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 Chapter 4

Silver Nanoparticles as Multi-Functional Drug Delivery

Systems

Nadezhda Ivanova, Viliana

Nadezhda Ivanova, Gugleva,
Viliana Gugleva,
Mirena Dobreva, Ivaylo Pehlivanov,
Mirena Dobreva, Ivaylo Pehlivanov, Stefan Stefanov
Stefan Stefanov and Velichka Andonova
and Velichka Andonova

Additional
Additional information is available
information is available at
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end of
of the
the chapter
chapter

http://dx.doi.org/10.5772/intechopen.80238

Abstract
Nanoparticles can surmount some essential problems of conventional small molecules
or biomacromolecules (e.g., DNA, RNA, and protein) used in some diseases by allowing
targeted delivery and overcome through biological barriers. Recently, silver nanopar-
ticles have been harnessed as delivery vehicles for therapeutic agents, including anti-
sense oligonucleotides, and other small molecules. Silver is the most profit-oriented
precious metal used in the preparation of nanoparticles and nanomaterials because of its
antibacterial, antiviral, antifungal, antioxidant and unusually enhanced physicochemi-
cal properties compared to the bulk material such as optical, thermal, electrical, and
catalytic properties. Small silver nanoparticles offer many advantages as drug carriers,
including adjustable size and shape, enhanced stability of surface-bound nucleic acids,
high-density surface ligand attachment, transmembrane delivery without harsh trans-
fection agents, protection of the attached therapeutics from degradation, and potential
for improved timed/controlled intracellular drug-delivery. Plant-mediated synthesis of
silver nanoparticles is gaining interest due to its inexpensiveness, providing a healthier
work environment, and protecting human health leading to lessening waste and safer
products. The chapter presents the essential physicochemical characteristics, antibacte-
rial, and anticancer properties which silver nanoparticles obtained by plant-mediated
methods possess, and their application as drug-delivery systems with a critical view on
the possible toxicity on the human body.

Keywords: antibacterial activity, anticancer activity, capping agents, plant extracts,

reducing agents, surface properties, toxicity

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons
© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative
Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,
Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
distribution, and reproduction in any medium, provided the original work is properly cited.
72 Nanomedicines

1. Introduction

Nanomedicine is a branch of medicine that uses nanomaterials and applies nanotechnolo-

gies in prevention, diagnostics, and treatment of diseases [1]. The broad definition of nano-
medicine involves nanoparticles (NPs) as drug delivery systems (DDSs), medical nanosensor,
biochips, insulin pumps, needleless injectors, etc. The unique properties of NPs are related to
their tiny size (generally between 1 and 100 nm), huge surface area, and surface characteris-
tics. “Nano” DDSs provide targeted delivery of optimal dose with reduced side effects and
toxicity. Moreover, NPs solve problems related to drug solubility and bioavailability. These
“nano” carriers can protect the drug from the hazardous environment as well as to overcome
the biological barriers to entry the drug to the targeted tissues and to deal with drug resis-
tance. They possess organic or inorganic origin and can be prepared from different polymers,
metals, ceramics, etc.

Silver is the most profit-oriented precious metal used in the preparation of NPs and nano-
materials. These are known because their antibacterial, antiviral, antifungal, antioxidant, and
unusually enhanced physicochemical properties compared to the bulk material such as opti-
cal, thermal, electrical, and catalytic properties [2–5]. About 500 tons of silver nanoparticles
(AgNPs), used in various industries and everyday life, are produced per year [6, 7]. Rising
demand for silver nanomaterials requires the development of eco-friendly synthesis methods.
In general, AgNPs can be produced by chemical, physical, and biological methods. Chemical
protocols are mainly based on the chemical reduction of Ag+ ions by organic and inorganic
agents, such as sodium borohydride, sodium citrate, sodium ascorbate, elemental hydrogen,
N, N-dimethylformamide, polymers, Tollens methods, etc. The reducing agent reduces Ag+
and leads to the formation of Ag0, metallic silver, which agglomerates into oligomeric clus-
ters. These clusters may form colloidal particles of metallic silver. Different surfactants and
polymers are used to prevent particles from further agglomeration and protect their shape
[8]. The most important physical methods are based on evaporation-condensation technique
and laser ablation of silver bulk material in solution. Both physical approaches did not use
chemical reagents that may be hazardous to the environment and the human body. Although
these require costly specialized equipment, physical methods provide an alternative to envi-
ronmentally unfriendly and time-consuming chemical protocols.

Biological or so-called “green” methods do not use toxic chemicals in the preparation tech-
niques. Moreover, these methods are based on the usage of bacteria, fungi, algae, and plants to
obtain AgNPs characterized by size and shape depending optical, electrical, and antimicrobial
properties [8, 9]. These are based on bioreduction of Ag+ ions in the aqueous medium where
the reducing agents are cited above biological sources. Synthesis of AgNPs using living micro-
organisms (bacteria and fungi) can be performed either intracellularly or extracellularly [10].
The extracellular synthesis is cheaper, less time-consuming, and requires simplified manufac-
turing technology compared to the intracellular synthesis. Studies used culture supernatants
of pathogenic and nonpathogenic microorganisms like A. flavus, B. indicus, B. cereus, Bacillus
strain CS 11, E. coli, P. proteolytica, P. meridiana, S. aureus, etc. [10–12]. The drawbacks of bacte-
rial synthesis of AgNPs are related to the selection and cultivation of suitable bacterial strain,
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 73
http://dx.doi.org/10.5772/intechopen.80238

a mandatory stage of purification, the poor understanding of the mechanisms governing the
nanoparticle formation which hinders scaling laboratory process in the industry as well as the
requirements of highly aseptic conditions and their maintenance [13].
Plant-mediated green synthesis of AgNPs is gaining immense popularity because of its eco-
friendly nature, accessibility, economy, execution-simplicity, and the possibility of large-scale
production. Many studies have used different plant extracts such as Azadirachta indica, Crocus
sativus L., Calliandra haematocephala, Neem leaves, Madhuca longifolia, grape seed extract,
Andean blackberry fruit extract, geranium leaf aqueous extract, marigold flower, etc. [7, 14–19].
The rich phytochemical composition of the extracts used implies its complex action, for exam-
ple, as reducing, stabilizing, and capping agents. The AgNPs thus obtained can be exploited
as DDSs for different active pharmaceutical ingredients.
The chapter presents the essential physicochemical characteristics, antibacterial, and antican-
cer properties, which AgNPs obtained by plant-mediated methods possess, and their applica-
tion as DDSs with a critical view on the possible toxicity on the human body.

2. Plant-mediated synthesis of silver nanoparticles

It is well known that plant extracts have a rich phytochemical composition including pheno-
lics, saponins, terpenoids, flavonoids, catechins, tannins, enzymes, proteins, polysaccharides,
etc. All of these biomolecules take place in a very complicated mechanism of reduction and
stabilization of Ag+ ions to form AgNPs. For example, Li et al. suggested a recognition-
reduction-limited nucleation and growth model to explain the possible formation mechanism
of AgNPs in Capsicum annuum L. extract [19]. According to the authors, the proteins which
have amine groups played a reducing and controlling role during the formation of AgNPs
in the solutions, and that the secondary structure of the proteins changed after reaction with
Ag+ ions. In another study, Mirgorod and Borodina, based on the surface-enhanced Raman
spectroscopy data, stated that the NPs were formed as a result of a redox reaction between
flavonoids and Ag+ ions as well as there are flavonoids near the surface of the AgNPs, which
react complexly with Ag+ ions and with the NPs [20]. Ahmed and co-workers described dif-
ferent approaches of syntheses of AgNPs and protocols employed for their synthesis in detail
[21].
It is important to note that technological parameters such as temperature, pH, the concentra-
tion of Ag+ ions, duration of the obtaining process, phytochemical composition of the extract
used, mechanical stirring, microwave assistance, etc., are crucial both for nanoparticle prepa-
ration and for their characteristics and fate [6, 7, 14–19]. These parameters affect not only the
process of reduction of Ag+ ion and formation of metallic silver but also its agglomeration
into oligomeric clusters which may form colloidal particles with specific features. Amin and
co-workers found that the time of reaction, temperature, and volume ratio of methanol extract
from Solanum xanthocarpum berry to AgNO3 could accelerate the reduction rate of Ag+ ions
and affect the AgNPs size and shape [22]. The NPs were found to be about 10 nm in size,
mono-dispersed in nature, and spherical in shape.
74 Nanomedicines

Surface functionality of nanomaterials is crucial for their applicability, compatibility, and

safety. Generally, surface behavior defines how a nano-entity will interact with biosystems,
environment, etc. [23]. AgNPs are characterized with variable morphology—size, shape,
surface area, purity/coating—and related electrochemical and electromagnetic properties—
charge, zeta potential, redox potential, surface plasmon resonance, and conductivity [24, 25].
A change or intentional attempt to control these essential characteristics is an essential tool
in tailoring AgNPs for specific purposes and might be highly sought on several accounts: (1)
increased stability; (2) increased selectivity; (3) increased therapeutic or diagnostic potency;
(4) enhanced catalytic activity; (5) reduced toxicity; and (6) reduced reactivity [23, 25]. Surface
functionalization of AgNPs may be determined by the synthesis pathway chosen (one-step
functionalization) or additional treatment after isolation (multi-step functionalization).

3. Surface properties of silver nanoparticles

3.1. Purity on the surface of “green” synthesized AgNPs

The “green” synthesis of AgNPs using plant extracts often results not only in deliberate, but
also inevitable surface functionalization because every component in the total aqueous plant
extract (being reducers, stabilizers, or concomitant constituents) has a certain affinity to the
silver surface [16, 24, 26]. After isolation and purification, surface remain only those compo-
nents which can bind the strongest is “attached” to the AgNPs. Sorption, or so-called “attach-
ment,” might occur due to chemical (chemisorption) or physical (physisorption) phenomenon.
Chemisorption, in the case of AgNPs, happens via ionic, covalent, or coordinate-covalent
chemical bonds. S-containing molecules (some amino acids, peptides, and proteins) possess
the highest affinity to the silver surface because of the strong Ag-S bond and hence are the first
to be considered for interaction [17, 26–28]. Next, N and O atoms from amide, amino, hydroxyl,
phenol, carboxyl, and carbonyl groups are targets for complex formation with Ag+ ions and
thus also very likely to be absorbed on the surface [7, 15, 16, 18, 25–30]. The latter exist in most
primary and secondary metabolites in plants (phenolic acids, polyphenols, flavanoids alka-
loids, glycosides, polysaccharides proteins, etc.) and are found to be present on AgNPs’ surface
by many researchers [7, 15–18, 24, 28, 31]. Physisorption arises due to Van der Waals forces,
and though is much weaker compared to chemisorption; it is non-specific and can affect every
polarized unit in the AgNP’s surrounding. Knowing that the electric potential of colloidal silver
can be considerable, this explains the significant role of physisorption for the surface function-
alization of “green” synthesized AgNPs. It has to be noted, that regardless the mechanism, bio-
molecules participating in Ag+ ion reduction, are more likely to enter in an interaction with the
silver surface because of their initial intimate contact with the arising particles [7, 15–18, 24, 31].
A question may arise whether this heterogenic and uncontrollable “impurity” on AgNPs’ sur-
face following “green” synthesis with plant extracts is only advantageous or does it have any
weak sides. In fact, this highly depends on the particles’ designation. The presence of tannins,
proteins, polysaccharides, flavonoids, and lipids has been proven to benefit stability, increases
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 75
http://dx.doi.org/10.5772/intechopen.80238

AgNPs’ catalytic, antibacterial, and antioxidant activity and reduces toxicity by passivation
of the surface [7, 14–16, 21, 31–34]. However, the “coating” of AgNPs reduces their size and
agglomeration rate, as well as some researchers, suggest that this may have an adverse effect
on cytotoxicity [33]. Furthermore, for surface-selective analytical techniques (such as surface-
enhanced Raman spectroscopy, SERS), where the use of AgNPs provides promising results
as enhancers, a “clear” surface is required that allows access to targeted analytes [26]. In this
regard, the use of pure natural reducers (e.g., the flavonoids quercetin, chrysin, apigenin,
luteolin, etc.) might be preferable instead of the total plant extract [25, 26, 28, 35]. However, if
the presence of multicomponent and unpredictable adherence on the AgNPs is unwelcome,
still the need for a “capping” agent exists. Sugars and polysaccharides, proteins and pro-
teoglycans as glucose, galactose, mannose, chitosan, sodium alginate, glucans, gelatin, and
others are commonly used as coatings for the purpose [17, 27, 35, 36]. These are most often
being included in the reduction media during synthesis, whereas the mechanisms of their
attachment to the surface follow the above-described principles [27, 35].

3.2. The surface area of AgNPs

The active surface area of AgNPs is determined by their size, shape, and agglomeration rate.
Reaction conditions as pH, temperature, extract volume and concentration, reactants ratio,
and time define the dimensions and degree of crystal growth and thus affect the size and
shape of the silver aggregates [16, 24, 25, 31].

The polydispersity of the resulting AgNPs is a disadvantage of the “green” synthesis with
plant extracts, which is likely due to the uncontrollable deposition of different compounds
on the surface and the heterogeneity of the reaction media. In this regard, the use of an o/w
microemulsion-upgraded method has shown good results [18]. Post-synthesis agglomeration
may lead to enlargement of the aggregates and eventually to colloid instability. Here is the
role of the “cap” on AgNPs’ surface, which is aimed to overcome the attractive forces between
the particles and increase physical stability. A large surface area is desirable because it pro-
vides greater catalytic and antibacterial efficacy due to the increased Ag+ release from the
surface which is a fundamental mechanism of AgNPs’ antibacterial action [25, 32]. However,
this precise mechanism, proven by many, is also related to increased oxidative stress and
cytotoxicity [33, 35]. Furthermore, AgNPs smaller than 10 nm can pass through the nuclear
pores and interact with chromosomes and DNA. Thus such particles are proper for gene
therapy and diagnostics, but dangerous regarding genotoxicity [33]. On the one hand, each
intervention leading to suppression of particles’ agglomeration and reducing their size is
welcome concerning stability and potency in catalysis, antibacterial therapy, and diagnostics.
On the other hand, the same intervention can be potentially hazardous concerning increased
toxicity of the NPs obtained [25, 33, 35].
The shape of AgNPs has also been demonstrated to have an impact on toxicity [34, 37]. For
example, wire-shaped AgNPs have shown higher toxicity compared to spherical NPs [37],
whereas another study testifies that plate-shaped AgNPs’ toxic potential exceeds those of
wires and spheres [34].
76 Nanomedicines

3.3. Electrochemical and electromagnetic properties of AgNPs

The charge and zeta potential of AgNPs occurring in suspension are main factors determin-
ing the stability of the colloidal system and depend highly on the synthesis of variables as
well. Among them, pH of the reaction media and the type of coating are crucial [31, 35]. Zeta
potential (ζ) is the potential occurring between the surface of AgNPs and the surrounding
liquid phase and is an important measure for the stability of colloidal systems. Values beyond
ζ = ± 30 mV are usually taken as a requirement for colloid’s endurance [31]. Adjustment of
pH during synthesis is considered an electrostatic approach for stabilization of colloids (by
changing the type and quantity of the electric charge), whereas the coating aims to diminish
the attractive forces in a steric way [31, 35]. AgNPs obtained by reduction with plant extracts
most often are negatively charged [7, 15, 17, 18, 27, 29, 35]. The negative zeta potential can be
considered an advantage because increased cellular uptake and subsequent cytotoxicity are
found for positively charged AgNPs [23, 33].

The presence of “capping” agents on the surface is essential for the stability of colloidal sys-
tems, but they also affect the so-called “redox potential” of AgNPs, that is, their ability to
acquire electrons and be reduced [38]. Low redox potential is needed for oxidation on the
surface and Ag+ release and therefore promotes higher antibacterial activity and toxicity [33].
In some cases, the immobilization of AgNPs in slightly permeable “cap” may lead to loss of
ability for oxidation and antibacterial properties [35].
The surface plasmon resonance (SPR) is a characteristic optical property of AgNPs due to
resonant oscillation of electrons on the surface caused by irradiation with light [39]. This elec-
tromagnetic phenomenon results in an intense peak in the violet-blue sector of the visible
spectrum [7, 15, 18, 24, 26]. The latter depends strongly on surface functionality (size, coating,
etc.) and is considered proof for successful AgNPs synthesis [7, 15, 18, 24, 26, 39].

3.4. Association of AgNPs in complexes and delivery vehicles

A few attempts to incorporate “green” synthesized AgNPs in the structure of liposomes, cyclo-
dextrins, nanoemulsions, and hydrogel beads are reported. Such approaches give the oppor-
tunity for targeted delivery, better compatibility, and lower toxicity [35, 40]. For example,
one-step synthesis of AgNPs-stabilized liposomes have shown improved stability, compat-
ibility, and antibacterial properties of resulting vesicles compared to AgNPs alone, also giv-
ing the opportunity for dermal delivery [40]. Other studies report that association of AgNPs
with β-cyclodextrin improves their catalytic activity [25], whereas kappa-carrageenan hydrogel
beads of “green” synthesized AgNPs have been found to deliver Ag+ in a desirable controlled
manner [41].

3.5. Functionalization by conjugation

Next level surface functionalization is the conjugation of AgNPs with bioactive molecules.
This approach, unlike all of the above mentioned, can not only change but also lead to entirely
new functions. The conjugation of oligonucleotides to metal nanoparticles’ surface is widely
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 77
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researched for targeted gene therapy and bio-diagnostics. However, the attachment of DNA
sequences on AgNPs surface has been challenging due to the lower stability of the complex.
Few successful reports are available from the past years with disulfide or sulfhydryl inserted
DNA [42, 43].
An exciting field of study is the AgNPs potential as drug-delivery carriers [29, 30, 44].
Hypotheses suggest that AgNPs can be used as vehicles to transport drug molecules to tar-
get zones and thereby improve therapeutical efficacy; furthermore, express synergism with
synthetic antibiotics regarding antibacterial properties. These assumptions have been tested
by several scientist in the field, who report successful conjugation of tetracycline (multiple
hydroxyl, phenol, and amide groups), glycopeptide antibiotic vancomycin (multiple amide,
phenol, and hydroxyl groups), and the immunosuppressant azathioprine (S-atom and basic
N-atoms in heterocycle) [29, 30, 44].

4. Antibacterial activity of silver nanoparticles

Since ancient times elemental silver and its compounds have been used as antimicrobial
agents. AgNPs synthesized by different methods were widely tested and had been proved
effective against over 650 microorganisms including bacteria (both Gram-positive and Gram-
negative), fungi, and viruses [21, 45]. Multiple mechanisms of antibacterial action of AgNPs
are considered, but most studies simplified to three primary mechanisms: (1) adhesion of
AgNPs onto the surface of cell wall and membrane; (2) penetration of AgNPs inside the cell
and damaging of intracellular structures (mitochondria, vacuoles, and ribosomes), and bio-
molecules (protein, lipids, and DNA); and (3) generation of reactive oxygen species (ROS),
leading to induced cellular toxicity, and oxidative stress [21, 45, 46]. According to Prabhu
et al. and Dakal et al., modulation of signal transduction pathways is also a distinct mecha-
nism of antimicrobial action of AgNPs [45, 47].

The adhesion of AgNPs onto the surface of the cell wall is facilitated by the positive
surface charge of the AgNPs, and the occurred electrostatic attraction between AgNPs
and the negatively charged cell membrane of microorganisms [48]. The interaction of
Ag+ ions with the proteins containing sulfur, presented in the bacteria cell wall, irre-
versibly disrupted the bacterial cell wall [49]. The damage of cell membranes by AgNPs
causing structural changes renders bacteria more permeable and disturbs the respiratory
function [45, 46]. Morones et al. demonstrated the existence of silver in the membranes of
treated bacteria as well as in the interior of it by transmission electron microscopy (TEM)
analysis [50]. The composition and thickness of the cell wall also influence the antimicro-
bial potential of AgNPs [45, 48]. In Gram-negative bacteria such as E. coli, Pseudomonas,
Salmonella, the cell wall consists of a layer of lipopolysaccharide, followed by a thin layer
of peptidoglycan (3–4 nm). The cell wall in Gram-positive bacteria such as Staphylococcus,
Streptococcus, Bacillus is mainly composed of a thick layer of peptidoglycan (30 nm thick-
ness) [48, 51]. Hence, AgNPs exhibit greater antimicrobial effect against Gram-negative
bacteria regardless of their resistance level as compared to Gram-positive bacteria [49].
78 Nanomedicines

It has also been proposed that Ag+ ion enters the cell and interacts with the sulfur and
phosphorus of the DNA, which can lead to problems in the DNA replication of the bacte-
ria and cell death [47].

The antibacterial potential of AgNPs has related also with the generation of free radicals and
ROS and consequent increase in oxidative stress in cells. Silver ion can interact with the thiol
groups of many vital enzymes, inactivate them and generate ROS. An excessive amount of
generated free radicals lead to direct damage to mitochondrial membrane causing necrosis
and eventually cell death [52].
The antimicrobial effect of AgNPs depends on various parameters including discussed above
size, shape, zeta potential, dose, and colloidal state [15, 46, 49]. AgNPs having a size in the
range of 10–100 nm showed strong bactericidal potential against both Gram-positive and
Gram-negative bacteria [50, 51]. Depending on the size of the NPs, the large surface area
comes in contact with the bacterial cells to provide a higher percentage of interaction than
bigger particles [51, 53].

The effect of shape on the antibacterial activity of AgNPs has been studied by Pal et al. [54].
The AgNPs of different shapes (triangular, spherical, and rod) were tested against E. coli.
According to the authors, triangular NPs are more active than spherical NPs, which are again
more active than rod-shaped AgNPs against E. coli. This could be due to their larger surface
area to volume ratios and their crystallographic surface structures [54]. Rout et al. synthesize
AgNPs of different shapes (i.e., spherical, triangular, and rod) by using Mulberry (Morus
rubra L.) leaves extract and studied their antibacterial activities against E. coli in both liquid
systems and on an agar plate. High reactivity of the truncated triangular NPs has also been
observed in comparison to spherical and rod-shaped particles [55].

Sondi and Salopeck-Sondi investigated the antibacterial activities of AgNPs against E. coli
on Luria-Bertani agar plates and reported that the antibacterial activity of AgNPs was dose-
dependent [56]. AgNPs in colloidal form, that is, suspended nano-sized Ag particles have
shown enhanced antimicrobial potential over AgNPs alone. Colloidal AgNPs produced by
green synthesis are characterized with controlled size, high stability, and improved anti-
bacterial activity which is examined in different studies by directly exposing bacteria to
AgNPs [45, 57].

Okafor et al. produced AgNPs by green synthesis from aloe, geranium, magnolia, and black
cohosh extracts and studied their antibacterial activity on different species of bacteria: three
Gram-negative and three Gram-positive bacteria [58]. The overall results indicated that the
AgNPs showed antibacterial activity at doses of 2 and 4 ppm towards the Gram-positive
and Gram-negative test bacteria. Aloe extract NPs showed the highest antibacterial activity,
followed by black cohosh and geranium NPs with the lowest inhibition. The high antimi-
crobial effect of the aloe produced AgNPs may be due to a combination of the AgNPs and
the aloe bioactive molecules (quinines and other aromatic compounds), which in combina-
tion enhanced the inactivation or growth inhibition of the bacteria species. In another study,
Zhang and co-workers also reported that aloe-produced NPs have a high inhibitory growth
in E. coli at low concentrations [59].
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 79
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Ahmed et al. synthesized AgNPs using Azadirachta indica aqueous leaf extract and studied their
antibacterial activity towards both Gram-positive (S. aureus) and Gram-negative (E. coli) bacte-
rial strains compared with control and plant extract alone [7]. According to the authors, AgNPs
showed effective antimicrobial properties compared to others due to their vast surface area pro-
viding better contact with the cell wall of microorganisms. Also, Bagherzade et al. synthesized
AgNPs using an extract of saffron (Crocus sativus L.) [14]. The biosynthesized AgNPs showed a
significant antibacterial effect against E. coli, P. aeruginosa, K. pneumonia, S. flexneri, and B. subtilis.

Gomathi et al. obtained spherical shaped AgNPs using Datura stramonium leaf extract and
studied their antibacterial activity against E. coli and S. aureus using well diffusion technique
[32]. The authors reported that AgNPs exhibited greater antibacterial activity against E. coli
than S. aureus, due to the variation in cell wall membrane of these bacteria. In another study,
spherical-shaped AgNPs with dimensions of 50–100 nm were observed using Alternanthera
dentate aqueous extract and were tested against E. coli, P. aeruginosa, K. pneumonia, and E. fae-
calis by agar diffusion method [60]. The authors reported that the antibacterial effect of AgNPs
was size- and dose-dependent and was more pronounced against Gram-negative bacteria
than Gram-positive bacteria.

Antimicrobial activity of AgNPs with various antibiotics has been studied, and the synergistic
antibacterial effect has been found. The bactericidal potential of AgNPs synthesized from
the leaf extract of Murraya koenigii singly and in combination with antibiotics (gentamycin,
ampicillin, and streptomycin) against the pathogenic bacteria, namely E. coli, S. aureus, and
P. aeruginosa was studied [61]. The authors reported that AgNPs in combination with genta-
mycin showed the maximum activity against E. coli with an increase in fold area 4.06, while
tetracycline combination with NPs showed maximum activity against S. aureus. The authors
concluded that the activity of standard antibiotics was significantly increased in the presence
of AgNPs and that can be used against antibiotic-resistant pathogens effectively.

5. Silver nanoparticles as anticancer drug delivery systems

Over the past years, nanomedicine created new horizon in the future development of antican-
cer strategies. Conventional cancer treatment such as chemotherapy, radiotherapy, or surgery
has its limitations associated with drugs toxicity, unpredictable side effects, drug resistance
problems, and lack of specificity. AgNPs overcome these disadvantages by reducing side
effects and enhancing the efficiency of cancer therapy. One of their distinguishing features is
the ability to cross various biological barriers and to provide targeted delivery of drugs. Green
synthesis of AgNPs together with specific delivery of anticancer drugs to tumor tissues offers
an innovative approach for improving cancer treatment [62].

5.1. Anticancer activity of biologically synthesized AgNPs

The anticancer activity of biologically synthesized AgNPs has been studied using both in
vitro and in vivo models. The reported results suggested that cytotoxicity of AgNPs may be
80 Nanomedicines

influenced by particle size, shape, and surface chemistry. Several authors have claimed that
increasing AgNPs concentration the viability of tumor cells decreases [63, 64].
The effect of time and concentration of AgNPs on inhibition of cell viability and membrane
leakage are evaluated with a variety of methods [65, 66]. Usually, MTT assay, quantification
of ROS, RT-PCR, and western blotting techniques are used for the assessment of AgNPs abil-
ity to inhibit cellular growth and mediate cell death [65–68]. In vitro cytotoxic activity in a
dose-dependent manner was estimated for green synthesized AgNPs from different plants—
Vitex negundro L., Acalypha indica, Euphorbia nivulia, and Premna serratifolia [63]. MCF-7 (human
breast adenocarcinoma) cell lines were treated with AgNPs obtained by the use of Erythrina
indica and Andrographis echioides extracts. In both cases, the growth of cancer cells was inhib-
ited following AgNPs concentration-response relationship [63]. Similar results were found in
other studies [65, 67]. The AgNPs obtained by use of Artemisia marshalliana Sprengel extract
and Ganoderma neo-japonicum Imazeki extract had a confirmed cytotoxic potential on human
gastric cancer AGS cell line and MDA-MB-231 human breast cancer cell. The authors found
that the cytotoxic activity of AgNPs was time- and dose-dependent as well as the size of NPs
and the temperature of the preparation process.
Dependence on anticancer activity of AgNPs on human cancer cell lines has been found,
according to the source for the synthesis of NPs as well as on the type of the cell lines [69].
Extracts from fruits, leaves, seeds, and roots of Citrullus colocynthis produced AgNPs with
different size and alteration in ID50 on various cell lines. The toxicity assay of biologically syn-
thesized AgNPs using seaweed Ulva lactuca showed potential cytotoxicity of AgNPs against
tumors. For human colon cancer, HT-29 cell lines ID50 was 49 μg/ml whereas its value reached
12.5 μg/ml in human liver cancer Hep G-2 cell lines.
One of the significant drawbacks of conventional anticancer therapy is drug-mediated toxic-
ity in healthy cells. AgNPs synthesized from plants have the potential to avoid this problem
by offering selective toxicity to cancer cells. AgNPs produced using leaf extract of Podophyllum
hexandrum Royle induced cytotoxicity to cervical carcinoma cells. The reported results proved
that AgNPs could selectively inhibit the cellular mechanism of HeLa by DNA damage and
caspase-mediated cell death [70]. In another study, the cytotoxicity of AgNPs towards cancer
cells was estimated comparing human myeloblastic leukemia cells HL60 and cervical cancer
cells HeLa to normal peripheral blood mononuclear cells (PBMC) [66]. Sargassum vulgare had
been used for the green synthesis of AgNPs. It was found that HL60 cells were affected by
AgNP-mediated toxicity while the normal PBMC suffered less damage.

It has been proven that biologically synthesized AgNPs show substantial anticancer activity
with less toxic manner compared to particles whose preparation involves some toxic and
expensive chemicals. The production AgNPs through green chemistry approach via Cleome
viscosa plant extract offers another solution for optimizing anticancer treatment. Anticancer
activity was in vitro evaluated against human cancer cell lines PA1 (Ovarian teratocarcinoma
cell line) and A549 (Human lung adenocarcinoma) [68]. The results concluded that green
synthesized AgNPs could inhibit cancer cells growth and provide great potential in the treat-
ment of cancer.
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 81
http://dx.doi.org/10.5772/intechopen.80238

To determinate the anticancer efficacy of biologically synthesized AgNPs and to fully appre-
hend the mode of programmed cell death three critical parameters need to be taken into
consideration: (1) DNA fragmentation; (2) structural changes in the cell morphology; and
(3) Annexin V binding and caspase activation. Upregulation of apoptosis is only one of the
possible mechanisms for antiproliferative activity of biosynthesized AgNPs that was proven
in many studies [67, 71, 72]. AgNPs could elicit cell death through ROS generation, membrane
leakage, activation of caspases, and DNA damage [65, 66, 72].

5.2. AgNPs for targeted drug delivery

AgNPs represent an alternative therapeutic strategy as DDSs in curing cancer because

these can provide passive or active targeting to tumor tissue. Accumulation of drugs
at desired sites increases the efficacy of anticancer therapy in vivo. Receptor-mediated
endocytosis can facilitate cellular uptake of drugs. This kind of active targeting relays on
molecular recognition. Suggested approach for optimizing biogenic AgNPs properties is
surface functionalization with specific targeting molecules or coating with biocompatible
and biodegradable polymers [73, 74]. For example, AgNPs obtained by use of various
concentrations of Setaria verticillata seed extract were loaded with hydrophilic anticancer
drugs, doxorubicin (DOX), and daunorubicin (DNR). The significant loading (80.50%) and
capacity (40.25%) efficiency of DOX-AgNPs and DNR-AgNPs presented them as future
novel DDSs [64].
Drug delivery into the cells by endocytosis depends on the size of NPs. Spherical-shaped
AgNPs were extracted from Aerva javanica plant and conjugated with the anti-cancer drug
gefitinib. Scanning transmission electron microscopy (STEM) images determinates average
size of 5.7 nm. The apoptotic potential of gefitinib-AgNPs has been compared to gefitinib
alone. Reduction of cell viability of breast cancer cells MCF-7 treated with conjugated gefi-
tinib-AgNPs was significant. Delivery of gefitinib using AgNPs optimizes its effectivity and
reduces side effects [75].
The variety of green synthesized AgNPs exhibiting anticancer activity offer new treatment
opportunities. Their specific features as nanocarriers benefit the development of DDSs with
unique properties and biocompatible profile.

6. Silver nanoparticles as photoactivated drug delivery vectors

Nanoparticles can surmount some essential problems of conventional small molecules or bio-
macromolecules (e.g., DNA, RNA, and protein) used at some diseases by allowing targeted
delivery and overcome through biological barriers [76]. Noble metal NPs have specific high
developed photophysical properties which contribute to their potential as photoactivated
drug delivery vectors [77]. AgNPs have been used extensively as biological sensors which
take advantage of plasmon resonance (PR) to enhance detection of specific targets. Noble
metal nanoparticle-based sensors benefit from the extreme sensitivity of localized surface
82 Nanomedicines

plasmon resonance (LSPR) spectra to environmental changes. Application of metal nanopar-

ticles is not limited to molecular detection. Recently, AgNPs have been harnessed as delivery
vehicles for therapeutic agents, including antisense oligonucleotides and other small mol-
ecules. Small metal NPs offer many advantages as drug carriers, including adjustable size and
shape, enhanced stability of surface-bound nucleic acids, high-density surface ligand attach-
ment, transmembrane delivery without harsh transfection agents, protection of the attached
therapeutic from degradation, and potential for improved timed/controlled intracellular
release. The photophysical properties of AgNPs may potentially bring these to the forefront
of drug delivery, enabling targeted delivery, spatiotemporally controlled (photo-)release, and
delivery confirmation via imaging [78].
AgNPs in the diameter range of ~2–100 nm exhibit SPR spectra in the visible region, which
are tunable and dependent on particle shape, size, environment, and interparticle distance.
AgNPs have unique properties which make them a desirable alternative particle type in many
cases. AgNPs are the strongest light scatterers of the noble metal particles, and it is reported
that the light scattering cross section of AgNP is ~10 times greater than that of a similarly
sized gold NPs. The extinction (light absorption and scattering) band of AgNPs is due to free
conduction electron oscillations, and bound electron movements also contribute to the opti-
cal spectra. Thus enhancement of absorption/emission of light by molecules near the AgNPs
surface is dependent on particle size and proximity or overlap of the resonance (SPR) spectra
with the absorption/emission bands of the molecular species [78].

Mie Theory has calculated the light absorption and scattering properties for AgNPs of differ-
ent sizes. For larger particle sizes (~50–60 nm), the scattering efficiency (Qsca) is higher (≈ 5).
The AgNPs in this size range scatter light at or above the solid metal surface, but the scattering
efficiency increases even higher to 5.8 for size 70–80 nm while maintaining surface PR in the
UV to the visible range. This characteristic is ideal for traditional and red-shifted photocleav-
able compounds typically used as photo-caging compounds [78].
The generality of current nanoscale delivery systems are polymeric in their essence. The stud-
ies of metallic NPs have shown their suitability for delivery of various therapeutic agents
including small molecules, antisense oligonucleotides, and siRNAs. Nanoscale silver is one of
the optically active surface-enhancing substrates available. AgNP-based single delivery plat-
forms incorporate solutions to both intracellular detection and external control over surface-
tethered drug release via chemical photothermal or photochemical triggers [77].

Light-responsive systems are of great interest in the field of drug delivery and gene therapy,
owing to the capability of external, spatiotemporal control over the delivery, and activation of
therapeutics coupled with such systems. Electromagnetic radiation triggers light-responsive
DDSs, typically in the UV, visible, and near-infrared (NIR) range. These systems are based upon
photosensitive compounds which can be incorporated into a drug delivery vehicle, or coupled
to the drug itself (“caging” compounds), and may switch to an active or inactive state upon
electromagnetic irradiation within a specific frequency range. Caged compounds are potent
tools for spatiotemporal control over drug activity in living systems. Photocleavable groups
have been used to the cage, or inactivate, various biomolecules, including nucleotides, pro-
teins, and nucleic acids, for controlled, on-site photo-activation. Uncaging via light irradiation
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 83
http://dx.doi.org/10.5772/intechopen.80238

allows rapid, spatially, and temporally defined release of a biomolecule at intended tissues or
even within a specific intracellular compartment [78].

AgNPs with the size of 60–80 nm decorated with thiol-terminated photolabile DNA oligo-
nucleotides were used as photo-activated drug delivery vectors [77]. In vitro assays showed
efficient photo-activation of surface-tethered caged ISIS2302 antisense oligonucleotides with
internal photo-cleavable linkers. These nanocarriers have several advantages such as protec-
tion against nucleases, efficient photorelease, and enhanced cellular uptake when compared
to commercial transfection agents. The light-induced release of anti-sense oligonucleotides for
silencing ICAM-1 (intracellular adhesion molecule-1) has potential application in the wound
healing, where inflammation is a significant criterion such as in Crohn’s disease.

7. Toxicity assessment of silver nanoparticles

Nanotechnology has been rapidly growing with utilization in a wide range of commercial
products throughout the world. However, there is still a lack of information concerning the
increase of human, animal, and environmental exposure to NPs including AgNPs and the
potential risks related to their short- and long-term toxicity. However, some studies have
already been made.

7.1. In vitro tests

AgNPs have emerged as an important class of nanomaterials for a wide range of industrial
and medical applications that have potential risks to human health. In vitro studies reported,
that AgNPs produced toxicity targeted a variety of organs including the lung, liver, brain,
vascular system, and reproductive organs. AgNPs induced the expression level of genes
involved in cell cycle progression and apoptosis. Possible mechanisms of AgNP toxicity
include induction of ROS, oxidative stress, DNA damage, and apoptosis [79].

To understand the toxicity of NPs in vitro different tests have been assessed. Testing silver
(Ag – 15 nm), molybdenum (MoO3–30 nm), and aluminum (Al – 30 nm) NPs on mouse sper-
matogonial cell line have been determined concentration-dependent toxicity for all types.
AgNPs were the most toxic (5–10 μg ml−1), and reduced mitochondrial function drastically
and increased membrane leakage [80]. Similar conclusions have been made testing the toxic
effects of the metal/metal oxides NPs mentioned above on rat liver-derived cell line (BRL
3A). Results showed that mitochondrial function decreases significantly in cells exposed to
AgNPs at (5–50 μg ml−1). Fe3O4, Al, MoO3, and TiO2 had no measurable effect at lower doses
(10–50 μg ml−1), while there was a significant effect at higher levels (100–250 μg ml−1) [81].

Generally, in in vitro tests, the mechanism of AgNPs-mediated cytotoxicity is mainly based

on the induction of ROS. Notably, exposure to AgNPs causes a reduction in GSH, elevated
ROS levels, lipid peroxidation, and increased expression of ROS responsive genes; it also
leads to DNA damage, apoptosis, and necrosis. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphe-
nyl tetrazolium bromide) reduction, Alamar Blue (Invitrogen, Carlsbad, CA) reduction, and
84 Nanomedicines

lactate dehydrogenase (LDH) leakage were used as parameters for cytotoxicity assessment.
Toxicity of different AgNPs was compared to that of various corresponding concentrations
of Ag+ ions. Based upon the IC50 values determined by three cytotoxicity assays, AgNPs and
Ag+ ions did not exhibit a dramatic difference in cytotoxicity [82]. The cytotoxicity and geno-
toxicity of AgNPs are size-, concentration-, and exposure time-dependent. The cell viability
was determined by MTT and CB assay in macrophage (RAW 264.7, J774.1), pulmonary epi-
thelial (A549), renal epithelial (A498), hepatic (Hep G2), and neuronal (Neuro 2A) cell lines.
AgNPs showed a concentration-dependent reduction in cell viability after 72 h incubation in
all cell lines. A498 and RAW 264.7 cells appeared to exhibit the highest sensitivity to the toxic
effects of AgNPs and showed significant reduction in cell viability at 1 and 3 g/ml AgNP-
concentration, respectively. On the other hand, A549 cells were least sensitive to cytotoxic
effects of AgNPs. The internalization of NPs can induce stress response(s) due to stimula-
tion of free radical production, which in turn, stimulates inflammatory signaling pathways.
Hence, the production of reactive nitrogen species (RNS), ROS, and cytokines following
AgNPs exposure was determined. AgNPs significantly increased nitrite release by RAW 264.7
cells at the highest concentration following 72 h incubation. AgNPs also stimulated ROS pro-
duction in a concentration-dependent manner after 24 h incubation. Inflammatory cytokine
(tumor necrosis factor-α [TNF-α] and interleukin-6 [IL-6]) production was significant at 10
and 100 g/ml while 1 g/ml showed no effect on cytokine production. Free radical production
has been demonstrated to bear a direct correlation with cytotoxicity of NPs. However, the
involvement of other mechanisms cannot be ruled out. Therefore, to determine the contribu-
tion of free radicals in AgNP cytotoxicity, cells were incubated with AgNPs in the presence
of various antioxidants. Surprisingly, the most potent antioxidants like superoxide dismutase
(SOD) and catalase showed no significant protection from AgNPs cytotoxicity. Therefore, two
cell membrane ROS scavengers—Trolox (water-soluble vitamin E analog) and tempol (broad-
spectrum antioxidant and SOD mimetic)—were investigated. In line with observations in
SOD and catalase-treated cells, Trolox and tempol also failed to protect cells from AgNPs
cytotoxicity. On the other hand, weak antioxidants like N-acetylcysteine (NAC), methionine
and cysteine abrogated the cytotoxic effect of AgNPs. The relative ineffectiveness of potent
antioxidants suggests that free radical-dependent mechanisms do not significantly influence
cytotoxicity of AgNPs [83]. Other studies showed that p53 protein expression level increased
within 4 h after the cells were exposed to AgNPs. The up-regulated expression patterns of
p53 protein in two types of mammalian cells by AgNPs exposure suggest that the p53 could
be an excellent molecule marker to assess the genetic nanotoxicity. The results suggest the
different surface chemistry of AgNPs have different effects on genotoxicity [84]. Beer et al.
concluded that free Ag+ ions in AgNPs preparations play a considerable role in the toxicity
of AgNPs suspensions [85]. While the contribution of the free Ag+ ion to the measured toxic-
ity of AgNPs suspensions is an essential determinant for the toxicity, a combined effect of
Ag+ ion and AgNPs appears for lower concentrations of Ag+ ions. These data indicate that
the amount of Ag+ ions in AgNPs preparations should be routinely measured and reported in
toxicological work. They advise that the supernatant of AgNPs suspensions should be used
as an additional standard control to make reliable statements of the toxicity of AgNPs and to
discriminate between Ag+ ions toxicity and AgNPs-induced toxicity [85].
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 85
http://dx.doi.org/10.5772/intechopen.80238

7.2. In vivo tests

The most significant problem to understand is the real impact of AgNPs on human health and
animals. There are several in vivo studies on cytotoxicity and genotoxicity of AgNPs reported.
Due to the ultra-small sizes of AgNPs, they have high mobility in different environments,
and humans are easily exposed via routes such as inhalation, ingestion, skin, etc. AgNPs can
translocate from the route of exposure to other vital organs and penetrate into cells.

Inhalation toxicity of AgNPs has been investigated on Sprague–Dawley rats over a period of
28 days. Results showed that the male and female rats did not show any significant changes in
body weight relative to the concentration of AgNPs during the 28-day experiment. There were
also no significant changes in the hematology and blood biochemical values in either the male
or female rats. Whereas, some investigators have reported that lungs are primary target tis-
sues affected by prolonged inhalation exposure to AgNPs [86]. Lee et al. have reported AgNPs
exposure modulated the expression of several genes associated with motor neuron disorders,
neurodegenerative disease, and immune cell function, indicating potential neurotoxicity and
immunotoxicity associated with AgNPs exposure [87]. Minimal pulmonary inflammation or
cytotoxicity of mice was found after 10 days of AgNPs exposure. Gastrointestinal toxicology
caused by AgNPs (60 nm) exposure via ingestion has also been tested over a period of 28 days
in Sprague–Dawley rats. Results showed that the male and female rats did not show any
significant changes in body weight relative to the doses of AgNPs during the 28-day experi-
ment. Some significant dose-dependent changes were found in the alkaline phosphatase and
cholesterol values in either the male or female rats, seeming to indicate that exposure to over
more than 300 mg of AgNPs may result in slight liver damage. Results suggested that AgNPs
do not induce genetic toxicity in male and female rat bone marrow in-vivo [88]. Ahamed et al.
indicated that AgNPs produce reproductive failure, developmental malformations, and mor-
phological deformities in some non-mammalian animal models. Common causes of AgNPs-
induced toxicity include oxidative stress, DNA damage, and apoptosis [79].
Generally, very few papers on the in vivo toxicology of AgNPs were found, so further inves-
tigation is needed in this field to evaluate precisely the real impact of AgNPs in commercial
products to humans and animals.

8. Conclusion

Plant-mediated synthesis of AgNPs has revealed new horizons in drug-delivery. On the one
hand, this approach of nanoparticle preparation is preferable due to its economic, accessible,
eco-friendly nature, and simplicity of execution. On the other hand, the rich phytochemi-
cal composition of plant extracts performs a multi-functional role in the synthesis process
of AgNPs as reducing, stabilizing, and the surface-active agent. The AgNPs thus obtained
are usually characterized by tiny sizes, monodispersity, and stability of colloidal system
because of the capping properties of some of the biomolecules in the extract. Despite their
excellent antibacterial, antiviral, antifungal, anticancer, antioxidant, and unusually enhanced
86 Nanomedicines

physicochemical properties compared to the bulk material, the AgNPs could be used as
vehicles to transport drug molecules (such as oligonucleotides, DNA, siRNA, etc.) to tar-
geted tissues and cells and thereby to improve therapeutic efficacy. Moreover, AgNPs could
express synergism with different antibiotics regarding enhanced antibacterial properties. In
this regard, the AgNPs might be used as multi-functional drug carriers having great poten-
tial in targeted drug-delivery, minimizing side effects, and improving therapeutic efficacy.
However, there is still a lack of information concerning the increase of human, animal, and
environmental exposure to AgNPs and the potential risks related to their short- and long-
term toxicity. Further profound investigations are needed for their safe inclusion as DDSs in
commercially available products for the prevention and treatment of life-threatening diseases.

Conflict of interest

The authors have declared no conflicts of interest for this article.

Author details

Nadezhda Ivanova, Viliana Gugleva, Mirena Dobreva, Ivaylo Pehlivanov, Stefan Stefanov
and Velichka Andonova*

*Address all correspondence to: andonova_v@abv.bg

Department of Pharmaceutical Technologies, Faculty of Pharmacy, Medical University –

Varna “Prof. Dr. Paraskev Stoyanov”, Varna, Bulgaria

References

[1] Satalkar P, Elger BS, Shaw DM. Defining nano, nanotechnology and nanomedicine: Why
should it matter? Science and Engineering Ethics. 2016;22(5):1255-1276

[2] Le Ouay B, Stellacci F. Antibacterial activity of silver nanoparticles: A surface science

insight. Nano Today. 2015;10(3):339-354

[3] Phull A-R, Abbas Q, Ali A, Raza H, Kim SJ, Zia M, Haq I. Antioxidant, cytotoxic and
antimicrobial activities of green synthesized silver nanoparticles from crude extract of
Bergenia ciliata. FJPS. 2016;2(1):31-36

[4] Tolaymat TM, El Badawy AM, Genaidy A, Scheckel KG, Luxton TP, Suidan M. An
evidence-based environmental perspective of manufactured silver nanoparticle in syn-
theses and applications: A systematic review and critical appraisal of peer-reviewed
scientific papers. Science of the Total Environment. 2010;408(5):999-1006

[5] CD Creative diagnostics®. CD Bioparticles products. Properties and Application of

Silver Nanoparticles. Available from: https://www.cd-bioparticles.com/t/Properties-
and-Applications-of-Silver-Nanoparticles_60.html [Accessed: 2018-06-16]
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 87
http://dx.doi.org/10.5772/intechopen.80238

[6] Larue C, Michel H, Sobanska S, Cécillon L, Bureau S, Barthès V, et al. Foliar exposure of
the crop Lactuca sativa to silver nanoparticles: Evidence for internalization and changes
in Ag speciation. Journal of Hazardous Materials. 2014;264:98-106

[7] Ahmed S, Saifullah, Ahmad M, Swami BL, Ikram S. Green synthesis of silver nanopar-
ticles using Azadirachta indica aqueous leaf extract. Journal of Radiation Research and
Applied Science. 2016;9:1-7

[8] Iravani S, Korbekandi H, Mirmohammadi SV, Zolfaghari B. Synthesis of silver nanopar-

ticles: Chemical, physical and biological methods. RPS. 2014;9(6):385-406

[9] Naseem T, Farrukh MA. Antibacterial activity of green synthesis of iron nanoparticles
using Lawsonia inermis and Gardenia jasminoides leaves extract. Journal of Chemistry.
2015;912342

[10] Das VL, Thomas R, Varghese RT, Soniya EV, Mathew J, Radhakrishnan EK. Extracellular
synthesis of silver nanoparticles by the Bacillus strain CS 11 isolated from industrialized
area. 3 Biotech. 2014;4(2):121-126

[11] Nanda A, Saravanan M. Biosynthesis of silver nanoparticles from Staphylococcus aureus

and its antimicrobial activity against MRSA and MRSE. Nanomedicine. 2009;5(4):452-456

[12] Shivaji S, Madhu S, Singh S. Extracellular synthesis of antibacterial silver nanoparticles

using psychrophilic bacteria. Process Biochemistry. 2011;49:830-837

[13] Iravani S. Bacteria in nanoparticle synthesis: Current status and future prospects.
International Scholarly Research Notices. Oct 29, 2014;2014:359316. DOI: 10.1155/2014/
359316

[14] Bagherzade G, Tavakoli MM, Namaei MH. Green synthesis of silver nanoparticles using
aqueous extract of saffron (Crocus sativus L.) wastages and its antibacterial activity
against six bacteria. Asian Pacific Journal of Tropical Biomedicine. 2017;7(3):227-233

[15] Raja S, Ramesh V, Thivaharan V. Green biosynthesis of silver nanoparticles using

Calliandra haematocephala leaf extract, their antibacterial activity and hydrogen peroxide
sensing capability. Arabian Journal of Chemistry. 2017;10(2):253-261

[16] Verma A, Mehata MS. Controllable synthesis of silver nanoparticles using Neem leaves
and their antimicrobial activity. Journal of Radiation Research and Applied Science.
2016;9(1):109-115

[17] Patil MP, Singh RD, Koli PB, Patil KT, Jagdale BS, Tipare AR, et al. Antibacterial potential
of silver nanoparticles synthesized using Madhuca longifolia flower extract as a green
resource. Microbial Pathogenesis. 2018;12:184-189

[18] Rivera-Rangel RD, González-Muñoz MP, Avila-Rodriguez M, Razo-Lazcano TA, Solans

C. Green synthesis of silver nanoparticles in oil-in-water microemulsion and nano-emul-
sion using geranium leaf aqueous extract as a reducing agent. Colloids and Surfaces A:
Physicochemical and Engineering Aspects. 2018;536:60-67

[19] Li S, Shen Y, Xie A, Yu X, Qiu L, Zhanga L, et al. Green synthesis of silver nanoparticles
using Capsicum annuum L. extract. Green Chemistry. 2007;9:852-858
88 Nanomedicines

[20] Mirgorod YA, Borodina VG. Preparation and bactericidal properties of silver nanopar-
ticles in aqueous tea leaf extract. Inorganic Materials. 2013;49(10):980-983
[21] Ahmed S, Ahmad M, Swami BL, Ikram S. A review on plants extract mediated synthe-
sis of silver nanoparticles for antimicrobial applications: A green expertise. Journal of
Advanced Research. 2016;7(1):17-28
[22] Amin M, Anwar F, Janjua MRSA, Iqbal MA, Rashid U. Green synthesis of silver nanopar-
ticles through reduction with Solanum xanthocarpum L. berry extract: Characterization,
antimicrobial and urease inhibitory activities against Helicobacter pylori. International
Journal of Molecular Sciences. 2012;13(8):9923-9941
[23] Kim ST, Saha K, Kim C, Rotello VM. The role of surface functionality in determining
nanoparticle cytotoxicity. Accounts of Chemical Research. 2013;46(3):681-691
[24] Islam NU, Jalil K, Shahid M, Rauf A, Muhammad N, Khan A, et al. Green synthesis and
biological activities of gold nanoparticles functionalized with Salix alba. Arabian Journal
of Chemistry. 2015:12 (in press). DOI: 10.1016/j.arabjc.2015.06.025
[25] Li P, Li S, Wang Y, Zhang Y, Han G. Green synthesis of β-CD-functionalized monodis-
persed silver nanoparticles with enhanced catalytic activity. Colloids and Surfaces A:
Physicochemical and Engineering Aspects. 2017;520:26-31
[26] Švecová M, Ulbrich P, Dendisová M, Matějka P. SERS study of riboflavin on green-
synthesized silver nanoparticles prepared by reduction using different flavonoids: What
is the role of flavonoid used? Spectrochimica Acta. Part A, Molecular and Biomolecular
Spectroscopy. 2018;195:236-245
[27] Shao Y, Wu C, Wu T, Yuan C, Chen S, Ding T, et al. Green synthesis of sodium alginate-
silver nanoparticles and their antibacterial activity. International Journal of Biological
Macromolecules. 2018;111:1281-1292
[28] Ikram F, Qayoom A, Shah MR. Synthesis of epicatechin coated silver nanoparticles for
selective recognition of gentamicin. Sensors and Actuators B: Chemical. 2018;257:897-905
[29] Buszewski B, Rafinska K, Pomastowski P, Walczak J, Rogowska A. Novel aspects of
silver nanoparticles functionalization. Colloids and Surfaces A: Physicochemical and
Engineering Aspects. 2016;506:170-178
[30] Kaur A, Goyal D, Kumar R. Surfactant mediated interaction of vancomycin with silver
nanoparticles. Applied Surface Science. 2018;449:23-30
[31] Sudha A, Jeyakanthan J, Srinivasan P. Green synthesis of silver nanoparticles using
Lippia nodiflora aerial extract and evaluation of their antioxidant, antibacterial and cyto-
toxic effects. Resource-Efficient Technologies. 2017;3:506-515
[32] Gomathi M, Rajkumar PV, Prakasam A, Ravichandran K. Green synthesis of silver
nanoparticles using Datura stramonium leaf extract and assessment of their antibacterial
activity. Resource-Efficient Technologies. 2017;3:280-284
[33] Ahmed KB, Nagy AM, Brown RP, Zhang Q, Malghan SG, Goering PL. Silver nanopar-
ticles: Significance of physicochemical properties and assay interference on the interpre-
tation of in vitro cytotoxicity studies. Toxicology In Vitro. 2017;38:179-192
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 89
http://dx.doi.org/10.5772/intechopen.80238

[34] George S, Lin S, Ji Z, Thomas C, Li L, Mecklenburg M, et al. Surface defects on plate-

shaped silver nanoparticles contribute to its hazard potential in a fish gill cell line and
zebrafish embryos. ACS Nano. 2012;6:3745-3759
[35] Kim S, Ryu D. Silver nanoparticle-induced oxidative stress, genotoxicity and apoptosis
in cultured cells and animal tissues. Journal of Applied Toxicology. 2013;33:78-89
[36] Vedelago J, Gomez CG, Valente M, Mattea F. Green synthesis of silver nanoparticles
aimed at improving theranostics. Radiation Physics and Chemistry. 2018;146:55-67
[37] Stoehr LC, Gonzalez E, Stampfl A, Casals E, Duschl A, Puntes V, et al. Shape matters:
Effects of silver nanospheres and wires on human alveolar epithelial cells. Particle and
Fibre Toxicology. 2011;8:36
[38] Quinsaat JEQ. Synthesis and functionalization of silver nanoparticles for the preparation
of high permittivity nanocomposites [thesis]. Lausanne: EPFL; 2015
[39] Smitha SL, Nissamudeen KM, Philip D, Gopchandran KG. Studies on surface plasmon
resonance and photoluminescence of silver nanoparticles. Spectrochimica Acta. Part A,
Molecular and Biomolecular Spectroscopy. 2008;71:186-190
[40] Castangia I, Marongiu F, Manca ML, Pompei R, Angius F, Ardu A, et al. Combination
of grape extract-silver nanoparticles and liposomes: A totally green approach. European
Journal of Pharmaceutical Sciences. 2017;97:62-69
[41] Azizi S, Mohamad R, Rahim RA, Mohammadinejad R, Ariff AB. Hydrogel beads bio-
nanocomposite based on kappa-carrageenan and green synthesized silver nanopar-
ticles for biomedical applications. International Journal of Biological Macromolecules.
2017;104:423-431
[42] Zhang X, Servos MR, Liu J. Fast pH-assisted functionalization of silver nanoparticles with
monothiolated DNA. Chemical Communications (Cambridge). 2012;48(81):10114-10116
[43] Lee J, Lytton-Jean A, Hurst SJ, Mirkin CA. Silver nanoparticle-oligonucleotide conjugates
based on DNA with triple cyclic disulfide moieties. Nano Letters. 2007;7(7):2112-2115
[44] Prasad SR, Elango K, Damayanthi D, Saranya JS. Formulation and evaluation of aza-
thioprine loaded silver nanopartilces for the treatment of rheumatoid arthritis. AJBPS.
2013;3(23):28-32
[45] Dakal TC, Kumar A, Majumdar RS, Yadav V. Mechanistic basis of antimicrobial actions
of silver nanoparticles. Frontiers in Microbiology. 2016;7:1831
[46] Duran N, Duran M, de Jesus MB, Seabra AB, Favaro WJ, Nakazato G. Silver nanopar-
ticles: A new view on mechanistic aspects on antimicrobial activity. Nanomedicine.
2016;12:789-799
[47] Prabhu S, Poulose EK. Silver nanoparticles: Mechanism of antimicrobial action, synthe-
sis, medical applications, and toxicity effects. International Nano Letters. 2012;2:32
[48] Abbaszadegan A, Ghahramani Y, Gholami A, Hemmateenejad B, Dorostkar S, Nabavizadeh
M, et al. The effect of charge at the surface of silver nanoparticles on antimicrobial activ-
ity against gram-positive and gram-negative bacteria: A preliminary study. Journal of
Nanomaterials. 2015;720654
90 Nanomedicines

[49] Keat CL, Aziz A, Eid AM, Elmarzugi NA. Biosynthesis of nanoparticles and silver
nanoparticles. Bioresour Bioprocess. 2015;2:47
[50] Morones JL, Elichiguerra A, Camacho K, Holt JB, Kouri JT, Ramirez MY. The bactericidal
effect of silver nanoparticles. Nanotechnology. 2005;16(10):2346-2353
[51] Rai MK, Deshmukh SD, Ingle AP, Gade AK. Silver nanoparticles: The powerful
nanoweapon against multidrug-resistant bacteria. Journal of Applied Microbiology.
2012;112:841-852

[52] Huang CC, Aronstam RS, Chen D, Huang Y. Oxidative stress, calcium homeostasis and
altered gene expression in human lung epithelial cells exposed to ZnO nanoparticles.
Toxicology In Vitro. 2010;24:45-55
[53] Kvitek L, Panacek A, Soukopova J, Kolar M, Vecerova R, Prucek R, et al. Effect of surfac-
tants and polymers on stability and antibacterial activity of silver nanoparticles (NPs).
Journal of Physical Chemistry C. 2008;112(15):5825-5834

[54] Pal S, Tak YK, Song JM. Does the antibacterial activity of silver nanoparticles depend on
the shape of the nanoparticle? A study of the gram-negative bacterium Escherichia coli.
Applied and Environmental Microbiology. 2007;73(6):1712-1720

[55] Rout A, Jena PK, Sahoo D, Bindhani BK. Green synthesis of silver nanoparticles of dif-
ferent shapes and its antibacterial activity against Escherichia coli. International Journal
of Current Microbiology and Applied Sciences. 2014;3(4):374-383
[56] Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: A case study on
E. coli as a model from gram-negative bacteria. Journal of Colloid and Interface Science.
2004;275(1):177-182

[57] Guzman M, Dille J, Godet S. Synthesis and antibacterial activity of silver nanoparticles
against gram-positive and gram-negative bacteria. Nanomedicine. 2012;8:37-45

[58] Okafor F, Janen A, Kukhtareva T, Edwards V, Curley M. Green synthesis of silver

nanoparticles, their characterization, application and antibacterial activity. International
Journal of Environmental Research and Public Health. 2013;10:5221-5238

[59] Zhang Y, Yang D, Kong Y, Wang X, Pandoli O, Gao G. Synergetic antibacterial effects of
silver nanoparticles @ aloe vera prepared via a green method. Nano Biomedicine and
Engineering. 2010;2:252-257
[60] Kumar DA, Palanichamy V, Roopan SM. Green synthesis of silver nanoparticles using
Alternanthera dentate leaf extract at room temperature and their antimicrobial activ-
ity. Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy. 2014;127:
168-171
[61] Bonde SR, Rathod DP, Ingle AP, Ade RB, Gade AK, Rai MK. Murraya koenigii-mediated
synthesis of silver nanoparticles and its activity again three human pathogenic bacteria.
Journal of Nanoscience Methods. 2012;1:25-36
 Silver Nanoparticles as Multi-Functional Drug Delivery Systems 91
http://dx.doi.org/10.5772/intechopen.80238

[62] Kajani A, Zarkesh-Esfahani S, Hamid S, Bordbar AK, Khosropour A, Razmjou A, et al.

Anticancer effects of silver nanoparticles encapsulated by Taxus baccata extracts. Journal
of Molecular Liquids. 2016;223:549-556
[63] Rao PV, Nallappan D, Madhavi K, Rahman S, Wei LJ, Gan SH. Phytochemical and
biogenic metallic nanoparticles as anticancer agents. Oxidative Medicine and Cellular
Longevity. 2016;2016:3685671. Epub 2016 Feb 23. DOI: 10.1155/2016/3685671
[64] Naz M, Nasiri N, Ikram M, Nafees M, Qureshi MZ, Ali S, et al. Eco-friendly biosynthesis,
anticancer drug loading and cytotoxic effect of capped Ag-nanoparticles against breast
cancer. Applied Nanoscience. 2017;7:793-802
[65] Gurunathan S, Raman J, Abd Malek SN, John PA, Vikineswary S. Green synthesis of sil-
ver nanoparticles using Ganoderma neo-japonicum Imazeki: A potential cytotoxic agent
against breast cancer cells. International Journal of Nanomedicine. 2013;8:4399-4413
[66] Govindaraju K, Krishnamoorthy K, Alsagaby S, Singaravelu G, Premanathan M. Green
synthesis of silver nanoparticles for selective toxicity towards cancer cells. IET Nano-
biotechnology. 2015;9(6):325-330
[67] Salehi S, Shandiz AS, Ghanbar F, Darvish MR, Ardestani M, Mirzaie A, et al.
Phytosynthesis of silver nanoparticles using Artemisia marschalliana sprengel aerial part
extract and assessment of their antioxidant, anticancer, and antibacterial properties.
International Journal of Nanomedicine. 2016;11:1835-1846
[68] Lakshmanan G, Sathiyaseelan A, Kalaichelvan PT, Murugesan K. Plant-mediated syn-
thesis of silver nanoparticles using fruit extract of Cleome viscosa L.: Assessment of their
antibacterial and anticancer activity. Karbala International Journal of Modern Science.
2018;4:61-68
[69] Singh J, Singh T, Rawat M. Green synthesis of silver nanoparticles via various plant
extracts for anti-cancer applications. GJN. 2017;2(3). DOI: 1019081/GJN.2017.02.555590
[70] Rath M, Panda SS, Dhal NK. Syntesis of silver nano particles from plant extract and its
application in cancer treatment: A review. IJPAES. 2014;4(3):137-145
[71] Khalifa KS, Hamouda RA, Hamz HA. Antitumor activity of silver nanoparticles biosyn-
thesized by micro algae. Journal of Chemical and Pharmaceutical Research. 2016;8(3):1-6
[72] Gopinath P, Gogoi SK, Chattopadhyay A, Ghosh S. Implications of silver nanoparticle
induced cell apoptosis for in vitro gene therapy. Nanotechnology. 2008;19(7):075104
[73] Jeyaraj M, Rajesh M, Arun R, MubarakAli D, Sathishkumar G, et al. An investigation
on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized
silver nanoparticles using Podophyllum hexandrum on human cervical carcinoma cells.
Colloids and Surfaces. B, Biointerfaces. 2013;102:708-717
[74] Blanco MD, Teijón C, Olmo RM, Teijón JM. Targeted nanoparticles for cancer therapy.
In: Sezer AD, editor. Recent Advances in Novel Drug Carrier Systems. Rijeka: InTech;
2012. pp. 241-248
92 Nanomedicines

[75] Khalid S, Hanif R. Green biosynthesis of silver nanoparticles conjugated to gefitinib as

delivery vehicle. IJASET. 2017;5(2):59-63

[76] Park W, Na K. Advances in the synthesis and application of nanoparticles for drug
delivery. Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology.
2015;7(4):494-508
[77] Brown PK, Qureshi AT, Moll AN, Hayes DJ, Monroe WT. Silver nanoscale antisense
drug delivery system for photoactivated gene silencing. ACS Nano. 2013;7(4):2948-2959

[78] Qureshi AT. Silver nanoparticles as drug delivery systems [thesis]. Louisiana State
University; 2013

[79] Ahamed M, AlSalhi MS, Siddiqui MKJ. Silver nanoparticle applications and human
health. Clinica Chimica Acta. 2010;411:1841-1848

[80] Braydich-Stolle L, Hussain S, Schlager JJ, Hofmann M-C. In vitro cytotoxicity of nanopar-
ticles in mammalian germline stem cells. Toxicological Sciences. 2005;88(2):412-419

[81] Hussain SM, Hess KL, Gearhart JM, Geiss KT, Schlager JJ. In vitro toxicity of nanopar-
ticles in BRL 3A rat liver cells. Toxicology In Vitro. 2005;19:975-983

[82] Kim S, Choi JE, Choi J, Chung K-H, Park K, Yi J, et al. Oxidative stress-dependent toxicity
of silver nanoparticles in human hepatoma cells. Toxicology In Vitro. 2009;23:1076-1084

[83] Singh R, Ramarao P. Cellular uptake, intracellular trafficking and cytotoxicity of silver
nanoparticles. Toxicology Letters. 2012;213:249-259

[84] Ahamed M, Karns M, Goodson M, Rowe J, Hussain SM, Schlager JJ, et al. DNA dam-
age response to different surface chemistry of silver nanoparticles in mammalian cells.
Toxicology and Applied Pharmacology. 2008;233:404-410

[85] Beer C, Foldbjerg R, Hayashi Y, Sutherland DS, Autrup H. Toxicity of silver nanopar-
ticles—Nanoparticle or silver ion? Toxicology Letters. 2012;208:286-292

[86] Ji JH, Jung JH, Kim SS, Yoon J-Y, Park JD, Choi BS, et al. Twenty-eight-day inhalation
toxicity study of silver nanoparticles in Sprague-Dawley rats. Inhalation Toxicology.
2007;19:857
[87] Lee H-Y, Choi Y-J, Jung E-J, Yin H-Q, Kwon J-T, Kim J-E, et al. Genomics-based screen-
ing of differentially expressed genes in the brains of mice exposed to silver nanoparticles
via inhalation. Journal of Nanoparticle Research. 2010;12:1567

[88] Kim YS, Kim JS, Cho HS, Rha DS, Kim JM, Park JD, et al. Twenty-eight-day oral toxicity,
genotoxicity, and gender-related tissue distribution of silver nanoparticles in Sprague-
Dawley rats. Inhalation Toxicology. 2008;20:575

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