Medicinal plants: Treasure trove for green synthesis of metallic nanoparticles and their biomedical applications

Medicinal plants: Treasure trove for green synthesis of metallic nanoparticles and their biomedical applications

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Journal Pre-proof Medicinal plants: Treasure trove for green synthesis of metallic nanoparticles and their biomedical applications Harish Chandra, Pragati Kumari, Elza Bontempi, Saurabh Yadav PII:

S1878-8181(19)31299-X

DOI:

https://doi.org/10.1016/j.bcab.2020.101518

Reference:

BCAB 101518

To appear in:

Biocatalysis and Agricultural Biotechnology

Received Date: 4 September 2019 Revised Date:

6 January 2020

Accepted Date: 28 January 2020

Please cite this article as: Chandra, H., Kumari, P., Bontempi, E., Yadav, S., Medicinal plants: Treasure trove for green synthesis of metallic nanoparticles and their biomedical applications, Biocatalysis and Agricultural Biotechnology (2020), doi: https://doi.org/10.1016/j.bcab.2020.101518. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2020 Published by Elsevier Ltd.

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Medicinal Plants : Treasure Trove for Green Synthesis of Metallic Nanoparticles and their

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biomedical applications

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Harish Chandra1, ¥, Pragati Kumari2,¥*, Elza Bontempi3, Saurabh Yadav4*

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249404, India

Department of Botany and Microbiology, Gurukula Kangri Vishwavidayalaya, Haridwar

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Scientist Hostel-S-02, Chauras campus, Srinagar Garhwal, Uttarakhand, 246174, India

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INSTM and Mechanical and Industrial Engineering Department, University of Brescia, Via

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Branze 38, 25123, Brescia, Italy.

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Srinagar Garhwal, Uttarakhand, 246174, India

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*Corresponding author, ¥ Contributed equally

Department of Biotechnology, Hemvati Nandan Bahuguna Garhwal (Central) University,

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Abstract

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The cornerstone of nanoscience and nanotechnology are nanoparticles which have immense

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power and functional ability in diverse fields. Nanoparticles are synthesized by physical,

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chemical methos but limitations are due to its toxicity. We have discussed few green synthesis

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routes which are eco friendly and less toxic methods, including alage, microorganisms, plants

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etc.. Expoiting the potential of medicinal plants, is one of the green synthesis routes and is

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significant because the current therapeutic approaches have toxicity problems and microbial

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multidrug resistance issues. As the metal nanoparticles have received great attention across the

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globe, so in this study we have discussed and focused many different metallic nanoparticles

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obtained by green synthesis using medicinal plants. We have also discussed the types, size and

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medicinal properties like antibacterial, antifungal, anticancer, antiviral activities of

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nanoparticales. The biomolecules, secondary metabolites and coenzymes present in the plants

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help in easy reduction of metal ions to nanoparticles. Such nanoparticles are considered as

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potential antioxidants and promising candidates in cancer treatment. The simplified model

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summarises the green synthesis, its characterization using physicochemical means and their

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biomedical applications. Succinctly, we have discussed the recent advances in green synthesis of

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metallic nanoparticles, milestones, therapeutic applications and future perspectives of

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biosynthesized nanoparticles from some important medicinal plants.

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Keywords: Green synthesis, Nanoparticle, Nanotechnology, Medicinal plants, Antibacterial,

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Antifungal

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1. Introduction The life expectancy of human beings has increased after the immense progress in medical

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sciences. Earlier the average life of a human being was about 50-60 years. Now it has been

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increased to approx 70 years and above (Roser 2016). This is possible due to immense

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contribution of applied research in medical sciences in the field of diagnostics, prevention and

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targeted treatment (Roser 2016). The most valuable contribution of microbiological sciences

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was the discovery of antibiotics. When first antibiotic penicillin was discovered and introduced

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in 1940, it saved the life of many soldiers who earlier died of septic or wound infection mainly

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caused by Staphylococcus spp (Tan and Tatsumura 2015). Soon after the introduction of

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penicillin, Staphylococcus became resistant and use of penicillin became less relevant to treat

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Staphylococcus infection.. During evolution, bacteria developed resistance towards

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antimicrobial agents (Fair and Tor 2014). Also the indiscriminate use of these antibiotics have

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added to the woes. Researchers across globe are quite aware that such irrational use of

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antibiotics, will make the treatment difficult as antibiotics will become toothless (Ventola, 2015).

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So the world is in the search of newer alternative of pharmaceuticals which are small and

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capable of targeted delivery (Taylor 2016). Thus, the scientists are keen to investigate smaller

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medicinally important molecules and nanotechnology seems to be the result of that curiosity.

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Nanotechnology is emerging science which deals with the study of small particles or

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nanoscale materials whose size ranges from 1-100 nm ( 1 nm = 10-9 m). Ancient system of

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medicine in India, Ayurveda deals with the different formulations which are suggestive about

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the nanosize range in for e.g. Swarna Bhashm, Muktashukti bhasma, Abhrak bhashma, Tamra

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bhasma, Louha Bhasma (Pal et al., 2015;Sharma and Prajapati 2016). Bhasmas are generally a

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metallic preparations having decoction of medicinally important herbs and it is widely used for 3

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the treatment of various diseases (Pal et al., 2015). These bhasmas were considered to be aseptic

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and free from toxicity in therapeutic doses which proved its enhanced efficacy (Chaudhary,

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2011). The term nanotechnology was first given by N. Taniguchi at the international conference

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on industrial production in Tokyo in 1974, in order to describe the very thin development of

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materials within nanometer range. This idea was taken up by Feynman and it was later developed

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by Drexler. In the year 2000 American President Mr. Bill Clinton showed interest in

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nanotechnology and started funding for cutting edge research. On the similar lines, President

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George W. Bush signed and introduced the Nanotechnology Research and Development Act.

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This legislation made nanotechnology research a national priority and of prime importance

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thereby forming National Technology Initiative (NNI). This nanotechnology based applied

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research has found importance in many research institutions around the world.

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2. Why Green synthesis ?

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In the last few years Indian subcontinent witnessed great progress in terms of herbal based

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products. This revolution is due to the trust of people towards Ayurvedic products or herbal based

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products as most of the people now aware of side effects of a chemically derived or synthetic

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antimicrobial compounds, which have adverse effects on human health. Another reason was the

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development of antibiotic resistance to multiple drugs (Srivastava et al., 2014; Chandra et al.,

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2017). In medical sciences the infection is controlled by the use of antibiotics, however due to

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the emergence of Multiple Drug Resistance (MDR) bacteria, it is very much difficult to treat

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such patients who are infected with MDR pathogenic bacteria. In the case of Urinary tract

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infection, E. coli is the most frequently encountered bacteria and it became resistant to most of

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the common drugs which is being used today to control the infection. This bacteria have shown

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good response towards Gentamicin but the problem associated with this drug is its side effects. It 4

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is nephrotoxic and could cause permanent kidney damage (Pazhayattil and Shirali 2014). So, to

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avoid such toxic and life threatening side effects, people are shifting their dependence towards

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herbal based antimicrobial agents which are effective, eco-friendly and relatively free from side

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effects. Towards this we performed searching by using the keywords

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“green AND synthesis AND nanoparticles AND plant” and results showed that India has more

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than 1400 published papers from 1996 to 2018 and is the most active region working in this field

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of research, followed by Iran which published about 260 papers (Fig. 1). With these keywords,

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the search result showed that the field of materials science is promising and stands second one

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after the Biochemistry, Genetics and Molecular Biology area containing the largest number of

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publications (Fig. 2). For the synthesis of nanoparticle, researcher have tried different methods

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like physical and chemical methods. However, these methods weretime consuming, expensive,

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require sophisticated electronic or electrical equipment. These methods use toxic chemicals that

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have many health hazards and it was not environment friendly (Chandra et al., 2019). So, green

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synthesis including important medicinal plants were proving helpful and promising in this regard

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(Elemike et al. 2019; Khatami et al. 2018).

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3. Synthesis of metallic nanoparticles

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3.1 Sol-Gel Method

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The Sol-gel method is well known method for the synthesis of metallic NPs in which there are

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certain essential preparation steps for the synthesis nanoparticles. Initially monomers are

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converted to the sol (a colloidal solution) which is the precursor for subsequent gel formation

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(Owens et al., 2016). This method has the following steps i.e. the metal undergoes hydrolysis and

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forms metal hydroxide solution and rapid condensation leads to three-dimensional gel formation.

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The gel is then dried and the product is rapidly converted to Xerogel or Aerogel. The 5

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nanostructures obtained are of high purity and homogenous structure. Sol-gel process is more

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preferred as it is economically feasible and involves low-temperature (Thiagarajan et al., 2017;

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Owens et al., 2016).

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3.2 Polyol Method

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In Polyol process there is synthesis of metallic nanoparticles by using polyols which

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acts as a reaction medium which has role of reducing agent, solvent and complexation

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agent simultaneously with dissolved stabilizing agents (Dhand et al., 2015). It is

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liquid phase synthesis in which polyol group such as ethylene glycol etc. reacts with

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low cost metal salts with different proportion following the addition of sulphuric acid

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and heating the solution upto boiling temperatures. After the reaction, solution was

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cooled to room temperature, and the particles were separated from the liquid by

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centrifugation and then repeatedly washed with ethanol. The resulting particles were

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dried at room temperature till further use (Kim et al., 2006).

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3.3 Microemulsion

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Microemulsion solutions contain at least three components viz. polar, non polar and

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surfactant. Microemulsions are homogeneous, isotropic and thermodynamically

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stable solutions (Malik et al., 2012). It is one of the inorganic nanoparticle synthesis

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method in which microemulsiom material and reactant are mixed together due to the

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collision of water droplets of microemulsions with reactant. The precipitation occurs

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in the nanodroplets followed by nucleation and coagulation. The resultant

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nanoparticles surrounded by water (Rane et al., 2018). There are namely two routes

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for synthesis of the nanoparticles viz. one micro emulsion method and two micro

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emulsion method.

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3.4 Hydrothermal synthesis

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Nowdays the potential of rapid and continuous techniques for controlled production of

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inorganic nanomaterials has been demonstrated (Darr and Poliakoff 1999). The

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promising green approach are mostly water-based viz. continuous hydrothermal synthesis

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process (Bartkowiak et al., 2018; Jaggessar and Yarlagadda, 2020). Hydrothermal

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process is a liquid phase technology that has gained importance during years. In the

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hydrothermal process, inside an engineered mixer having preheated water under high

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temperature and high pressure gets reacted with aqueous metal salt solution under

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continuous flow and nanoparticle metal oxide is the product (Darr and Poliakoff 1999).

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The process varies and the product is dependent upon the process conditions and reagents

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used.

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4. Green synthesis routes of metal nanoparticles

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There are different routes of green synthesis methods to synthesize nanoparticles exploiting

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microbes and plants etc. which have been proven safe, efficient and cost-effective (Gowramma

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et al., 2015). The different routes for the synthesis of metallic nanoparticles uses biological

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organisms like bacteria, viruses, actinomycetes, fungi (including yeast), plant extracts etc. (Shah

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et al., 2015). These green synthesis methods involve an eco-friendly approach using green

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chemistry. In comparison to the above-mentioned biological entities working as an efficient

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factories for nanoparticle synthesis, the NPs formed via plants is comparatively a straight

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forward method which is more advantageous approach (Iravani 2011). Microbes have the ability

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to help synthesize inorganic materials and this is a bottom-up approach. Each microbe acts in a

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different manner and interacts differently with particular metallic ions. The formation of metallic

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nanoparticles depends on cellular milieu (pH and temperature) in specific microorganisms and

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the biochemical processing in respective microbes determines with a particular size and

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morphology (Iravani 2014). There are different routes of green synthesis exploiting the potential

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of the following organisms-

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4.1 Bacteria

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The different bacteria possesses inherent ability to reduce the heavy metals and can be

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considered potential candidates for synthesis of nanoparticles. This reduction is a combination of

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many factors nanoparticles and few organic functional groups present at bacterial cell wall seems

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to be an important factor which induces reduction. In this category, the bacterial species helpful

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in the synthesis of metallic nanoparticles are Escherichia coli, Bacillus cereus, Klebsiella

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pneumoniae, Actinobacter sp., Lactobacillus spp., Corynebacterium sp., Pseudomonas sp. etc.

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(Iravani 2014; Sunkar and Nachiyar 2012). Both intracellular and/or extracellular mechanisms

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are used by bacterial cells involved in the synthesis of metallic nanoparticles. Facile biosynthesis

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of zirconium dioxide crystalline nanoparticles were synthesized using Acinetobacter sp. KCSI1

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(Suriyaraj et al., 2019). The gold (Au) ions were effectively reduced by using Pseudomonas

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aeruginosa and it resulted in the extracellular synthesis of gold nanoparticles (Husseiny et al.,

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2007). Also the reduction of palladium and its biocrystallization was observed by Desulfovibrio

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desulfuricans (Yong et al., 2002). Regarding the yield of nanoparticles and their proper

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synthesis, selection of correct culture media for a specific bacterial populations and the desired

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metallic salt is very important (Roh et al., 2001; Yong et al., 2002)

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4.2. Actinomycetes

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There are reports about the biogenic synthesis of metal nanoparticles from actinomycetes and

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their biomedical and therapeutic applications (Golinska et al., 2014). In the actinomycetes

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culture, the enzymes secreted from the cell wall and cell membrane helps in the reduction of

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silver and gold ions and the proteins help in capping and stabilization of the nanoparticle

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formation (Sukanya et al., 2013).

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4.3. Algae

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Algae offers a quick and cheap route of green synthesis (Baker et al., 2013). Algae came in use

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for NP synthesis because the nucleation and growth of crystal was accelerated due to the

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presence of negative charge on the surface of algal cells and also the industrial synthesis is at

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very low cost. The metabolites secreted by the culture medium of the Chaetomorpha linum algae

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causes the reduction of silver nitrate. Flavonoids and terpenoids in the extract acted as effective

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capping and stabilizing agents and also they helped in the formation of NPs which were useful in

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medical fields (Kannan et al., 2013). Spirulina platensis mediated biosynthesis of gold

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nanoparticles and the bio functionality was tested by its antibacterial activity against S. aureus

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and B. subtilis (Suganya et al., 2015). Polysaccharides of algal species helps in controlling the

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size and shape of silver nanoparticles. Few marine algae are efficient in nanoparticle synthesis

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and the NPs are useful also (El-Rafie et al., 2013).

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4.4. Fungi

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Another group of organisms like fungi have enzymes and proteins which can reduce metal ions

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into nanoparticles and also stabilize the resulting nanoparticles (Khandel and Shahi 2018).

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Plethora of different proteins in fungi convert the metallic salts into corresponding nanoparticles 9

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and the process is very fast. There are many fungi viz. Aspergillus sp., Fusarium sp., and

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Penicillium sp. which have their biosynthetic ability to create both silver and gold nanoparticles

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(Vigneshwaran et al., 2007; Philip 2009; Kathiresan 2009). Trichothecium sp, an alkali tolerant

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fungus were used in biosynthesis of gold nanoparticles (Ahmad et al., 2005). Green synthesis of,

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silver nanoparticle from soil fungus Penicillium italicum was carried out and tested effectively

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against multi drug resistance (MDR) pathogens (Nayak et al., 2018). In comparison of bacteria,

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fungi contains and efficiently secrete different proteins and several enzymes per unit biomass for

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proper synthesis of metallic nanoparticles due to which yield of nanoparticles is increased

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(Narayanan and Sakthivel 2010). Yeasts are simple eukaryotes and have proper post-translational

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methods which helps proteins and enzymes for efficient function. Silver nanoparticles were used

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in the biotransformation using Saccharomyces cerevisiae (Korbekandi et al., 2016). The

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synthesis of nanoparticles were in the cells, inside cell membrane and attached to cell membrane

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and also outside of the yeast cells (Korbekandi et al., 2016). A silver tolerant yeast strain MKY3

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was used in green synthesis of silver nanoparticles extracellularly (Kowshik et al., 2003). Thus

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they are preferred in comparison to bacteria due to mass production of nanoparticles and easy

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handling and control in laboratory using simple nutrients (Moghaddam et al., 2015).

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4.5. Plants

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The routes via plants especially medicinal plants offers extra advantages and also it does not

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require any complex protocols or methodologies. In the case of microbes, there are multi-step

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process including isolation of potential microbe, specific culture preparation and sub-

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culturing, maintenance of culture etc. Furthermore, the process via plants is comparatively

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easy for scaling up, for large scale production of nanoparticles (Jha et al., 2009; Bar et al.,

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2009). In green synthesis method involving the plant materials such as leaves, root, stem, 10

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bark, flower, fruit etc. (Akhtar et al., 2013) which act as reducing and stabilizing agent are

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mixed with desired metal solution such as Silver nitrate (AgNO3), Titanium oxide (TiO2),

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Zinc Oxide (ZnO), Zinc acetate (Zn(CH3COO).2H2O), Hydrogen tetrachloroaurate

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(HAuCl4), hexachloroplantic acid (H2PtCl6.6(H2O), Copper (II) sulfate pentahydrate salt

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(CuSO4.5H2O) and other metals (Figure 3). Gopinath et al. (2012) synthesized silver

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nanoparticles from the medicinal plant, Tribulus terrestris L. fruit bodies and reported the

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spherical shaped NPs having size between 16-28 nm. These green synthesized AgNPs were

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tested against Staphylococcus aureus, Escherichia coli, Streptococcus pyogens,

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Pseudomonas aeruginosa, Bacillus subtilis and and found to have bactericidal activity

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against all tested isolates of multi-drug resistant bacteria. Also the silver nanoparticles from

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leaf of Tribulus terrestris bearing medicinal properties displayed antibacterial activity

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(Gopinath et al. 2015). The phytomediated synthesis of silver and gold nanoparticle from

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Mentha piperita leaves and their antimicrobial activity was evaluated against human

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pathogens E. coli and Staphylococcus aureus. The gold nanoparticle was found superior than

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silver nanoparticles (Mubarakali et al., 2011). The silver nanoparticle synthesized from the

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leaves of Podophyllum hexandrum inhibited the cellular mechanism of HeLa by DNA

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damage and caspase-mediated cell death. (Jeyaraj et al., 2013). Titanium dioxide

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nanoparticles ( TiO2NPs) were synthesized from Cissus quadrangularis extract was reported

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to have significant bactericidal activity against E. coli and Staphylococcus (Priyadarshani et

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al., 2019). Cinnamon was used as cinnamon as reductant and stabilizer in green synthesis of

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silver NPs and antibacterial activity was observed (Premkumar et al., 2018). The

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biosynthesized zinc oxide nanoparticles from leaf of Justicia wynaadensis which is a

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medicinal herb showed antimitotic and DNA-binding potential (Hemanth Kumar et al.,

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2019). The extract of common herbal plant, Caesalpinia pulcherrima was used for silver

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nanoparticle synthesis and proved cytotoxic on HCT116 cell lines (Deepika et al., 2020). The

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phytomediated synthesis of Zinc oxide nanoparticle was described by Chandra et al., (2019)

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in which Berberis aristata leaves was used for the synthesis of ZnONPs. Francis et al.(2017)

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synthesisized and characterized gold and silver nanoparticle from the leaf extract of

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Mussaenda glabrata by using green synthesis method. A medicinal plant, Cissus

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quadrangularis was used for facile biosynthesis of copper oxide nanoparticles which were

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used as potential antifungal agents against Aspergillus niger, and Aspergillus

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flavus (Devipriya & Roopan 2017). The green biosynthesis of zinc oxide nanoparticles by

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the use of extract from the flower of Nyctanthes arbor-tristis and their antifungal potential

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against many phytopathogens (Jamdagni et al., 2018). A medicinal herb Rosmarinus

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officinalis L. (rosemary) was used for green synthesis of iron nanoparticles. Rosemary-

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FeNPs proved to have cytotoxicity on 4T1 and C26 cancer cell lines (Farshchi et al., 2018).

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The facile biosynthesis of nanoparticles by using plants especially medicinal plants is safe,

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less time needed and economically viable in comparison to other biological organisms like

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microbes etc. (Shah et al., 2015; Mittal et al., 2013). The green synthesized nanoparticles

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possess increased antimicrobial activity in comparison to the other nanoparticles. This

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activated antimicrobial activity may be the result synergistic action of few proteins which

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function in capping and subsequently stabilizing the biosynthesized nanoparticles (Roy et al.,

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2019).

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5. Characterization of nanoparticle

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For nanoparticle synthesis, reaction mixture is exposed to sunlight directly and the color change

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is observed to determine nanoparticle formation.The agglomeration of NPs is avoided after 12

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removing from sunlight once the color intensities are stored in dark. This nanopowder is further

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subjected to characterization using physicochemical means. After synthesis of metal nanoparticle

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it is essential to characterize the synthesized NPs (Figure 3) to know whether the synthesized

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metal nanoparticle are in the nano range and also for shape and chemical nature. Various

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techniques are used for the study of synthesized nanoparticle. X ray diffraction (XRD) analysis

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of the green synthesized nanopowders is carried out and the average crystallite size of

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nanocrystals was calculated using Scherrer's equation (Scherrer 1918). The maximum

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absorbance of the synthesized nanoparticle was measured by UV-Vis Spectrometry and Fourier

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transform- infra red (FTIR) spectroscopy helps in identifying the functional groups involved.

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The shape of the nanopowders were analysed using Scanning electron microscopy and the

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elemental compositions of the nanoparticle was analysed using Energy dispersive X-ray

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spectroscopy (EDX). A noninvasive method is dynamic light scattering (DLS) which is used to

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measure the size distribution of the synthesized nanoparticles. DLS method depends on the

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nanoparticle interaction with light and relies on Rayleigh scattering. These above mentioned

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techniques were used for the characterization of the following biosynthesized nanoparticles from

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medicinal plants e.g. silver nanoparticles from cinnamon (Premkumar et al., 2018), Caesalpinia

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pulcherrima (Deepika et al., 2020), zinc oxide nanoparticles from Justicia wynaadensis

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(Hemanth Kumar et al., 2019), Berberis aristata (Chandra et al., 2019), Nyctanthes arbor-

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tristis (Jamdagni et al., 2018), Solanum nigrum (Ramesh et al., 2015), gold and silver

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nanoparticle from Mussaenda glabrata (Francis et al., 2017), Mentha piperita (Mubarakali et al.,

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2011), copper oxide nanoparticles from Cissus quadrangularis (Devipriya & Roopan 2017) and

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iron nanoparticles from Rosmarinus officinalis (Farshchi et al., 2018). The confirmation of the

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biosynthesized crystals were done by the physico-chemical techniques mentioned above and

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later the biofunctionality was ascertained.

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6. Mechanism of action of nanoparticles

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The modern medicine system is more popular among all form of medicines due its

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immediate action and life saving effects. However, traditional system of medicines takes

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longer time but it acts on the root cause of the diseases.. The Homeopathic medicines are

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based on the nanoparticles and they consist of highly reactive nanoparticles (Bell and

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Koithan, 2012). The nanoparticles display their cytotoxicity by inducing apoptosis or by

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inducing cell cycle arrest (de Stefano et al. 2012). The smaller size of selenium NPs

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enters the cancerous cells via nucleus and does DNA breakage which ultimately leads to

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cell death. The selenium NPs does cytotoxic effects by the disruption of glutathione and

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thioredoxin systems by generation of reactive oxygen species (Menon et al., 2018).

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Mechanism of action of antibacterial activity by the biosynthesized nanoparticles are due

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to the following (i) Reactive Oxygen Species (ROS) generation (ii) bacterial protein

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denaturation (iii) entry in the bacterial cell wall and causes cell death. (iv) disrupting

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bacterial respiratory chain that leads to cell death (v) formation of asymmetrically shaped

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pits in the cell membrane and changes permeability of membrane, thereby causing release

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of lipopolysaccharides and membrane proteins (Rajeshkumar and Bharath, 2017). These

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may be the putative methods by which the nanoparticles exhibit their action and

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biofunctionality. Zinc nanoparticles disrupts plasma membrane of bacteria but the cause

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of interference in permeability of cell is not well established. Some hypothesis may be

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ascribed for this phenomenon viz. the nanoparticles exhibit their antibacterial activity via

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induction of oxidative stress and release of metal ion etc. (Chandra et al. 2019) and also

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formation of hydrogen peroxide from the surface of zinc oxide NPs (Rai et al. 2009).

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7. Medicinal value of Plants

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The burgeoning incidence of diseases caused by MDR pathogenic bacteria in developed and

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developing countries put immense pressure on medical fraternity to search the alternate treatment

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of these resistant microorganisms. In India, people are now showing faith to alternate systems of

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medicines such as Ayurveda, Homoeopathic and Unani practices. All these systems are directly

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or indirectly use medicine which is already described in ancient literature. As mentioned in the

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previous section that different kinds of Bhasmas as prescribed for the treatment of various

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ailments fall within the permissible limit of nanosize (Sharma and Prajapati 2016). The

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researchers from different countries reported the synthesis of different metal nanoparticles from

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medicinal plants and have shown important therapeutic properties such as antioxidant activity,

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antimicrobial activity, wound healing properties, insecticidal activity, anticancer activity,

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immunomodulatory activity, antidiabetic activity, hepatoprotective activity etc. (Table 1). Thus,

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a new era of nanomedicine (Muhammad et al. 2019) is on the rise and use of medicinal plants

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will immensely benefit through their use in formation of metallic nanoparticles..

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7.1. Applications of medicinal plants as nanomedicine

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The application in pharmaceutical and biotechnology industries by the use of nanomedicine has

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seen profound effects (Wagner et al., 2006; Shi et al., 2011). Approximately 100 products of

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nanomedicines clinically approved are finding prominence in the field ranging from drug

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delivery, bio-imaging, biomaterials and diagnostic or other medical devices (Etheridge et al.,

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2013). The drugs which we ingest remains available in a very low amount in the blood streams

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and is termed as bioavailabilty. So, to achieve that bioavailability level, dose optimization is

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necessary and the drug which we take should have a specific target. So, to get appropriate

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bioavailability, theoretically heavy quantity of drugs must be taken but that will result in more

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side effects. The nanotechnology approach is promising technology and has proved that it has

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site specific drug delivery (Hanafy et al. 2019; Wang et al. 2017). Due to this approach the

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required dose of drug will be used and side-effects can be reduced substantially. The use of drugs

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in small quantities and the specific target can reduce the cost of drugs and even reduce pain to

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the patients (Nikalje, 2015). The synthesized polyethylene glycol (PEG) nanoparticles carrying

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antibiotics and termed as “PEGylation” helps in improving drug efficiency and helps delivery of

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genes to target cells. The coatings by PEG helps administration of nanoparticle formulations in

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efficient manner. These formulations were evaluated for the control of bacterial infection, and it

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was found to efficiently target bacterial infection more precisely inside the body (Suk et al.,

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2016). The nano delivery of particles, containing a sub-layer of pH sensitive chains of the amino

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acid histidine, is used to destroy bacteria that have developed resistance to antibiotics because of

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the large dose and sustained release of the drug. Applications of nanotechnology is nowdays

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used efficiently to treat various infectious diseases (Radovic-Moreno et al. 2012). The resin

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based composites having an inorganic residues as filler particles of different sizes in the range of

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supra micron, submicron and nano level are used in dentistry (Schmalz and Arenholt 2009). In

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dental implants and surgery, the resin content of the composites is reduced by filling it with

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nanoparticles. Nanohybrids are the composites and they are a mix of small and large particles in

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a resin based composites but the nano-composites are the ones having only nano particles

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(Albers 2002). The filler particles in a resin based composites vary differently in composition of

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nanoparticles. Earlier the resin-based nanocomposites used to contain the quartz particles (Albers

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2002). The filler nanoparticles gave new dimension to the research in dentistry. With the advent

360

of nanotechnology, silver nanoparticles have proved to be efficient antimicrobial agents due to

361

its interactions with bacteria (Rai et al., 2009; Lok et al., 2006). Silver nanoparticles (AgNPs)

362

were used in filling dental caries from the last few years (Correa et al., 2015; Priyadarsini et al.,

363

2018) for preventing bacterial infections and colonization (Manikandan et al., 2017). These

364

nanoparticles have been used in many dentistry areas like endodontics (Samiei et al. 2013; Lotfi

365

et al. 2011), implantology (Zhao et al., 2011), restorative dentistry (Cheng et al., 2011) and

366

dental prosthetics (Nam 2011). In dental implants also the silver nanoparticles have been

367

successfully used (Flores et al. 2010; Zhao et al., 2011). Durner et al. (2011) and Cheng et al.

368

(2012) have reported the use of silver NPs in restorative dentistry. The nanocomposites

369

containing acrylic resins and silver nanoparticles possess antimicrobial effect against bacteria

370

and also provided better strength in dental treatment (Kassaee et al., 2008).The data on the

371

release of silver nanoparticlesfrom dental implants or fillings are rare.

372

The oral bacteria Streptococcus mutans was inhibited by the antibacterial action of the

373

silver particles which were released from the resin-based composites having silver ion-implants

374

as fillers (Yamamoto et al. 1996). Therefore, from all these studies it can be assumed that either

375

silver nanoparticles as such are released or the silver ions from these particles have beneficial

376

effects. The silver nanoparticle incorporation were aimed for avoiding the microbial colonization

377

thereby, increasing overall oral health.

378

7.2. Nanoparticles in Urinary Tract Infection

379

UTI are most common infection in human beings mainly caused by bacteria and Candida spp. It

380

is most commonly encountered in females and affecting people of all ages. The worldwide

17

381

incidence rate of UTI was 150 million people per year (Swify et al., 2015; Flores-Mireles et al.,

382

2015). E. coli is responsible for approx. 80% of urinary tract infections (UTIs) and while

383

choosing appropriate antibiotics such bacteria should be targeted (Kang et al., 2018).

384

The UTI affects urinary tract including urethra, ureter and kidney. The most frequent

385

microorganism encountered in urine examination is E. coli, followed by Klebsiella, enterococci,

386

groupB streptococci, Pseudomonas, and Proteus mirabilis (Srinivas et al., 2014). UTI is

387

experienced by about 12% of men and 40% women at least once during their lifetime but 40%

388

women display recurrent UTI (Foxman, 2010; Ikäheimo et al., 1996). UTIs are prevalent during

389

the medical complications of pregnancy and this increased incidence during pregnancy is due to

390

the physiological changes that occurs in the genitourinary tract of females during pregnancy

391

(Abdullah and Al-Moslih, 2005; Jeyabalan A, Lain, 2007).

392

Most of the antibiotics used for the control of UTI infections are now ineffective due to

393

development of resistance against these antibiotics (Lee et al., 2018). These urinary tract

394

pathogens acquire resistance due to acquistation of resistance gene acquired through horizontal

395

gene transfer or by other means such as irrational administration of antibiotic without knowing

396

the sensitive pattern of causal microorganisms (Lee et al., 2018). Chandra et al. (2019),

397

synthesized zinc oxide nanoparticle from Berberis aristata and evaluated its antibacterial activity

398

against urinary tract pathogens isolated from urine sample of patients and found that it has

399

significant antibacterial activity against E. coli, Staphylococcus aureus, Klebsiella pneumonia

400

but it has not shown positive response against Proteus spp and S. typhi. The green synthesized

401

copper oxide nanoparticles from indian medicinal plant, Tabernaemontana divaricate leaf, has

402

effective antibacterial activity against the urinary tract pathogens (Sivaraj et al. 2014).

18

403

Ravikumar et al. (2012) also observed in vitro antibacterial activity of Al2O3 nanoparticle as

404

antibacterial agents against urinary bacterial pathogens.

405

7.3. Antiparasitic activity of nanoparticles

406

Malaria is one of the most prevalent parasitic diseases and spread through the female Anopheles

407

mosquito. As per the data given by National Vector Borne Disease control program number of

408

death has been reduced tremendously (NVBDCP, 2012). In India incidence of chloroquine

409

resistance was reported from various places of India and that resistance was due to the K767

410

mutations in pfcrt gene of P. falciparum (Vinayak et al., 2003). So, there is need of some

411

alternate and effective strategy to control malaria in resistant cases. There are several reports

412

which describes the effectiveness of silver nanoparticle (AgNPs). The AgNPs synthesized from

413

the leaf of Andrographis paniculata was investigated for antiplasmodial activity against P.

414

falciparum at 100 µg/mL concentration and it showed 83% inhibition (Paneerselvam et al.,

415

2011). AgNPs synthesized from the leaf extract of Euphorbia hirta also showed the total

416

reduction of P. falciparum (Paneerselvam et al., 2015). Generally, researchers have found

417

different botanicals to control mosquitoes by acting as larvicidal and pupicidal (Prabhu et al.,

418

2011). The green synthesis nanoparticle from plants are reported to have oviposition deterrent,

419

mosquitocidal and inhibitory effect on adult stage of mosquitoes (Soni et al., 2014; Benelli,

420

2016). Silver nanoparticles biosynthesized from Feronia elephantum plant had larvicidal activity

421

against third instar larvae of Anopheles stephensi, Culex quinquefasciatus and Aedes Aegypti

422

(Veerakumar et al., 2014). Green synthesized gold nanoparticles from the Cymbopogon citratus

423

plant of medicinal importance, was observed to be effective against various mosquitos vector

424

(Murugan et al., 2015). The fungal-mediated green synthesis of silver and gold nanoparticles

425

(AgNPs and AuNPs) were effective as larvicidal agents and their efficiency was shown against 19

426

the Anopheles stephensi, Culex quinquefasciatus and Aedes Aegypti (Soni et al., 2012). Dengue

427

fever reduces platelet counts and is fatal to the patient. However, no efficient treatment apart

428

from the platelet transfusion is possible. However, herbs are known to have potential to increase

429

the platelet counts and that can be exployed by green synthesis. Recently, gold nanoparticles

430

from Artemisia vulgaris (Sundararajan et al., 2017) and Cymbopogon citratus (Murugan et al.,

431

2015) were shown to have inhibitory action against dengue fever vector Aedes aegypti.

432

7.4. Antiviral activity of nanoparticles

433

In recent years, there is a burgeoning incidence of a viral infection such as viral hepatitis, HIV,

434

Swine flu and SARS virus, , West Nile virus, monkey pox virus, Hantavirus, Nipah virus,

435

Hendravirus, chikungunya virus are causal agents for other viral borne infection (von Overbeck

436

2003; Daszak et al., 2013). The treatment available in allopathic practice is an administration of

437

the antiviral drug e.g. acyclovir which inhibits viral multiplication but the treatment suffers from

438

many side effects and fail to complelely remove the virus particles from the patient’s body. The

439

life cycle and multiplication of virus follows steps i.e. virus adsorption, virus penetration,

440

uncoating, expression of the virus genome, replication of the genome, protein synthesis,

441

assembly and release of the virus. The drugs developed for viral infection have action on one of

442

the above mentioned steps. The metal nanoparticle e.g. silver nanoparticles has shown promising

443

response against viral infection by inhibiting any of the steps listed above (Galdiero et al., 2011).

444

The silver nanoparticle has antiviral activity against HIV infected cells (Sun et al., 2005; Lara et

445

al., 2010), Hepatitis B Virus (Lu et al., 2008), Herpes simplex virus type I (Baram-Pinto et al.,

446

2009), monkey pox virus (Rogers et al., 2008) and chikungunya virus (Sharma et al., 2019).

447 448

7.5. Antiproliferative/anticancer activity/cytotoxic effect of metallic nanoparticles 20

449

Cancer stands out to be one of the greatest scourge to human life. The standards in treatment and

450

diagnosis of the disease have been elevated by the nanotechnology and considered as the most

451

encouraging introduction for cancer therapy. In the modern system of medicine, some effective

452

treatments available to control or remove the cancerous cells from the body includes

453

chemotherapy, radiation and surgical removal of cancerous tissues. The phytosynthesized

454

nanoparticle has shown some hope for certain type of cancers. Silver nanoparticles from leaf

455

extract of Melia azedarach were active against the HeLa cervical cancer cell lines (Sukirtha et

456

al., 2011). The silver nanoparticle synthesized from Nepeta deflersiana plant have shown

457

anticancer properties against cervical cancer (i.e. HeLa cells) (Al-Sheddi et al., 2018). The MTT

458

assay of silver nanoparticle synthesized from the aqueous leaf extract of pomegranate (Punica

459

granatum) shown anticancer properties against cervical cancer (Sarkar and Kotteeswaran, 2018).

460

Copper oxide nanoparticles were formed from Phaseolus vulgaris (Black bean) and can induce

461

apoptosis and acts as inhibitory action of HeLa cells (Nagajyothi et al., 2017) Mousavi et al.

462

(2018) reported that phytosynthesized silver nanoparticles from Artemisia turcomanica leaf

463

have to cytotoxic activity against gastric cancer cells (AGS) as well as normal fibroblast cells

464

(L–929) by MTT assay. The silver nanoparticles from Piper longum showed cytotoxic effects on

465

HEp-2 cancer cells (Jacob et al., 2011). Gold nanoparticles Allium cepa were useful against the

466

MCF-7 breast cancer cells (Parida et al., 2011). The silver nanoparticles synthesized from the

467

plant have now proven to have potential to control cancer (Abdel-Fattah and Ali, 2018).

468

8. Conclusions

469

The green synthesis of nanoparticles poses research interest to scientists across the globe, due to

470

its applications. Medicinal plants contain phytometabolites of therapeutic values and are

471

currently exploited for benefit to mankind. This method is found to be promising and eco21

472

friendly for the synthesis of metal nanoparticles. Our review article discussed the biosynthesized

473

metallic nanoparticles from various medicinal plants including their pharmaceutical and

474

therapeutic applications in different sectors like biomedical, drug delivery, nanomedicine and,

475

diagnostics. Since the nanoparticles from medicinal plants are free from toxic contaminants, they

476

are well suited in medical sciences and therapeutics. The biosynthesized nanoparticles from

477

medicinal plants have gained the level of competency or biofunctionality as compared to their

478

physically and chemically synthesized nanoparticles. The potential of nanotechnology is

479

emerging in newer fields like DNA nanotechnology which may help to reduce drug toxicity and

480

enhance the efficiency of drug targeting.

481

Acknowledgements- The authors are thankful to Dr. Hemant Ritturaj Kushwaha, Jawaharlal

482

Nehru University, New Delhi, India for critically reading the manuscript.

483

Funding- This research did not receive any specific grant from funding agencies in the public,

484

commercial, or not-for-profit sectors.

485

Legends

486

Figure 1- (Upper) Number of papers reporting the keyword search

487

“green AND synthesis AND nanoparticles AND plant” from SCOPUS (data were analyzed

488

form 1996 to 2018) (Lower) Geographical map depicting India, with more than 1400 published

489

papers rom 1996 to 2018 followed by Iran that published about 260 papers, as the most active

490

region working in this field of research.

491

Figure 2- The field of materials science is second one after the Biochemistry, Genetics and

492

Molecular Biology area containing the largest amount of publications.

22

493

Figure 3- Green synthesis of metallic nanoparticles, their physico-chemical characterization and

494

biomedical applications.

495

Table 1- Metallic nanoparticles synthesized from different medicinal plants, plant part used, size

496

in nm, and their medicinal properties.

497

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498

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40

S.No

Medicinal Plant

1. 2. 3. 4.

Syzygium cumini Ixora coccinea Ananas comosus L. Memecylon umbellatum Malva parviflora

5.

6. 7. 8. 9. 10. 11. 12. 13. 14. 15.

Jatropha curcas Jatropha gossypifolia Pedilanthus tithymaloides Euphorbia milii Vitex Negundo Artocarpus heterophyllus Euphorbia hirta Coleus aromaticus Cycas Iresine herbstii

16. Melia azedarach 17. Eclipta prostrata 18. Tinospora cordifolia Miers 19. Boswellia ovalifoliolata 20. Andrographis paniculata 21. Nelumbo nucifera

Types of Nanoparticles (NPs) AgNP AgNP AgNP AgNP

Plant part used Seed Leaves Fruit Leaf

Size of NPs (in nm) 93 13-57 12 15-20

AgNps

Leaf

19-25

AgNP AgNP AgNP

Latex Latex Latex

73 62 123

AgNP AgNP AgNP

Latex Leaf Seed

AgNP AgNP AgNP AgNP

Medicinal Properties

References

Antioxidant activity ND Antioxidant activity Antimicrobial activity

Banerjee and Narendhirakannan, (2011) Muthu and Rangasamy, (2013) Ahmed and Sharma (2012) Arunachalam et al., 2013

Inflammation, abscesses, pimples, kidney infections, vaginal inflammation, fever, headache, spider stings, bronchitis, pharingitis, tuberculosis Antibacterial Antibacterial Antibacterial

Zayed et al., 2012

105 18.2 10.78

Antibacterial Antibacterial Antibacterial

Patil et al., 2012 Zargar et al., 2011 Jagtap and Bapat (2013)

Leaf Leaf Leaf Leaf

20-30 40-50 2-6 44-64

Karthikeyan et al. (2012) Mahendran and Gurusamy (2013) Jha and Prasad (2013) Dipankar and Murigun (2012)

AgNP AgNP AgNP

Leaf Leaf Leaf

78 35-65 55-80

Larvicidal and Pupicidal Bactericidal Antioxidant Antioxidant, Antimicrobial and antitumor activity Antitumor activity Larvicidal activity Pediculocidal and Larvicidal

AgNP

Stem

30-40

Rheumatic pains

Ankanna et al. (2010)

AgNP

Leaf

35-55

Anti-plasmodial Activity

Panneerselvam et al. (2011)

AgNP

Leaf

45

Larvicidal activity against

Santhoshkumar et al., 2011

Patil et al., 2012 Patil et al., 2012 Patil et al., 2012

Sukirtha et al. (2012) Rajakumar and Rahuman (2011) Jayaseelan et al. (2011)

22. Ocimum

AgNP

Leaf

3-20

Stem, Root 23. 24. 25. 26.

malaria and filariasis vectors Treatment of headaches, coughs, diarrhea, constipation, worms and kidney malfunctions

Mallikarjuna et al. (2011)

Catharanthus roseus Azadirachta indica Olive Hibiscus rosa sinensis 27. Aloe vera 28. Acalypha indica

AgNP AgNP AgNP AgNP

Leaf Leaf Leaf Leaf

35-55 43 20-25 5000

Antiplasmodial activity Antimicrobial Antibacterial Fish pathogen A. hydrophila

Ponarulselvam et al. (2012) Gavhane et al. (2012) Khalil et al. (2014) Philip (2010)

AgNP AgNP

Leaf Leaf

70-192 20-30

Chandran et al. 2006 Krishnaraj et al., 2010

29. Garcinia mangostana 30. Phyllanthus niruri

AgNP

Leaf

35

Antibacterial Antibacterial against fish pathogens MDR Human pathogens

AgNP

Leaf

32-53

Antimicrobial and cytotoxic effect

Krishnamoorthy and Jayalakshmi (2012)

31. Argemone Mexicana 32. Hibiscus subdariffa

AgNP ZnONPs

Leaf Leaf

20 12-46

33. Aloe barbadensis miller ZnONPs

Leaf

25-40

34. Trifolium pratense

ZnONPs 35. Nyctanthes arbor-tristis ZnONPs

Flower

Veerasamy et al., 2011

Antimicrobial Singh et al. (2010) Antibacterial and Antidiabetic Bala et al. (2015) Sangeetha et al. (2011) Cosmetics

100-190 Antibacterial

Dobrucka and Długaszewska, ( 2016)

Flower

12-32

Antifungal

Jamdagni et al. (2018)

36. Solanum nigrum

ZnONPs

Leaf

29.79

Antibacterial

Ramesh et al. (2015)

37. Azadirachta indica

ZnONPs

Leaf

18

Antimicrobial

Elumalai and Velmurugan (2015)

38. Passiflora caerulea

ZnONPs

Leaf

37.67

Antibacterial

Santhoshkumar, et al. (2017)

39. Murraya koenigii

ZnONPs

Leaf

30-35

Antimicrobial

Divyapriya et al. (2014)

40. Hibiscus subdariffa

ZnONPs

Leaf

16-60

Antibacterial and Antidiabetic Bala et al. (2015)

41. Berberis aristata

ZnO NPs

Roots

20-40

Urinary tract pathogens

Chandra et al. (2019)

Highlights • • • •

Green synthesis involves eco-friendly, less toxic methods exploiting medicinal and aromatic plants etc. Biosynthesized metallic nanoparticles have shown potential role as antibacterial agents. The nanoparticles are considered as potential antioxidants and promising candidates in cancer treatment. They will open new avenues in Biomedical applications like antiparasitic, dentistry, urinary tract infections etc.