Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність
Medicinal plants can be used as reducing agents in the preparation of metal nanoparticles by green synthesis because of the chemotherapeutic and anti-infectious properties of natural compounds. Therefore, this paper reports the green synthesis of silver and iron nanoparticles from leaf and flower ex...
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| Дата: | 2021 |
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2021
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Plant Introduction| _version_ | 1860145113537183744 |
|---|---|
| author | Shawuti, Shalima Bairam, Chasan Beyatlı, Ahmet Kariper, İshak Afşin Korkut, Isık Neslişah Aktaş, Zerrin Öncül, Mustafa Oral Kuruca, Serap Erdem |
| author_facet | Shawuti, Shalima Bairam, Chasan Beyatlı, Ahmet Kariper, İshak Afşin Korkut, Isık Neslişah Aktaş, Zerrin Öncül, Mustafa Oral Kuruca, Serap Erdem |
| author_sort | Shawuti, Shalima |
| baseUrl_str | https://www.plantintroduction.org/index.php/pi/oai |
| collection | OJS |
| datestamp_date | 2023-08-26T20:39:08Z |
| description | Medicinal plants can be used as reducing agents in the preparation of metal nanoparticles by green synthesis because of the chemotherapeutic and anti-infectious properties of natural compounds. Therefore, this paper reports the green synthesis of silver and iron nanoparticles from leaf and flower extracts of Nerium oleander and their capacity as anticancer and antimicrobial agents. Nanoparticle manufacturing and structural characterization of silver and iron nanoparticles are reported. The formation of nanoparticles is characterized by scanning electron microscopy with energy dispersive X-ray spectroscopy, UV-Vis and Fourier transform infrared (FTIR) spectroscopy. Nanoparticles formation was also investigated the surface charge, particle size, and distribution using zeta sizer analysis by DLS. Green synthesis of silver and iron nanoparticles using N. oleander showed different levels of selective cytotoxicity against K562 (human chronic myeloid leukemia cells) in low concentrations and were not cytotoxic to the HUVEC (human umbilical vein endothelial cells) in the same concentrations. Silver nanoparticles showed antibacterial activity against multidrug pathogens, while iron nanoparticles failed to show such activity. Results of the present research demonstrate the potential use of green synthesized nanoparticles in various biomedicine and pharmaceuticals fields in the future. |
| doi_str_mv | 10.46341/PI2021010 |
| first_indexed | 2025-07-17T12:53:58Z |
| format | Article |
| fulltext |
© The Authors. This content is provided under CC BY 4.0 license.
Plant Introduction, 91/92, 36–49 (2021)
RESEARCH ARTICLE
Green synthesis and characterization of silver and iron nanoparticles
using Nerium oleander extracts and their antibacterial and anticancer
activities
Shalima Shawuti 1, Chasan Bairam 2, Ahmet Beyatlı 3, İshak Afşin Kariper 4, Isık Neslişah
Korkut 1, Zerrin Aktaş 5, Mustafa Oral Öncül 6, Serap Erdem Kuruca 1
1 Department of Physiology, Faculty of Medicine, Istanbul University, Turgut Özal Millet str., 34093 Istanbul, Turkey
2 Renewable Energy and Oxide Hybrid Systems Laboratory, Department of Physic, Faculty of Science, Istanbul University,
Şehzadebaşı str., 34134 Istanbul, Turkey
3 Department of Medicinal and Aromatic Plants, Hamidiye Vocational School of Health Services, University of Health Sciences,
Tıbbiye str. 38, 34668 Istanbul, Turkey
4 Faculty of Education, Erciyes University, Yenidoğan Mahallesi, Turhan Baytop Sokak str. 1, 38280 Kayseri, Turkey
5 Department of Microbiology, Faculty of Medicine, Istanbul University, Turgut Özal Millet str., 34093 Istanbul, Turkey
6 Department of Infectious Diseases, Faculty of Medicine, Istanbul University, Turgut Özal Millet str., 34093 Istanbul, Turkey
Received: 19.08.2021 | Accepted: 21.11.2021 | Published online: 28.11.2021
Abstract
Medicinal plants can be used as reducing agents in the preparation of metal nanoparticles by green synthesis
because of the chemotherapeutic and anti-infectious properties of natural compounds. Therefore, this
paper reports the green synthesis of silver and iron nanoparticles from leaf and flower extracts of Nerium
oleander and their capacity as anticancer and antimicrobial agents. Nanoparticle manufacturing and
structural characterization of silver and iron nanoparticles are reported. The formation of nanoparticles
is characterized by scanning electron microscopy with energy dispersive X-ray spectroscopy, UV-Vis and
Fourier transform infrared (FTIR) spectroscopy. Nanoparticles formation was also investigated the surface
charge, particle size, and distribution using zeta sizer analysis by DLS. Green synthesis of silver and iron
nanoparticles using N. oleander showed different levels of selective cytotoxicity against K562 (human
chronic myeloid leukemia cells) in low concentrations and were not cytotoxic to the HUVEC (human
umbilical vein endothelial cells) in the same concentrations. Silver nanoparticles showed antibacterial
activity against multidrug pathogens, while iron nanoparticles failed to show such activity. Results of the
present research demonstrate the potential use of green synthesized nanoparticles in various biomedicine
and pharmaceuticals fields in the future.
Keywords: Nerium oleander, green synthesis, Ag-nanoparticles, Fe-nanoparticles, cellular cytotoxicity, antibacterial effect
https://doi.org/10.46341/PI2021010
UDC 58.04 : 615.322
Authors’ contributions: Shalima Shawuti is principle investigator. Chasan Bairam is master student who carried out XRD and UV-Vis
analysis. Ahmet Beyatlı is researcher collect and prepare leafs extract; also did phytochemical screening test, İshak Afşin Kariper is
researcher who perform SEM and DLS analysis. Isık Neslişah Korkut is master student who investigate the cytotoxic test, Zerrin Aktaş
and Mustafa Oral Öncül are collaborators who perform antibacterial tests and write evaluation on those part. Finally, Serap Erdem
Kuruca is co-principle investigator and supervisor.
Funding: This work was also supported by the Scientific Research Projects Coordination Unit of Istanbul University.
Competing Interests: We wish to confirm that there are no known conflicts of interest with this publication and there has been no
significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and
approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. We
further confirm that the order of authors listed in the manuscript has been approved by all of us.
https://creativecommons.org/licenses/by/4.0/
https://orcid.org/0000-0002-9118-8680
https://orcid.org/0000-0002-8676-6597
https://orcid.org/0000-0001-5225-6217
https://orcid.org/0000-0001-9127-301X
https://orcid.org/0000-0001-8550-5400
https://orcid.org/0000-0002-5998-0440
https://orcid.org/0000-0002-1681-1866
https://orcid.org/0000-0001-7878-9994
Plant Introduction • 91/92 37
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Introduction
Nanoparticles (NPs) are synthesized generally
by expensive chemical synthesis methods
that require the use of toxic chemicals. Thus,
using biomolecules (i.e., bacteria, fungi, or
plants) for NPs synthesis became a common
method in past years that is safe, low-cost,
and ecofriendly. Different plant extracts can
be act as safe natural capping, reducing, and
stabilizing agents without being a source of
thermal or chemical hazards (Fedlheim & Foss,
2001; Arya, 2010). Nanoparticles formation
can be a glimmer of hope for the production
of drugs that can be used against infectious
diseases and cancer.
Nerium oleander L. (Apocynaceae), grown in
wetlands of the Mediterranean region, can be
seen wild as well cultivated as an ornamental
plant in parks and gardens. It is an evergreen
shrub with pink and white flowers (Baytop,
1999). The plant grows up to 2–6 m tall. The
leaves are in pairs or whorls of three, thick
and leathery, dark-green, narrowly lanceolate,
5–21 cm long, and with an entire margin.
Flowers develop in clusters at the end of
branches, the diameter of each flower range
about 2.5–5 cm with a deeply 5-lobed fringed
corolla around the central corolla tube. The
fruit is a long narrow capsule 5–23 cm long,
which splits open at maturity to release
numerous downy seeds (Baytop, 1999; Kiran &
Prasad, 2014).
The leaves and flowers of N. oleander used
in folkloric medicine among people in Turkey
for rheumatism and urticaria (Bulut & Tuzlaci,
2013; Sağıroğlu et al., 2013). The latex of plant
used for eczema (Gürdal & Kültür, 2013). Over
the world, different parts of this plant are
used traditionally for the treatment of various
human ailments, including dermatitis, eczema,
herpes, skin cancer, asthma, epilepsy, malaria,
and tumors (Santhi, 2011). Nerium oleander is
considered one of the most poisonous plants
in the world which leads annually to the
death of many people and animals (Rubini
et al., 2019). This toxicity is due to toxins like
oleandrin, oleandrigenin, and nerine, which
belong to cardiac glycosides (Al-Badrani et al.,
2008; Zibbu & Batra, 2010). In addition, the
plant contains terpenoids and steroids (Santhi,
2011).
Despite the toxicity of plant, different
scientific studies conducted on various parts of
N. oleander showed its antibacterial (Chauhan
et al., 2017), hepatoprotective and antioxidant
(Singhal & Gupta, 2012), antiproliferative
(Wong et al., 2011), antidiabetic (Sikarwar et al.,
2009), anti-inflammatory (Erdemoglu et al.,
2003), and anticancer (Pathak et al., 2000;
Turan et al., 2006) activities. This study was
aimed to synthesize Ag and Fe nanoparticles
using leaves and flowers of N. oleander and
then evaluate its antibacterial and anticancer
activity against human chronic myeloid
leukemia cell line.
Material and methods
Chemicals and reagents
An anhydrous FeCl3 with 98 % purity (Merck,
Germany), AgNO3 with 99.5 % purity (Sigma-
Aldrich, USA) were used as metal sources.
Dulbecco’s modified Eagle medium (DMEM)
and fetal bovine serum (FBS) (Gibco, UK)
and MTT (3-[4,5-dimethylthiazol-2-yl]-2,4-
diphenyltetrazolium bromide) were applied.
All reagents used were of analytical grades.
Plant material
Nerium oleander leaves and flowers were
collected from Servetiye Village, Sakarya
Province, Turkey in June 2020. Plant was
identified at the Herbarium of Faculty of
Pharmacy, Istanbul University (voucher
number – ISTE-117270).
Preparation of extracts
Collected dried leaves and flowers of
N. oleander were washed thoroughly (three
times) in distilled water and homogenized
using a mortar and pestle. The shade dried
leaves and flowers of N. oleander were
powdered and then 10 g of both leaves and
flowers were suspended in 100 ml of distilled
water. Mixtures stirred for 20 min at 60 °C,
then allowed to cool at room temperature,
and then filtered using a Whatman no. 42
filter paper and centrifuged at × 2000 rpm
for 20 min (Byrne et al., 2016). The extracts
prepared were then transferred to a sterile
container. The extracts were stored at 4 °C and
freshly used.
Preliminary phytochemicals screening
Nerium oleander extracts were subjected
to qualitative screening for the presence
38 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
of various phytochemicals using standard
procedures (Tyler, 1993; Harborne, 1998).
Preparation of Nerium oleander silver
nanoparticles (NO-AgNPs)
2.5 mL from the leaf (L) and flower (F) extract
solutions were taken and mixed with 1 mM
AgNO3 in 47.5 mL deionized water and a
solution of 50 mL in amount was obtained. The
pH values of crude leaf and flower extracts, pH
values of samples just after mixing with AgNO3
metal source and after 24 h were measured
(Table 1). The pH of the prepared AgNO3
solution was 5.28.
Preparation of Nerium oleander iron
nanoparticles (NO-FeNPs)
Same sample preparation steps were followed
for iron nanoparticles. Briefly, 5 mL from the
leaf and flower extract solutions was taken
and mixed with 0.2 M FeCl3 in 45 mL deionized
water and a solution of 50 mL in amount was
obtained. The pH values of crude leaf and
flower extracts, pH values of samples just
after mixing with FeCl3 and after 24 h were
measured (Table 1). The pH of the prepared
FeCl3 solution was 2.25.
Characterization of Nerium oleander AgNPs
and FeNPs
The synthesized nanoparticles were
characterized through a UV-Vis
spectrophotometer Shimadzu 2600. The
reduction of nanoparticles was monitored by
UV-spectrophotometer range of absorbance
from 250–480 nm. The crystalline structures
of the green-synthesized N. oleander (AgNP)
and (FeNP) were examined by XRD Rigaku
Flex 600 (600 models, with λ = 1.5406 and
with a step size of 0.02 Å) at speed of 3 °
min-1. Particle size and zeta potential were
measured by Malvern Nano ZS. Morphology
and elemental metal mapping were recorded
using a high-resolution scanning electron
microscope (SEM, Carl Zeiss Ultra Plus
Gemini Fesem) were used to investigate 2D
surface morphologies. The composition
analyses of the samples were performed by
EDX (EDX spectrometer attached to SEM).
Fourier transformed infrared (FTIR) analyses
were carried out on a liquid sample with
Bruker Alpha FTIR spectrometer in the range
from 4000–500 cm-1. The device had a DTGS
detector and ten scans were conducted for
each spectrum with resolution four.
Cytotoxic assay
K562 (human chronic myeloid leukemia
cells) and HUVEC (human umbilical vein
endothelial cells) cell lines were obtained
from American Type Culture Collection
(ATCC). Cells were cultured in DMEM with
10 % FBS and 1 % penicillin/streptomycin in
a 5 % CO2 humidified incubator, maintained
at 37 °C. First, N. oleander nanoparticles were
sterilized and diluted with DMEM to prepare
four different dilutions which are 1, 1/2, 1/5,
and 1/10. MTT assays were performed in
96-well plates. The plant extract and metal
concentrations found in these nanoparticle
dilutions are also shown in Table 2. K562
cells (about 105 cells per well) were seeded
and incubated for 72 h. Then, supernatants
were removed, and 10 µL (MTT – 5 mg/mL)
solution was added to each well. Following
incubation at 37 °C for 3.5 h and kept dark in
a humidified atmosphere at 5 % CO2 in the air.
Subsequently, the supernatant was discarded,
and the precipitated formazan was dissolved
in dimethyl sulfoxide (100 µL per well). The
optical density of the solution was evaluated
using a microplate spectrophotometer at
a wavelength of 570 nm (Mosmann, 1983).
GraphPad Prism was used to calculate cell
viability and IC50 values.
Formation Extract pH Extract+Metal NO3 (0h) Extract+metal NO3 (24h)
NOL-AgNPs 6.78 6.56 4.21
NOF-AgNPs 5.42 6.12 3.74
NOL-FeNPs 6.78 2.32 2.17
NOF-FeNPs 5.42 2.33 2.25
Table 1. pH values of the NO-AgNPs and NO-FeNPs. The pH values were measured on leaf extract, after
mixing with a metal salt, and after 24 hours of mixing.
Plant Introduction • 91/92 39
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Antibacterial activity
The antibacterial potential was tested
against 14 different human pathogenic
bacteria including three Gram-negative
bacteria (Escherichia coli ATCC 35218, clinical
isolates of carbapenem-resistant Klebsiella
pneumoniae (CRKpn) and carbapenem-
resistant E. coli (CREc)) and 11 Gram-positive
bacteria (Staphylococcus aureus ATCC 29213
and ATCC 25923, inducible-clindamycin-
resistant S. aureus (ICRSa) BAA976-1, hetero-
resistant S. aureus (hVISA), clinical isolates
of methicillin-resistant S. aureus (MRSA),
methicillin-resistant coagulase-negative
S. aureus (MR-CoNS:2), vancomycin-resistant
Enterococcus faecium (VREf), E. faecalis ATCC
29212 and 51279, and vancomycin susceptible
E. faecalis (VSEf)). The species were identified
by using the Vitek 2 system (bioMerieux
Vitek Inc.).
Antibacterial activity was detected by
minimum inhibition concentrations (MICs),
which were determined by serial microdilution
method (MIC ranges – 2.5–0.0012 mg/L
for NOL-AgNPs and NOF-AgNPs, and 5.0–
0.0024 mM for AgNO3) following CLSI (2018).
Briefly, 100 μL of each concentration were
added to a well (96-well microplate) containing
100 μL of Mueller Hinton Broth (MHB) and
10 μL of inoculum (0.5 McFarland; 1.5 × 108
colony forming units/mL). Plates were then
incubated at 37 °C for 24 h. Bacterial growth
was determined by absorbance at 600 nm.
List of applied abbreviations
ATCC – American Type Culture Collection
CREc – carbapenem-resistant Escherichia coli
CRKpn – carbapenem-resistant Klebsiella
pneumoniae
DLS – dynamic light scattering
DMEM – Dulbecco’s modified Eagle medium
FBS – fetal bovine serum
FTIR – Fourier transform infrared
HUVEC – human umbilical vein endothelial
cells
hVISA – heteroresistant Staphylococcus aureus
IC50 – half maximal inhibitory concentration
ICRSa – inducible clindamycin-resistant
S. aureus
K562 – human chronic myeloid leukemia
cells
MIC – minimum inhibition concentrations
MR-CoNS:2 – methicillin-resistant coagulase-
negative S. aureus
MRSA – methicillin-resistant S. aureus
NO – Nerium oleander
NOF – N. oleander flowers extract
NOL – N. oleander leaves extract
NPs – nanoparticles
SEM – scanning electron microscopy
UV-Vis – Uv-Vis spectroscopy
VREf – vancomycin-resistant Enterococcus
faecalis
VSEf – vancomycin-susceptible E. faecalis
XRD – X-ray spectroscopy
Results and discussion
Preliminary phytochemicals screening
Qualitative phytochemical analysis of
N. oleander extracts showed the presence
of different active components in the
aqueous extracts (Table 3). Previous works
on N. oleander revealed the presence of
alkaloids, flavonoids, glycosides, tannins,
terpenoids, and saponins in this plant
Dilutions NOF-Ag NOL-Ag NOF-Fe NOL-Fe
1 500 ug/mL 500 ug/mL 1000 ug/mL 1000 ug/mL
20 uM/mL 20 uM/mL 1 mM/mL 1 mM/mL
1/2 250 ug/mL 250 ug/mL 500 ug/mL 500 ug/mL
10 uM/mL 10 uM/mL 0.5 mM/mL 0.5 mM/mL
1/5 100 ug/mL 100 ug/mL 200 ug/mL 200 ug/mL
4 uM/mL 4 uM/mL 0.02 mM/mL 0.02 mM/mL
1/10 50 ug/mL 50 ug/mL 100 ug/mL 100 ug/mL
2 uM/mL 2 uM/mL 0.01 mM/mL 0.01 mM/mL
Table 2. The nanoparticle dilutions used in cytotoxicity tests.
40 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
(Chaudhary et al., 2015; Saranya et al., 2017).
The existence of these constituents can be
the main reason behind the biological activity.
Synthesis of Fe/Ag NPs by visual inspection
After 24 h of reaction, the reaction solution
color changed from light to dark color, which
can be seen in Fig. 1. The reduction of Fe3+
ions exhibits a dark color due to the excitation
of surface plasmon vibration in a metal
nanoparticle. Similarly, in the reduction of
Ag+ ions, the solution color change from light
pink to light yellow. Visual photo images of the
NOF-NPs are not reported due to similarity in
colors with NOL-NPs.
SEM and DLS measurement
The scanning electron microscopy (SEM)
technique was used to evaluate the morphology
and size of the green synthesized NOL-AgNPs.
Fig. 2 represents the surface images and
DLS size distribution of green synthesized
nanoparticles (NO-AgNPs). Specifically, the
nanoparticles appear aggregated and spread
uniform shapes. Iron nanoparticles seem
also spherical with 70 nm average diameters.
In another study, silver nanoparticles from
N. oleander flowers were synthesized (Bharathi
& Shanthi, 2017). Silver particles of about 10 μm
by SEM analysis are very small compared to
ours, but we do not know their effectiveness
as authors did not repot their bioactivity.
Besides, such small particles are not suitable
for clinical use as they will be much easier to
eliminate by the immune system (Bharathi &
Shanthi, 2017).
The zeta potential is an indicator of
surface charge potential, which is an
important parameter for understanding
the stability of nanoparticles in aqueous
suspensions. Table 4 summarizes DLS size
distribution measurements carried out on
green synthesized NPs. For the NOF-FeNPs,
the average particle size was 1872 nm with a
polydispersity index of 0.69 (zeta potential
– +5.3 mV). On the other hand, particle
sizes of NOF-AgNPs were found 76 nm size
with relatively homogenous distribution
(polydispersity index – 0.266, zeta potential
– +8.1 mV). Secondly, For the NOL-FeNPs,
the average particle size was 609 nm with a
polydispersity index of 0.54 (zeta potential
– +7.4 mV). On the other hand, particle
sizes of NOL-AgNPs were found 93 nm size
with relatively homogenous distribution
(polydispersity index – 0.364, zeta potential
– +8.8 mV). No aggregation of the colloidal
was observed for several months. Therefore,
it may suggest that all synthesized NOF-NPs
and NOL-NPs were highly stable when stored
at the required temperature. It was also
observed that produced NPs had positively
charged on their surface with zeta potential
values above 5 mV.
UV spectroscopy
The characterization of silver and iron
nanoparticles by UV-spectrophotometer from
the range of 350–900 nm was performed to
monitor the reduction of metal ions and their
Phytochemicals Leaves Flowers
Alkaloids + +
Flavonoids – +
Saponins + +
Terpenoids – –
Steroids – –
Tannins + +
Glycosides + +
Table 3. Preliminary phytochemical screening of
leaves and flowers of Nerium oleander extract (plus
and minus indicate the presence and absence of
the chemicals, respectively).
Figure 1. Synthesis of Ag and Fe nanoparticles
exhibit color change. First row – solutions fabricated
with leaf extracts (1 – NOL-Ag; 2 – NOL-Ag after 24 h;
3 – crude NOL extract; 4 – NOL-Fe; 5 – NOL-Fe after
24 h). Second row – solutions fabricated with flower
extracts (6 – NOF-Ag; 7 NOF-Ag after 24 h; 8 – crude
NOF extract; 9 – NOF-Fe; 10 – NOF-Fe after 24 h).
Plant Introduction • 91/92 41
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Figure 2. SEM image and DLS size distribution of Nerium oleander related NPs. The SEM images were
obtained on dried powder samples.
42 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
stability. The broad absorption peaks in the
range from 326 to 432 nm were represented
in Fig. 3. UV-Vis spectra were performed for
NO-AgNPs and NO-FeNPs fabricated with leaf
and flower extracts. To observe any shift, both
crude NOL and NOF were also investigated.
The absorption peaks of a plant extract with an
organic mixture were not able to record since
they are belonging to C-C and C-H electronic
transitions (below 250 nm wavelength). The
absorption peaks for NOF at 326 and 384 nm
wavelengths belong to the n-π* and π-π*
transitions (Wang et al., 2014). It is because
this plant extract contains carbon-carbon
double bonds, nitrogen-oxygen bonds, or
cyclic aromatic structures. This absorption
peak was only seen at 366 nm for NOL. While
Ag nanoparticle synthesized with NOF gives
an absorption peak at 432 nm as expected.
The synthesized Fe NP continued to interact
with functional groups in the plant extract,
and while a specific 435 nm absorption peak
was observed in the region belonging to a
typical metal nanoparticle, an absorption
peak appeared at 377 nm due to its interaction
with functional groups. Another possible
explanation for this phenomenon is that
the Fe nanoparticle can be in Fe2O3 or Fe3O4
structure types (Wang et al., 2014). UV-Vis
spectra with the NOL nanoparticles showed
much clearer absorption peaks of metal
nanoparticles compared to a case in NOF.
The Ag NPs absorption peak was observed at
437 nm whereas the Fe NPs was detected at
408 nm.
XRD analysis
The crystalline structures of the green-
synthesized NOL-FeNPs, NOF-FeNPs NOF-
AgNPs, and NOF-AgNPs were furtherly
examined by XRD analysis. The obtained
patterns were demonstrated in Fig. 4 with
labeled indices together with two theta values.
The obtained diffraction peaks at 2θ values
of 23.5 °, 26.6 °, 35.8 °, 39.1 °, and 46.2 ° were
assigned to (012), (120), (110), (113) and (202)
lattice planes, respectively. Those sets of lattice
planes were identical to those reported for
standard iron metal. For NOL-FeNPs, the iron
nanoparticles are FCC and crystalline (cubic
crystalline structure, a = 4.07100 Å; JCPDS files
no. 84-0713 and 04-0783). NO-AgNPs are FCC
Formation Average diameter (nm) Polydispersity index Zeta potential (mV ± SD)
NOF-FeNPs 130.0 0.690 +5.3 ± 9.3
NOF-AgNPs 76.1 0.266 +8.1 ± 0.3
NOL-FeNPs 39.1 0.251 +7.4 ± 0.4
NOL-AgNPs 92.9 0.364 +8.8 ± 0.8
Table 4. The DLS, polydispersity index, and zeta potential of NO-NPs.
Figure 3. UV-Vis of NOL and NOF extracts and NO-FeNPs and NO-AgNPs.
Plant Introduction • 91/92 43
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Figure 4. XRD graphs of NOL-Fe, NOF-Fe, NOL-Ag, and NOF-Ag.
and crystalline (cubic crystalline structure,
a = 4.07100 Å; JCPDS files no. 84-0713 and 04-
0783). The obtained diffraction peaks at 2θ
values of 26.50 °, 36.78 °, and 38.40 ° were
assigned to (220), (111), and (111) lattice planes,
respectively. Those sets of lattice planes were
identical to those reported for standard silver
metal (JCPDS files no. 84-0713 and 04-0783).
FTIR analysis
FTIR spectra of NOF and NOF-NPs are
presented in Fig. 5. The absorption band
at 3300 cm-1 was mainly attributed to
OH vibration. The absorption peaks were
assigned to the stretching vibration of C=C
(1645 cm-1). Compared to NOF extract’s FTIR,
the disappearance of the most functional
group is due to the successful reduction
of metal ions. Three main bands were
demonstrated in the FTIR spectrum of both
NOF-FeNPs and NOF-AgNPs The presence of
OH bonds and C=O functional groups on the
NOF-AgNPs and NOF-AgNPs were presented
at 3244 cm-1 and 1633 cm-1, respectively. It was
reported in the literature that FeNPs exhibit
a characteristic stretching Fe-O vibration
peak at 576 cm-1 (Wang et al., 2014). For NOF-
AgNPs, stretching vibrations at 631 cm-1 can
also be attributed to the reduction of Ag+ to
Ag. In similar green synthesis studies also
reported observation of reduction of Ag+ to
Ag peak at around 538 cm-1 (Erdogan et al.,
2019). The FTIR spectrums of NOL-Ag and
NOL-Fe are similar to the FTIR spectrums of
NOF-Ag and NOF-Fe. Therefore, there was
no need to reinterpret the NOL-Ag and NOL-
Fe spectrums.
Cytotoxicity assay
NOL-AgNPs are effective in the K562 cancer
cell line (IC50 – 2.3 uM). However, NOF-AgNPs
are variable (IC50 – 10 uM). Similarly, NOF-
44 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
FeNPs (IC50 – 7 uM) are more variable than
the NOL-FeNPs (IC50 – 48 uM). Our results
suggested that NOL-Ag, NOF-Ag, NOF-Fe,
and NOL-Fe are effective on the K562 cell
line in low concentrations. Furthermore, we
may conclude that NOF-Ag and NOF-Fe NPs
have a cytotoxic effect on the K562 cell line in
similar concentrations. However, NOL-Fe was
cytotoxic at concentrations approximately 20
times higher than NOL-Ag (Fig. 6).
In other studies, N. oleander conjugated
gold nanoparticles were synthesized to
investigate in vitro anticancer activity
on MCF-7 cell lines. IC50 values of these
nanoparticles were found between 74.04 and
130.87 μg/mL. These values are much higher
than ours (Barai et al., 2018).
HUVECs were used in this study as a
control. Ag and Fe NPs were not effective
on HUVEC cells at the same concentrations.
NOL-Ag (IC50 – 100 uM), NOF-Ag (IC50 –
100 uM), NOF-Fe (IC50 – 390 uM), and NOL-Fe
(IC50 – 430 uM). The concentrations of Ag and
Fe NPs, which are cytotoxic on HUVEC cells,
are more than ten-fold higher compared
to K562 cells. These results show that
Figure 5. FTIR of NOL-Ag, NOL-Fe, NOF-Ag, NOF-Fe,
and crude NOL and NOF extracts.
nanoparticles are harmless to normal cells
when used at low doses, which are cytotoxic
to leukemia cells (Fig. 6).
Antibacterial activity
Based on the results in Table 4, the tested
bacteria were able to be killed at a low
concentration of AgNO3 (< 0.00976 mM). The
green-synthesized NOL-Ag and NOF-Ag were
able to inhibit bacteria including multidrug
pathogens. As showed in Table 5, the MIC
(mg/mL | mg/mM) values of NOL-Ag and
NOF-Ag against Gram-negative bacteria were
ranged from 0.019 | 0.039 to 0.3125 | 0.625
and 0.078 | 0.156, respectively. While the
MIC (mg/mL | mg/mM) values of NOL-Ag
and NOF-Ag against Gram-positive bacteria
ranged from 0.078 | 0.156 to 0.3125 | 0.625 and
0.078 | 0.156 to 0.625 | 1.25, respectively. There
is no significant difference observed between
Gram-negative and Gram-positive bacteria
including multi-resistant bacteria. The result
of Fe-NPs was not given because the results
were not effective.
Plant-derived essential oils and extracts
have an antimicrobial effect with low
toxicity and can be recommended as
potential natural preservatives. According
to Ríos & Recio (2005), extracts can be
classified as significant (MIC < 100 mg/L),
moderate (100 < MIC ≤ 512 mg/L), or
weak (MIC > 512 mg/L) depending on
their respective activities against the
corresponding pathogens. An important
advantage for the used metallic ions is that
silver ions have relatively low toxicity to
human cells while adversely affecting bacteria
and fungi by different mechanisms, including
binding to the thiol groups of protein and
denaturing them, programmed cell death
(apoptosis), and causing the DNA to be in the
condensed form (Lansdown, 2006; Mohamed
et al., 2020).
Several studies documented the green
synthesis of AgNPs using plant extracts.
Also, antimicrobial effects of AgNPs against
multidrug-resistant bacteria including
E. coli, P. aeruginosa, and MRSA have been
studied by many researchers (Rai et al., 2012;
Paredes et al., 2014; Malik et al., 2015; Kar
et al., 2016; Chauhan et al., 2017; Nagababu &
Rao, 2017).
Plant Introduction • 91/92 45
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Figure 6. Cytotoxic activity of synthesized NOF-Ag, NOL-Ag, and NOF-Fe, NOL-Fe NPs against K562 and
HUVEC cell lines.
Conclusions
Synthesized NPs have been successfully
implemented in the fields of medicine
and environmental remediation. The
green synthesis of silver NPs was not only
demonstrated by visual inspection and but also
by performing systematic spectral techniques
(UV-Vis absorption, FTIR spectroscopy,
and SEM analysis). FTIR results proved that
bioactive compounds responsible for silver
bio-reduction could be proteins and flavonoids
presumed to act as reducing and capping
agents for the silver and iron nanoparticles.
This research supports the idea that the
total pH of the solution should be considered
when making a medical evaluation. The SEM
particle size for both NPs matches with DLS
46 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
No Bacteria NOL-Ag NP
(mg/mL | mg/mM)
AgNO3
(mM)
NOF-AgNP
(mg/mL | mg/mM)
1 E. coli 0.313 | 0.625 < 0.00976 0.078 | 0.156
2 CRKpn 0.078 | 0.156 < 0.00976 0.078 | 0.156
3 CREc 0.019 | 0.039 < 0.00976 0.078 | 0.156
4 S. aureus (ATCC 29213) 0.156 | 0.313 < 0.00976 0.625 | 1.250
5 S. aureus (ATCC 25923) 0.156 | 0.313 < 0.00976 0.313 | 0.625
6 ICRSa 0.313 | 0.625 < 0.00976 0.313 | 0.625
7 hVISA 0.313 | 0.625 < 0.00976 0.313 | 0.625
8 MRSA 0.313 | 0.625 < 0.00976 0.313 | 0.625
9 MR-CoNS 0.039 | 0.078 < 0.00976 0.078 | 0.156
10 MR-CoNS 0.039 | 0.078 < 0.00976 0.078 | 0.156
11 VREf 0.156 | 0.313 < 0.00976 0.156 | 0.313
12 E. faecalis (ATCC 51279) 0.078 | 0.156 < 0.00976 0.156 | 0.313
13 VSEf 0.039 | 0.078 < 0.00976 0.078 | 0.156
14 E. faecalis (ATCC 29212) 0.3125 | 0.625 < 0.00976 0.313 | 0.625
Table 5. The MICs of AgNO3, NOL-AgNPs, and NOF-AgNPs against Gram-negative and Gram-positive
bacteria.
Note. CRKpn – clinical isolates of carbapenem-resistant Klebsiella pneumoniae; CREc – carbapenem-resistant
Escherichia coli; ICRSa – inducible clindamycin-resistant Staphylococcus aureus BAA976-1; hVISA – hetero-
resistant S. aureus; MRSA – clinical isolates of methicillin-resistant S. aureus; MR-CoNS:2 – methicillin-
resistant coagulase-negative S. aureus; VREf – vancomycin-resistant Enterococcus faecium; VSEf – vancomycin-
susceptible E. faecalis.
analysis, which was around 100 nm. The
green synthesized NO-AgNPs and NO-FeNPs
are cytotoxic to the human chronic myeloid
leukemia cells in low concentrations and not
cytotoxic to the HUVEC cell line in the same
concentrations. The tested bacteria were able
to be killed at a low concentration of AgNO3
(< 0.00976 mM). The green synthesized NOL-
AgNPs and NOF-AgNPs were able to inhibit
bacteria including multidrug pathogens. We
can hypothesize here that green synthesis
AgNPs can be decreased the cytotoxic effects
of AgNO3 in vivo and the possible use of high
doses of AgNO3 as antimicrobial drugs.
Acknowledgements
The authors would like to thank Professor
Musa Mutlu Can from Istanbul University,
Faculty of Science, Department of Physic,
Renewable Energy and Oxide Hybrid Systems
Laboratory for allowing us to use his laboratory
and equipment.
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48 Plant Introduction • 91/92
S. Shawuti, C. Bairam, A. Beyatlı, İ. A. Kariper, I. N. Korkut, Z. Aktaş, M. O. Öncül et al.
Зелений синтез та характеристика наночастинок срібла та заліза отриманих
з використанням екстрактів Nerium oleander та їх антибактеріальна та
протипухлинна активність
Шаліма Шавуті 1, Часан Байрам 2, Ахмет Беятли 3, Ішак Афшін Каріпер 4, Исик Неслішах Коркут 1,
Зеррін Акташ 5, Мустафа Орал Онцюл 6, Серап Ердем Куруца 1
1 Кафедра фізіології, медичний факультет Стамбульського університету, вул. Тургута Озала Міллета,
Стамбул, 34093, Туреччина
2 Лабораторія відновлюваних джерел енергії та оксидних гібридних систем, кафедра фізики,
факультет природничих наук Стамбульського університету, вул. Шехзадебаші, Стамбул, 34134,
Туреччина
3 Кафедра лікарських та ароматичних рослин Гамідійського професійно-технічного училища
охорони здоров'я Університету медико-санітарних наук, вул. Тібійє, 38, Стамбул, 34668, Туреччина
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Plant Introduction • 91/92 49
Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander
Лікарські рослини можуть використовуватися як відновники при одержанні наночастинок металів
шляхом зеленого синтезу. Отримані наночастинки характеризуються хіміотерапевтичним та
протиінфекційним властивостям природних сполук. Зокрема, у цій праці йдеться про зелений
синтез наночастинок срібла та заліза з використанням екстрактів листя та квіток Nerium oleander, а
також аналізується їх властивості як протипухлинних та протимікробних засобів. Повідомляється
про особливості виготовлення наночастинок загалом та структурну характеристику наночастинок
срібла та заліза зокрема. Формування наночастинок досліджено за допомогою сканувальної
електронної мікроскопії та енергодисперсійної рентгенівської спектроскопії, UV-Vis та інфрачервоної
спектроскопії з перетворенням Фур’є (FTIR). Окрім того, також було досліджено поверхневим заряд,
розміри і розподілом наночастинок за допомогою DLS аналізу. Зелений синтез наночастинок срібла
та заліза з використанням N. oleander показав різні рівні селективної цитотоксичності щодо K562
(клітини хронічної мієлоїдної лейкемії людини) у низьких концентраціях і не був цитотоксичним для
HUVEC (ендотеліальні клітини пупкової вени людини) у тих же концентраціях. Наночастинки срібла
виявляли антибактеріальну активність по відношенню до мультирезистентних патогенів, тоді як
наночастинки заліза не виявляли такої активності. Результати цього дослідження підтверджують
потенціал використання наночастинок зеленого синтезу у різних сферах біомедицини та
фармацевтики в майбутньому.
Ключові слова: Nerium oleander, зелений синтез, Ag-наночастинки, Fe-наночастинки, клітинна цитотоксичність,
антибактеріальний ефект
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| id | oai:ojs2.plantintroduction.org:article-1592 |
| institution | Plant Introduction |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2025-07-17T12:53:58Z |
| publishDate | 2021 |
| publisher | M.M. Gryshko National Botanical Garden of the NAS of Ukraine |
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| resource_txt_mv | wwwplantintroductionorg/b6/b7652803a65ce780611f7e0f2a5b45b6.pdf |
| spelling | oai:ojs2.plantintroduction.org:article-15922023-08-26T20:39:08Z Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander extracts and their antibacterial and anticancer activities Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність Shawuti, Shalima Bairam, Chasan Beyatlı, Ahmet Kariper, İshak Afşin Korkut, Isık Neslişah Aktaş, Zerrin Öncül, Mustafa Oral Kuruca, Serap Erdem Medicinal plants can be used as reducing agents in the preparation of metal nanoparticles by green synthesis because of the chemotherapeutic and anti-infectious properties of natural compounds. Therefore, this paper reports the green synthesis of silver and iron nanoparticles from leaf and flower extracts of Nerium oleander and their capacity as anticancer and antimicrobial agents. Nanoparticle manufacturing and structural characterization of silver and iron nanoparticles are reported. The formation of nanoparticles is characterized by scanning electron microscopy with energy dispersive X-ray spectroscopy, UV-Vis and Fourier transform infrared (FTIR) spectroscopy. Nanoparticles formation was also investigated the surface charge, particle size, and distribution using zeta sizer analysis by DLS. Green synthesis of silver and iron nanoparticles using N. oleander showed different levels of selective cytotoxicity against K562 (human chronic myeloid leukemia cells) in low concentrations and were not cytotoxic to the HUVEC (human umbilical vein endothelial cells) in the same concentrations. Silver nanoparticles showed antibacterial activity against multidrug pathogens, while iron nanoparticles failed to show such activity. Results of the present research demonstrate the potential use of green synthesized nanoparticles in various biomedicine and pharmaceuticals fields in the future. Лікарські рослини можуть використовуватися як відновники при одержанні наночастинок металів шляхом зеленого синтезу. Отримані наночастинки характеризуються хіміотерапевтичним та протиінфекційним властивостям природних сполук. Зокрема, у цій праці йдеться про зелений синтез наночастинок срібла та заліза з використанням екстрактів листя та квіток Nerium oleander, а також аналізується їх властивості як протипухлинних та протимікробних засобів. Повідомляється про особливості виготовлення наночастинок загалом та структурну характеристику наночастинок срібла та заліза зокрема. Формування наночастинок досліджено за допомогою сканувальної електронної мікроскопії та енергодисперсійної рентгенівської спектроскопії, UV-Vis та інфрачервоної спектроскопії з перетворенням Фур’є (FTIR). Окрім того, також було досліджено поверхневим заряд, розміри і розподілом наночастинок за допомогою DLS аналізу. Зелений синтез наночастинок срібла та заліза з використанням N. oleander показав різні рівні селективної цитотоксичності щодо K562 (клітини хронічної мієлоїдної лейкемії людини) у низьких концентраціях і не був цитотоксичним для HUVEC (ендотеліальні клітини пупкової вени людини) у тих же концентраціях. Наночастинки срібла виявляли антибактеріальну активність по відношенню до мультирезистентних патогенів, тоді як наночастинки заліза не виявляли такої активності. Результати цього дослідження підтверджують потенціал використання наночастинок зеленого синтезу у різних сферах біомедицини та фармацевтики в майбутньому. M.M. Gryshko National Botanical Garden of the NAS of Ukraine 2021-11-28 Article Article application/pdf https://www.plantintroduction.org/index.php/pi/article/view/1592 10.46341/PI2021010 Plant Introduction; No 91/92 (2021); 36-49 Інтродукція Рослин; № 91/92 (2021); 36-49 2663-290X 1605-6574 10.46341/PI91-92 en https://www.plantintroduction.org/index.php/pi/article/view/1592/1519 Copyright (c) 2021 Shalima Shawuti, Chasan Bairam, Ahmet Beyatlı, İshak Afşin Kariper, Isık Neslişah Korkut, Zerrin Aktaş, Mustafa Oral Öncül, Serap Erdem Kuruca http://creativecommons.org/licenses/by/4.0 |
| spellingShingle | Shawuti, Shalima Bairam, Chasan Beyatlı, Ahmet Kariper, İshak Afşin Korkut, Isık Neslişah Aktaş, Zerrin Öncül, Mustafa Oral Kuruca, Serap Erdem Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title | Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title_alt | Green synthesis and characterization of silver and iron nanoparticles using Nerium oleander extracts and their antibacterial and anticancer activities |
| title_full | Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title_fullStr | Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title_full_unstemmed | Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title_short | Зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів Nerium oleander та їх антибактеріальна та протипухлинна активність |
| title_sort | зелений синтез та характеристика наночастинок срібла та заліза отриманих з використанням екстрактів nerium oleander та їх антибактеріальна та протипухлинна активність |
| url | https://www.plantintroduction.org/index.php/pi/article/view/1592 |
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