CHARACTERIZATION AND BIOMEDICAL APPLICATIONS OF

SILVER NANOPARTICLES USING ADIANTUM INCISUM FORSSK.

By
MUHAMMAD AIZAZ SHUAIB
Synopsis submitted to the Abdul Wali Khan University Mardan in the partial fulfillment
of the requirements for the degree of

MASTER OF PHILOSOPHY
IN
BOTANY

DEPARTMENT OF BOTANY
FACULTY OF CHEMICAL AND LIFE SCIENCES
ABDUL WALI KHAN UNIVERSITY MARDAN
SESSION (2021-2023)
 CHARACTERIZATION AND BIOMEDICAL APPLICATIONS OF
SILVER NANOPARTICLES USING ADIANTUM INCISUM FORSSK.

By
MUHAMMAD AIZAZ SHUAIB
REGISTRATION NO: AWKUM-682250

Approved by

Supervisory Committee

__________________________ Supervisor
Prof. Dr. Waheed Murad
Department of Botany

__________________________ Co supervisor
Prof. Dr. Muhammad Irshad
Department of Botany

__________________________ Member
Prof. Dr. Anwar Hussain
Department of Botany

__________________________ Convener, Graduate Studies Committee

Prof. Dr. Muhammad Hamayun

__________________________ Dean, Faculty of Chemical and Life Sciences

Prof. Dr. Sultan Ayaz

____________________
Prof. Dr. Saeed Islam Director Academics/ASRB

DEPARTMENT OF BOTANY
FACULTY OF CHEMICAL AND LIFE SCIENCES
ABDUL WALI KHAN UNIVERSITY MARDAN
SESSION (2021-23)
 RESEARCH PROPOSAL
FOR
MASTER OF PHILOSOPHY IN BOTANY
TOPIC:

CHARACTERIZATION AND BIOMEDICAL APPLICATIONS OF

SILVER NANOPARTICLES USING ADIANTUM INCISUM FORSSK.

Submitted by: Muhammad Aizaz Shuaib

Supervisor Prof. Dr. Waheed Murad
Registration No: AWKUM-682250

Session: 2021-23

Current Semester: 3rd

Co Supervisor Prof. Dr. Muhammad Irshad
Departmental test score: 64

Approval of GSC Recommended for submission to ASRB in 36th meeting of GSC held
on 08/11/2022.

Approval of ASRB

Remarks

DEPARTMENT OF BOTANY
FACULTY OF CHEMICAL AND LIFE SCIENCES
ABDUL WALI KHAN UNIVERSITY MARDAN
SESSION (2021-2023)
1. INTRODUCTION
1.1 Nanotechnology
The term "Nano" is used as the one-billionth (10-9) prefix. Nanoparticles are particles of two or
more dimensions, size ranges between 1- 100 nm in length (Alahazi et al., 2010).
Nanotechnology is a modern scientific advancement that has only been adopted in this century.
Its use in agricultural and food production is essentially modern compared to that of drugs and
medicinal delivery. As science has advanced, nanotechnology has emerged to expand and change
the entire agrifood sector, having the ability to increase the world's food production, in addition
to the nutrient content, food quality, and safety (Thiruvengadam et al., 2018).
In the early 21st century, nanotechnology has emerged as one of the most important scientific
fields, as researchers take advantage of the distinctive properties of atomic and molecule
assemblies made at the nanometer scale. Our ability to control the chemical, biological and
physical characteristics of these particles enables researchers’ capability to logically synthesize
and apply nanoparticles for diagnostic applications and for drug administration (McNeil, 2005).
More than anything else, nanotechnology is a facilitating technology, allowing us to do new
things in almost every conceivable technological discipline. Additionally, its applications will
result in cleaner, superior, quicker, intelligent and affordable products and production methods.
Just as other enabling technologies, e.g., electricity and microelectronics, have changed lives,
Nanotechnology is likely to have a comparable effect, and the transformation could happen much
faster due to the development of robust computers, international connectivity, and many other
technological advancements (Zhu et al., 2004).
In a period of about 50 years, nanotechnology has become the foundation for remarkable
exponential growth and industrial applications. For example, medical equipment has been
significantly impacted by nanotechnology in the pharmaceutical communities of practice, such as
drug delivery systems, imaging probes and diagnostic biosensors. The usage of nanomaterials
has drastically grown for manufacturing advancements in the cosmetics, food industries, shelf
life, packaging and bioavailability (Hulla et al., 2004).
1.2 Nanoparticles
Nanoparticles (NPs) is defined by International Union of Pure and Applied Chemistry as
microscopic particles that range in size from 1 to 100 nm (Batista et al., 2017). These particles
possess unique physical characteristics include conductivity, stability, and optical characteristics,
these properties make them a great choice for materials science and biology. Because NPs have
numerous uses in optics, biology, and electronics. They have been shown to be a successful
option for cellular engineering, tissue engineering, biomolecules, protein diagnosis, environment,
energy, and other applications (Maleki, 2019).
Nanoparticles (NPs) hold a special place in nanoscience and nanotechnology, not only because
of their unique properties that result from their reduced dimensions, but also because they are
potential components for developing more complex nanostructures in nanotechnology
(Benelmekki, 2021).
Because of rapid development of different diagnostic tools and therapy approaches, cancer's
fatality rate, known as one of the deadliest diseases has been significantly inhibited, has been
significantly inhibited. Unfortunately, none of the available cancer treatments can be made to
selectively target and bind cancer cells in order to minimize toxicity and adverse effects. To
overcome this situation, the synthesis of nanoparticles emerges as a new additive. To resolve this
issue, the synthesis of nanoparticles emerges as a new contribution (Yesilot & Aydin, 2019).
Nanoparticles present a new perspective for the protection, detection and treatment of tumors.
Due to their widespread use, the demands of commercial nanoparticle have been increased in
many fields, plant-mediated green synthesis offers a simple and affordable step as an additional
process, which is sustainable and friendly to the environment (Kulkarni et al., 2011).
1.3 Silver Nanoparticles
Silver (Ag) is an important chemical element with atomic number 47 and atomic weight is about
107.8681 g/mol. It is soft, white, lustrous transition metal. Its melting point is 961.8C and its
Boiling point is 2,162C. Silver nanoparticles (Ag NPs) are essential in the field of nanomaterials
and efficient against drug-resistant microbes (Rai et al., 2012).
For thousands of years silver and its compounds have been used for therapeutic and antibacterial
purposes. Silverware was used by the ancient Greeks and Romans to preserve drink, wine, and
food. Hippocrates employed silver-based remedies to treat ulcers and speed up the healing of
wounds (Xu et al., 2020).
Recently, silver nanoparticles (AgNPs) have received a lot of attention, because of their
exceptional physical, chemical and biological properties, and their superiority stems mainly from
the size, shape, composition, structure and crystallinity of AgNPs compared to their bulk forms
(Lee & Jun, 2019). At present, silver nanoparticles have attracted a lot of attention because of
their remarkable biological activities. They are used in cosmetics, bio-labeling, drug delivery,
preservation of food, sensing, water purifications and wound healings. Moreover, silver
nanoparticles are used in different fascinating products, including paints, textiles and catalysis
(Ahmed & Mustafa, 2019).
Silver nanoparticles are used most frequently in the medical industry in topical ointments to treat
open wounds and prevent infection in burn (Lui et al., 2006).
1.4 Plant Description
Pteridophytes are one of the earliest plant groups on earth. The study of pteridophytes which
inhibit a special niche between non seedless and seed-bearing plants. This group of plants is
incredibly diverse in nature having ecological, medicinal, evolutionary, environmental,
ethnobotanical and ornamental significance. In view of these, Microsorum membraneceum,
Lepisorus nudus, Aspenium indicum, Microsorum membraneceum ferns was studied
thoroughly for its potential phytochemical components to demonstrate its therapeutic
applications (Jadhav, 2019).
Adiantum species, commonly known as maidenhair ferns, have a long history of use for
respiratory problems like coughing, colds, pneumonia, fever, and mucus development (Reddy et
al., 2001). Additionally, they have been used as anticatarrhal, demulcent, astringent,
emmenagogue, diuretic, tonic, expectorant, febrifuge, and in the treatment of erysipelas and
diabetes. (Rastogi et al., 2018).
Adiantum incisum Forssk. is an evergreen fern belonging to family Adiantaceae, which is
commonly found in the hilly range of the Swat, Murree and Kashmir region of Pakistan and is
prevalent around the world. A. incisum is known locally by the names of Sumble, Mayursikha,
Raja hans and Hansraj (Bilal et al., 2022).
Adiantum incisum is locally used to cure a variety of diseases, including vomiting, urinary
infections, ulcer, dysentery, hair fall, sterility and healing of wounds (Irfan et al., 2021). A.
Adiantum incisum Forssk (Pteridaceae) is a fern and it is used to treat internal body burning,
jaundice, cough, cold, hemicrania, fever, skin conditions, and liver problems. Additionally, it is
said that this plant has anti-diabetic properties (Kazmi et al., 2019).
The antidiabetic activitiy of Adiantum incisum extract have been evaluated in streptozotocin-
induced diabetic rats. The juice and fronds powder of A. incisum have been used traditionally to
treat Type2 Diabetes Mellitus (TII DM); however, there is no scientific evidence to support its
multipurpose folkloric claim as given in previous paragraph (Bilal et al., 2022).
2. OBJECTIVES
i. To utilize A. incisum plant extracts as reducing agents for the synthesis of silver
nanoparticles.
ii. To evaluate various characterizations analysis i.e., UV-vis spectrophotometry, FTIR and
SEM of the prepared silver nanoparticles.
iii. To evaluate the in vitro antimicrobial, phytotoxic, insecticidal and antioxidant
significance of A. incisum.
iv. To evaluate the in vivo analgesic properties of silver nanoparticles.

3. REVIEW OF LITERATURE
Wei et al. (2015) studied procedures of synthesized silver nanoparticles and assess their
physiochemical, phytotoxicity, and their impact on human life, they emphasized the use of
AgNPs against photo and radio sensitizers, viruses, evaluated anti-cancer properties, breast and
lungs cancer, therapeutic effect on leukemia, skin diseases.
Dakal et al. (2016) Some biogenic AgNPs demonstrated better antibacterial activity than others,
depending on the plant extract and component employed for synthesis. According to reports on
the mechanism of antimicrobial AgNPs activity, DNA loses its capacity to reproduce, and
membrane rupture prevents bacterial development.
Shah et al. (2017) studied the main constituents of the Sapindus genus are saponins, which
have the ability to treat inflammatory diseases. In this study, the effects of soap nut (Sapindus
mukorossi) stem bark methanol extract and its fractions were examined for their antioxidant,
analgesic, anti-inflammatory, and anti-inflammatory properties.
Khan et al. (2018) For the synthesis of silver nanoparticles, experimented with fungus.
AgNPs were discovered to have a small size range, a particular composition, and a high mono
dispersity, all of which were identified for various pharmacological, biological, and
immunomodulatory uses.
Adepoju et al. (2019) Due to the presence of secondary metabolites, synthesized silver
nanoparticles using green methods have also been reported to have biological activities including
antifungal, anti-cancer, anti-inflammatory, antiplasmodial, antifungal, antidiabetic, and
antimicrobial activity against various bacterial strains.
 Das et al. (2020) AgNPs have been synthesized using green techniques from variety of
useful medicinal plants materials including, Azadirachta indica, Ocimum tenuiflorum and
Ocimum sanctum (Jain and Mehta, 2017). Equisitum aevense is a perennial medicinal fern
belong to family Equisetaceae. Pharmacological research has revealed that E. arvense has
multiple biological potentials including, anti-cancer, anti-inflammatory, antibacterial, antifungal,
and anti-proliferative properties.
4. RESEARCH METHODOLOGY
4.1 Collection of plant
Adiantum incisum plant will be collected from swat, Khyber Pakhtunkhwa and will be identified
with the help of flora of Pakistan and matched with an existing specimen present in the
herbarium, Department of Botany, AWKUM (Abdul Wali Khan University Mardan) (Zeb et al.,
2020).
4.2 Drying and powdering
The selected plant will be washed with tap water to remove includes soil particles, dust, and
other silicates. Then the plant specimen will be placed for shade dryness to avoide loss of
volatile constituents. The completely dried plant sample will be then grinded with electric
grinder. The obtained powder will be weight and will be kept in air tight bags till further use
(Das et al., 2010).
4.3 Preparation of ethanolic extract
To make plant extract, 100g of powdered plant A, incisum will be transferred to flask and added
to 500mL of ethanol and will be placed at room temperature for 7 days with occasional shaking.
The sample will be filtered through Whatman filter paper, After 7 days. Then the filtrate will be
placed on water-bath at 75±37oC for evaporation of ethanol. After complete drying the resulting
ethanolic extract of A. incisum will be used as decreasing agent in the nanoparticle’s synthesis
(Leon et al., 2013).
4.4 Preparation of methanolic extract
For making plant extract, 50g of powdered plant of A. incisum will be transferred to flask and
added 250ml methanol. The sample will be placed for 7 days in a shaker. After 7 days, the
sample will be filtered, and the filtrate will be placed in water-bath at 64±7oC for evaporation of
methanol. After complete drying the resulting methanolic extract of A, incisum will be used as
reducing agent in the nanoparticle’s synthesis (Leon et al., 2013).
4.5 Biosynthesis of Silver nanoparticles
Biosynthesis of Silver nanoparticles (AgNPs) from A. incisum, 200 µg/ml aqueous solution of
plant extract will be mixed with 1mM aqueous solution of silver nitrate in particular ratios (1:1,
1:2, 1:3, 1:4, 1:5, 1:6) and will be put on magnetic stirrer for 24 hours with constant stirring so
that brown coloration occurs as silver ions reduced through phytometabolites indicating
formation of AgNPs (Noruzia et al., 2011). The same process will be repeated for methanolic
extract.
4.5.1 Characterization of Silver nanoparticles
Bio fabricated AgNPs will be characterized by UV-spectroscopy. Further, the nanoparticles will
be centrifuged at 4800 rpm for 15 minutes, and the final dry pellet will be washed in distilled
water and filtered. The filtrate containing silver nanoparticles will be used for characterization
studies (Durga et al., 2014).
4.6 Effects of temperature, salt and pH on silver nanoparticles
Various effects of different concentration of temperature, salt and pH on silver nanoparticles will
be done (Santos and Marzan, 2000).
4.7 Biological Activities
4.7.1 Antimicrobial activities
Plant extract and silver nanoparticles will be screened for its antibacterial potential by disc
diffusion technique (Aziz et al., 2014). Anti-fungal screening of AgNPs and extract of A. incisum
will be performed using agar dilution method (Parekh and Chanda, 2008).
4.7.2 Phytotoxicity
Phytotoxic activity of plant extract and silver nanoparticles of A. incisum will be carried out
against lamina minor (Dastagir and Hussain 2013).
4.7.3 Antioxidant activity
Free radical scavenging assay (Antioxidant potential) for plant extract and silver nanoparticles
derived from Adiantum incisum will be evaluated by using 1, 1-diphenyl- 2-picrylhydrazyl
(DPPH) (Aziz et al., 2014).
4.7.4 Insecticidal activity
Silver nanoparticles prepared of plant extract will be tested for insecticidal activity against
Tribolium castaneum (Dastagir and Hussain, 2013).
4.7.5 Analgesic Activity
A comparative analgesic potential of plant extract and prepared nanoparticles will be carried out
using swiss albino mice as test animals (Rauf et al., 2021).
4.7.5 Statistical analysis
The data will be analyzed by using SPSS and Origin 7.5 Software (Ilyas et al., 2018).
4.8 Tentative time of research project

S. No Activity Duration

1 Literature Study 2 months

2 Field and Laboratory work 4 months

3 Thesis Writing 6 months

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