cost and in a
short amount of time via a facile sol-gel autocombustion method at 700oC. The formtion of
spinel chromite nanosamples was confirmed by XRD and FTIR investigations. The cubic
shape of the synthesized NPs was demonstrated through morphological analyses utilizing
FESEM and HR-TEM. The elemental and crystalline nature was confirmed by EDS spectra
and SAED pattern respectively. Using the UV- DRS technique, the optical behavior of
NCNPs was investigated, and the band gap was found to be as 2.7 eV. The luminescent
feature of the synthesized nanomaterial was shown by the strong emission band at 701 nm.
The bactericidal and anticarcinogenic abilities of NCNPs were also investigated. Among the
selected microbes, S. aureus exhibited the highest level of sensitivity (9 mm) against NCNPs,
while E. coli exhibited the lowest (5 mm). MTT model showed significant cytotoxic potential
of the NCNPs against HeLa and MCF-7 cells with the IC50 concentrations at 170.04 μg/ml
and 174.96 μg/ml respectively. The results revealed the effective performance of NCNPs against
all of the selected cancer cell lines by causing a decrease in cell-viability percentage in the
dose-dependent trend. Thus our work supports the fact that synthesized Ni-chromite NPs can
act as promising candidates for bactericidal agents and therapeutic agents for human cervical
and breast carcinoma treatment.

Approval for Ethics: None of the writers in this paper have ever done any research on either
humans or animals.

Participant Consent: All individual participants in the study provided informed consent.

Consent to Publish: All authors agree that the manuscript's copyright will be transferred to the
journal upon approval for publication.
Competing interests: The authors state that they do not have any competing interests.

References

[1] 1. Hu, Z. Q., Liu, X. Q., Huang, D. F., Gao, H., & Liu, J. X. (2012). Synthesis and
characterization of spinel-type NiCr2O4 nanocrystalline. Key Engineering
Materials, 512, 86-90.
[2] 2. Choudhary, P., & Varshney, D. (2018). Elucidation of structural, vibrational and
dielectric properties of transition metal (Co2+) doped spinel Mg-Zn
chromites. Journal of Magnetism and Magnetic Materials, 454, 274-288.
[3] 3. Josephine, B. A., Manikandan, A., Teresita, V. M., & Antony, S. A. (2016).
Fundamental study of LaMgxCr1−xO3−δ perovskites nano-photocatalysts: Sol-gel
synthesis, characterization and humidity sensing. Korean Journal of Chemical
Engineering, 33, 1590-1598.
[4] 4. Hemmati-Eslamlu, P., Habibi-Yangjeh, A., & Khataee, A. (2023). Anchoring
spinel NiCr2O4 nanoparticles on tubular g-C3N4: Efficacious pn heterojunction
photocatalysts for removal of tetracycline hydrochloride under visible light. Journal
of Alloys and Compounds, 932, 167571.
[5] 5. Dumitru, R., Manea, F., Păcurariu, C., Lupa, L., Pop, A., Cioablă, A., ... &
Ianculescu, A. (2018). Synthesis, characterization of nanosized ZnCr2O4 and its
photocatalytic performance in the degradation of humic acid from drinking
water. Catalysts, 8(5), 210.Dumitru, R., Manea, F., Păcurariu, C., Lupa, L., Pop, A.,
Cioablă, A., ... & Ianculescu, A. (2018). Synthesis, characterization of nanosized
ZnCr2O4 and its photocatalytic performance in the degradation of humic acid from
drinking water. Catalysts, 8(5), 210.
[6] 6. Zhang, D., Li, Q., Li, R., Li, H., Gao, H., Zhao, F., ... & Jiang, W. (2021).
Significantly enhanced thermal decomposition of mechanically activated ammonium
perchlorate coupling with nano copper chromite. ACS omega, 6(24), 16110-16118.
[7] 7. Patil, S. P., Sanadi, K. R., & Helavi, V. B. (2017). Impact of Ni substitution on
structural, electrical and thermoelectrical properties of zinc alluminium chromites
synthesized by sol-gel route and their photocatalytic investigation. Materials
Research, 20, 1445-1453.
[8] 8. Xie, K., Fang, J., Li, L., Deng, J., & Chen, F. (2022). Progress of graphite carbon
nitride with different dimensions in the photocatalytic degradation of dyes: A
review. Journal of Alloys and Compounds, 901, 163589.
[9] 9. Yazdanbakhsh, M., Khosravi, I., Goharshadi, E. K., & Youssefi, A. (2010).
Fabrication of nanospinel ZnCr2O4 using sol–gel method and its application on
removal of azo dye from aqueous solution. Journal of hazardous materials, 184(1-
 3), 684-689.
[10] 10. Noshahi, N. A., Nadeem, K., & Kamran, M. (2021). Role of Mn doping on
magnetic properties of multiferroic NiCr2O4 nanoparticles. Ceramics
International, 47(8), 10643-10649.
[11] 11. Vickers, A. (2004). Alternative cancer cures:“unproven” or “disproven”?. CA: A
Cancer Journal for Clinicians, 54(2), 110-118.
[12] 12. Li, X., Cui, R., Liu, W., Sun, L., Yu, B., Fan, Y., ... & Watari, F. (2013). The use
of nanoscaled fibers or tubes to improve biocompatibility and bioactivity of
biomedical materials. Journal of Nanomaterials, 2013, 14-14.
[13] 13. Aziz, N., Faraz, M., Sherwani, M. A., Fatma, T., & Prasad, R. (2019).
Illuminating the anticancerous efficacy of a new fungal chassis for silver
nanoparticle synthesis. Frontiers in chemistry, 7, 65.
[14] 14. Sun, Y., Du, Y., Lan, J., Zhan, W., & Zhang, T. C. (2021). A new method (ball
milling and sodium sulfide) for mechanochemical treatment of soda ash chromite
ore processing residue. Journal of Hazardous Materials, 415, 125601.
[15] 15. Zhang, D., Li, Q., Li, R., Li, H., Gao, H., Zhao, F., ... & Jiang, W. (2021).
Significantly enhanced thermal decomposition of mechanically activated
ammonium perchlorate coupling with nano copper chromite. ACS omega, 6(24),
16110-16118.
[16] 16. Kumar, D. R., Ahmad, S. I., Lincoln, C. A., & Ravinder, D. (2019). Structural,
optical, room-temperature and low-temperature magnetic properties of Mg–Zn
nanoferrite ceramics. Journal of Asian Ceramic Societies, 7(1), 53-68.
[17] 17. Patil, S. P., Sanadi, K. R., & Helavi, V. B. (2017). Impact of Ni substitution on
structural, electrical and thermoelectrical properties of zinc alluminium chromites
synthesized by sol-gel route and their photocatalytic investigation. Materials
Research, 20, 1445-1453.
[18] 18. Rashad, M. M., & El-Sheikh, S. M. (2011). Magnetic properties of nano-clusters
lanthanum chromite powders doped with samarium and strontium ions synthesized
via a novel combustion method. Materials Research Bulletin, 46(3), 469-477.
[19] 19. Yazdanbakhsh, M., Khosravi, I., Goharshadi, E. K., & Youssefi, A. (2010).
Fabrication of nanospinel ZnCr2O4 using sol–gel method and its application on
removal of azo dye from aqueous solution. Journal of hazardous materials, 184(1-
3), 684-689.
[20] 20. Nagajyothi, P. C., Deyarayapalli, K. C., Tettey, C. O., Vattikuti, S. P., & Shim,
J. (2019). Eco-friendly green synthesis: catalytic activity of nickel hydroxide
nanoparticles. Materials Research Express, 6(5), 055036.
[21] 21. Noshahi, N. A., Nadeem, K., & Kamran, M. (2021). Role of Mn doping on
magnetic properties of multiferroic NiCr2O4 nanoparticles. Ceramics
 International, 47(8), 10643-10649.
[22] 22. Saxena, P., Choudhary, P., Yadav, A., Rai, V. N., Varshney, M., & Mishra, A.
(2019). Effect of transition metal substitution on structural and dielectric properties
of Mg0.5Zn0.5−xCrxCo2O4 (0.0≤ x≤ 0.5) cobaltite. Journal of Materials Science:
Materials in Electronics, 30, 7292-7300.
[23] 23. Becheri, A., Dürr, M., Lo Nostro, P., & Baglioni, P. (2008). Synthesis and
characterization of zinc oxide nanoparticles: application to textiles as UV-
absorbers. Journal of Nanoparticle Research, 10, 679-689.
[24] 24. Kamran, M., Ullah, A., Rahman, S., Tahir, A., Nadeem, K., ur Rehman, M. A.,
& Hussain, S. (2017). Structural, magnetic, and dielectric properties of multiferroic
Co1−xMgxCr2O4 nanoparticles. Journal of Magnetism and Magnetic Materials, 433,
178-186.
[25] 25. Wani, T. A., & Suresh, G. (2021). A comprehensive review of LiMnPO4 based
cathode materials for lithium-ion batteries: current strategies to improve its
performance. Journal of Energy Storage, 44, 103307.
[26] 26. Donga, S., & Chanda, S. (2022). Caesalpinia crista seeds mediated green
synthesis of zinc oxide nanoparticles for antibacterial, antioxidant, and anticancer
activities. BioNanoScience, 12(2), 451-462
[27] 27. Wani, T. A., Suresh, G., Masrour, R., Batoo, K. M., & Rasool, A. (2023). A
structural, morphological, optical and magnetic study of nickel-substituted zinc (Ni–
Zn) ferrite nanoparticles synthesized via glycine assisted gel autocombustion
synthesis route. Materials Chemistry and Physics, 307, 128169.
[28] 28. Kamran, M., Ullah, A., Rahman, S., Tahir, A., Nadeem, K., ur Rehman, M. A.,
& Hussain, S. (2017). Structural, magnetic, and dielectric properties of multiferroic
Co1−xMgxCr2O4 nanoparticles. Journal of Magnetism and Magnetic Materials, 433,
178-186.
[29] 29. Callister Jr, W. D. (2007). Materials science and engineering an introduction.
[30] 30. Aly, S. A., Akl, A. A., & Mahmoud, D. H. (2015). Microstructural and electrical
characteristics of sprayed Tungsten oxide thin films. Int. J. New. Hor. Phys, 2(2),
47-52.
[31] 31. Govindarajan, D., Uma Shankar, V., & Gopalakrishnan, R. (2019).
Supercapacitor behavior and characterization of RGO anchored V2O5
nanorods. Journal of Materials Science: Materials in Electronics, 30, 16142-16155.
[32] 32. Kong, X. Y., Ng, B. J., Tan, K. H., Chen, X., Wang, H., Mohamed, A. R., &
Chai, S. P. (2018). Simultaneous generation of oxygen vacancies on ultrathin BiOBr
nanosheets during visible-light-driven CO2 photoreduction evoked superior activity
and long-term stability. Catalysis Today, 314, 20-27.
[33] 33. Othman, A. A., Ali, M. A., Ibrahim, E. M. M., & Osman, M. A. (2016).
Influence of Cu doping on structural, morphological, photoluminescence, and
electrical properties of ZnO nanostructures synthesized by ice-bath assisted
sonochemical method. Journal of Alloys and Compounds, 683, 399-411.
[34] 34. Periasamy, C., Prakash, R., & Chakrabarti, P. (2010). Effect of post annealing on
structural and optical properties of ZnO thin films deposited by vacuum coating
technique. Journal of Materials Science: Materials in Electronics, 21, 309-315.
[35] 35. Kumar, R. A., Hata, S., & Gopchandran, K. G. (2013). Diethylene glycol
mediated synthesis of Gd2O3: Eu3+ nanophosphor and its Judd–Ofelt
analysis. Ceramics International, 39(8), 9125-9136.
[36] 36. Samanta, P. K., & Chaudhuri, P. R. (2012). Understanding the transition levels
of photoluminescence of ZnO quantum dots under weak confinement. Journal of
Optics, 41(2), 75-80.
[37] 37. Bindu, P., & Thomas, S. (2014). Estimation of lattice strain in ZnO
nanoparticles: X-ray peak profile analysis. Journal of Theoretical and Applied
Physics, 8, 123-134.
[38] 38. Wang, J., Wang, G., Wei, X., Liu, G., & Li, J. (2018). ZnO nanoparticles
implanted in TiO2 macrochannels as an effective direct Z-scheme heterojunction
photocatalyst for degradation of RhB. Applied Surface Science, 456, 666-675.
[39] 39. Salari, H. (2019). Kinetics and mechanism of enhanced photocatalytic activity
under visible light irradiation using Cr2O3/Fe2O3 nanostructure derived from
bimetallic metal organic framework. Journal of Environmental Chemical
Engineering, 7(3), 103092.
[40] 40. Liang, X., Zhang, X. J., You, T. T., Wang, G. S., Yin, P. G., & Guo, L. (2016).
Controlled assembly of one-dimensional MoO3@ Au hybrid nanostructures as SERS
substrates for sensitive melamine detection. CrystEngComm, 18(40), 7805-7813.
[41] 41. Al-Nafiey, A., Kumar, A., Kumar, M., Addad, A., Sieber, B., Szunerits, S., ... &
Jain, S. L. (2017). Nickel oxide nanoparticles grafted on reduced graphene oxide
(rGO/NiO) as efficient photocatalyst for reduction of nitroaromatics under visible
light irradiation. Journal of Photochemistry and Photobiology A: Chemistry, 336,
198-207.
[42] 42. Mikhailova, E. O. (2020). Silver nanoparticles: Mechanism of action and
probable bio-application. Journal of functional biomaterials, 11(4), 84.
[43] 43. Mendes, C. R., Dilarri, G., Forsan, C. F., Sapata, V. D. M. R., Lopes, P. R. M.,
de Moraes, P. B., ... & Bidoia, E. D. (2022). Antibacterial action and target
mechanisms of zinc oxide nanoparticles against bacterial pathogens. Scientific
Reports, 12(1), 2658.
[44] 44. Yusof, N. A. A., Zain, N. M., & Pauzi, N. (2019). Synthesis of ZnO
nanoparticles with chitosan as stabilizing agent and their antibacterial properties
against Gram-positive and Gram-negative bacteria. International journal of
biological macromolecules, 124, 1132-1136.
[45] 45. Karthik, K., Madhukara Naik, M., Shashank, M., Vinuth, M., & Revathi, V.
(2019). Microwave-assisted ZrO2 nanoparticles and its photocatalytic and
antibacterial studies. Journal of Cluster Science, 30, 311-318.
[46] 46. Meikle, T. G., Dyett, B. P., Strachan, J. B., White, J., Drummond, C. J., & Conn,
C. E. (2020). Preparation, characterization, and antimicrobial activity of cubosome
encapsulated metal nanocrystals. ACS applied materials & interfaces, 12(6), 6944-
6954.
[47] 47. Sirelkhatim, A., Mahmud, S., Seeni, A., Kaus, N. H. M., Ann, L. C., Bakhori, S.
K. M., ... & Mohamad, D. (2015). Review on zinc oxide nanoparticles: antibacterial
activity and toxicity mechanism. Nano-micro letters, 7, 219-242.
[48] 48. Islam, F., Shohag, S., Uddin, M. J., Islam, M. R., Nafady, M. H., Akter, A., ... &
Cavalu, S. (2022). Exploring the journey of zinc oxide nanoparticles (ZnO-NPs)
toward biomedical applications. Materials, 15(6), 2160.
[49] 49. Donga, S., & Chanda, S. (2022). Caesalpinia crista seeds mediated green
synthesis of zinc oxide nanoparticles for antibacterial, antioxidant, and anticancer
activities. BioNanoScience, 12(2), 451-462.
[50] 50. Wahab, R., Siddiqui, M. A., Saquib, Q., Dwivedi, S., Ahmad, J., Musarrat, J., ...
& Shin, H. S. (2014). ZnO nanoparticles induced oxidative stress and apoptosis in
HepG2 and MCF-7 cancer cells and their antibacterial activity. Colloids and
surfaces B: Biointerfaces, 117, 267-276.
[51] 51. Shawkey, A. M., Rabeh, M. A., Abdulall, A. K., & Abdellatif, A. O. (2013).
Green nanotechnology: anticancer activity of silver nanoparticles using Citrullus
colocynthis aqueous extracts. Adv. Life Sci. Technol, 13, 60-70.
[52] 52. Parasuraman, P., Anju, V. T., Lal, S. S., Sharan, A., Busi, S., Kaviyarasu, K., ...
& Syed, A. (2019). Synthesis and antimicrobial photodynamic effect of methylene
blue conjugated carbon nanotubes on E. coli and S. aureus. Photochemical &
Photobiological Sciences, 18, 563-576.
[53] 53. Gliga, A. R., Skoglund, S., Odnevall Wallinder, I., Fadeel, B., & Karlsson, H. L.
(2014). Size-dependent cytotoxicity of silver nanoparticles in human lung cells: the
role of cellular uptake, agglomeration and Ag release. Particle and fibre
toxicology, 11, 1-17.
[54] 54. Kanagesan, S., Hashim, M., AB Aziz, S., Ismail, I., Tamilselvan, S., Alitheen, N.
B., ... & Purna Chandra Rao, B. (2016). Evaluation of antioxidant and cytotoxicity
activities of copper ferrite (CuFe2O4) and zinc ferrite (ZnFe2O4) nanoparticles
synthesized by sol-gel self-combustion method. Applied Sciences, 6(9), 184.
[55] 55. Zafar, A., Singh, S., Satija, Y. K., Saluja, D., & Naseem, I. (2018). Deciphering
the molecular mechanism underlying anticancer activity of coumestrol in triple-
negative breast cancer cells. Toxicology in Vitro, 46, 19-28.
[56] 56. Karthik, K., Shashank, M., Revathi, V., & Tatarchuk, T. (2019). Facile
microwave-assisted green synthesis of NiO nanoparticles from Andrographis
paniculata leaf extract and evaluation of their photocatalytic and anticancer
activities. Molecular crystals and liquid crystals.
ToC Graphical Abstract Information Click here to access/download;ToC Graphical Abstract
Information;TOC NiCr2O4.docx

TOC

Spinel type NiCr2O4 nanoparticles (NCNPs) have been

successfully prepared by facile solgel auto-combustion route to
study their antibacterial and anticancer activities. After
characterizations, it was found that the synthesized NCNPs were
effective against selected microorganisms. S. aureus exhibited
the highest level of sensitivity (9 mm), while E. coli had the
lowest (5 mm). Moreover, the MTT test revealed the high
cytotoxicity of NCNPs against HeLa and MCF-7 cell lines.
These results leads us to the fact that NCNPs can be considered
as prospective candidates for bactericidal and anti-carcinogenic
agents.
Supplementary Data

Supplementary Data

Figure 19: Light microscopic image of HeLa cell line treated with prepared NiCr2O4 NPs. a. Normal
HeLa cell line b.toxicity,1000µg/ml; c. toxicity,250µg/ml; d.toxicity,62.5µg/ml; and e.
toxicity,31.2µg/ml
Figure 20: Light microscopic image of MCF-7 cell line treated with prepared NiCr2O4 NPs. a. Normal
HeLa cell line b. toxicity,1000µg/ml; c. toxicity,250µg/ml; d. toxicity,62.5µg/ml; and e.
toxicity,31.2µg/ml