Journal of Sol-Gel Science and Technology (2024) 110:828–841

https://doi.org/10.1007/s10971-024-06409-6

ORIGINAL PAPER

Solution-gelation synthesis of silver nanoparticles utilizing Justicia

tranquebariensis extract for antibacterial, antioxidant, antifungal
and anticancer activity
B. Mary Dayana1 J. Thomas Joseph Prakash2 Jothi Vinoth Kumar3 Merum Dhananjaya4 Sang Woo Joo4
● ● ● ● ●

Mir Waqas Alam5

Received: 21 February 2024 / Accepted: 29 April 2024 / Published online: 20 May 2024
© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2024

Abstract
The distinctive biological properties exhibited by silver nanoparticles (AgNPs) have propelled this material to the forefront of
biological property candidates. Solution-gelation synthesis emerges as a highly promising method among preparation
techniques to compensate for the shortcomings inherent in both chemical and physical approaches. The objective of this
1234567890();,:

investigation was to synthesize JT-AgNPs utilizing Justicia tranquebariensis (JT) extract as a stabilizing and reducing agent
1234567890();,:

and AgNO3 solution as the Ag precursor. The obtained JT-AgNPs underwent physicochemical characterization employing
various analytical techniques. In XRD analysis revealed the crystal structure of the JT-AgNPs. The JT-AgNPs with a spherical
shape were identified through TEM, and exhibit a size in 22.3 nm. The zeta potential value of −21.6 mV indicates excellent
stability for green synthesized JT-Ag NPs. The antibacterial activity of synthesized JT-AgNPs was assessed. When compared
to the plant extract, JT-AgNPs exhibit effective inhibition of fungal strains, at a dosage of 10 μL/mL. The antioxidant
properties of JT-AgNPs showed a proton radical scavenging efficiency of 77.64% at 100 μg/mL (IC50 value of 44.41 μg/mL).
At a concentration of 15 μg/mL, the synthesized JT-AgNPs demonstrated remarkable efficacy against MDA-MB-231 cancer
cells, (IC50 value of 11.02 μg/mL). These results suggest a promising future herbal medication in nanomaterials form.

Graphical Abstract

* J. Thomas Joseph Prakash 2

PG & Research Department of Physics, Government Arts College,
armyjpr1@yahoo.co.in
(Affiliated to Bharathidasan University, Trichy-24),
* Merum Dhananjaya Tiruchirappalli, Tamil Nadu 620 022, India
msdhananjaya51@gmail.com 3
Bio Nanocomposite Research Center, Department of Food and
* Sang Woo Joo Nutrition, Kyung Hee University, Seoul 02447, Republic of Korea
swjoo1@gmail.com
4
School of Mechanical Engineering, Yeungnam University, 280
1
Daehak-ro, Gyeongsan, Gyeongbuk 38541, Republic of Korea
Department of Physics, St. Joseph’s College, (Affiliated to 5
Bharathidasan University, Trichy-24), Tiruchirappalli, Tamil Nadu Department of Physics, College of Science, King Faisal
620 002, India University, Al-Ahsa 31982, Saudi Arabia
Journal of Sol-Gel Science and Technology (2024) 110:828–841 829

Keywords Justicia tranquebariensis Silver nanoparticles Antibacterial activity Antioxidant properties

● ● ●

Highlights
● Silver nanoparticles (Ag NPs) were produced by aqueous leaf extract of Justicia tranquebariensis.
● The antioxidant properties of JT-Ag NPs showed a proton radical scavenging efficiency of 77.64% at 100 μg/mL (IC50
value of 44.41 μg/mL).
● The generated nanoparticles underwent characterization, and their impact on MDA-MB-231 cancer cell lines was
investigated.

1 Introduction of antimicrobial and anticancer pharmaceuticals. Various

metal-based nanoparticles including gold, silver, zinc, iron,
Cancer, which occurs by uncontrolled development or titanium, magnesium, and alginate [7], exhibit diverse
reproduction of body cells and frequently results in death, properties. Among them, silver possesses the ability to
kills almost 7 million people each year, even in indus- modulate cell systems, serving both as a regulator and a
trialized countries [1]. According to some estimates, cancer medium for detecting and diagnosing various bodily dis-
killed more than 9.5 million individuals in 2017, making it orders [8]. The eco-friendly synthesis of metal nanoparticles
the leading cause death after cardiovascular disease [2]. through a green approach is a notable method that empha-
There are numerous varieties of cancer. Lung disease, one of sizes environmentally conscious practices [9]. The synthesis
the cancers, is rather prevalent. According to the 2020 of nanoparticles using greener methods marks a significant
Cancer Report, lung cancer affects 18 people out of every advancement compared to other approaches, as it is
100,000 people and is the second leading cause of death straightforward, cost-effective, relatively reproducible, and
among cancer disorders. Breast cancer is another type of frequently results in more stable materials. Microorganisms
cancer that often kills people [3]. Based to the 2020 can be employed for the synthesis of nanoparticles [10].
Worldwide Cancer Reports, tumors in the breast is the sixth The green synthesis process eliminates the need for ele-
leading cause of death. Plants are widely used to treat a vated temperatures, pressures, energies, or hazardous sub-
variety of ailments. A variety of plant parts can be utilized to stances. Consequently, numerous researchers are now shifting
treat ailments. Extracts from various plant sections can be away from conventional synthetic methods. In comparison to
obtained and employed in a variety of medical treatments alternative methods, plants produce nanoparticles that are more
[4]. The majority of cancer medications (about 60%) are stable and are relatively easy to scale up. Moreover, the risk of
derived from plants, and plants are commonly used in the contamination is reduced. Preserving the biocompatibility of
design and development of novel drugs. One of these flora is synthetic nanomaterials is crucial, as chemically synthesized
the walnuts tree. The common walnut tree grows throughout materials pose various drawbacks to human health and the
Asia and Europe, as well as in Africa and America. The environment [11]. Due to their distinctive characteristics, silver
walnut tree’s Latin name is Nux Gallica, and it can grow to nanoparticles, stand out as one of the most promising metal
be 25–35 m long and 2 m in diameter. The walnut tree’s nanoparticles utilized in the field of nanomedicine. AgNPs are
roots, stems, and leaves are all incredibly different, and each commonly employed in topical ointments plays a crucial role
portion is medically essential [5]. Various parts of the walnut in preventing infections in injury [11], microbicidal [12],
tree have been utilized to treat a wide range of medical antimycotic [13], anti-inflammatory agent [14], counteract the
conditions. The bark, leaves, and stem of walnuts are used in activity of viruses [15], angiogenesis inhibitor [16], and anti-
beauty products, medicine, pesticides, and other industries. platelet effect [17]. There are different methods available for
In recent years, the exploration and application of nano- the synthesis of AgNPs, which include chemical reduction
materials have been burgeoning due to the unique properties synthesis [18], photochemical and chemical reactions in
exhibited by nano-sized objects, stemming from alterations reverse micelles [19], thermolysis of silver compounds [20],
in their surface-to-volume ratio compared to bulk materials. radiation enhanced [21], electrolysis [22], ultrasonic [23] and
The predominant share of nanomaterials has showcased microwave supported process [24]. Nevertheless, aside from
exceptional quantum confinement and distinctive catalytic necessitating the use of organic solvents and toxic reducing
attributes across a myriad of biochemical reactions, thereby agents, these approaches come with drawbacks, including low
enhancing methodologies for electronic, environmental, and yield, high-energy demands, and the necessity for challenging
biomedical applications [6]. In the realm of biotechnology, purification processes [25]. Biosynthesis of AgNPs using
nanomaterials find application in bio-imaging, diagnostics, microbes [26], mycelium [27], fungus [28], seaweeds [29, 30],
bio-sensing, and gene therapy, as well as in the development and plants are well documented.
830 Journal of Sol-Gel Science and Technology (2024) 110:828–841

Table 1 Phytochemical constituents of Justicia tranquebariensis distilled water. Subsequently, the leaf samples were dried in
aqueous leaf extracts
the shade and finely powdered. Silver Nitrate (AgNO3-
S. No. Phytoconstituents Reagents Aqueous leaf extract of 98.3%) and ammonia solution (NH3- 97%) were purchased
Justicia from Merck Ltd. Co.
tranquebariensis

1 Alkaloids Mayer’s + 2.2 Preparation of Justicia tranquebariensis leaves

2 Tannins Lead acetate + aqueous extract
Ferry chloride
Wagner’s For phytochemical extraction, 2 g of finely powdered Jus-
3 Carbohydrates Wagner’s − ticia tranquebariensis leaves were boiled in 100 ml of
Molisch’s
water. The resulting extract was filtered using Whatman
4 Steroid Liebermann +
Burchard’s
No.1 filter paper.
5 Saponins Gelatin +
6 Fixed oils Spot test −
2.3 Phytochemical analysis of the plant extract
7 Glycosides Legal’s +
Bontrager In the Tiruchirappalli district of Tamil Nadu, close to Mettur
8 Protein Millon’s − village, Justicia tranquebariensis leaves were collected
Biuret from their native environment. The leaves went through
9 Flavonoids Alkaline + cleaning, washing in double-distilled water, fine grinding,
Reagent and filtration. Several qualitative chemical analysis proce-
Shinoda’s dures were used to determine the chemical composition of
10 Terpenoids Thionyl + the extract. Qualitative phytochemical analyses were per-
chloride
formed using established protocols. Using qualitative ana-
lysis, phytochemical components, including alkaloids,
The Justicia tranquebariensis, a subshrub belonging to flavonoids, terpenoids, steroids, glycosides, tannins, and
the Acanthaceae family, is recognized as a significant carbohydrates, were discovered in the aqueous extracts of
reservoir of various bioactive phytochemicals, such as the Lantana camara leaf sample, as shown in Table 1.
phenolic compounds and tannins [31]. The leaf extract from
Justicia tranquebariensis exhibits diverse physiological 2.4 Green synthesis of JT-Ag NPs
attributes, encompassing anti-inflammatory, antibacterial,
antioxidant, cardioprotective, and antioxidative activities To fabricate the JT-Ag NPs, we commenced by combining
[32–35]. We employed a nanotechnological approach for 10 mL of silver nitrate (5 mM) with 50 mL of Justicia tran-
the initial examination of the anticancer capabilities of silver quebariensis extract in separate 100 mL containers. By vig-
NPs derived from Justicia tranquebariensis. orously agitating these mixtures with a magnetic stirrer, they
The present study concentrates on creating silver NPs were reduced to a homogeneous state as liquids. The pH of
through the utilization of an aqueous leaf extract from the resulting solutions was then adjusted with care to 7
Justicia tranquebariensis. Following the synthesis of NPs, through the drop-by-drop addition of ammonia solution.
their properties were analyzed, and their antibacterial, Following that, the reaction solution was stirred continuously
antifungal, and antioxidant capacities were evaluated. The while being heated to 90 °C. With the progression of the
MDA-MB-231 cells were employed in the investigation of evaporation process, the solution underwent a massive gel-
the anticancer properties of JT-AgNPs synthesized from like transformation. In pursuit of achieving a dried gel, the gel
Justicia tranquebariensis. underwent an additional heating process to 120 °C. The
results were then transferred to porcelain receptacles and
subjected to a two-hour heating process at 500 °C. The JT-
2 Experimental methods AgNPs were subsequently cooled for twenty-four hours in the
oven, which led to the formation of JT-Ag NPs powder. The
2.1 Materials subsequent procedure entailed the analysis of the structural,
morphological, and optical characteristics of JT-Ag NPs.
The plant was collected in October 2023 from Tiruchir-
appalli District, Tamil Nadu, India. The plant’s identity was 2.5 Characterization of JT-Ag NPs
verified at Rapinet Herbarium, St. Joseph’s College, Tir-
uchirappalli, Tamil Nadu, India. To eliminate dust, soil, and The identification of the synthesized JT-Ag NPs was con-
other extraneous materials, leaves samples were treated with ducted using a UV-Vis spectrometer (Perkin Elmer Lambda
Journal of Sol-Gel Science and Technology (2024) 110:828–841 831

35) covering a wavelength range from 200 nm to 1000 nm. were quantified to verify their approximate concentration of
FT-IR Spectroscopy (Perkin Elmer) was employed to 105 CFU/ml.
examine the attachment of functional groups and their
associated bonds within the material sample, covering a 2.7.3 Fungal strains used
wavelength range from 4000 cm−¹ to 500 cm−¹. The
hydrodynamic size of the nanoparticles in the examined The fungal test organisms, Aspergillus flavus (MTCC 356)
sample was determined using Dynamic Light Scattering and Aspergillus niger (MTCC 227), utilized in this inves-
(DLS). The crystalline structure of nanoparticles synthe- tigation were obtained from the National Chemical
sized through a green process was examined using X-ray Laboratory (NCL) in Pune, Maharashtra, India. The anti-
diffraction. The identification of powdered Ag NPs was fungal activity of the JT-AgNPs sample was determined
carried out through XRD, utilizing CuKα radiation with a using the disc diffusion method. Sterile petri dishes were
wavelength (λ) of 1.54 nm and scanning the angle 2θ in the coated with Sabouraud’s dextrose agar (SDA), and the test
range of 20° to 80°. For the examination of surface mor- organisms were inoculated. A 6mm-wide sterile disc was
phology, HR-SEM (FEI 250 FEG) with an EDAX setup loaded with 10 μL each of Fluconazole, AgNO3, pure plant
was employed. To analyze particle size, HR-TEM (JEOL- extract, and JT-Ag NPs. These prepared discs were placed
2100 + model) was utilized. on the agar plates, allowing the compounds to diffuse for
30 min at room temperature. A positive control was estab-
2.6 Antibacterial activities lished using a standard antibiotic disc containing 10 μL of
fluconazole. The plates were placed in an incubator at 37 °C
2.6.1 Collection of test organisms for 24 h, and the diameter of the inhibitory zone was
determined in millimeters.
Test organisms, including two Gram-negative bacterial
strains (Escherichia coli MTCC 25922 and Streptococcus 2.8 Antioxidant activity (DPPH assay method)
aureus MTCC 2451) and two Gram-positive bacterial
strains (Staphylococcus aureus MTCC 25923 and Bacillus In order to determine the antioxidant power of the sample,
subtilis MTCC 2451), were acquired from the Microbes its stable DPPH, or free radical activity, was measured. The
Type culture and Collection (MTCC) in Chandigarh, India. antioxidant capacity of the sample was evaluated through
The antibacterial effectiveness of JT-Ag NPs against spe- stable DPPH free radical activity. Varying concentrations
cific microorganisms was assessed using the disc diffusion (20–100 μL) of the synthesized NPs (1000 μL) were blen-
method. Sterile Petri dishes with a diameter of 60 mm were ded with 500 μL of an ethanolic DPPH solution (0.05 mM).
coated with Muller Hinton Agar and inoculated with the test The freshly prepared DPPH solution, stored at 4 °C in the
organisms. Discs of 6 mm width were impregnated with dark, underwent thorough shaking. Subsequently, 96%
10 μL each of Amoxicillin, AgNO3, plant extract, and JT- ethanol (2.7 mL) was added to the mixture. After allowing
Ag NPs using the sterile discs. These prepared discs were the combination to stand for five minutes at 540 nm, the
then positioned on the agar plates, allowing the compounds absorbance was determined using spectrophotometry. The
to diffuse for 30 min at room temperature. A positive con- absorbance was adjusted to zero using ethanol. The radical
trol was established using a standard antibiotic disc con- activity of the investigated samples was calculated and
taining 10 μL of amoxicillin. The experiment was repeated expressed as a percentage of inhibition.
three times. The plates were incubated for 24 h at 37 °C, and  
AB
the zone of inhibition was measured in millimeters. Percentð%Þinhibition of DPPH activity ¼ x100
A
2.7 Antifungal activities The absorbance values of the blank and sample are
denoted as A and B, respectively. The concentration
2.7.1 Culture media necessary to achieve 50% inhibition was calculated using
the concentration versus percentage of inhibition curve.
The antifungal assessment was carried out using Sabour-
aud’s dextrose agar from Hi Media Pvt. Ltd., 2.9 Cytotoxicity against cancer cells
Mumbai, India.
2.9.1 Source of chemical and reagents
2.7.2 Inoculum
Ploughed saline, streptomycin (C21H39N7O12 99.9%),
Following a six-hour incubation of each fungal strain penicillin-G (C16H17KN2O4S 97.31%), L-glutamine
individually in Sabouraud’s dextrose broth, the suspensions (H2NCOCH2CH2CH(NH2)CO2H 97%), Dulbecco’s
832 Journal of Sol-Gel Science and Technology (2024) 110:828–841

Fig. 1 a Synthesized Justicia

tranquebariensis JT-AgNPs,
b Uv -Vis spectra of JT-AgNPs

Modified Eagle’s Medium, Bovine serum albumin, ethylene contingent upon both the particle size and the dielectric
diamine tetraacetic acid (C14H20N2O10Na4 93.9%), 3-(4,5- constant of the medium [36]. Absorption of visible radiation
dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide, is due to the induced polarization in conduction electrons of
2'7’diacetyl dichloro fluorescein, trypan blue metal NPs concerning their immobile nucleus. When a
(C34H24N6Na4O14S4 92.89%), trypsin-EDTA, ethanol specific wavelength aligns with the size of JT-AgNPs,
(CH3CH2OH 96%), dimethyl sulfoxide ((CH3)2SO 99.9%), dipole oscillation is initiated in a compensated form of
and acridine orange (C17H19N3 98%) were all obtained from induced polarization. Subsequently, the electrons within the
Sigma Aldrich Chemicals Pvt. Ltd. and Triton X-100, JT-AgNPs resonate, leading to the absorption of radiation.
respectively. All additional compounds utilized were pro- The peak at 450 nm is due to the excitation of longitudinal
cured from Hi Media Laboratories Pvt. Ltd., India, and were plasmon vibrations Fig. 1b [37]. Hence, based on this out-
of analytical grade. come, it can be inferred that the synthesized JT-AgNPs
exhibit notable aqueous stability, and the phytochemicals
2.9.2 Cell culture maintenance found in Justicia tranquebariensis effectively serve as both
reducing and capping agents [38].
They were collected from the cell repository of the National
Centre for Cell Sciences (NCCS), which is located in Pune, 3.2 X-Ray diffraction technique
India. MDA-MB-231 breast cancer cells were obtained. In
order to prevent bacterial contamination, the cell line was The confirmation of the structure of JT-AgNPs was
grown in Dulbecco’s Modified Eagle Media (DMEM) that achieved through the identification of characteristic peaks in
was supplemented with 10% Fetal Bovine Serum (FBS). the XRD pattern, Fig. 2. The peaks were disclosed in the
Additionally, antibacterial drugs, specifically penicillin XRD pattern at 24.31°, 64.47°, 44.52°, and 38.20° assigned
(100 μg/mL) and streptomycin (100 μg/mL), were added to the (210), (220), (200), and (111) planes for the JT-AgNPs
the medium. In a humidified environment with 5% carbon respectively, signifying a crystalline structure [39]. The
dioxide and 37 degrees Celsius, the culture that included the diffraction peak at 78.44° corresponds to the (311) plane
cell lines was kept alive. and has the face-centered cubic structure of JT-AgNPs. The
diffraction line intensities and peak positions matched the
standard reference of silver (ICDD No. 03–065–2781)
3 Results and discussion effectively. It was evident from the XRD pattern that the JT-
AgNPs were crystalline. Green synthesized JT-AgNPs
3.1 UV-Vis spectroscopic technique exhibited analogous diffraction peaks to JT-AgNPs, indi-
cating the presence of a cubic face-centered crystal (FCC)
Uv-Vis spectroscopy plays a crucial role in confirming the structure [40, 41]. This outcome demonstrated that JT-
formation of JT-AgNPs. The bio-reduction of silver ions AgNPs formed in a crystalline nature. The JT-AgNPs were
was identified through the observable color change. When found to have an average crystallographic size of about
Justicia tranquebariensis aqueous extract was combined 24.7 nm from the Debye-Scherrer equation.
with silver nitrate, the color transformed from a pale light
yellow to reddish brown, indicating the successful forma- 3.3 Fourier transform infrared spectroscopy
tion of JT-AgNPs. This alteration in color is attributed to
surface plasmon vibrations in JT-AgNPs, as illustrated in In order to ascertain the potential bio-molecules that serve
the accompanying Fig. 1a. The absorption patterns of metal as capping and reducing agents for JT-AgNPs produced via
JT-AgNPs in Surface Plasmon Resonance (SPR) are Justicia tranquebariensis plant extract, FTIR spectroscopy
Journal of Sol-Gel Science and Technology (2024) 110:828–841 833

Fig. 2 XRD pattern of JT-AgNPs Fig. 3 FTIR spectrum of synthesized JT-AgNPs

was utilized. A versatile method for assessing the functional light scattering intensity and particle dimension determines the
groups in a sample is Fourier transform infrared spectro- signal pattern, which is dominated by larger molecules [38].
scopy, as different functional groups absorb different fre- Following that, DLS estimates the hydrodynamic diameter of
quencies of infrared light. Additionally, each molecule the particle, ligands, ions, and molecules that are connected to
possesses a unique spectrum referred to as a fingerprint that the outside and move with the particles in solution.
enables fast identification of one molecule from another.
Both the plant extract and the JT-AgNPs exhibited com- 3.5 SEM & TEM analysis
parable spectra (Fig. 3). The O–H vibrations are responsible
for the peak that occurs at 3389 cm−1 and 3167 cm−1. The SEM demonstrates the occurrence of synthesized JT-AgNPs
C–H stretching is responsible for the other peak, which may with clarity as shown in Fig. 5a, the NPs have a brick
be found at 2867 cm−1. When it comes to the plant extract, a spherical shape. The TEM analysis was utilized to assess the
few more peaks occurred in the region of 2000–2500 cm−1. dimensions and structure of the green JT-AgNPs that were
These peaks, which all represented the stretching vibration synthesized; the results are illustrated in Fig. 5b. The TEM
of C–H, were all present. The peak represents the C=O image clearly illustrates the existence of a capping agent
stretching at 1635 cm−1. The C–H stretching vibrations, within Justicia tranquebariensis, which serves as a protective
N–H bending vibrations, −CH3 wagging vibrations, and coating. The TEM analysis was employed to evaluate the
C–OH stretching vibrations are the vibrations that correlate dimensions and structure of the environmentally produced JT-
to the bands that come in the range of 1500–1200 cm−1. At AgNPs. It is spherical in shape and has a diameter of 22.3 nm
1029 cm−1 and 1214 cm−1, the C–N stretching of amines (Fig. 5c). As a result of (111) reflections, the SAED pattern of
can be observed. As a result of C–H stretching (aromatic), produced JT-AgNPs exhibits a more intense spherical ring in
the peaks that occur at 803 cm−1, and 850 cm−1are closer proximity to the center. Refractions at (220) and (200)
explained. The JT-AgNPs exhibited a loss of intensity in the give rise to both the second and third rings, respectively. The
peaks at 3167 cm−1, 2313 cm−1, 2124 cm−1, and 1214 cm−1 FCC structure and polycrystalline character of the produced
that were present in the extract. This loss of intensity JT-Ag NPs are indicated by the SAED pattern (Fig. 5d) [42].
showed the involvement of these homologous groups dur-
ing the reduction of silver to silver nanoparticles. 3.6 Energy dispersive Xray spectroscopy analysis

3.4 Dynamic light scattering technique The EDX signal confirmed the presence of elemental silver.
The strong peaks between 0–3 kV Fig. 6a in the EDAX
The hydrodynamic particle size scattering of produced JT- spectrum of synthesized JT-AgNPs are right related to K
AgNPs was evaluated using the dynamic light scattering and L lines of silver. To determine the elemental arrange-
(DLS) technique. The hydrodynamic particle size graph ment on the outside of the green synthesized Ag NPs,
revealed that the average diameter of the manufactured JT- EDAX was used. It was established that the silver compo-
AgNPs was around 90.1 nm, with a low polydispersity index nent (66.73%) was the important component, and Fig. 6b
(0.316), as shown in Fig. 4. The power relationship between displays the prominent peak obtained at 3KeV. In addition
834 Journal of Sol-Gel Science and Technology (2024) 110:828–841

Fig. 4 DLS analysis of synthesized JT-AgNps

Fig. 5 a SEM image of

synthesized JT-AgNPs, b TEM
image of JT-AgNps, c Average
diameter of JT-AgNPs, d SAED
pattern of JT-AgNps

to Ag (66.73%), additional elements discovered in the acquire their negative surface charges as a result of phy-
EDAX study included carbon (3.9%), chlorine (4.3%), and tochemicals. The relatively negative charge on the surface
potassium (11.3%). These components may have come of JT-AgNPs facilitates the manifestation of a stable
from the plant extract’s phytochemicals, which capped the structure, as it prevents detrimental conditions such as
JT-AgNPs [42]. agglomeration and fluctuation from occurring. Further-
more, the exclusive possession of a negative charge on the
3.7 Zeta potential of JT-AgNPs surface enables efficient interaction with biological
organisms, including bacteria and fungi, that are posi-
Zeta potential analysis data for the synthesized JT-AgNPs tively charged. As a result, their biocompatibility is
showed a surface charge of −21.6 mV Fig. 7. Ag NPs enhanced [39].
Journal of Sol-Gel Science and Technology (2024) 110:828–841 835

Fig. 6 a, b EDAX spectrum of synthesized JT-AgNPs

3.8 Biological activity of the synthesized JT-Ag NPs against two-Gram negative bacterial strains (e.g., Escher-
ichia coli (MTCC 25922) Fig. 8c, Streptococcus aureus
3.8.1 Antimicrobial activities (MTCC 2451) Fig. 8d, and two-Gram positive bacterial
strains (a, b) Staphylococcus aureus (MTCC 25923) were
Using the disc diffusion method, the antibacterial activity of examined. The zones of inhibition against Bacillus subtilis
green synthesized NPs was investigated. Plants typically (MTCC 2451) and Escherichia coli (MTCC 25922) were
include a variety of antioxidant enzymes and metabolites. found to be comparable. Table 1 reveals that the produced
They greatly aid in the examination of oxidative damage to JT-Ag NPs had high and comparable antibacterial activity
cellular constituents. This plant is rich in fiber, manganese, against Gram-negative and Gram-positive microorganisms.
vitamin C, and other essential elements. Antioxidative Due to their size, JT-AgNPs may easily access the nucleus
complexes are present in it. This study used the disc dif- content of bacteria, and they have a huge and outstanding
fusion method to investigate the JT-AgNps antibacterial and surface area, allowing for widespread contact with bacteria.
antifungal activities against specific microorganisms. This could explain why they have the best antimicrobial
effect. The diameters of the inhibitory zones on the plate of
3.8.1.1 Antibacterial activity of JT-AgNPs The results for agar are reported in millimeters. The tests were performed
the antibacterial activity of synthesized JT-AgNPs are three times on each treatment sample, and the outcomes are
shown in Table 2. The inhibitory effects of JT-AgNPs shown in Table 2. Streptococcus aureus has a higher
836 Journal of Sol-Gel Science and Technology (2024) 110:828–841

Fig. 7 Zeta potential technique

of synthesized JT-AgNPs

Fig. 8 Antibacterial activity of

synthesized JT-AgNPs against
(a) Staphylococcus aureus, (b)
Bacillus subtilis, (c) Escherichia
coli, (d) Streptococcus aureus

suspended antibacterial activity than Staphylococcus aur- 3.8.1.2 Antifungal activity of JT-Ag NPs Antifungal activity
eus. The proposed possible mechanism of antibacterial of as-prepared JT-AgNPs against Aspergillus flavus (MTCC
capability using JT-AgNPs is depicted in Fig. 9 [43]. 356) Fig. 10a and Aspergillus niger (MTCC 227) Fig. 10b
Journal of Sol-Gel Science and Technology (2024) 110:828–841 837

Fig. 9 Possible antimicrobial

mechanism using JT-AgNPs

Fig. 10 Antifungal activity of

synthesized Justicia
tranquebariensis JT-AgNPs
against (a) Aspergillus flavus (b)
Aspergillus niger

as tested fungal strains have been investigated using the disc of concentrations, including 2.5, 5, 7.5, 10, and 15 μg/mL,
diffusion method. The results showed that Aspergillus niger following a period of incubation lasting 24 h, as shown in
is to be more susceptible to JT-AgNPs than Aspergillus Fig. 12. Higher concentrations of nanoparticles and
flavus fungal isolate with an inhibition zone of about 7 mm nanocomposites (15 μg/mL) resulted in 38.81% cell via-
at an appropriate volume of 10 μL of as-prepared JT- bility, indicating anticancer efficacy against breast cancer
AgNPs solution [44, 45]. MDA-MB-231 cells. The IC50 value for JT-AgNPs was
11.02 μg/mL, as shown in Fig. 13a. Figure 13b depicts the
3.8.1.3 Antioxidant activity of JT-AgNPs The findings cell viability at various concentrations. One of the primary
indicate that, in comparison to the standard ascorbic acid, advantages of combination medicines is their potential for
the samples demonstrate antioxidant activities at elevated providing synergistic effects [48]. In combination therapy,
concentrations Fig. 11a. The aqueous extract has 77.64% the combined therapeutic benefit of the medications was
antioxidant activity at a concentration of 500 μg/mL, while found to be greater than the sum of their separate benefits.
the ascorbic acid has 89.63% at the same concentration. The These incentives have shifted drug research efforts toward
capacity for proton radical scavenging is attributed to anti- the hunt for combination medicines. The optimal ther-
oxidants as assessed through the DPPH radical scavenging apeutic combination with the highest antitumor activity
assay Fig. 11b, c [46, 47]. can be estimated using combination index (CI) iso-
bologram analysis/multiple drug effect, which is an
3.8.1.4 Cytotoxicity against cancer cells In order to effective technique to demonstrate that medications work
determine the cytotoxicity (MTT assay) of the produced synergistically [49]. The JT-AgNPs are considered phy-
JT-AgNPs, the viability of breast cancer MDA-MB-231 tocompound transporters and may have anticancer prop-
cells was examined. During the current investigation, the erties. Cell membranes include negatively charged
JT-AgNPs that were synthesized were obtained in a range elements such as lipids, whereas nanocomposites have
838 Journal of Sol-Gel Science and Technology (2024) 110:828–841

Fig. 11 a Antioxidant activity of

samples, b JT-AgNPs, c DPPH
assay method

Fig. 12 Morphological changes

in control and JT-AgNPs treated
Breast cancer MDA-MB-231
cells for 24 h

positive charges with opposing charges and are respon- 4 Conclusion

sible for nanoparticle uptake and internalization [50]. The
JT-AgNPs’ surface area is the most essential determinant The synthesis of JT-AgNPs occurred through the utilization of
in cell internalization. The cytotoxic activity is caused by an aqueous extract derived from Justicia tranquebariensis. The
the creation of ROS, which activates the caspase cascade characterization of these nanoparticles was subsequently ver-
of apoptosis, DNA damage, and mitochondrial malfunc- ified through the application of analytical techniques, including
tion. Figure 14 depicts the possible mechanisms of JT- UV–visible spectrophotometry(UV), X-ray diffraction (XRD)
AgNPs’ anticancer activity. and Fourier Infrared Spectroscopy (FT-IR). Biological
Journal of Sol-Gel Science and Technology (2024) 110:828–841 839

Fig. 13 a IC50 value for the synthesized JT-AgNPs, b The cell viability for different concentrations

Fig. 14 The possible mechanism

of anticancer activity of JT-
AgNPs

Table 2 Zone of inhibition for

Samples Concentrations Organisms/Zone of inhibition (mm)
synthesized JT-AgNPs against
(μL/mL)
Gram-positive and Gram- Staphylococcus Bacillus Escherichia coli Streptococcus
negative bacterial strains aureus subtilis aureus

A (Amoxicillin) 10 7 8 12 11
B (silver) 10 0 0 0 0
C (Plant extract) 10 1 0 2 3
D (NPs) 10 4 7 7 5

methodologies are being suggested in this context, notwith- substances, operates with reduced energy usage, and does not
standing the utilization of conventional ways for producing JT- necessitate trained labor. The combination of these qualities,
AgNPs. The utilization of a biological methodology in align- together with the fundamental principles of green chemistry,
ment with the principles of green chemistry is an ecologically enhances the profitability, biocompatibility, and reproducibility
sustainable strategy that yields reduced waste generation and of biological fabrication as a synthesis method for producing
energy consumption. The utilization of plant resources for the extremely stable JT-AgNPs. The prepared JT-AgNPs under-
synthesis of JT-AgNPs is seen as a sophisticated approach went characterization, and their impact on MDA-MB-231
towards a safer future, as it eliminates the need for costly toxic cancer cell lines was investigated. According to the
840 Journal of Sol-Gel Science and Technology (2024) 110:828–841

characterization studies, the produced JT-AgNPs exhibited an Ni-Cu-Zn based nanosized metal oxides for photocatalytic and
average particle diameter of 22 nm and a crystallite size of sensor applications. Crystals 11(12):1467
8. Alam MW, BaQais A, Mir TA, Nahvi I, Zaidi N, Yasin A (2023)
24 nm. Notably, we are the first to synthesize JT-AgNPs from
Effect of Mo doping in NiO nanoparticles for structural mod-
this plant, demonstrating enhanced inhibition of cell growth. ification and its efficiency for antioxidant, antibacterial applica-
Importantly, our process avoids the use of artificial or hazar- tions. Sci Rep. 13(1):1328
dous NPs. The materials exhibit significant potential in com- 9. Renuka R, Devi KR, Sivakami M, Thilagavathi T, Uthrakumar R,
Kaviyarasu K (2020) Biosynthesis of silver nanoparticles using
bating cancer cells. The findings, obtained through exposure to
Phyllanthus emblica fruit extract for antimicrobial application.
solid tumors and various cancer cells, provide valuable insights Biocatalysis Agric Biotechnol 24:101567
for our forthcoming studies. 10. Alam MW, Allag N, Utami M, Waheed-Ur-Rehman M, Al Saleh
Al-Othoum M, Sadaf S (2024) Facile Green Synthesis of α-Bis-
Acknowledgements The authors were like to thank PG & Research muth Oxide Nanoparticles: Its Photocatalytic and Electrochemical
Department of Physics, Government Arts College, Thiruchirappalli- Sensing of Glucose and Uric Acid in an Acidic Medium. J
620 022 for provide the opportunity to carry out this research work. Compos Sci 8(2):47
This work was supported by the Deanship of Scientific Research, Vice 11. Nithyabalaji R, Ramya RM, Kavitha R, Radhakrishnan K, Kumar
Presidency for Graduate Studies and Scientific Research, King Faisal JV, Al-Asbahi BA, Joo SW (2024) Molecular structure, character-
University, Saudi Arabia [Project No: GRANT6,159]. ization, in vitro and in-silico studies of N, N-dimethyl aminophenyl
schiff’s base-chalcone hybrid. Chem Phys Impact 8:100422
12. Sriram MI, Kanth SBM, Kalishwaralal K, Gurunathan S (2010)
Author contributions B. Mary Dayana: Conceptualization, Methodology, Antitumor activity of silver nanoparticles in Dalton’s lymphoma
Writing - Original Draft Mir Waqas Alam; Formal analysis, Jothi Vinoth ascites tumor model. Int J Nanomed 5:753–762
Kumar: data acquisition, revision, and tabulation. Merum Dhananjaya, 13. Panáček A, Kolář M, Večeřová R, Prucek R, Soukupová J,
Sang Woo Joo and J. Thomas Joseph Prakash: supervision, review, and Kryštof V, Kvítek L (2009) Antifungal activity of silver nano-
proofreading. particles against Candida spp. Biomaterials 30(31):6333–6340
14. Nadworny PL, Wang J, Tredget EE, Burrell RE (2008) Anti-
inflammatory activity of nanocrystalline silver in a porcine contact
Compliance with ethical standards dermatitis model. Nanomed Nanotechnol Biol Med 4(3):241–251
15. Rogers JV, Parkinson CV, Choi YW, Speshock JL, Hussain SM
(2008) A preliminary assessment of silver nanoparticle inhibition of
Conflict of interest The authors declare no competing interests.
monkeypox virus plaque formation. Nanoscale Res Lett 3:129–133
16. Gurunathan S, Lee KJ, Kalishwaralal K, Sheikpranbabu S, Vai-
Ethical approval Furthermore, we affirm that the research was con- dyanathan R, Eom SH (2009) Antiangiogenic properties of silver
ducted in adherence to international, national, and institutional reg- nanoparticles. Biomaterials 30(31):6341–6350
ulations pertaining to the protection of biodiversity rights, animal 17. Safaepour M, Shahverdi AR, Shahverdi HR, Khorramizadeh MR,
experimentation, and clinical research. Additionally, no animals or Gohari AR (2009) Green synthesis of small silver nanoparticles
humans were injured during this investigation. using geraniol and its cytotoxicity against fibrosarcoma-wehi 164.
Avicenna J Med Biotechnol 1(2):111
18. Alam MW, Khalid NR, Naeem S, Niaz NA, Ahmad Mir T, Nahvi
I, Zaidi N (2022) Novel Nd-N/TiO2 nanoparticles for photo-
References catalytic and antioxidant applications using hydrothermal
approach. Materials 15(19):6658
1. Seçme A, Bozer BM, Kocaman AY, Erenler R, Calimli MH 19. Alam MW, Najeeb J, Naeem S, Usman SM, Nahvi I, Alismail F,
(2024) Synthesis, characterization, and anticancer properties of Ag Nawaz A (2022) Electrochemical methodologies for investigating
nanoparticles derived from walnut leaves tested on cells of L929, the antioxidant potential of plant and fruit extracts: A review.
MCF-7 and H1299. J Drug Deliv Sci Technol 94:105478 Antioxidants 11(6):1205
2. Vardhana J, Kathiravan G (2015) Biosynthesis of silver nanoparticles 20. Navaladian S, Viswanathan B, Viswanath RP, Varadarajan TK
by endophytic fungi Pestaloptiopsis pauciseta isolated from the leaves (2007) Thermal decomposition as route for silver nanoparticles.
of Psidium guajava Linn. Int J Pharm Sci Rev Res 31(1):29–31 Nanoscale Res Lett 2:44–48
3. Nagarajaiah S, Shivanna Giresha A, Gopala Krishna P, Manikrao 21. Kumar JV, Tammina SK, Rhim JW (2024) One-step hydro-
Gadewar M, Praveen M, Nanda N, Venkatesh Yatish K (2024) thermal synthesis of sulfur quantum dots for detection of Hg2+
Anti‐oncogenic Potential and Inflammation Modulatory Response ions and latent fingerprints. Colloids Surfaces A 690:133682
of Green Synthesized Biocompatible Silver Nanoparticles. Chem 22. Kumar JV, Alam MW, Selvaraj M, Almutairi HH, Albuhulayqah
Biodivers 21:e202301533 M, Sadaf S, Joo SW (2024) Fluorescent carbon dots for biodiesel
4. Zarfeshany A, Asgary S, Javanmard SH (2014) Potent health production: A Comprehensive review (2019–2024). Inorganic
effects of pomegranate. Adv Biomed Res 3(1):100 Chem Commun 162:112247
5. Barakat MA, Anjum M, Kumar R, Alafif ZO, Oves M, Ansari MO 23. Zhu J, Liu S, Palchik O, Koltypin Y, Gedanken A (2000) Shape-
(2020) Design of ternary Ni (OH)2/graphene oxide/TiO2 nano- controlled synthesis of silver nanoparticles by pulse sonoelec-
composite for enhanced photocatalytic degradation of organic, trochemical methods. Langmuir 16(16):6396–6399
microbial contaminants, and aerobic digestion of dairy waste- 24. Alam MW, Al Qahtani HS, Souayeh B, Ahmed W, Albalawi H,
water. J Clean Prod 258:120588 Farhan M, Naeem S (2022) Novel copper-zinc-manganese ternary
6. Tammina SK, Jyothi L, Kumar JV, Srivastava H, Naraharisetty metal oxide nanocomposite as heterogeneous catalyst for glucose
SRG (2024) Sensing, detoxification and bactericidal applications sensor and antibacterial activity. Antioxidants 11(6):1064
of nitrogen-doped carbon dots. Diamond Rel Mater 144:111013 25. Mukunthan KS, Elumalai EK, Patel TN, Murty VR (2011)
7. Alam MW, Aamir M, Farhan M, Albuhulayqah M, Ahmad MM, Catharanthus roseus: a natural source for the synthesis of silver
Ravikumar CR, Ananda Murthy HC (2021) Green synthesis of nanoparticles. Asian Pac J Tropical Biomed 1(4):270–274
Journal of Sol-Gel Science and Technology (2024) 110:828–841 841

26. Sreekanth TVM, Pandurangan M, Kim DH, Lee YR (2016) Green trend applications: Synthesis, properties, and future directions. J
synthesis: in-vitro anticancer activity of silver nanoparticles on Mol Liquids, 394:123787
human cervical cancer cells. J Clust Sci 27:671–681 41. Jores K, Mehnert W, Drechsler M, Bunjes H, Johann C, Mäder K
27. Sachin S, Sundaram SJ, Franklin JB, Raj AD, Kumar JV, Alam (2004) Investigations on the structure of solid lipid nanoparticles
MW (2024) Synthesis of Zinc Oxide nano bars incorporated with (SLN) and oil-loaded solid lipid nanoparticles by photon corre-
activated Carbon (ZnO NBs/AC) nanocomposites for high specific lation spectroscopy, field-flow fractionation and transmission
capacitance value. J Sol Gel Sci Technol 109:896–904 electron microscopy. J Controlled Rel 95(2):217–227
28. Krishnaraj C, Jagan EG, Rajasekar S, Selvakumar P, Kalaichelvan 42. Sooraj MP, Nair AS, Vineetha DJCP (2021) Sunlight-mediated
PT, Mohan NJCSBB (2010) Synthesis of silver nanoparticles using green synthesis of silver nanoparticles using Sida retusa leaf
Acalypha indica leaf extracts and its antibacterial activity against extract and assessment of its antimicrobial and catalytic activities.
water borne pathogens. Colloids Surf B: Biointerfaces 76(1):50–56 Chem Pap 75(1):351–363
29. Kumar JV, Rhim JW (2024) Fluorescent carbon quantum dots for 43. Dogiparthi LK, Sana SS, Shaik SZ, Kalvapalli MR, Kurupati G,
food contaminants detection applications. J Environ Chem Eng Kumar GS, Gangadhar L (2021) Phytochemical mediated synth-
12:111999. esis of silver nanoparticles and their antibacterial activity. SN
30. Govindaraju K, Kiruthiga V, Kumar VG, Singaravelu G (2009) Appl Sci 3(6):631
Extracellular synthesis of silver nanoparticles by a marine alga, 44. Kumar VN, Tamilanban T, Sultana TS, Manasa K, Ragulkumar E,
Sargassum wightii Grevilli and their antibacterial effects. J Kumar JV, Arul K (2023) In silico study of traditional Chinese
Nanosci Nanotechnol 9(9):5497–5501 medicinal compounds targeting alzheimer’s disease amyloid beta-
31. Nima P, Astalkshmi A, Ganesan V (2013) A comparative account peptide (1–42). Chem Phys Impact 7:100383
on the green synthesis of silver nanoparticles using leaves and 45. Ghandehari S, Tabrizi MH, Ardalan P, Neamati A, Shali R (2019)
pods of Peltophorumpterocarpum (DC.) Backer ex Heyne. J Green synthesis of silver nanoparticles using Rubia tinctorum
Biopro Tech 98:308–317 extract and evaluation the anti-cancer properties in vitro. IET
32. Jadhav K, Dhamecha D, Bhattacharya D, Patil M (2016) Green Nanobiotechnol 13(3):269–274
and ecofriendly synthesis of silver nanoparticles: characterization, 46. Acharya K, Samui K, Rai M, Dutta BB, Acharya R (2004)
biocompatibility studies and gel formulation for treatment of Antioxidant and nitric oxide synthase activation properties of
infections in burns. J Photochem Photobiol B Biol 155:109–115 Auricularia auricula. Indian J Exp Biol 42(5):538–540
33. Heydari R, Rashidipour M (2015) Green synthesis of silver nano- 47. Stefanovits-Bányai É (2003) Antioxidant effect of various
particles using extract of oak fruit hull (Jaft): synthesis and in vitro rosemary (Rosmarinus officinalis L.) clones. Acta Biol Szege-
cytotoxic effect on MCF-7 cells. Int J Breast Cancer 2015:846743 diensis 47(1-4):111–113
34. Johnson I, Prabu HJ (2015) Green synthesis and characterization 48. Abad MHK, Nadaf M, Yazdi MET (2023) Biosynthesis of ZnO.
of silver nanoparticles by leaf extracts of Cycas circinalis, Ficus Ag2O3 using aqueous extract of Haplophyllum obtusifolium:
amplissima, Commelina benghalensis and Lippia nodiflora. Int Characterization and cell toxicity activity against liver carcinoma
Nano Lett 5(1):43–51 cells. Micro Nano Lett 18(6):NA–NA
35. Kumar B, Smita K, Cumbal L, Debut A (2017) Green synthesis of 49. Eslamieh-Ei FM, Sharifimoghaddammood N, Poustchi Tousi SA,
silver nanoparticles using Andean blackberry fruit extract. Saudi J Basharkhah S, Mottaghipisheh J, Es-Haghi A, Iriti M (2023)
Biol Sci 24(1):45–50 Synthesis and its characterisation of selenium/silver/chitosan and
36. Alahmad A, Feldhoff A, Bigall NC, Rusch P, Scheper T, Walter cellular toxicity against liver carcinoma cells studies. Nat Product
JG (2021) Hypericum perforatum L.-mediated green synthesis of Res 1–9
silver nanoparticles exhibiting antioxidant and anticancer activ- 50. Rahimi E, Asefi F, Afzalinia A, Khezri S, Zare-Zardini H,
ities. Nanomaterials 11(2):487 Ghorani-Azam A, Yazdi MET (2023) Chitosan coated copper/
37. Gebru H, Taddesse A, Kaushal J, Yadav OP (2013) Green silver oxide nanoparticles as carriers of breast anticancer drug:
synthesis of silver nanoparticles and their antibacterial activity. J cyclin D1/P53 expressions and cytotoxicity studies. Inorg Chem
Surf Sci Technol 29(1-2):47–66 Commun 158:111581
38. Bharadwaj KK, Rabha B, Pati S, Choudhury BK, Sarkar T, Gogoi
SK, Edinur HA (2021) Green synthesis of silver nanoparticles Publisher’s note Springer Nature remains neutral with regard to
using Diospyros malabarica fruit extract and assessments of their jurisdictional claims in published maps and institutional affiliations.
antimicrobial, anticancer and catalytic reduction of 4-nitrophenol
(4-NP). Nanomaterials 11(8):1999 Springer Nature or its licensor (e.g. a society or other partner) holds
39. Tho NTM, An TNM, Tri MD, Sreekanth TVM, Lee JS, Naga- exclusive rights to this article under a publishing agreement with the
jyothi PC, Lee KD (2013) Green synthesis of silver nanoparticles author(s) or other rightsholder(s); author self-archiving of the accepted
using Nelumbo nucifera seed extract and its antibacterial activity. manuscript version of this article is solely governed by the terms of
Acta Chim Slovenica 60(3):673–678 such publishing agreement and applicable law.
40. Kumar JV, Karthika D, Radhakrishnan K, Arul V, Alam MW,
Rosaiah P, Joo SW (2023) MXene nanocomposites for current