BioNanoSci.
(2018) 8:5–16
DOI 10.1007/s12668-017-0413-3
Recent Advances in Green Synthesis of Silver Nanoparticles
and Their Applications: About Future Directions. A Review
T. M. Abdelghany 1,2 & Aisha M. H. Al-Rajhi 3 & Mohamed A. Al Abboud 2 &
M. M. Alawlaqi 2 & A. Ganash Magdah 4 & Eman A. M. Helmy 5 & Ahmed S. Mabrouk 2
Published online: 26 April 2017
# Springer Science+Business Media New York 2017
Abstract Nanoparticle biosynthetic discipline is still under
development and is known to have a big impact on numerous
manufactures for a long time. Nowadays, biosynthesis of sil-
ver nanoparticles (AgNPs) had gained so much attention in
developed countries due to development demand of environ-
mental friendly technology for material synthesis. The use of
green chemistry is environmental friendly, non-toxic, and
cheap. This review focused on the recent scientific publica-
tions in the green synthesis field of AgNPs and its applica-
tions. A number of microorganisms including bacteria, fungi,
yeasts, algae, and plants either intra- or extracellular have been
found to be capable of synthesizing AgNPs. All scientific
reports reflect the unique properties AgNPs possess that find
myriad applications such as antibacterial, antifungal, antivi-
rus, and anticancer drugs, larvicidal excellent catalytic natural
action towards degradation of dyes, very good antioxidants,
treatment of diabetes-related complications, and wound
healing activities. The recent strategy for improving the effi-
cacy of antibiotics is to combine them with AgNPs in order to
control the microbial infections as confirmed by the damage
* T. M. Abdelghany
tabdelghany@yahoo.com
1
Department of Botany and Microbiology, Faculty of Science,
Al-Azhar University, Cairo, Egypt
2
Department of Biology, Faculty of Science, Jazan University,
Jazan, Saudi Arabia
3
Department of Biology, Faculty of Science, Princess Nora Bent
Abdularahman University, Riyadh, Saudi Arabia
4
Department of Biology, Faculty of Science, King Abdulaziz
University, Jeddah, Saudi Arabia
5
Regional Center for Mycology and Biotechnology, Al-Azhar
University, Cairo, Egypt
action of AgNPs on microbial deoxyribonucleic acid. This
review describes also the microorganism/plant extract and
the reaction parameters used in synthesis of the AgNPs, which
hold prominent impact on their size, shape, and application.
Recently published information on AgNP synthesis and its
applications are summarized in this review.
Keywords Applications . Biosynthesis . Bacteria . Fungi .
Algae . Plants . Silver nanoparticles
1 Introduction
Nanotechnology deals with the nanoparticles having a size of
1–100 nm in one dimension. Nowadays, nanoparticles have
been used commercially for a wide variety of coating areas
including electronics, energy contact actions, and medicines.
AgNPs play a major part in the commercial applications of
these nanoparticles in the field of pharmaceutical and other
medical sciences (Fig. 1). Biosynthesized AgNPs are more
acceptable for medical applications due to superior biocom-
patibility than chemically synthesized ones. The use of green
chemistry involving plant extract and microbes is environ-
mental friendly, non-toxic, and cheap [1]. The recent strategy
to improve the efficacy of drugs is to combine them with metal
nanoparticles for the control of microbial infections [2].
Interaction of AgNPs with heavy metal in the biological re-
moval process remains poorly understood. Yanan et al. [3]
showed that the biological removal electrical capacity of
heavy metals increased with the concentration of AgNPs, sug-
gesting that an appropriate concentration of AgNPs has a stim-
ulating effect on the removal of heavy metals by
Phanerochaete chrysosporium. AgNPs possess unique attri-
butes which are breakthrough myriad applications such as
antimicrobial, anticancer, larvicidal, catalytic, and wound
6 BioNanoSci. (2018) 8:5–16

Fig. 1 Various applications of

silver nanoparticle (AGNPs)

healing activities [4]. During the last 5 years, many efforts
were put into developing new greener and cheaper methods
for the synthesis of nanoparticles [5]. The biosynthesis of
nanoparticles has been proposed as a cost-effective and envi-
ronmental friendly alternative to chemical and physical
methods [6]. A number of microorganisms such as bacteria,
fungus, yeasts, and plants either intra- or extracellular which
are of higher crop yields and lower expenses have been dis-
covered to be capable of synthesizing nanoparticles [7]. The
purpose of our investigation is to open new perspectives and
the future applications of nanomaterial biosynthesis as being
potential antimicrobial agents.
human pathogens, Escherichia coli, Klebsiella sp.,
Salmonella sp., Pseudomonas sp. and Staphylococcus aureus,
revealed that the maximum antimicrobial natural process was
for S. aureus [12]. Endophytic fungus Fusarium sp. was sub-
jected to the extracellular biosynthesis of AgNPs [13], and the
TEM revealed the formation of small sized spherical shaped
AgNps ranging between 12 and 20 nm. Antibacterial activity
of AgNPs against E. coli, Salmonella typhi, and S. aureus
showed 26, 26, and 28 mm, respectively, maximum zones of
inhibition at 60 μl concentration of AgNPs. Also, AgNPs are
biosynthesized extracellularly using the mold Fusarium
graminearum with average diameter 45.5 nm and exhibited
antimicrobial activity against Pseudomonas aeruginosa,
Salmonella sp., C. albicans, and E. coli [14].

2 Biosynthesis by Fungi

Fungi and other microorganisms are ideal candidates in the

synthesis of metal nanoparticles (Fig. 2) with different sizes
(Table 1), because of their ability to secrete large amount of
enzymes [7–9]. Different reducing agents excreted by mi-
crobes (Fig. 3). Salvadori et al. [10] reported that the use of
dead biomass of the fungus Hypocrea lixii as a biological
system is a new, effective, and environmental friendly
bioprocess for the product of nanomaterials. Biological syn-
thesis of AgNPs was found to be between 123 and 195 nm [7]
Fig. 2 Conversion silver nitrate to AgNPs(A, silver nitrate solution
using the fungus Pestaloptiopsis pauciseta, 50–100 nm [11] inoculated with biomass of F. moniliforme; B, distilled water inoculated
and 20–80 nm [12] using Candida albicans. Comparing with biomass of F. moniliforme; C, silver nitrate solution without biomass
C. albicans antimicrobic effect against multidrug-resistant of F. moniliforme [9]
BioNanoSci. (2018) 8:5–16 7

Table 1 Fungal and bacterial biosynthesis AgNPs with different sizes was 9 and the shaking velocity adjusted at 150 rpm was the
Producer organism Size (nm) Reference most suitable for synthesis of the investigated AgNPs. They
are spherical, monodispersed with average size of 12.2 nm.
Candida albicans 50–100 [11] These AgNPs have good antibacterial activity against Gr+ve
Fusarium sp. 12–20 [13] and Gr-ve bacteria. According to Abd El-Aziz [16], produc-
Trichoderma harzianum 19–63 [17] tion of AgNPs was safe and economically viable by success-
Fusarium solani 5–30 [16] fully synthesized culture filtrates of Fusarium solani with high
Cunninghamella phaeospora 12.2 [15] stability and size ranges from 5 to 30 nm. The cell filtrate of
Aspergillus versicolor 15.5 [21] Trichoderma harzianum was used as a producer of AgNPs,
Sclerotinia sclerotiorum 25–30 [23] resulting in its formation within 3 h, and the TEM analysis
Colletotrichum sp. 20–50 [26] showed polydisperse spherical and occasionally ellipsoid
Aspergillus clavatus 25–145 [22] nanoparticles in the size range of 19–63 nm with average size
Aspergillus niger 25–175 [22] of 34.77 nm [17]; AgNPs have biological assay used in agri-
Aspergillus flavus 45–185 [22] cultural purposes to increase the viability of seeds. The TEM
Aspergillus fumigatus 5–95 [22] analysis has revealed that the size of AgNPs synthesized by
Trichoderma viride 15.5 [20] Penicillium expansum and Aspergillus terreus, ranged be-
Penicillium expansum 14–25 [18] tween 14–25 nm and 10–18 nm, respectively [18]. These re-
Aspergillus terreus 10–18 [18] sults have also indicated that the AgNPs synthesized by
Cyanobacterial aqueous 38–88 [27] A. terreus and P. expansum at the concentration of 220 μg/
Nocardiopsis valliformis 5–50 [32] 100 ml media gave the highest reduction of ochratoxin A,
Bacillus pumilus, B. persicus 77–92 [39] where the percentages of reductions were 58.87 and 52.18%,
Pilimelia columellifera 12.7 [38] respectively. Three endophytic fungi Aspergillus tamarii
Bacillus subtilis 40–60 [36]
PFL2, Aspergillus niger PFR6, and Penicillium ochrochloron
Bacillus safensis 5–30 [35]
PFR8 isolated from an ethno-medicinal plant Potentilla
Corynebacterium glutamicum 15 [34]
fulgens L. were used for the biosynthesis of AgNPs [19].
The nanoparticles synthesized using the fungus A. tamarii
Pseudomonas mandelii 1.9–10 [33]
PFL2 were found to have the smallest average particle size
(3.5 ± 3 nm) as compared to the nanoparticles biosynthesized
Culture supernatant (CS) from the fungus Cunninghamella using the other two fungi A. niger and P. ochrochloron which
phaeospora is proved as good new biological source for the produced average particle sizes of 8.7 ± 6 and 7.7 ± 4.3 nm,
biosynthesis of AgNPs [15]. Moreover, the CS formed after respectively.
96 h at 30 °C when the initial pH of the fermentation medium The AgNPs synthesized by Trichoderma viride [20] were
in sizes ranging from 1 to 50 nm. The biogenic AgNPs sig-
nificantly inhibited the growth of all tested pathogenic bacte-
ria. Biosynthesis of AgNPs (with the average size of 15.5 nm)
using extracellular filtrate of Aspergillus versicolor ENT7 as
reducing agent has been reported and exhibited a very good
antioxidant and antimicrobial activity [21]. Many species of
Aspergillus were tested for AgNP synthesis [22]; Aspergillus
fumigatus as the most efficient species showed the highest
nitrate reductase activity among the studied species while
Aspergillus flavus exhibited the lowest capacity in the biosyn-
thesis of AgNPs which was in accord with its low nitrate
reductase activity. Sizes of AgNPs were 5–95, 25–145, 25–
175, and 45–185 nm when using A. fumigatus, A. clavatus,
A. niger, and A. flavus, respectively. Juhi et al. [23] utilize
plant pathogenic fungi Sclerotinia sclerotiorum MTCC 8785
strain for synthesis and optimization of AgNPs production as
well as evaluation of antibacterial properties. TEM images
showed the spherical shape of AgNPs with average particle
size ranges from 25 to 30 nm. Antimicrobial activity of
AgNPs may be due to lysis of AgNPs to the deoxyribonucleic
Fig. 3 Reducing agents within microbial synthesis of silver nanoparticles acid [24]. In another study A. niger also used for biosynthesis
8 BioNanoSci. (2018) 8:5–16
of AgNPs with size range of 11.19–36.83 nm [25].
Mycosynthesis of AgNPs with size ranging from 20 to
50 nm was achieved by endophytic Colletotrichum sp. [26].
DNA of E. coli treated with the AgNPs synthesized by
Colletotrichum sp. showed deformed and damaged deoxyri-
bonucleic acid indicating the action of AgNPs.
3 Biosynthesis by Bacteria
Biosynthesis of AgNPs using 30 cyanobacteria was investi-
gated [27], where Cyanobacterial aqueous extracts were sub-
jected to AgNP synthesis at 30 °C. Scanning of these aqueous
extracts containing AgNP in UV-Visible range showed single
peak. SEM micrographs of AgNP from cyanobacterial ex-
tracts showed that though synthesis of nanoparticles occurred
in all strains, their reaction time (30 to 360 h), shape, and size
(38 to 88 nm) varied (Table 1). Extracellular biosynthesis of
AgNP using a P. aeruginosa [28] and E. coli [29, 30] was
reported. Also, an evaluation of the antimicrobial activity of
the biosynthesized AgNPs against human pathogenic and op-
p o r t u n i s t i c m i c r o o rg a n i s m s , n a m e l y, S . a u re u s ,
Staphylococcus epidermidis, Enterococcus faecalis, Proteus
mirabilis, Acinetobacter baumannii, E. coli, P. aeruginosa,
Klebsiella pneumoniae, S. typhi, Vibrio cholerae, and
Bacillus subtilis was provided. Antimicrobial activity of bac-
terial biosynthesized AgNPs was evaluated for their combined
effects with antibiotics against microbes [30]. Action mecha-
nism of AgNPs and their relation to the generation of oxida-
tive stress in bacteria was studied. Quinteros et al. [31] dem-
onstrated that AgNPs generated oxidative stress in S. aureus,
E. coli, and P. aeruginosa mediated by the increment of reac-
tive oxygen species and that was correlated with a better anti-
microbial activity. Rathod et al. [32] reported the biological
synthesis of spherical AgNPs within the size range of 5–
50 nm by alkaliphilic actinobacterium Nocardiopsis
valliformis, and demonstrated their antibacterial and cytotox-
icity activity. Pseudomonas mandelii synthesized the least-
sized AgNPs with an average diameter of 1.9–10 nm, at tem-
perature as low as 12 °C. On studying their larvicidal activity,
LC90 (lethal concentration) values against Anopheles
subpictus and Culex tritaeniorhynchus larvae were at 31.7
and 35.6 mg/l, respectively [33]. Synthesis, characterization,
and the antibacterial activity of AgNPs from native isolate of
Corynebacterium glutamicum has been reported [34] with
particle size about 15 nm and were found to have enhanced
antimicrobial activity against selected pathogenic strains.
According to Lateef et al. [35], crude extracellular keratinase
obtained from a novel keratin-degrading bacterial strain,
Bacillus safensis, was used for the synthesis of AgNPs.
They are spherical in shape with a size range of 5–30 nm.
The particles showed effective inhibitory activity against five
clinical isolates of E. coli. The SEM graph revealed spherical
shaped AgNPs in the range of 40–60 mm synthesized by
B. subtilis [36]. Amorphous, spherical shapes with sizes under
100 nm AgNP production by Bacillus thuringiensis and
Enterobacter cloacae was performed by Parastoo et al. [37];
AgNPs showed better wound healing efficacy than the control
groups with histological analysis. Biological synthesis of
AgNPs (spherical and 12.7 nm sizes) by acidophilic actino-
mycetes Pilimelia columellifera subsp. pallida was reported
by Patrycja et al. [38] and showed that activity of antibiotics
was enhanced, when tested in combination with AgNPs.
Bacillus pumilus, Bacillus persicus, and Bacillus
licheniformis were able to catalyze the synthesis of AgNPs
with sizes of 77–92 nm [39]. These nanoparticles, especially
those synthesized by B. licheniformis, were stable and showed
an excellent in vitro antimicrobial activity against important
human pathogens and a considerable antiviral activity against
the Bean Yellow Mosaic Virus. Anasane et al. [40] reported
extracellular biosynthesis of AgNPs by acidophilic
P. columellifera subsp. pallida and its in vitro antifungal ac-
tivity against fungi-causing superficial mycoses including
Malassezia furfur, Trichophyton rubrum, C. albicans, and
Candida tropicalis. Results by Baker et al. [41] showed
K. pneumoniae (MTCC 7407) and Xanthomonas campestris
to be more sensitive among the test human pathogen and
phytopathogen, respectively, to AgNPs synthesized by
Pseudomonas fluorescens. These study also reported syner-
gistic effect of AgNPsin combination with kanamycin which
displayed increased fold activity up to 58.3% against
K. pneumoniae. The antibacterial activity mechanisms are
clarified in Fig. 4.
4 Biosynthesis by Plants
The use of plants as the production assembly of AgNPs has
drawn attention, because of its rapid, ecofriendly, non-patho-
genic, economical protocol, providing a single step technique
for the biosynthetic processes and containing important phy-
tochemicals used as reducing agents for AgNP synthesis
(Fig. 5). A large number of plants are reported to facilitate
AgNP syntheses with different sizes are mentioned (Table 2)
and are discussed briefly in the presented review. Biogenic
synthesis of AgNPs was achieved using extracts from sixteen
commonly available plants by sonication method [42] and
their antibacterial assay carried out for the biosynthesized
AgNPs against bacteria (E. coli, Salmonella paratyphi,
S. aureus, and B. subtilis) disclosed excellent antibacterial
activity. Synthesis of AgNPs has been demonstrated using
extracts of Chrysophyllum oliviforme reducing aqueous silver
nitrate [6]. AgNPs were prepared through green route with the
aid of Momordica charantia leaf extract as both reductant and
stabilizer [43]. Plant extracts from Ocimum tenuiflorum,
Solanum trilobatum, Syzygium cumini, Centella asiatica,
BioNanoSci. (2018) 8:5–16 9

Fig. 4 Antibacterial activity

mechanisms of silver
nanoparticles

and Citrus sinensis were used for the synthesis of AgNPs with spherical in shape with an average particle size of 25.24 nm.
an average size of 28, 26.5, 65, 22.3, and 28.4 nm, respective- The H. indicus mediated synthesis of AgNPs showed rapid
ly, using atomic force microscope [44]; their antimicrobial synthesis and higher inhibitory activity (34 ± 0.2 mm) against
activity was performed against S. aureus, P. aeruginosa, isolated bacteria Shigella sonnei at 40 μg/ml. Lantana camara
E. coli, and K. pneumoniae. The highest antimicrobial activity
of AgNPs synthesized by S. trilobatum and O. tenuiflorum Table 2 Plant and algal biosynthesis AgNPs with different sizes
extracts was found against S. aureus (30 mm) and E. coli
Producer organism Size (nm) Reference
(30 mm), respectively. Green synthesis of AgNPs employing
the leaf extract of Justicia glauca [45] produced AgNPs of Ocimum tenuiflorum 28 [44]
10–20 nm in dimensions and showed significant antibacterial Solanum trilobatum 26.5 [44]
and antifungal activity as Streptococcus mutans, S. aureus, Syzygium cumini 65 [44]
Lactobacillus acidophilus, Micrococcus luteus, B. subtilis, Centella asiatica 22.3 [44]
E. coli, P. aeruginosa, and C. albicans. Latha et al. [46] also Citrus sinensis 28.4 [44]
studied leaf extract of Hemidesmus indicus for AgNP synthe- Justicia glauca 10–20 [45]
sis. TEM images proved that the synthesized AgNPswere Hemidesmus indicus 25.24 [46]
Banana peel extract (BPE) 23.7 [49]
Potentilla fulgens 10–15 [51]
Abutilon indicum 5–25 [57]
Pistacia atlantica 27 [58]
Ziziphus jujuba 20–30 [59]
Emblica officinalis 15 [62]
Prosopis farcta 10.8 [63]
Indoneesiella echioides 29 [81]
Rauvolfia serpentina 7–10 [82]
Achillea biebersteinii 12 [83]
Descurainia sophia 1–35 [84]
Artemia salina 2–50 [85]
Pithophora oedogonia 34.05 [86]
Botryococcus braunii 15.67 [87]
Coelastrum sp. 19.28 [87]
Spirulina sp. 13.85 [87]
Limnothix sp. 25.65 [87]
Caulerpa racemosa 25 [90]
Spirulina platensis 11.5 [95]
Nostoc sp. 20.3 [95]
Fig. 5 Bioreductant phytochemicals
10
mediated AgNPs with different leaf extract quantity for the
evaluation of efficient bactericidal activity [47]. AgNPs were
successfully synthesized using AgNO3 via an eco-friendly
and simple green route using Abelmoschus esculentus (L.)
pulp extract at room temperature [48]. The efficacy of
AgNPs against Jurkat cells in vitro was reported and showed
that the IC50 dose of AgNPs leads to the increase in intracel-
lular reactive oxygen species and significantly diminished mi-
tochondrial membrane potential, indicating the effective in-
volvement of apoptosis in cell death, and also exhibited good
antimicrobial activity against different gram class bacteria
[48]. Banana peel extract (BPE) as a reducing and capping
agent was used for AgNP synthesis under optimum conditions
including silver nitrate (1.75 mM), BPE (20.4 mg dry weight),
pH (4.5), and incubation time (72 h) [49]; the average size of
nanoparticles was 23.7 nm as determined by dynamic light
scattering. AgNPs showed effective antibacterial activity
against representative pathogens of bacteria and yeast, and
showed synergistic effect with levofloxacin antibiotic, the an-
timicrobial activity increased by 1.16- to 1.32-fold. In another
study [50], AgNPs were synthesized from Cavendish banana
peel extract with diameter that was in the range of 23–30 nm.
AgNPs in these study showed antibacterial activity against
Gr+ve and Gr-ve bacteria, and also showed a strong DPPH
radical and ABTS scavengers compared to the aqueous peel
extract of C. banana. Biosynthesis of AgNPs using various
parts of medicinal plant P. fulgens. The root extract was found
to have the highest potentiality for the synthesis of nanoparti-
cles. A scanning electron microscopy (SEM) study showed
that the size of the nanoparticle was in the range from 10 to
15 nm and spherical in shape [51]. Furthermore, AgNPs were
evaluated as cytotoxic against various cancer cell lines, and
showed good toxicity at 0.2 to 12 μg/ml AgNPs showed good
toxicity. AgNPs also exhibited comparable antimicrobial ac-
tivity against both Gr+ve and Gr-ve bacteria. Synthesis of
AgNPs using root extract of Erythrina indica was reported
[52]; synthesized AgNPs were found to be spherical in shape
with size in the range of 20–118 nm. These AgNPs showed
potent antibacterial activity against Gr+ve and Gr-ve bacteria
and also exhibited excellent cytotoxic effect on breast and
lung cancer cell lines. Palaniyandi et al. [53] mixed gum ex-
tract of neem (Azadirachta indica) with silver nitrate solution
for AgNPs formation. The synthesized AgNPs exhibited anti-
bacterial activity against clinical isolates of Salmonella
enteritidis and Bacillus cereus. Also, Shakeel et al. [54] syn-
thesized AgNPs using A. indica aqueous leaf extract.
Pugazhendhi et al. [55] stated that synthesis of AgNPs
using Alpinia calcarata root possesses very good
antimicrobial activity which was confirmed by resazurin dye
reduction assay method, and thus, it is a potential source of
antimicrobial agent. Ramar et al. [56] used ethanolic extract of
rose (Rosa indica) petals for AgNP synthesis and evaluated
their potential antibacterial activity using selective human
BioNanoSci. (2018) 8:5–16
pathogenic microbes and anticancer activity using human
colon adenocarcinoma cancer cell line. The characterized
AgNPs showed an effective antibacterial activity against Gr-
ve (E. coli and K. pneumoniae) than Gr+ve (S. mutans and E.
faecalis) bacteria. Rani et al. [57] reported the biological syn-
thesis of Abutilon indicum silver nanoparticles (AIAgNPs)
using aqueous A. indicum leaf extract (AILE) and tested their
biological applications. TEM analysis revealed the spherical
biogenic AIAgNPs of 5 to 25 nm in size. These AIAgNPs
were found to exhibit good free radical scavenging activities,
and the intense zone of inhibition displayed by them in six
different pathogenic species indicates their potential antibac-
terial effect. Synthesis of AgNPs using seed aqueous extract of
Pistacia atlantica and its antibacterial activity were described
by Babak et al. [58]. It was observed that the growths of
Ag-NPs are stopped within 35 min of reaction time. XRD
confirmed the crystalline nature of the 27 nm nanoparticles.
Antibacterial activities of the AgNPs tested against the growth
of Gr+ve (S. aureus) using SEM indicated that the most strains
of S. aureus were damaged and extensively disappeared by
addition of AgNPs. Ziziphus jujuba leaf extract as a reducing
and stabilizing agent was used for AgNPs formation [59].
TEM images showed that the synthesized nanoparticles are
of different shapes with 20–30 nm size range. Synthesized
AgNPs also exhibited good antimicrobial activity against E.
coli. Manikandan et al. [60] reported that the aqueous silver
ions when exposed to unripe fruits of S. trilobatum extract
were reduced and stabilized over long time resulting in bio-
synthesis of surface functionalized AgNPs. These AgNPs
were tested for its antibacterial activity against human patho-
genic bacteria (S. mutans, E. faecalis, E. coli, K. pneumoniae)
and demonstrated anticancer activity in vitro against human
breast cancer cell line. Ashokkumar et al. [61] observed that
A. indicum leaf extract can reduce silver ions into AgNPs
within 15 min of reaction time. The obtained AgNPs showed
highly potent antibacterial activity towards Gr+ve (S. aureus
and B. subtilis) and Gr-ve (S. typhi and E. coli) microorgan-
isms. Synthesizing AgNPs in an aqueous medium was de-
signed [62] using Emblica officinalis fruit extract as stabilizer
and reducer. The formation of AgNPs (average size of about
15 nm) depends on the effect of extract concentration and pH.
AgNPs had significant antibacterial against both Gr+ve and
Gr-ve bacterial strains. Spherical shape AgNPs with mean
diameter 10.8 nm were synthesized also from extract of
Prosopis farcta at room temperature. These AgNPs showed
higher antibacterial activity against multidrug-resistant clini-
cal isolates [63]. Cassia tora leaves extract as reducing agent
was used for AgNP production [64], and AgNP antibacterial
activity against E. coli, P. aeruginosa, S. aureus, and B.
subtilis in different extents was tested, and the highest activity
was observed in E. coli. Efficiency of Andrographis echioides
for green synthesis of AgNPs was studied by Elangovan et al.
[65]. Biosynthesized AgNPs inhibited proliferation of human
BioNanoSci. (2018) 8:5–16
breast adenocarcinoma cancer cell line (MCF-7) with 31.5 μg/
ml at 24 h incubation. Results obtained by Afrah [66]
indicated that extracellular synthesis of AgNPs mediated by
Eucalyptus camaldulensis leaf extract had an efficient
bactericidal activity. Satheesh and Kathireswari [67]
synthesized the AgNPs from silver nitrate using the aqueous
extract of Lawsonia inermis (Henna) plant with much prom-
ising positive results of antibacterial activity. The results of
Ashraf et al. [68] illustrated that biosynthesized AgNPs from
Aloe vera leaf extract have potential antiglycating ability that
may enable their use as therapeutics in the treatment of
diabetes-related complications.
AgNPs were biosynthesized by using leaf extract of
Convolvulus pluricaulis (Shankapushpi, bindweed) and ex-
hibited phenol removal, catalytic and electrocatalytic proper-
ties [69]. Biosynthesis of AgNPs was synthesized from silver
ion using Casuarina equisetifolia L., leaf extract [70], Prunus
amygdalus [71], Aloysia triphylla leaves extract [72], aqueous
fruit extract of Syzygium alternifolium [73], aqueous stem
bark extract of S. alternifolium [74], aqueous extract of
Vigna radiata [75], kenaf (Hibiscus cannabinus) cellulose ex-
tract and sucrose [76], leaf extract of L. camara [43], Vitis
vinifera extract [77], aqueous seed extract of
Dracocephalum moldavica [78], and from aqueous leaf ex-
tracts of Eichhornia crassipes [79] as reducing agents.
Pugazhendhi et al. [80] successfully used Dioscorea alata as
mediator for AgNP syntheses. The synthesized AgNPs
possessed very good antimicrobial activity. AgNPs (29 nm)
were synthesized by using Indoneesiella echioides (L.) leaf
extract as a reducing and stabilizing agent [81] and showed
antioxidant effects and anticancer activity. Sudipta et al. [82]
reported that the synthesized spherical shaped AgNPs (7–
10 nm) from the leaf extract of Rauvolfia serpentina Benth
have antimicrobial and larvicidal activities as well as
cytotoxicity against HeLa and MCF-7 cell lines.
Cytotoxic properties of AgNPs (12 nm) synthesized
using Achillea biebersteinii flower extract were detected
on a human breast cancer cell line [83]. The AgNPs
caused a dose-dependent decrease in cell viability, frag-
mentation in nucleic acid, inhibition of the proliferation
and induction of apoptosis on MCF-7 by suppressing spe-
cific cell cycle genes, and simulation of programmed cell
dead genes. Khatamiet al. [84] concluded that
Descurainia sophia can be used as an effective method
for biosynthesis of AgNPs with size 1–35 nm and con-
centration of 25 μg/ml of the AgNPs with maximum in-
hibitory effect on mycelium growth of Rhizoctonia solani.
They also concluded that the minimum inhibitory, bacte-
ricidal concentrations of AgNPs against Agrobacterium
tumefaciens (strain GV3850) and A. rhizogenes (strain
15843) were 4 and 8 μg/ml, respectively. The toxicity
effect of biosynthesized AgNPs (2–50 nm) from
Syringodium isoetifoliuma against the brine shrimp,
11
Artemia salina exhibited significantly increasing mortality
with magnificent antibacterial activity against thirteen
clinical bacterial pathogens [85].
5 Biosynthesis by Algae
Sinha et al. [86] noted that synthesis process was considerably
rapid, and AgNPs (size of 34.03 nm) were generated within
few minutes of silver ions coming in contact with the algal
extract of Pithophora oedogonia. According to Vijay et al.
[87], AgNPs produced by different strains of microalgae in-
cluding Botryococcus braunii, Coelastrum sp., Spirulina sp.,
and Limnothix sp. showed diameters of 15.67, 19.28, 13.85,
and 25.65 nm, respectively. AgNPs of sizes between 15 and
47 nm were biosynthesized using aqueous extract of Chlorella
vulgaris as reducing agent [88]. Khaled et al. [89] evaluated
the antitumor efficiency of different concentrations of AgNPs
biosynthesized by the blue green algae, Anabaena oryzae,
Nostoc muscorum, and Calothrix marchic, on Ehrlich
Ascites Carcinoma in vitro. Red marine macroalgae,
Amphiroa fragilissima aqueous extract was used as a reducing
agent for the synthesis of AgNPs [90]. AgNPs exhibited anti-
bacterial activity against E. coli, B. subtilis, K. pneumoniae,
S. aureus, and P. aeruginosa. The synthesized AgNPs (25 nm)
using aqueous extract of green alga Caulerpa racemosa [90]
showed antibacterial activity. Similar study on this alga
showed excellent catalytic activity for AgNPs towards degra-
dation of methylene blue [91]. Red algae Laurencia
aldingensis and Laurenciella sp. extracts were used to prepare
AgNPs [92].
High toxicities of the AgNPs on the P4 human foreskin
fibroblast cell line, as well as on human uterine sarcoma
(MES-SA) and the corresponding doxorubicin-resistant mu-
tant MES-SA/Dx5 cells were detected. AgNPs with size of
17.6 nm were synthesized through bioreduction of silver ions
using the Spirogyra varians [93] and can act as a powerful
antibacterial agent against various pathogenic bacteria.
According to Vijay et al. [87], AgNPs produced by different
strains of microalgae including B. braunii, Coelastrum sp.,
Spirulina sp., and Limnothix sp. showed particle sizes of
15.67, 19.28, 13.85, and 25.65 nm, respectively. Also,
AgNPs with size ranged between 15 and 47 nm were
biosynthesized using aqueous extract of C. vulgaris [88].
Internalization of AgNPs by Chlamydomonas reinhardtii
was evidenced for the first time by a combination of high-
resolution imaging techniques [94], Theses techniques pro-
vide sufficient evidence for cellular internalization and bio-
transformation of AgNPs in C. reinhardtii, which are very
helpful for understanding the behavior and fate of AgNPs in
the aquatic environment. Synthesis of AgNPs using Spirulina
platensis (average size of most particles was of 11.5 nm) and
Nostoc sp. (average size of most particles was of 20.3 nm) at
12
room temperature was studied by Entesar et al. [95]. The
AgNPs showed potential antibacterial activity against human
pathogens like S. aureus, S. epidermidis, K. pneumoniae, and
E. coli. Also, AgNPs (3.2–7.0 nm) biosynthesized by
S. platensis, as powder algal biomass [96], showed consider-
able antimicrobial and antioxidant activities for AgNPs, giv-
ing as such a promising approach in the pharmaceutical indus-
try. Nafe et al. [97] reported that Chlorella pyrenoidosa, offers
a high degree of consistency in morphology of synthesized
AgNPs, and also demonstrated the antibacterial and photocat-
alytic properties of these AgNPs. Synthesis of AgNPs using
aqueous extract of Sargassum polycystum C. Agardh and
evaluation of their antibacterial and cytotoxic potentials were
reported [98]. Generally speaking, the size of AgNPs differed
according to producer (Table 2).
6 Biological Methods Advantages of Silver
Nanoparticle Synthesis
Earlier reports cite the use of microorganism such as al-
gae, bacteria, yeast, and fungi for the biosynthesis of
AgNPs. Recently, several plant extracts have been
emerged as novel resources for their ability to produce
safe and non-toxic nanoparticles [11–13, 29–33]. In fact,
biosynthesis results in low-energy use and environmental
impact, with respect to conventional chemical synthesis
methods. Further, the high specificity of biomolecules in-
volved in the biosynthesis process may enable an efficient
control of AgNPs size and shape, whose tight control is
critical to optimize applications. The use of plants as sus-
tainable and renewable resources in the synthesis of
AgNPs is more advantageous over microorganisms,
which need expensive methodologies for maintaining mi-
crobial cultures and more time for synthesis. Benefits of
synthesis of AgNPs using plant extracts are that it is eco-
nomical, energy efficient, cost-effective; provide healthier
work places and communities; and protect human health
and environment leading to lesser waste and safer prod-
ucts [99]. Also, previous studies reported that the
bioreduction potential of the plant extracts is comparative-
ly higher than the microbial culture [45]. Waste products
that resulted from the microbial-based method is likely to
be more harmful to the environment depending on the
type of microbes involved in the synthesis. Hence,
plant-mediated synthesis brings less contamination and
so reducing the impact on the environment. With all the
aforementioned advantages and outstanding features over
other methods, the biosynthetic method employing plant
extracts has viable technique as well as a good alternative
to conventional chemical and physical nanoparticle prep-
aration methods, and even microbial methods [100].
BioNanoSci. (2018) 8:5–16
7 Characterization and Properties of Silver
Nanoparticles
Silver nanoparticles characterization after synthesis such as
size, shape, size distribution, surface area, shape, solubility,
aggregation, etc. is necessary, because the physicochemical
properties of a particle could have a significant impact on their
biological properties and need to be evaluated before
assessing toxicity or biocompatibility. Seung et al. [101] stated
that a variety of physiochemical properties such as size distri-
bution, electrostatics, surface area, general morphology, and
aggregation may significantly affect physiological interac-
tions between nanomaterials and target biological areas.
Therapeutic effects of nanoparticles on brain and retinal dis-
eases depend on size, surface charge, and shape of nanoparti-
cles [102]. To evaluate the synthesized nanomaterials, many
analytical techniques have been used, including ultraviolet
visible spectroscopy (UV-vis spectroscopy), X-ray diffrac-
tometry (XRD), Fourier transform infrared spectroscopy
(FTIR), X-ray photoelectron spectroscopy (XPS), dynamic
light scattering (DLS), scanning electron microscopy (SEM),
transmission electron microscopy (TEM), atomic force mi-
croscopy (AFM), and zeta potential and antibacterial activities
[103]. The spectral response of AgNP in the visible and infra-
red region of the electromagnetic spectrum has attracted even
more attention. The phenomenon known as the surface plas-
mon resonance (SPR) is localized on the surface of the nano-
structure. These resonances stem from the collective oscilla-
tion of surface conduction electrons driven by the incident
electromagnetic field. The SPR properties of metal nanoparti-
cles are strongly size and shape dependent in addition to cap-
ping effects and dielectric constant of the medium. X-ray dif-
fraction is a conventional technique for determination of crys-
tallographic structure and morphology. There is increase or
decrease in intensity with the amount of constituent. This
technique is used to establish the metallic nature of particles
gives information on translational symmetry size and shape of
the unit cell from peak positions and information on electron
density inside the unit cell, namely where the atoms are locat-
ed from peak intensities. DLS is employed for sizing nanopar-
ticles and determining their state of aggregation in suspen-
sions. FTIR is used to determine the nature of associated func-
tional groups and structural features of biological extracts with
nanoparticles. The calculated spectra clearly reflect the well-
known dependence of nanoparticle optical properties. SEM
and TEM are mainly used for morphological studies of nano-
particles. The characterization of scanning electron micro-
scope analysis that is employed to determine the size, shape,
and morphologies of formed nanoparticle SEM that gives
high-resolution images of the surface of a sample is desired
[104]. Gel-electrophoresis (GE) has been demonstrated in re-
cent decades to successfully sort a great variety of nanoparti-
cles according to their size, charge, surface chemistry, and
BioNanoSci. (2018) 8:5–16
corona architecture [105]. Maria et al. [106] evaluated the use
of agarose GE for characterization of Ag nanoparticles (NPs)
in an industrial product.
8 Conclusions and Future Directions
An increasing awareness towards green chemistry and use of
green route for synthesis of AgNPs leads to a desire to develop
environmental friendly techniques. In this review, we provid-
ed an account of the biological sources for AgNP synthesis, as
well as their most promising applications in medical devices
and in environmental processes. Use of microorganisms for
green synthesis of AgNPs is an emerging and exciting area of
nanotechnology and may have significant impact on further
advances in nanoscience. Survey of scientific papers reveals
that these studies on AgNPs applications have been carried
out at in vitro, whereas there are no reports on in vivo appli-
cations. Also, compared with in vitro studies, significantly less
information are available on the potential mechanisms of tox-
icity of AgNPs from in vivo studies. It is expected that the
applications for AgNPs will continue to grow, but there are
more that still need to be understood with respect to their
accumulation in the environment and their potential long-
term effects on humans and animals. We strongly believe that
biosynthesized AgNPs will open a new direction towards var-
ious biomedical applications as Bnanodrugs^ in near future.
There are also hope to that AgNPs can be used in water dis-
infection, nanoweapon against phytopathogens and as surface
plasma resonance enhancers. Also, these AgNPs would pro-
vide a potential solution for the present energy crisis by find-
ing their use as energy-driven devices. Further studies are still
needed to understand the precise molecular mechanism lead-
ing to the formation of AgNPs by microorganisms in order to
have a better control over their size and shape.
Acknowledgments The authors acknowledge Al-Azhar University,
Jazan University, Princess Nora Bent Abdularahman University, and
King Abdulaziz University.
Compliance with Ethical Standards
Conflict of Interest The authors declare that they have no conflict of
interest.
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