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Artificial Cells, Nanomedicine, and Biotechnology

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Microwave assisted green synthesis of silver


nanoparticles using leaf extract of elephantopus
scaber and its environmental and biological
applications

Sijo Francis, Siby Joseph, Ebey P. Koshy & Beena Mathew

To cite this article: Sijo Francis, Siby Joseph, Ebey P. Koshy & Beena Mathew (2018) Microwave
assisted green synthesis of silver nanoparticles using leaf extract of elephantopus�scaber and its
environmental and biological applications, Artificial Cells, Nanomedicine, and Biotechnology, 46:4,
795-804, DOI: 10.1080/21691401.2017.1345921

To link to this article: https://doi.org/10.1080/21691401.2017.1345921

Published online: 06 Jul 2017.

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ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY, 2018
VOL. 46, NO. 4, 795–804
https://doi.org/10.1080/21691401.2017.1345921

Microwave assisted green synthesis of silver nanoparticles using leaf extract of


elephantopus scaber and its environmental and biological applications
Sijo Francisa, Siby Josephb, Ebey P. Koshya and Beena Mathewc
a
Department of Chemistry, St. Joseph’s College, Moolamattom, Idukki, India; bDepartment of Chemistry, St. George’s College, Aruvithura,
Kottayam, India; cSchool of Chemical Sciences, Mahatma Gandhi University, Kottayam, India

ABSTRACT ARTICLE HISTORY


The fabrication of spherical silver nanoparticles using the phytoreducing agent Elephantopus scaber is Received 8 February 2017
reported here. Irradiation of the reaction mixture under a domestic microwave oven enabled the forma- Revised 14 June 2017
tion of stable silver nanoparticles and was confirmed by UV-vis spectral portrait. Chemical components Accepted 19 June 2017
inherent in the aqueous leaf extract which reduces the monovalent silver were identified by FT-IR spec-
troscopy. The crystal structure of the synthesized nanoparticles was established as face centred cube by KEYWORDS
the powder XRD analysis. The TEM images suggest an average particle size of 37.86 nm to the silver Microwave; Elephantopus
nanoparticles. The prepared silver nanocatalysts can successfully reduce various organic nitro com- scaber; silver nanoparticles;
pounds, namely, 4-nitrophenol, 2-nitroaniline and 4-nitroaniline. The environmental pollution caused by nitro compounds;
dyes like eosin Y is effectively wiped off within a short span of time using the prepared nanocatalysts. antimicrobial; A375
The free radical quenching efficacy of the plant extract and the silver nanoparticles were checked by
employing DPPH assay bestowing ascorbic acid reference. The potential of the nanoparticles as antimi-
crobials against six human disease causing pathogens were tested through the well diffusion pathway.
The newly developed silver nanoparticles produced IC50 value 15.68 ± 0.15 lg/mL on human skin carcin-
oma cells, A375 and 65.49 ± 0.40 lg/mL on fibroblast cells, L929 when the cytotoxicity is studied
employing MTT assay. Elephantopus scaber showed IC50 value 50.55 ± 0.17 lg/mL against A375 cells.

Introduction
because of its applications in medicine and catalysis.
Phytochemical-mediated nanoparticle synthesis is a green
Silver nanoparticles are proved as antibacterial and antifungal
approach which reduces the extensive and excessive use of
agents against various human disease causing pathogens and
toxic chemicals. Nanomaterials wonderfully act as good
are the best remedy for various cancerous tissues [7–9].
catalysts in various chemical reactions like adsorption, poly-
Nanosilver is an ultimate solution for the efficient removal of
merization, photodegradation, etc. [1–4]. The reduction of
monovalent silver using plant phytochemicals under micro- dyes from various industries [10–14].
wave irradiation is a rapid method of silver nanoparticles’ Elephantopus scaber (E. scaber), an aromatic tropical herb
synthesis [5,6]. These nanoparticles are of unique interest which belongs to the family Asteraceae, has an admirable
traditional medicinal value. The plant contains many bioactive

CONTACT Beena Mathew beenamscs@gmail.com School of Chemical Sciences, Mahatma Gandhi University, Kottayam-686560, Kerala, India
ß 2017 Informa UK Limited, trading as Taylor & Francis Group
796 S. FRANCIS ET AL.

compounds like sesquiterpene lactones, flavonoids, steroids, centrifuge with a speed of 12,000 rpm. The capsule obtained
terpenoids and phenolic compounds [15]. It has been is dried in air and used for analysis.
reported that the plant E. scaber has possessed wonderful
wound healing and antimicrobial capacities [16]. Previous Characterization techniques
reports showed that the plant extract and the silver nanopar-
Electronic spectroscopic studies were done by a Shimadzu
ticles derived from it have good antioxidant power [17].
UV-2450 spectrophotometer. FT-IR spectrum was recorded
The anticancer activity of E. scaber has been attributed to
using Perkin Elmer 400 spectrometer. Powder XRD data was
the flavonoids [18] and the sesquiterpene lactones-deoxyele- drawn from a PANalytic X’PERT-PRO X-ray spectrometer.
phantopin and isodeoxyelephantopin [19] present in the Morphological investigations were obtained from High
plant extracts. The hepatoprotective activity of the ethanolic Resolution-Transmission Electron Microscopic (HR-TEM)
extract of E. scaber was proved in mice against ethanol- images using a JEOL JEM-2100 microscope equipped with
induced liver damage [20]. EDX attachment. Atomic Force Microscopic studies were con-
In the current work the leaf extract of E. scaber is used as ducted using WI Tec Alpha 300 RA instrument in the tapping
the talented reducing agent for the reduction of AgNO3 to mode. Photoluminescence spectral data were drawn using
silver nanoparticles by microwave irradiation method. The Fluorolog 3 TCSPC fluorescence spectrometer.
leaf extract served as an expert capping agent which pre-
vents agglomeration of the nanoparticles. Antimicrobial, anti- Reduction of nitro compounds
oxidant and in-vitro anticancer scopes of the synthesized
The hydrogenation of 4-nitrophenol to 4-aminophenol,
silver nanoparticles were investigated. For cytotoxicity studies
2-nitroaniline to 1, 2-phenylenediamine and 4-nitroniline to 1,
using MTT assay the human skin cancer cell lines A375 and
4-phenylenediamine by NaBH4 were taken as model reactions
fibroblast cell lines L929 were used. Incentive of the present
and performed in the presence of the newly synthesized sil-
work is the affirmation of the catalytic activity of the synthe-
ver nanocatalyst. 2 ml of 4-nitrophenol (8  105 M) was
sized nanosilver for the reduction of various nitrocompounds
placed in a quartz cuvette along with 0.5 ml freshly prepared
and organic dyestuffs by NaBH4 in a rapid rate.
NaBH4 (0.06 M) and 0.5 ml of AgNP-E. scaber (0.04 mg/mL).
UV-vis spectra were continuously recorded till the completion
Materials and methods of the reaction which was indicated by the disappearance of
the initial reactant peaks in the electronic spectra. The reac-
Silver nitrate (AgNO3), sodium borohydride (NaBH4), tion was conducted with 2-/4-nitroaniline (6  105 M) instead
2, 2-diphenyl-1-picrylhydrazyl (DPPH), 4-nitrophenol, 2-nitroa- of 4-nitrophenol. The kinetics of the reduction reactions were
niline, 4-nitroaniline and Eosin Y were used without further examined by recording the absorbance at wavelength 400,
purification. All materials used were of analytical grade and 412 and 380 nm for 4-nitrophenol, 2-nitroaniline and 4-nitroa-
are purchased from Merck India Ltd and the solutions were niline respectively, using UV-vis spectrophotometer in the
prepared in Millipore water. range 200–800 nm.

Preparation of E. Scaber leaf extract Catalytic degradation


Fresh leaves of E. scaber were collected and identified taxo- The synthesized silver nanoparticles were utilized in the
nomically. They were washed thoroughly in tap water fol- removal of organic dyes, causing environmental pollution. To
lowed by distilled water. 5 gm of 2 day air dried leaves were perform the degradation reaction, an interesting fluorescent
taken in a round bottom flask fitted with a condenser and cationic dye eosin Y (EY) was selected. 2 ml of (1  104 M) of
heated for 30 min with 100 ml of distilled water at 40  C. It is EY, 0.5 ml freshly prepared NaBH4 (0.06 M) and 0.5 ml of
cooled and filtered through Whatman No.1 filter paper and AgNP-E. scaber (0.02 mg/mL) were taken in a quartz cuvette
kept at 4  C in refrigeration for further use. of 1 cm path length. The UV-vis absorption spectra were
recorded at definite time intervals in the wavelength range of
200–800 nm. Complete disappearance of the colour of the
Synthesis of silver nanoparticles reaction medium was the direct indication of taking off of
In the microwave assisted synthesis of silver nanoparticles, the dye. The reaction kinetics was examined by measuring
90 ml of 1 mM AgNO3 and 10 ml of leaf extract were taken in absorbance at 515 nm of the reaction mixture at definite
a 250 ml beaker and stirred well. It was irradiated in a domes- time gaps.
tic microwave oven (Sharp R-219 T (W)) operating at a power
of 800 W and frequency 2450 MHz. The generation of silver
Antioxidant capacity using the DPPH assay
nanoparticles was visualized by the onset of brownish yellow
colour to the reaction mixture. The monitoring and confirm- Capacity of AgNP-E. scaber and E. scaber to scavenge free radi-
ation of the formation of silver nanoparticles were done by cals was ascertained using DPPH method [21]. The DPPH free
taking the UV-vis spectra after different microwave irradiation radical has red colour which turns yellow on its conversion to
time. The synthesized silver nanoparticles (AgNP-E. scaber) hydrazine molecule. Different concentrations of AgNP-E.
were purified by repeated centrifugation in a refrigerated scaber and E. scaber (12.5, 25, 50, 100 and 200 lg/mL) were
ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY 797

mixed with 0.1 mM DPPH solution in DMSO under dark condi- MTT assay. The cells were purchased from National Centre for
tion. After 20 min of incubation at room temperature the Cell Sciences (NCCS), Pune, India and was maintained in
reduction in colour intensity at 517 nm was measured and it Dulbecco’s modified Eagles medium (Gibco, Invitrogen).
was a direct measurement of the scavenging potential of the Monolayer of two day old confluent cells was trypsinized.
AgNP-E. scaber/E. scaber in terms of hydrogen donating ability. Hundred microlitres of cultured cell suspension (5  104 cells/
Ascorbic acid (10 mg/mL DMSO) was used as the reference. wells) was seeded in 96 well plates and were kept at 37  C in
Three millilitres of DPPH was taken as control. Inhibition (%) of a humidified 5% CO2 incubator (NBS Eppendorf, Germany).
the DPPH radical was calculated by the formula Various amounts of the plant extract and silver nanoparticles
 (6.25, 12.5, 25, 50 and 100 lg/mL) were added to the grown
Inhibition ð%Þ ¼ AbsControl – AbsSample =AbsControl  100 (1)
cells and incubated for 24 h. The untreated cells constitute
The experiment was repeated thrice and IC50 values were the control medium. The experiment was performed thrice.
estimated using Graph pad Prism software. The cytotoxicity was directly observed in an inverted phase
contrast tissue culture microscope (Olympus CKX41 with
Antimicrobial potential evaluation Optika Pro5 CCD camera) and the microscopic observations
were recorded as images. Any detectable changes in the
Biological significance of the synthesized silver nanoparticles morphology of the cells, such as rounding or shrinking of
as antibacterial and antifungal agents was accessed through cells, granulation and vocalization in the cytoplasm of the
the agar well diffusion method [22]. Microorganisms were cells were considered as indications of cytotoxicity. The viabil-
procured originally from Microbial Type Culture Collection, ity of cells were also evaluated by MTT assay [23]. The
Institute of Microbial Technology, Chandigarh, India. Sterile absorbance at 570 nm was measured by using a micro-plate
and solidified Muller Hinton agar (20 ml) plates were swabbed reader. Percentage inhibition was calculated by Equation (2).
with Bacillus subtilis (MTCC 441), Lactococcus lactis (MTCC The percentage of growth inhibition,
3041), Pseudomonas fluorescens (MTCC 2421) and
Mean OD of Samples  100
Pseudomonas aeruginosa (MTCC 424). Potato Dextrose agar % viability ¼ (2)
plates was swabbed by overnight grown Aspergillus flavus Mean OD of control
(MTCC 277) and Aspergillus penicillioides (ITCC 430). Five wells The IC50 Values were calculated for each set of values
of 6 mm were bored using a sterile well cutter and the wells using Graph pad Prism software and Mean ± standard devi-
A, B and C were loaded with E. scaber aqueous extract ation was identified.
(0.05 mg/mL, 50 lL), AgNP-E. scaber (1 mg/mL, 50 lL) and
AgNP-E. scaber (1 mg/mL, 70 lL), respectively. Chemical anti-
biotic streptomycin and antimycotic griseofulvin (10 mg/mL, Statistical analysis
50 lL) were used as the positive control for bacterial and fun- Biological data was expressed as mean ± standard deviation
gal plates respectively (well D). Millipore water (50 lL) added
and was analyzed by one way ANOVA followed by Tukey’s
to well E constituted the negative control since all the solu-
Post hoc analysis using Graph pad Prism software. A value of
tions were prepared in it. After 24 h and 1 week of incubation
p < .05 was considered as statistically important.
span the zone of inhibition was measured in mm for bacterial
and fungal stains, respectively.
Results and discussions
In-vitro anticancer activity determined by MTT assay Synthesis of silver nanoparticles from leaf of E. Scaber
In-vitro anticancer effect of E. scaber extract and the synthe- The aqueous leaf extract of E. scaber is used for the phytosyn-
sized silver nanoparticles were experimented using human thesis of silver nanoparticles from AgNO3. Upon microwave
skin cancer cell lines A375 and fibroblast cell lines L929 using irradiation of 1 min duration the silver nanoparticle formation

Figure 1. (a) UV-vis spectral plot of AgNP-E. scaber synthesised by microwave irradiation, the photograph in the inset shows the colour change on nanoparticles for-
mation, (b) FT-IR spectra of (1) E. scaber and (2) AgNP- E. scaber.
798 S. FRANCIS ET AL.

Figure 2. The photoluminescence spectra of AgNP-E. scaber (a) excitation spectrum, (b) emission spectrum under an excitation of 360 nm, (c) emission spectrum
under an excitation of 450 nm and (d) The powder XRD pattern of AgNP-E. scaber.

immediately starts functioning. The preliminary observation and AgNP-E. scaber. So it is proved that E. scaber leaf extract
to detect the formation of the silver nanoparticles is the rapid is a powerful reducing agent in the silver nanoparticles syn-
colour change of the colourless reaction mixture to brownish thesis. These functional groups of various organic compounds
yellow. The photograph of the reaction mixture before and present in the leaf extract play a key role in the stabilization
after microwave irradiation is given in the inset of Figure 1(a). the AgNP-E. scaber too. The broad peak formed between
The room temperature analogue of the same reaction was 3250–3350 cm1 is due to stretching vibrations of O–H.
performed by keeping the reaction mixture for 4 h at room The peak at 1738.4 cm1 arose from C ¼ Ostretching of the
temperature. No characteristic colour change was observed, plant constituents. The medium peak at 1590.82 cm1 is from
indicating no nanoparticles formation at this temperature. C ¼ Cstretching of aromatic rings. All predominant peaks in the
plant leaf extract were present in the AgNP-E. scaber showing
the capping and stabilizing effect of the plant extract. It is
Characterization of silver nanoparticles observed that most of the IR bands in AgNP-E. scaber have
undergone a slight shift in wave number from that of plant
UV-vis spectral analysis
extract because of its surface modification. The exact mech-
The most common technique used to elucidate and confirm
anism of the reduction process needs separation, purification
the formation of silver nanoparticles is UV-vis spectroscopy.
and identification of phytocomponents present in the aque-
We can detect a strong peak at 420 nm and the Surface
ous leaf extract of E. scaber. The reduction reaction can be
Plasmon Resonance peak arises from the oscillations of sur-
written as follows
face electrons of silver metal in an electromagnetic environ-
ment. As the time of irradiation advances, both the colour of
the reaction mixture and the intensity of the absorption max-
imum increases. The time dependent UV-vis spectral plot of
AgNP-E. scaber synthesized by microwave irradiation is
shown in Figure 1(a). The microwave assisted nanoparticle
synthesis is an economic rapid strategy and here it works rea- Photoluminescence and crystallographic analyses
sonably well. The synthesized silver nanoparticles were observed to be
photoluminescent in aqueous medium. Photoluminescent
emission shown by silver nanoparticles greatly dependent on
FT-IR spectral analysis excitation wavelength [24]. Figure 2(a) represents the excita-
The FT-IR spectrum of E. scaber and AgNP-E. scaber are tion spectrum of silver nanoparticles, measured in such a way
shown in Figure 1(b). It is seen that the same phytochemicals that the wavelength of emission was maximum [25].
and functional groups are present in both the leaf extract Photo excitation of AgNP-E. scaber using excitation
ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY 799

Figure 3. (a–e) TEM images of different magnifications, (f) HR-TEM image, (g) SAED pattern, (h) EDX spectrum, (i) particles’ size distribution, (j) and (k) AFM images
of AgNP-E. scaber.

wavelength 360 nm produced a very intense fluorescent particle size of 37.86 nm. The well separated particles noticed
emission peak at 467 nm (Figure 2(b)), while excitation in TEM images supported the vital role of phytochemicals in
through 450 nm produced fluorescence emission peak at E. scaber in the stabilization and capping actions of the nano-
523 nm with reduced intensity (Figure 2(c)). Silver nanopar- particles. The lattice fringes were clearly seen in the HR-TEM
ticles prepared and stabilized using chemical methods pro- portrait (Figure 3(f)). The Selected Area Electron Diffraction
duced fluorescent emission peaks in harmony with our (SAED) pattern (Figure 3(g)) with bright circular rings offered
findings which foster the optoelectronic value of microwave additional evidence to the crystalline nature of AgNP-E.
assisted and plant reduced silver nanoparticles [24,26]. In the scaber along with its preferred orientation towards (111)
case of silver nanoparticles the luminescence emission arises plane. The strong peak at 3 keV in Figure 3(h) confirmed the
from the electron-hole recombination processes, the electron elemental presence of silver. Particle size histogram
from sp conduction band above the Fermi level and hole (Figure 3(i)) provided the distribution of particles’ size; the
from d-band below the Fermi level [26,27]. Optical properties diameter of spheres mainly ranged between 20 and 60 nm.
shown by silver nanoparticles open a new coridore in laser The surface topography and shape of the synthesized sil-
technology and bioimaging processes [28]. ver nanometal was accustomed by the AFM images [29]. It
The crystalline nature of the silver nanoparticles is excel- clearly validated its spherical shape and nano scale diameter
lently proved by XRD analysis (Figure 2(d)). The 2h values (Figure 3(j,k)). The AFM images agreed with the results
were 38.19 , 44.19 , 64.70 and 76.92 , and they correspond obtained by the TEM analysis.
to (111), (200), (220) and (311) planes of fcc lattice of AgNP-E.
scaber.
Catalytic activity of silver nanoparticles
Microscopic and EDX analyses Reduction of nitro compounds catalyzed by AgNP-
Transmission electron microscopy is a powerful tool to eluci- E. Scaber
date the surface morphology and size of nanoparticles. Catalytic Reduction of 4-nitrophenol to 4-nitroaniline by
Figure 3(a–e) proved that the synthesized nanoparticles pos- AgNP-E. scaber: 4-nitrophenol is an important starting mater-
sess more or less spherical geometry and has an average ial for the preparation of many organic compounds and is
800 S. FRANCIS ET AL.

Figure 4. Time-based UV-vis spectral images for the reduction of (a) 4-nitrophenol (b) 2-nitroaniline and (c) 4-nitro anilines by NaBH4 catalyzed by AgNP-E. scaber
(0.02 mg/mL). ln [a] against time plots are shown in the inset.

inevitable in pesticides and leather industries. Incisive inhal-


ation of 4-nitrophenol causes headache, nausea, dullness,
etc., in human beings. The catalytic activity of the green syn-
thesized noble silver metal was authenticated by the reduc-
tion of 4-nitrophenol by NaBH4. When the reducing agent
NaBH4 was added to 4-nitrophenol the corresponding pheno-
late ion was formed by hydrogen ion abstraction and the
kmax shifted from 317 nm to 400 nm.
To manifest the catalytic activity of AgNP-E. scaber
(0.02 mg/mL), UV-vis spectra of the reaction mixture were
recorded at regular intervals and displayed in Figure 4(a).
From the figure it was evident that within a short period of
time the reduction process is completed and the peak corre-
sponding to amino group of 4-aminophenol is strengthened
at 295 nm. The observed isobestic points 228, 242, Figure 5. UV- vis spectra and ln [A] versus time plot for the degradation of
278 and 312 nm proved that the only product formed was eosin Y using NaBH4 catalyzed by AgNP-E. scaber (0.02 mg/mL).
4-aminophenol [12]. The kinetics of the reaction was assessed
as pseudo first order from the linearity of the graph between shifted to longer wavelengths that is 380 and 412 nm,
ln[A] and time. The pseudo first order kinetics is justified respectively [30,31].
since the concentration of reductant used was excess in com- In the present catalytic study the reduction reactions
parison with the reactant. The kinetic analysis showed that were continuously monitored using time dependent UV-vis
the rate constant was 0.2794 min1. spectral analysis. It is observed from Figure 4(b,c) that the
Reduction of 2- or 4-nitro anilines By NaBH4: Nitroanilines depletion of peak intensity at 380 and 412 nm is the direct
are important organic compounds used for the preparation indication of progress of the reduction reaction. The peak
of azo dyes. When freshly prepared NaBH4 is added to aque- around 300 nm in the case of phenylenediamines was
ous solution of 2- or 4-nitroanilines, the yellow color of the found to augment in its intensity with time. This was
reaction mixtures get amplified. As a result of charge transfer attributed to the formation of the corresponding diamino
transition the absorption maxima in the UV-vis spectra were derivatives. The completion of the reduction processes
ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY 801

were confirmed by the absolute discoloration of the reac-


tion mixtures. The nanocatalyst functioned as a relay sta-
tion between the electron donor (BH 4 ) and the organic
electron acceptor moiety. It was evident from the graphs
that 2-nitroaniline undergoes reduction at a slower rate
(25 min) compared to 4-nitroaniline (7 min). This may be
due to the effect of the substituent on the ortho position.
The reduction of 2- or 4-nitroanilines produced the corre-
sponding phenylenediamines. The pseudo-first order reduc-
tion kinetics were asserted [32] for 2- and 4-nitroanilines;
from the linear plots of ln[A] against time at absorbance
380 and 412 nm. The rate constants obtained from the
slope of the graph were 0.0751 and 0.2788 min1 and cor-
Figure 6. Antioxidant capacity of E. scaber and AgNP-E. scaber (12.5, 25, 50, 100 relation coefficients were 0.9965 and 0.9923, respectively
and 200 lg/mL) evaluated using DPPH assay. The values are given as the for the reduction of 2- and 4-nitroanilines.
mean ± SD (n ¼ 3).

Figure 7. (a) Photographs of the tested antimicrobial plates, where A¼ 50 lL of E. scaber (0.05 mg/mL), B ¼ 50 lL of AgNP-E. scaber (1 mg/mL), C¼ 70 lL of AgNP-E.
scaber (1 mg/mL), D¼ 50 lL of Streptomycin/Griseofulvin (10 mg/mL), E ¼ 50 lL of Millipore water (b) Zone of inhibition in mm for the antibacterial and antifungal
studies. The values are expressed as mean ± SD (n ¼ 6).
802 S. FRANCIS ET AL.

Figure 8. Morphological changes induced on (a) treated A375 cells by aqueous leaf extract of E. scaber, (b) treated L929 cells by AgNP-E. scaber (c) treated A375
cells by AgNP-E. scaber observed by inverted phase contrast tissue culture microscope after an incubation period of 48 h.

Reduction of fluorescent dye - eosin Y (k) and co-relation coefficient (r2) calculated from the graph
Eosin Y is an acidic xanthene dye which shows kmax at were 0.2173 min1 and 0.9977, respectively [33].
515 nm. The catalytic efficacy of the synthesized silver nano-
particles was also evaluated using the reduction of eosin
Antioxidant power evaluation – DPPH assay
Y by NaBH4. UV-vis spectrometric scans were recorded con-
tinuously one after another till the reaction was finished Free radical scavenging capacity of E. scaber and AgNP-E.
(Figure 5) indicated by decoloration of the reaction mixture. scaber were compared with the standard antioxidant ascorbic
A linear graph was obtained when ln[A] was plotted against acid (Figure 6). The IC50 values for E. scaber and AgNP-E.
time, where A is the absorbance at 515 nm. The rate constant scaber obtained using Graph pad Prism were 73.47 and
ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY 803

morphological changes induced on treated A375 cells by


AgNP-E. scaber (Figure 8(a)) and E. scaber (Figure 8(c))
observed by inverted phase contrast tissue culture micro-
scope after 48 h incubation period. Figure 8(b) showed the
effect on treated L929 cells by AgNP-E. scaber and Figure 9
displayed the cell viability (%) curve. IC50 values obtained for
AgNP-E. scaber and E. scaber when A375 cells were used are
15.68 ± 0.15 lg/mL and 50.55 ± 0.17 respectively. IC50 value
corresponding to AgNP-E. scaber when L929 cell lines were
used is 65.49 ± 0.40 lg/mL. The synthesized AgNP-E. scaber
showed a superior anticancer property than plant E. scaber
towards the skin carcinoma cell line A375 [39].

Conclusions
Figure 9. Cell viability (%) plot of E. scaber leaf extract and AgNP-E. scaber
towards A375 and L929 cells. The plotted values are the mean ± SD (n ¼ 3). The microwave assisted synthetic strategies of nanoparticles
preparation were exploited in the case of AgNP-E. scaber. The
6.629 lg/mL, respectively. Ascorbic acid produced an IC50 XRD studies showed that the synthesized AgNP-E. scaber has
value of 15.07 lg/mL. AgNP-E. scaber exhibited significantly a face centered cubic lattice. The spherical shape and the
higher antioxidant activity than the plant extract E. scaber for nano-regime size of the silver nanoparticles were well proved
all concentrations. The plant displayed an appreciable by TEM images. The adverse environmental impacts of
amount of antioxidant potential, since it is rich in flavonoids organic nitrocompounds can be easily rectified by applying
and phenols [17]. The antioxidant activity gets increased with AgNP-E. scaber nanocatalyst. The dye eosin Y can be rapidly
increasing the concentration of AgNP-E. scaber. Antioxidant and successfully degraded using AgNP-E. scaber catalyst. The
performance shown by E. scaber and AgNP-E. scaber offered synthesized silver nanoparticles are beneficial to mankind by
a new hope in medical and pharmaceutical industry. their appreciable antimicrobial and antioxidant potential.
Further, we have succeeded in establishing their ability to
manage skin cancer cell lines A375, in-vitro manner. The
Antimicrobial efficacy evaluation
green synthetic pathway makes the AgNP-E. scaber more
The effectiveness of AgNP-E. scaber as antimicrobial agents adequate for many therapeutic and biological applications.
towards various pathogenic organisms was displayed in In-vivo studies and mores investigations are needed for the
Figure 7(a). High zone of inhibition shown by AgNP-E. scaber clinical adaptation of the results.
(Figure 7(b)) affirmed its superior antimicrobial power. AgNP-
E. scaber hindered the growth of both gram negative and Acknowledgements
gram positive microorganisms. The inhibitory zone was
accredited to the phytochemicals flavonoids, alkaloids and The financial assistance to Sijo Francis from University Grants
Commission (under UGC-FDP), Government of India is gratefully
polyphenols and they were present in the aqueous plant
acknowledged.
extract that capped the silver nanoparticles. A dose-depend-
ent increase in the inhibition (%) was observed in the experi-
ment [34]. The antibacterial power has a prominent value Disclosure statement
than the antifungal activity [35]. Several explanations for the The authors’ report states that they have no conflict of interest and they
toxic activity of the silver nanoparticles were known. They are alone responsible for content and write up of the paper.
can penetrate through the cell wall and cause structural
changes by interacting with sulphur and phosphorous con-
taining biomolecules [36]. Another studies involving EPR Funding
spectra suggested that the generation of free radicals by the The financial assistance to Sijo Francis from University Grants
nanoparticles when it contact with the bacteria was respon- Commission (under UGC-FDP), Government of India is gratefully
acknowledged.
sible for the damage of cell membranes [37,38]. Antimicrobial
properties of E. scaber and AgNP-E. scaber can effectively be
tapped in many medical applications and consumer product References
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