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Current Understanding of Synthesis and Pharmacological Aspects of Silver
Nanoparticles
Article · October 2013
DOI: 10.13140/2.1.3744.0964
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2. American Journal of Phytomedicine and Clinical Therapeutics www.ajpct.org
Review Article
Current Understanding of Synthesis and
Pharmacological Aspects of Silver
Nanoparticles
Shyam Baboo Prasad1
, Vidhu Aeri2
, Yashwant1*
1
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara (Punjab).
2
Faculty of Pharmacy, Jamia Hamdard, Hamdard Nagar, (New Delhi).
ABSTRACT
Silver nanoparticles (SNPs) are one of the most frequently used
nanomaterials because of their antimicrobial properties. It can be
prepared by physicochemical and biological method. Silver ions has
toxic effects on many pathogens, including bacteria, viruses, and
fungi. Because of relatively low toxicity of SNPs in humans, it has
been used in various medical applications. Decrease in particle size
of silver nanoparticles may leads to increase in antibacterial activity
which has been associated with the increasing surface area to mass
ratio. As various diseases is triggered by these microorganism so
pharmacological evaluation of silver nanoparticles may be fruitful in
those disease which occurs due to bacteria, fungi and virus. The
present review deals with various method of preparation and reported
preclinical activity of silver and its derivatives.
Keywords: Silver nanoparticles, Nano crystalline silver,
Nanomedicine, Metallic nanoparticles, Silver.
INTRODUCTION
Nanotechnology is emerged as a
fastest growing field with numerous
applications in science and technology for
manufacturing new materials.
Nanotechnology is defined as the design,
characterization and application of
structures, devices and systems by
controlling shape and size at 1 to 100 nm1
.
Modern era is of nanomedicine owing to
their various therapeutic applications with
more efficacies and lesser side effects. The
popularity is due to their potential for
achieving specific process and selectivity in
pharmacological action2
. Metallic
nanoparticles, including gold, silver, iron,
zinc and metal oxide nanoparticles, have
shown great promise in biomedical
application, due to their large surface area to
volume ratio2
.SNPs or nanosilver (NS) are
emerging as one of the fastest growing
product in nanotechnology industry. In daily
life NS is used in room spray, wall paints,
water purifier and laundry detergent.SNPs
are also incorporated in textiles for
manufacturing of cloth, vests, underwear
and socks. It is estimated that of all nano
Address for
Correspondence
School of
Pharmaceutical
Sciences, Lovely
Professional
University, Phagwara
(Punjab)
E-mail: yash99yash
@gmail.com

3. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
materials in medical and healthcare sector,
NS application has higher degree of
commercialization. A wide category of
product is already available in market. In
medical sector they are used in wound
dressing, contraceptive devices, surgical
instrument and bone prostheses. SNPs or NS
are being used increasingly in wound
dressings, catheters, and various household
products due to their antimicrobial
activity2
.The antibacterial property of silver
has been known for thousands of years with
the ancient Greeks cooking from silver pot.
The antimicrobial properties of silver were
utilized as early as 1000 BC to keep water
safe. This is recently attributed to the anti-
microbial activities of released Ag+ ions.
The first recorded medicinal use of silver
goes back to 8th century3
. Silver nitrate was
used to treat ulcers in 17th and 18th century.
More recently silver is used as a biocide to
prevent infection in burns, traumatic wounds
and diabetic ulcers 4
. Silver interacts with
structural proteins and preferentially binds
with DNA bases to inhibit replication.
Furthermore, bactericidal effect of silver has
also been attributed to inactivation of the
enzyme phosphomannoseisomerase5
.
Presently silver is considered a non-essential
accumulative element. Silver is widely
distributed in human body fluid and tissues
including heart, lungs, aorta, blood,
erythrocytes, plasma, bones, brain, breast,
caecum, oesophagus, colon, diaphragm,
duodenum, hair, ileum, larynx, kidney,
urinary bladder, urine, liver, pancreas,
adrenal gland, thyroid gland, lymph nodes,
muscles, nails ovary, prostate gland,
rectum, serum, skin, spleen, testes, teeth
(dentine and enamel), trachea, uterus etc.
Such wide distribution in the human body
suggests that this metal could have some
specific functions which are not clear at
present6
.
There has been a resurgence of the
promotion of silver (as colloidal silver) as an
alternative medicine since 1990’s. Colloidal
silver has been marketed with claims that it
can treat various diseases being an essential
mineral supplement7,8
. Although colloidal
silver products are legally available as health
supplements, it is illegal in the U.S. to make
such claims of medical effectiveness for
colloidal silver. The commercial product
referred to as colloidal silver includes
solutions that contain various concentrations
of ionic silver compounds. Unlike other
modern medicine, the manufacturing of
colloidal silver is not standardized and thus
results in various concentrations and particle
sizes. At present, there are no evidence-
based medical uses for ingested colloidal
silver. Indeed, the U.S. National Center for
Complementary and Alternative Medicine
has issued an advisory indicating that the
marketing claims made about colloidal
silver are scientifically unsupported7
.
Method of preparation of SNPs
SNPs can be prepared by traditional
Ayurvedic literature, physico-chemical
method and biological method.
Traditional Ayurvedic method
Bhasma is the calcination product of
inorganic or organic substances and claimed
to be biologically produced nanoparticles.
Silver nanomedicine of ancient Ayurveda is
known as raupyabhama. It is prepared by
methods described an Ayurvedic text in
RasendrasaraSamagraha.Pure silver leaves
are mixed with equal quantity of sulphur (by
weight) and one half quantity of arsenic
trisulphide, soaked in lemon juice and
subjected to calcination process in sealed
earthen containers. The material is scraped
after cooling, pulverized, mixed with half its
weight of sulphur and arsenic trisulphide,
soaked in lemon juice and calcined again. On
cooling the scraped material is triturated with
lemon juice and subjected to calcination. This
process is repeated fourteen times to procure

4. Yashwant et al______________________________________________ ISSN 2321 – 2748
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the Bhasma. Thorough trituration at each
stage is necessary to ensure proper quality of
Bhasma9
.
Physicochemical syntheses of SNPs
Nanoparticles are mainly prepared by
physical and chemical methods. Silver
nanoparticles can be prepared by both
methods so-called ‘top-down’ and ‘bottom-
up’ methods. The top-down method deals
with the mechanical grinding of bulk metals
and subsequent stabilization of the
subsequentnanosized metal particles by the
addition of colloidal protecting agents10,11
. On
the other hand the bottom-up methods deals
with reduction of metals by electrochemical
methods, and sonodecomposition. There are
numerous physical and chemical approaches
for synthesis of silver nanoparticles reported
in Table 1.
Biological synthesis of SNPs
SNPs formed by chemical and
physical methods are very expensive and also
include the use of toxic chemicals. Moreover
it must be available at lower cost for their
effective utilization; thus, there is a need for
an environmentally and economically feasible
way to synthesize these nanoparticles. Apart
from chemical and physical methods, SNPs
can also be synthesized by biological
methods.Biological synthesis of SNPs is
possible with help of bacteria, fungi, and plant
extracts. Biological synthesis of silver
nanoparticles is a bottom-up method that
includes reduction/oxidation reactions. The
microbial enzymes or phytochemicals with
antioxidant or reducing properties act on the
corresponding compounds and give the
anticipated nanoparticles17,18
.
Synthesis of SNPs using bacteria
The mechanism behind synthesis of
SNPs is the presence of the nitrate reductase
enzyme which converts nitrate into nitrite.
The alpha-nicotinamide adenine dinucleotide
phosphate reduced form (NADPH) dependent
nitrate reductase present in bacteria
converted nitrate into nitrite and the electron
is transferred to the silver ion. So, the silver
ion is reduced to silver (Ag+ to Ag0
)19
.SNPs
synthesized using bacteria is shown in table 2.
Synthesis of SNPs using Fungi
The exact mechanism involved in
synthesis of SNPs using fungi is not clear. But
it is believed that the mechanism involve in
synthesis of SNPs by fungi is trapping of Ag+
ions at the surface of the fungal cells and the
subsequent reduction of the silver ions by the
enzymes present in the fungal system. Fungi
can produce larger amounts of nanoparticles
as compared to bacteria as they can secrete
larger amounts of proteins which directly
translate to higher yield of nanoparticles.
Considering the example of F. oxysporum, it
is believed that the NADPH-dependent nitrate
reductaseprocess are responsible for
nanoparticle formation30,31,32
. Different fungi
helpful in synthesis of SNPs are reported in
Table 3.
Synthesis of SNPs using Plant
Synthesize SNPs using microbes is
very slow process when in comparison with
plant extracts.
The major advantage of using plant
extracts for silver nanoparticle synthesis is
that they are easily available, safe, and
nontoxic in most cases, have a broad variety
of metabolites that can aid in the reduction of
silver ions, and are quicker than microbes in
the synthesis. Though the exact mechanism
involved in each plant varies as the
phytochemical involved varies. The major
mechanism involved is the reduction of the
ions. The main phytochemicals involved are
terpenoids, flavones, ketones, aldehydes,
amides, and carboxylic acids. Flavones,
organic acids, and quinones are water-soluble
phytochemicals that are responsible for the
immediate reduction of the ions. Studies have

5. Yashwant et al______________________________________________ ISSN 2321 – 2748
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revealed that xerophytes contain emodin, an
anthraquinone that undergoes
tautomerization, leading to the formation of
the silver nanoparticles. In the case of
mesophytes, it was found that they contain
three types of benzoquinones: cyperoquinone,
dietchequinone, and remirin. It was suggested
that the phytochemicals are involved directly
in the reduction of the ions and formation of
silver nanoparticles41
. SNPs synthesized using
plant is shown in Table 4.
Pharmacological Aspects
Silver NPs interact with HIV-I
Recently it was reported by
Elechiguerra et al. that silver NPs in a size
range 1–10 nm bind with HIV-I in a size
dependent fashion. For NPs preparation with
various surface modifications, they have
shown that silver concentrations over 25
µg/ml significantly inhibited HIV-1 infection.
Bare silver NPs showed superior effect,
whereas surface modification with BSA and
PVP showed moderate effect. This is a
promising study that explores potential use of
silver NPs towards millions of people
suffering from AIDs52,53
.
Antifungal
NS showed potent activity against clinical
isolates and ATCC strains of Trichophyton
mentagrophytes and Candida species (IC80,
1-7 µg/ml). The activity of nano-Ag was
comparable to that of amphotericin B, but
superior to that of fluconazole (amphotericin
B IC80, 1-5 µg/ml; fluconazole IC80, 10- 30
µg/ml). Additionally, author investigated their
effects on the dimorphism of Candida
albicans. The results showed nano-Ag
exerted activity on the mycelia54
.
Assessment of Antioxidant Activity
The rats were sacrificed on 15th day
after collection of blood samples and their
livers were excised immediately, washed in
ice cold Phosphate buffered saline (pH 7.4),
blotted dry, and weighed. A 10% w/v of liver
homogenate was prepared in 0.15 M Tris-HCl
buffer (pH: 7.4). The homogenate was
centrifuged at 2000×g for 20 min at 4 °C to
remove the cell debris and then the
supernatant was centrifuged (REMI C-24) at
12,000×g for 1 h at 4 °C. The supernatant
obtained were used for the determination of
lipid peroxidation23
, reduced glutathione
(GSH)24
,superoxide dismutase (SOD)25
and
catalase (CAT)26
.
Antibacterial activity
In order to achieve better antibacterial
activity, water insoluble nano-composites of
Ag, Ag/SiO2 hybrid and Ag colloid
nanoparticles were studied. Antibacterial
effectiveness of the Ag/SiO2 nanoparticles
was tested against general Escherichia coli (E.
coli ATCC 25922) and E. coli O157:H7 by
measuring the growth based through optical
density and digital counting of live-dead cells
using a fluorescent microscope, and a field
emission scanning electron microscope.
Minimum inhibitory concentrations values
against four representative bacteria along with
E. coli O157:H7. Results showed that Ag NPs
of 6.6±4.5 nm were attached to the surface of
SiO2 nanoparticles (74±13.5 nm) and the Ag-
colloid NPs (3.5±2 nm) showed excellent
antibacterial properties55
.
Antiplatelet activity
Thrombotic disorders have emerged
as serious threat to society. As anticoagulant
and thrombolytic therapies are usually
associated with serious bleeding
complications, the focus has now shifted to
regulating and maintaining platelets in an
inactive state. In a study ShrivastavaS et-al.
showed that NS has antiplatelet property and
effectively prevents integrin-mediated platelet
responses, both in vivo and in vitro, in a
concentration-dependent manner.
Ultrastructural studies show that NS

6. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
accumulates within platelet granules and
reduces interplatelet proximity. Our findings
further suggest that these nanoparticles do not
confer any lytic effect on platelets and thus
hold potential to be promoted as
antiplatelet/antithrombotic agents after careful
evaluation of toxic effects56
.
Anti-proliferative activity
Normal human lung fibroblasts (IMR-
90) and human glioblastoma cells (U251)
were exposed to different doses of SNPs in
vitro. Uptake of SNPsoccurred mainly
through endocytosis, accompanied by a time
dependent increase in exocytosis rate. The
electron micrographs revealed a uniform
intracellular distribution of SNPsboth in
cytoplasm and nucleus. SNPstreated cells
exhibited chromosome instability and mitotic
arrest in human cells. There was efficient
recovery from arrest in normal human
fibroblasts whereas the cancer cells ceased to
proliferate. Toxicity of SNPsis mediated
through intracellular calcium (Ca2+)
transients along with significant alterations in
cell morphology and spreading and surface
ruffling. Down regulation of major actin
binding protein, filamin was observed after
SNPsexposure. SNPsinduced stress resulted
in the up regulation of metallothionein and
hemeoxygenase -1 genes57
.
Cytotoxicity and genotoxicity of silver NPs in
human cells
The toxicity of starch-coated SNPs
was studied using normal human lung
fibroblast cells (IMR-90) and human
glioblastoma cells (U251). The toxicity was
evaluated using changes in cell morphology,
cell viability, metabolic activity, and
oxidative stress. SNPsreduced ATP content of
the cell caused damage to mitochondria and
increased production of reactive oxygen
species (ROS) in a dose-dependent manner.
DNA damage, as measured by single cell gel
electrophoresis (SCGE) and cytokinesis
blocked micronucleus assay (CBMN), was
also dose-dependent and more prominent in
the cancer cells. The SNPs treatment caused
cell cycle arrest in G(2)/M phase, possibly
due to repair of damaged DNA. Annexin-V
propidium iodide (PI) staining showed no
massive apoptosis or necrosis. The
transmission electron microscopic (TEM)
analysis indicated the presence of Ag-NPs
inside the mitochondria and nucleus,
implicating their direct involvement in the
mitochondrial toxicity and DNA damage. A
possible mechanism of toxicity is proposed
which involves disruption of the
mitochondrial respiratory chain by Ag-NPs
leading to production of ROS and interruption
of ATP synthesis, which in turn cause DNA
damage. It is anticipated that DNA damage is
augmented by deposition, followed by
interactions of Ag-NPs to the DNA leading to
cell cycle arrest in the G(2)/M phase58
.
Antiangiogenic activity
Silver and gold NPs were found to
display unique physical and biological
properties that have been extensively studied
for biological and medical applications. Gold
and silver NPs were prepared by chemical
reductants that utilize excess toxic reactants,
which need to be removed for biological
purposes. Author utilized an ideal method
involving a single synthetic step to prepare
metal NPs for evaluating potential effects on
angiogenesis modulation. These NPs were
prepared by reducing silver nitrate and gold
chloride with diaminopyridinyl (DAP)
derivatized heparin (HP) polysaccharides.
Both gold nanopartclea and SNPs reduced
with DAPHP exhibited effective inhibition of
basic fibroblast growth factor (FGF-2)-
induced angiogenesis, with an enhanced anti-
angiogenesis efficacy with the conjugation to
DAPHP (P<0.01) as compared to glucose
conjugation. These results suggest that
DAPHP-reduced silver NPs and gold NPs
have potential in pathological angiogenesis

7. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
accelerated disorders such as cancer and
inflammatory diseases59
.
Analgesic activity
Investigations were carried out by
Nadeem et al against four types of noxious
stimuli: mechanical (tail clip), chemical
(acetic acid-induced writhing), electrical
(pododolorimeter) and thermal (Eddys hot
plate and analgesiometer) in rats and mice.
Effects following naloxone pre-treatment and
maximum tolerated dose (MTD) were also
studied. A perusal of the results show that the
test drugs exhibited analgesic activity against
chemical, thermal and electrical stimuli but
such effects were not discernible against the
mechanical stimulus at the doses used. The
all-or-none criteria in the tail clip test were
used. Further the analgesic effects were
abolished / reduced in naloxone pre-treated
animals. These facts point to the involvement
of opiodergic receptors in analgesic actions of
silver preparations60
.
Hypolipidemic activity of silver preparations
in chicks
Three silver preparations (Varak or
foil, ash or Raupyabhasma and sol or
colloidal solution) were fed to three groups of
young, male chicks for 10 days. There was
significant fall in all the plasma lipid
fractions--total lipids, phospholipids,
triglycerides and total cholesterol. There was
a marked rise in silver content of plasma and
whole blood, ranging from 4 to 13 times,
suggesting that the observed hypolipidemic
action may be due to silver. The
administration of the three silver preparations
did not cause any retardation in growth, toxic
manifestation, side effect or any untoward
reaction61
.
Neuropsychobehavioural effects
Silver preparations used in Ayurveda
and Unani-Tibb showed anticataleptic and
growth
promoting effects without gross or subtle
toxicities, weight loss, sedation, motor deficit,
aggression or ill effects on cognitive
functions. The test drugs (50 mg/kg, p.o.)
caused significant reduction of haloperidol-
induced catalepsy in rats. Incorporation in the
diet of rat pups (1% w/w for 6 weeks) lead to
significantly higher growth rate when
compared to control animals. No appreciable
effects were discernible on other parameters6
.
Anti-inflammatory activity
Nanocrystalline silver had a direct
anti-inflammatory effect in the porcine
contact dermatitis model that improved the
overall outcome of the healing process62
.
Ulcerative colitis
Nano crystalline silver in dose of 4
mg/kg intracolonically or 40 mg/kg orally,
significantly reduced colonic inflammation
compared to the placeboandno-
treatmentgroups. Sulfasalazine (100mg/kg),
either intracolonically or orally, also reduced
colonic inflammation. Nano crystalline silver
significantly suppressed the expression of
matrix metalloproteinase (MMP)-9, tumour
necrosis factor (TNF)- α, interleukin (IL)-1 β,
and IL-12, whereas sulfasalazine suppressed
MMP-9, IL-1 β, and TNF- α, but not IL-12,
compared to placebo. Nano crystalline silver
administered intracolonically or orally
decreases ulcerative colitis in a rat model and
is as effective as sulfasalazine63
.
TOXICITY
In minute concentration silver is
consider to be non-toxic in normal use. One
of the most important side effect reported for
silver product is argyria. Argyria is
irreversible grey to black colouration of skin
due to deposition of silver in sub dermal
layer. Argyria is just a cosmetic problem it do
not cause any physical harm2
.

8. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
CONCLUSION
Synthesis of SNPs is possible due to
reduction of ion by enzyme present in
microorganism and antioxidant principle
present in plant. The ancient system of
medicine (Ayurveda) has reported silver
based nanomedicine 7th
century BC which is
biologically produced silver based
nanomedicine. SNPs has lots of therapeutic,
biomedical and social value due to its
antimicrobial properties. As microorganism
play important role in spreading of various
disease so special consideration should be
given on SNPs for prophylaxis and treatment
of disease.
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Table 1. SNPs synthesized by different physicochemical method
S.no Methods Size Reference
1
Chemical method of reduction of the metal salt AgBF4 by NaBH4
in water.
3‐40 nm 12
2
Electrochemical method which involves the electro reduction of
AgNO3 in aqueous solution in the presence of polyethylene glycol
10 nm 13
3
Sonodecomposition of an aqueous silver nitrate solution in an
atmosphere of argon‐hydrogen
20 14
4
Electrostatically complexing silver ions with an anionic surfactant
aerosol in extremely stable liquid foam. The foam is then drained
off and reduced by introducing sodium borohydride. These silver
nanoparticles are very stable in solution, suggesting that the
aerosol stabilizes them.
5‐40nm 15
5
Reduction of silver nanoparticles using variable fre‐ quency
microwave radiation.
15‐25 16

12. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
Table 2. Different bacteria for synthesis of SNPs
Organism Size (nm) Reference
B. licheniformis 50 20
Bacillus megaterium 46.9 21
Bravibacteriumcasei 50 22
Escherichia coli 5‐25 23
Enterobacter cloacae 50‐100 24
Klebsiella pneumonia 50 25
Lactobacillus fermentum 11.2 26
Proteus mirabilis 10‐20 27
Plectonemaboryanum 1 to 200 28
P. stutzeriAG259 200 29
Table 3. Different fungi for synthesis of SNPs
Organism Size (nm) Reference
Aspergillusclavatus 10 to 25 33
Aspergillusflavus 7 to 10 34
Aspergillusfumigatus 5 to 25 35
Coriolusversicolor 25 36
F. oxysporum 20 to 50 37
Fusariumsolani 5 to 35 38
Phanerochaetechrysosporium 100 39
Phoma sp. 3.2883 70 40

13. Yashwant et al______________________________________________ ISSN 2321 – 2748
AJPCT[1][7][2013]536‐547
Table 4. Different Plant used in synthesis of SNPs
Plant Size (nm) Reference
Aloe vera 15 to 20 42
Azadirachtaindica 50 43
Carica papaya 15 44
Cinnamomumcamphora leaf 55 to 80 45
Cinnamomumzeylanicum bark 50 to 100 46
Coriandrumsativum leaf 26 47
Desmodiumtriflorum 5 to 20 48
Jatrophacurcas 10 to 20 49
Medicagosativa 2 to 20 50
Piper betle leaf 3 to 37 51
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