doi.org/10.1002/cbdv.
202301533 Research Article
Anti-oncogenic Potential and Inflammation Modulatory
Response of Green Synthesized Biocompatible Silver
Nanoparticles
Shobha Nagarajaiah,[a] Aladahalli Shivanna Giresha,[b] Prashanth Gopala Krishna,*[c]
Manoj Manikrao Gadewar,[d] Manjappa Praveen,[e] Nagappa Nanda,[f] Deepadarshan Urs,[g]
Kattepura Krishnappa Dharmappa,[g] Bhangi Mutta Nagabhushana,[h] Srilatha Rao,[i]
Mallanna Mahadeva Swamy,[j] and Kalanakoppal Venkatesh Yatish[k]
This study presents a novel approach to synthesizing silver
nanoparticles (Ag NPs) using a solution combustion synthesis
(SCS) method with Catharanthus roseus (C. roseus) leaf extract.
The NPs were thoroughly characterized through X-ray diffrac-
tion (XRD), Scanning electron microscopy (SEM), Energy dis-
persive X-ray (EDX), Transmission electron microscopy (TEM),
and Selected area electron diffraction (SAED), elucidating their
crystal structure. Notably, the synthesized Ag NPs exhibited a
significant dose-dependent decline in viability of the MDA-MB
231 breast cancer cell line, with an IC50 value of 13.3 μg/mL,
underscoring their potential as potent anticancer agent. Beyond
cytotoxicity, the study pioneers an investigation into the
Introduction
In recent years, nanotechnology has witnessed remarkable
advancements, particularly in the synthesis of nanoparticles
(NPs). A noteworthy innovation in this dynamic field is the
emergence of green synthesis methods. These methods employ
plant extracts or natural substances to catalyze NP formation,
presenting a promising alternative to conventional
techniques.[1–5] Notably, green synthesis is distinguished by its
reduced toxicity and heightened environmental friendliness
compared to traditional methods.[4,6,7] This eco-friendly ap-
[a] Dr. S. Nagarajaiah
Department of Chemistry, Maharani’s Science College for Women, Mahara-
ni Cluster University, 560 001 Bengaluru, India
[b] Dr. A. Shivanna Giresha
Department of Biochemistry, Jain (Deemed-to-be University), School of
Science, JC Road, 560 027 Bangalore, India
[c] Dr. P. Gopala Krishna
Research and Development Center, Department of Chemistry, Sir M.
Visvesvaraya Institute of Technology, 562 157 Bengaluru, India
E-mail: prashanth_chem@sirmvit.edu
prashaanthgk@gmail.com
[d] Dr. M. Manikrao Gadewar
Department of Pharmacology, School of Medical and Allied Sciences, KR
Mangalam University, 122 103 Gurgaon, India
[e] Dr. M. Praveen
Centre for Advanced Materials Technology (CAMT), M.S Ramaiah Institute
of Technology, 560 054 Bengaluru, India
Chem. Biodiversity 2024, e202301533 (1 of 10)
www.cb.wiley.com
biocompatibility of Ag NPs by blood hemolsysis, providing
critical insights into their safety and biomedical applicability.
Furthermore, this research uncovers a distinctive facet of Ag
NPs, revealing their inhibitory effects on the inflammatory
enzyme secretory phospholipase A2 (sPLA2), a recognized
biomarker for breast cancer. The demonstrated in vitro and
in vivo inhibition of sPLA2 highlights the multifaceted potential
of Ag NPs in not only targeting cancer cells but also modulating
inflammatory responses associated with breast cancer, position-
ing the study at the forefront of advancements in nanomedicine
and cancer therapeutics.
proach involves using sustainable materials and techniques,
offering not only a more environmentally conscious alternative
but also a cost-effective and accessible one. Moreover, the
resulting NPs often exhibit superior properties compared to
those synthesized through alternative means.[8,9] Consequently,
green synthesis of NPs stands as a burgeoning area of research,
holding vast potential across diverse fields.
Among the array of NPs, Ag NPs have emerged as
particularly captivating entities in recent times. Their versatile
applications range from antibacterial agents to the creation of
robust materials, with a particularly exciting role in the medical
[f] Dr. N. Nanda
Department of Chemistry, BMS College of Engineering, 560 019 Bengaluru,
India
[g] D. Urs, Dr. K. Krishnappa Dharmappa
Inflammation Research Laboratory, Department of Studies and Research in
Biochemistry, Mangalore University, Jnana Kaveri Post Graduate Centre,
Chikka Aluvara, 571 232 Kodagu, India
[h] Dr. B. Mutta Nagabhushana
Department of Chemistry, M.S Ramaiah Institute of Technology, 560 054
Bengaluru, India
[i] Dr. S. Rao
Department of Chemistry, Nitte Meenakshi Institute of Technology, 560 064
Bengaluru, India
[j] Dr. M. Mahadeva Swamy
Department of PG Chemistry, JSS College of Arts Commerce and Science,
570 025 Mysuru, India
[k] Dr. K. Venkatesh Yatish
Department of Chemistry, Navkis College of Engineering, 573 217 Hassan,
India
© 2023 Wiley-VHCA AG, Zurich, Switzerland
 Research Article
doi.org/10.1002/cbdv.202301533
realm.[10,11] Renowned for their antimicrobial properties, Ag NPs
derived from plant extracts showcase enhanced antimicrobial
activity.[12] Their potential as coatings for medical devices,
reducing infection risks,[13] and as topical treatments for wound
care, which promote bacterial elimination and accelerate
healing, underscore their versatility. Furthermore, Ag NPs
demonstrate potential industrial applications, such as fortifying
materials and imbuing antibacterial properties in products like
clothing and carpeting.[14] Prospects even extend to applications
in solar panels and other energy-related technologies.[15]
Beyond medical applications, Ag NPs also hold promise in
water purification and air pollution control.[16] Research indi-
cates their efficacy in removing harmful bacteria and viruses
from water sources.[17] Additionally, materials based on Ag NPs
exhibit the capacity to capture and eliminate pollutants from
the air.[18]
Given the association between inflammation and cancer
onset, particularly in breast cancer, the study delves into the
role of phospholipase A2 (PLA2) in inflammatory processes and
its potential as a therapeutic target.[19,20] Notably, secretory
phospholipase A2 (sPLA2) is implicated in various malignancies,
including breast cancer, promoting carcinogenesis, cell prolifer-
ation, and local inflammation.[22] The inhibition of sPLA2
emerges as a pertinent aspect of cancer therapy, a facet
underscored by the anti-inflammatory activity demonstrated by
green-synthesized Ag NPs, which inhibit 15-LOX, COX-2, and
pro-inflammatory cytokines.[24,25] Moreover, the study explores
the intriguing connection between Ag NPs and snake venom,
revealing their shared potential to suppress sPLA2, suggesting
implications in anti-inflammatory and snake bite remedies.
Looking ahead, Ag NPs have the potential to revolutionize
disease treatment, water purification, and air quality
enhancement. The versatile Solution Combustion Synthesis
(SCS) technique, known for producing various nanostructured
materials, adds an innovative dimension to this exploration.[28–31]
The study introduces a swift and uncomplicated Ag NPs
production method utilizing the aqueous leaf extract of
C. roseus, renowned for its antioxidant, antiviral, antibacterial,
antibiotic, antifungal, and wound healing properties.[38–40] The
synthesized Ag NPs undergo comprehensive physicochemical
characterization, in vitro and in vivo anti-inflammatory studies,
and cytotoxicity assessments on MDA-MB-231, complemented
by an examination of its biocompatibility.
This research reveals the remarkable potential of green-
synthesized Ag NPs in the dynamic field of nanotechnology.
Situated at the nexus of innovation and environmental
awareness, the study explores the diverse uses of Ag NPs,
ranging from transforming medical procedures to addressing
issues related to water purification and air pollution mitigation.
In order to explore possible therapeutic pathways, the study
explores the anti-inflammatory characteristics of Ag NPs and
reveals how they interact with phospholipase A2 (PLA2). The
research not only advances nanotechnology but also explores
biocompatibility and cytotoxicity by utilising the therapeutic
properties of C. roseus and introducing a quick and novel way
to produce Ag NPs using the SCS process. This research
contributes to the evolving landscape of nanotechnology,
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offering insights into the synthesis, applications, and potential
therapeutic implications of Ag NPs, particularly in the context of
cancer and inflammation.
Experimental
Materials and Methods
All chemicals and reagents used in this study were of analytical
grade and sourced from Sigma-Aldrich. C. roseus leaves were
obtained from the vicinity of Bengaluru, India were used as fuel.
Plant Extract Preparation
Fresh C. roseus plant leaves, weighing 50g, were cleaned with
deionized water and then cut into small pieces. Soxhlet extraction
was performed using 250 mL of deionized water. The resulting
extract was filtered through Whatman No. 1 filter paper, and
synthesis was carried out using the filtered extract.
The fresh leaves of the plant C. roseus Linn were collected from the
vicinity of Bengaluru city in the month of February 2020. The
collected plant material was identified and authenticated by a
taxonomist in the Department of Botany at Bangalore University,
Bengaluru. The voucher specimen (14203) was deposited in the
Department of Botany at Bangalore University, Bengaluru, India, for
future correspondence
Synthesis of Ag NPs
A fixed range of plant extract volumes, ranging from 1 to 10 mL,
was introduced to 0.1 g of AgNO3. Deionized water was added as
necessary to achieve a total volume of 10 mL. The mixture was then
placed in a petri dish and positioned within a preheated muffle
furnace. Notably, this NP synthesis process eliminated the need for
supplementary reducing agents or surfactants.
Upon reaching 500 °C, the mixture underwent boiling, and ignition
occurred within just 5 minutes. This pattern continued within the
temperature range of 500 � 10 °C, ultimately yielding Ag NPs. In the
concluding phase, following a calcination period of 40 minutes at
500 � 10 °C, the resultant product was gradually cooled to ambient
temperature. This technique facilitated the creation of noble metal
Ag NPs. Remarkably, the plant leaf extract derived from C. roseus
served as both a proficient reducing agent and a stabilizer for these
Ag NPs.
Characterization
Many analytical techniques were used to characterize the produced
Ag NPs. PXRD (powder X-ray diffraction) pattern was obtained
using a PanAlytical X-pert diffractometer with Cu Kα radiation (λ =
1.5418 Å) as the source. The morphology of the Ag NPs was studied
by Scanning Electron Microscope (SEM, FEI Quanta 200 ESEM). The
developed phase was validated again using a SAED pattern from a
TEM (JOEL-TEM-2100).
Cell culture
The ATCC (American Type Culture Collection) provided the cell line
of breast cancer (MDA-MB-231). The ATTC methodology was
followed to culture, expand, and maintain the cells in Dulbecco’s
Modified Eagle’s Medium (DMEM) media under conventional cell
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cultures conditions like 100 μg/mL streptomycin and 100 units/mL ½ðControl diameter Test diameterÞ=Control diameter� � 100 (2)
penicillin; added 10 % heat to inactivated calf serum (Gibco BRL,
Life Technologies, Grand-Island, NY, USA) under 5 % carbon dioxide
at 37 °C (Binder, Germany).
Neutralization of edema-inducing activity
Inhibition of cell growth assay The method of Yamakawa et al., adapted by Vishwanath et al. was
followed.[50,51] The VRV-PLA2 (5 μg) alone or with different concen-
The MTT assay was used to estimate the antiproliferative effect of
trations of Ag NPs (5 to 20 μg) in a total volume of 20 μL was
Ag NPs on MDA-MB-231 cancer cells, as discussed in our earlier
injected into the intraplantar surface of the right hind footpad of
studies.[36,41–43] MDA-MB-231 cancer cells were seeded in a 96-well
mice. 20 μL saline was injected into the respective left footpad for
microplate at a density of 5×105 cells/mL. After 24 h of incubation
control. The animals were euthanized after 45 min by administering
at 37 °C and 5 % CO2, each well was treated with different doses of
anesthesia (30 mg/kg of pentobarbital i. p.), and both the hind
NPs. This was followed by another 24 h incubation at 37 °C and 5 %
limbs were cut at the ankle joint and weighed separately. The
CO2. After the initial incubation, 20 μL of MTT solution was added
percentage of edema was calculated by the following formula.
to each well, and cells were incubated at 37 °C for 4 hours in the
dark. Subsequently, 100 μL of DMSO was used to dissolve the
purple, water-soluble formazan crystals (Merck; Mumbai; India). Weight of the edematous leg
Edema ratio ¼ x 100 (3)
Cytotoxicity towards cancer cells was determined at an absorbance Weight of normal leg saline injected
of 590 nm using an ELISA reader (Tecan; Austria). Untreated media
with 0.1 % medium DMSO was used as a negative control. The
antiproliferative potential of Ag NPs was measured by determining
Results and Discussion
their IC50 value, which represents the concentration at which cell
growth is inhibited by 50 %. XRD

The PXRD pattern of Ag NPs is presented in Figure 1. By

Biocompatibility assay
Approximately 5 mL of fresh blood collected from a sheep was
centrifuged at 1000 rpm for 10 minutes. The red blood cells (RBCs)
were separated and washed twice with phosphate-buffered saline
(PBS) and then diluted at a 1 : 4 ratio. PBS served as the negative
control, while 1 % sodium dodecyl sulfate (SDS) was used as the
positive control. Different concentrations of Ag NPs were mixed
with the diluted erythrocyte suspension and incubated at 37 °C for
1 hour. Subsequently, the tubes were centrifuged at 300 rpm for
5 minutes. Using a microplate reader (Tecan, Austria), the super-
natant was collected, and absorbance was measured at 540 nm.
The proportion of hemolysis was determined using
equation (1).[44–46] A sample is considered hemocompatible if the
hemolysis rate is less than 10 %, indicating biocompatibility
evaluating the XRD pattern, it was possible to determine that
the Ag NPs were crystalline. The (111); (200); (220); and (311)
Bragg’s reflections of the face-centered cubic structure of Ag,
respectively were visible peaks in the XRD spectrum at 2θ =
38.12°; 46.28°; 64.04° and 776.84° and it matched with the
database of Joint Committee on Powder Diffraction Standards
(JCPDS) file No. 04-0783. This indicated that the diffraction
peaks were broad and the crystallite size was small.[52] This
pattern led to calculating the lattice constant, which was 3.87.
The Scherrer formula, which reads
d ¼ kl=bcosq (4)

[where d = crystallite size in Å; k = 1 (constant); λ = 0.1541 nm

(1) (wavelength); β = FWHM (full width at half maximum) and θ =
angle], was used to calculate the average crystallite size of the

Purification and inhibition of sPLA2

The secretory phospholipase A2 from Vipera ruselli snake venom
(VRV-PLA2) was purified according to previous work.[47] The
inhibition of PLA2 activity by Ag NPs was assessed using a solid
medium as described by Gutiérrez et al., (198[48] with slight
modification.[49] In brief, 100 mL of 0.1 M Tris HCl (pH 7.4 containing
5 mM CaCl2) was used to boil 1g of agarose. After it had cooled to
room temperature, 6 drops of egg yolk were added. Agitated and
transferred into sterile petri plates and allowed to solidify. Wells
were made using a gel puncture, and 20 μg of VRV-PLA2 was added.
The plates were incubated at 37 °C for 24 hours. For the test, Ag
nanoparticles pre-incubated with VRV-PLA2 (10, 20, and 30 μg) were
added. The results were expressed as percentages and the controls
containing only VRV-PLA2 (20 μg) were considered as 100 % of
phospholipase activity. Whereas, well without VRV-PLA2 served as a
negative control. Halo diameter (Zone of clearance) was measured
using a standard scale and the percentage of PLA2 inhibition was
determined using the formula:

Figure 1. PXRD pattern of Ag NPs.

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 Research Article
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Ag NPs used in the biogenic reduction process.[52–54] The
average crystallite size of Ag NPs was estimated to be 8 � 1 nm.
Figure 2. (a–d) SEM and (e) EDX image of the Ag NPs.
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SEM and EDX
EDX (Energy Dispersive X-ray) and SEM analyses were employed
to study the chemical purity and surface morphology of Ag
NPs, as shown in Figure 2(a–d). The particles were found to be
spherical with an uneven size distribution. According to the
EDX study, the presence of Ag was confirmed, with the
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 Research Article
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spectrum showing a prominent signal in the Ag area, approving
the establishment of Ag NPs. The EDX spectrum, presented in
Figure 2(e), indicated a strong presence of Ag with a weak
oxygen peak, possibly originating from organic molecules
bound to the surface of Ag NPs, representing the reduction of
silver ions to silver. This finding confirmed the complete
reduction of the Ag compound to Ag NPs, as revealed in the
spectrum
TEM and SAED
The TEM images at different amplifications and SAED outlines
are presented in Figure 3 (a–d). The TEM provided additional
insight into the size and morphology details of the formed Ag
NPs The TEM also observed the sphere-shaped nature of the Ag
NPs, with diameter ranging from 5–8 nm. The results derived
from the XRD pattern were found to be in good agreement
with the SAED pattern, confirming the polycrystalline nature of
Ag NPs (Figure 3d). TEM was utilized to examine particle size
and crystallinity, and the SAED patterns further validated the
polycrystalline nature of Ag NPs. A typical SAED pattern
recorded with the largest aperture is shown in Figure 3d.
Figure 3. (a–c) TEM and (d) SAED pattern images of Ag NPs.
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Anti-cancer studies Ag NPs
The anticarcinogenic activity of Ag NPs on MDA-MB231 cells
was assessed using the MTT assay with different concentrations
of 0.3, 0.6, 1.25, 2.5, 5, 10, and 20 μg/mL Ag NPs for 24 hours.
Cell proliferation was inhibited in a concentration-dependent
manner upon treatment, with a maximum proliferation inhib-
ition of 90 % observed at a concentration of 20 μg/mL. The IC50
value of Ag NPs on MDA-MB-231 was determined to be
13.3 μg/mL. The test sample, Ag NPs treatments, exhibited
significant dose-dependent inhibition of the growth of MDA-
MB-231 cells, indicating cytotoxicity.
MTT solution was primarily used to assess cell viability,
leading to mitochondrial dysfunction.[55] Figure 4 illustrates the
cytotoxic impact of Ag NPs on the MDA-MB231 cell line, and
Figure 5 presents a visual graphic of the toxicity of Ag NPs
towards cancer cells. These antiproliferative results unequivo-
cally show that Ag NP treatment sensitizes cancer cells, with
cell viability decreasing in a dose-dependent manner. The
findings suggest that Ag NPs exhibited 49 % cell viability at
their highest measured concentration of 20 μg/mL.
The literature indicates that Ag NPs cause cytotoxicity in a
cell-specific and proliferation-dependent manner, rapidly differ-
entiating cancer cells, which are the most vulnerable, from
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 Research Article
doi.org/10.1002/cbdv.202301533
Figure 4. Cytotoxic effect of Ag NPs in MDA MB 231 cell lines. Cells were
treated with various concentrations (0, 0.3, 0.6, 1.25, 2.5,5,10, and 20 μg/mL
of NPs) for 24 h.
Figure 5. Percent cell viability of Ag NPs in MDA MB 231 cell lines. Cells were
treated with various concentrations (0, 0.3, 0.6, 1.25, 2.5,5,10, and 20 μg/mL)
of NPs for 24 h.
quiescent cells, which are the least sensitive.[56–58] The IC50 value
of Ag NPs on MDA-MB-231 was determined to be 13.3 μg/mL.
Microscope images of untreated MDA-MB-231 cancer cells
and cells treated with Ag NPs are shown in Figure 6. It is
noteworthy that there was significant cell death at 20 μg/mL.
Figure 6. Phase images of untreated MDA-MB-231 cancer cells and cells treated with Ag NPs. MDA-MB-231 cancer cells treated with (a) untreated or control,
(b) Ag NPs (20 μg/mL).
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This may be attributed to pure Ag NPs penetrating the cell
membrane more efficiently due to their small particle size,
facilitating the formation of vesicles that can be invaded by
lysosomes, resulting in necrosis or mitochondrial damage.
Interactions between different types of nanoparticles and cells
have been discovered to be dependent on the nanomaterials’
size, shape, and surface contact.[59] Charged NPs are shown to
be more toxic compared to charged and neutral NPs[60]
Biocompatibility of Ag NPs
The application of Ag NPs in a biological system as an anti-
cancer agent necessitates determining its biocompatibility.
Hemolysis of erythrocytes in PBS, 1 % SDS, and upon Ag NPs
treatment was examined by measuring turbidity and erythro-
cyte lysis at 540 nm. The hemolysis rates at concentrations of
0.3125, 0.625, 1.25, 2.5, 5, 10, and 20 μg/mL of Ag NPs treatment
showed significantly less hemolysis compared to the negative
control (2 %) and positive control (100 %), as presented in
Table 1.
This study suggests that Ag NPs are biocompatible with no
detectable hemolysis at the concentrations tested. Ag NPs
demonstrated a less toxic effect on human RBC (hemolysis) at
0.3125–20 μg/mL, as shown in Figure 7 and Table 1. These
findings are in good agreement with the literature.[61]
Based on these results, it can be inferred that these Ag NPs
may not adversely affect normal cells. Consequently, they can
be considered in further studies, such as apoptosis and
clonogenic assays, cell cycle analysis, and DNA fragmentation.
Mechanism of Ag NPs on the target cells
The simple mechanism involved in cancerous cells and anti-
cancer activities is the action of ROS (reactive oxygen
species)[62,63] and also the formation of superoxide formation
anion (*O2 ) by recent studies. Due to the release of silver ions
from the nanoparticles, Ag NPs induce cancer cell death; hence
the anti-cancer activity of solution combustion-synthesized Ag
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Table 1. Hemolytic activity of Ag NPs.

Sample Conc. μg/mL % Inhibition

Control (PBS) 0 0
Positive control (1 % SDS) 10000 77.88
1.25 1.58
2.5 4.06
5 4.59
10 7.08
20 7.61

NPs towards MDB MA 231 cancer cells may be considered, and

the effect of silver ions is dose-dependent, that is, concen-
tration-dependent and changes from normal to cancer cells.[64,65]
Through both chemical and biological methods, the multifunc-
tional activities of our NPs have been compared with
commercially available NPs of different sizes and published
literature. In comparison, these Ag NPs demonstrated superior
anti-cancer ability when compared to previously reported and
commercially available Ag NPs. The toxicity for selective cancer
cells, attributed to the formation of silver ions and colloidal
silver and gold nanoparticles, exerts anti-tumoral effects against
cancer cells by inducing cellular ROS production, ultimately
responsible for cellular damage.[66–68]
Secretory PLA2 Inhibition Activity
Many Ag NPs synthesized from natural sources were reported
for anti-inflammatory activity by inhibiting, sPLA2, COX-1&2,[69]
edema, cytokines levels[70] and snake venom sPLA2.[27] In this
current work, different concentrations of the NPs were pre-
incubated at 370C for 30 min and subjected for in vitro and
Figure 8. sPLA2 inhibition by Ag NPs in egg yolk solid medium.
in vivo PLA2 inhibition. The PLA2 inhibition potential of NPs was
evaluated after 18h of incubation of the plates in an incubator
at the same temperature. The formation of a clear halo around
the well in the gel characterized phospholipase activity (i. e.,
breakdown of phospholipid). At a concentration of 30 μg,
43.3 percent VRV-PLA2 was inhibited (Figure 8), and inhibition
was concentration-dependent (Table 2).
In in vivo edema neutralization activity, the sPLA2 (VRV-
PLA2) injected footpad showed an edema of greater than 171 �
1.6 % compared with the saline-injected leg. The NPs inhibited
edema formation in a dose-dependent manner when co-
injected with VRV-PLA2. The edema ratio was decreased to
136 � 2.1 % at 20 μg concentration of Ag NPs (Figure 9).

Figure 7. Hemolytic activity of Ag NPs.

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Table 2. sPLA2 inhibition by Ag NPs in egg yolk solid medium.
Well component Halo diameter (mm)
sPLA2 Alone (20 μg) 3.0
sPLA2 + AgNP (10 μg) 2.8
sPLA2 + AgNP (20 μg) 2.6
sPLA2 + AgNP (30 μg) 1.7
Figure 9. Neutralization of edema-inducing activity by Ag NPs.
Discussion
Platinum drugs, while widely used in cancer treatment, come
with serious side effects, including gastrointestinal and hemor-
rhagic damage.[71] Moreover, resistance to platinum-based drugs
has been observed in many cancer cells.[72] This has led to
increased research interest in novel transition metal conjugates.
For instance, Taraxacum officinale, a herbal dandelion, demon-
strated potent cytotoxicity against HepG2 (human liver cancer
cells).[73] Additionally, Ag NPs synthesized using various plant
extracts have shown promising anti-cancer effects against
different cancer cell lines.[74–76]
In vitro studies have revealed that Ag NPs possess anti-
cancer, antiproliferative, and antiangiogenic activities by affect-
ing the apoptotic pathway through the formation of free
oxygen radicals.[76] Compounds with antiangiogenic potential,
such as Ag NPs, can inhibit the expression of abnormally
expressed signaling proteins and exhibit consistent anti-cancer
effects.[76] Previous research by Asharani et al. (2009) demon-
strated significant antiproliferative effects of silver nanoparticles
on human glioblastoma cells.[77] Moreover, studies by Tran H. V.
et al. (2010)[78] and Sanpui et al. (2011)[79] highlighted that
nanosilver interferes with normal cellular function, affecting
membrane integrity and inducing apoptotic signaling in
mammalian cells.
Nanosilver, according to Hsin et al. (2008), can cause
mitochondria-dependent apoptosis in various cell lines.[80] Intra-
venous administration of aqueous silver oxide nanoparticles has
shown anti-cancer properties in transplanted lymphosarcoma
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tumor models.[81] Shawkey et al. (2013) reported anti-cancer
activity of Ag NPs against various malignancy cell lines,
% Inhibition
including colon carcinoma (HCT116), breast carcinoma (MCF7),
– liver carcinoma (HepG2), and intestine carcinoma (CaCo2).[82]
6.66 The cytotoxicity of nanosilver is attributed to dynamic phys-
13.33 icochemical interactions with intracellular proteins and DNA.[83]
43.33 Ag NPs act as anti-cancer agents by inhibiting tumor cell
growth, possibly through the inhibition of signaling cascades
involved in cancer development. They can promote cytotoxicity
against cancer cells while minimizing harm to healthy cells.
Specifically, Ag NPs have shown anti-inflammatory activity by
inhibiting sPLA2 both in vitro and in vivo conditions.[84] This
versatility has led to their extensive use in research, cancer
treatment, and as drug carriers[85–87]
Conclusions
In summary, the utilization of plant-assisted SCS allows for the
straightforward creation of highly porous Ag NPs, offering a
practical and single-step approach. Analysis of the PXRD pattern
confirmed the crystalline nature and identified a pure cubic
phase without impurity peaks. Our investigation demonstrated
the potent anti-cancer efficacy of Ag NPs against MDA-MB-231
cells. The hemolytic assay indicated a negligible impact on
erythrocytes by Ag NPs. Additionally, both in vitro and in vivo
experiments revealed the inhibitory effect of Ag NPs on the
inflammatory enzyme secretory phospholipase A2 (sPLA2), a
significant biomarker for breast cancer. This highlights the
promising therapeutic potential of metal NPs synthesized
through SCS with plant-derived sources. Insights from similar
studies hold substantial promise for the development of
innovative anti-cancer therapeutic agent.
Experimental Section
Ag NPs were synthesized through an environmentally friendly
and self-sustaining SCS method using C. roseus leaf aqueous
extract. The resulting sample underwent thorough character-
ization using various techniques, PXRD, SEM, EDS, and TEM. The
synthesized Ag NPs were then evaluated for their anticancer
activity against the human breast cancer cell line MDA-MB-231
through a MTT assay. In addition to assessing their anticancer
properties, the anti-inflammatory activity of Ag NPs was
investigated both in vivo and in vitro. Furthermore, the bio-
compatibility of Ag NPs was examined through a blood
hemolysis assay, ensuring a comprehensive evaluation of their
potential applications.
Funding
NIL
© 2023 Wiley-VHCA AG, Zurich, Switzerland
 Research Article
doi.org/10.1002/cbdv.202301533
Ethical approval
Experiments were conducted as per the protocols of the
Institutional Animal Ethical Committee (IAEC), Mangalore Uni-
versity India (No: IAECMU-RP-15).
Author Contributions
Conceptualization, generation of the hypothesis, experimenta-
tion, and manuscript writing: Shobha Nagarajaih, Prashanth GK,
Giresha, Dharmappa, Deepdarshan, and M. Praveen; Software
analysis: Shobha Nagarajaih, Praveen, Nanda N, Manoj M,
Srilatha Rao, K.V. Yatish, M. Mahadeva Swamy; Review and
editing: Prashanth GK, Giresha, Shobha Nagarajaih, Nanda N,
Revision plagiarism errors and proofread: Prashanth GK, K.V.
Yatish, M. Mahadeva Swamy, and Bhangi Mutta Nagabhushana.
All authors have read and agreed to the published version of
the manuscript.
Conflict of Interests
The authors declare no conflict of interest.
Data Availability Statement
Data available on request from the authors.
Keywords: Anti-inflammatory · Blood hemolysis · Catharanthus
roseus · Cytotoxicity · in vivo
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