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Green synthesis of silver nanoparticles from Guava (Psidium guajava Linn.) leaf for antibacterial, antioxidant and cytotoxic activity on HT-29 cells

(Colon cancer)

Sandhiya V1, Gomathy B1, Sivasankaran M.Rm2, Thirunavukkarasu P1*, Mugip Rahaman A1, Asha S3

1Department of Biotechnology, Dr. M.G.R Educational & Research Institute, Deemed to be University, Chennai, Tamil Nadu, India.

2Department of Biotechnology, Dwaraka Doss Goverdhan Doss Vaishnav College, Chennai, Tamil Nadu, India.

3Department of Biochemistry, D.M.K College for Women, Vellore-632001, Tamil Nadu, India.

Corresponding Author: Dr. P. Thirunavukkarasu E-mail address: [email protected]

ABSTRACT

In this study, biocompatible silver nanoparticles synthesized from aqueous leaf extract of Psidium guajava Linn. The green synthesized AgNPs were characterized by various spectroscopic and microscopic techniques (UV Spectroscopy, FTIR, XRD and SEM). The UV-visible spectroscopic analysis showed the absorbance peak at 418nm, which confirmed the synthesis of silver nanoparticles. Identification of functional groups present in the plant extract, which involved in the reduction of silver ion to AgNPs was analyzed by Fourier transform infrared spectroscopy (FTIR). X-ray diffraction (XRD) analysis confirmed crystalline nature of the synthesized nanoparticles. The synthesized AgNPs were round and spherical shape and 20nm size were confirmed by scanning electron microscopy (SEM). Initial phytochemical analysis revealed the presence of secondary metabolites in the aqueous guava leaf extract. The antibacterial activity of guava leaf extract and AgNPs was dose-dependent manner and it was revealed synthesized silver nanoparticles showed potential antibacterial activity than guava leaf extract. The antioxidant activity of synthesized AgNPs showed highest free radical inhibition (80.2%) at 250 µg/ml as compared to guava leaf extract (62.21 %). The biosynthesized silver nanoparticles were observed to possess high cytotoxicity against colon (HT-29) cancer cells with IC50- 68.26 µg/ml. In conclusion, the results obtained in the present study revealed the green synthesized silver nanoparticles from guava leaf extract exhibited considerable antioxidant, antibacterial and anticancer potential.

KEYWORDS: Silver nanoparticles; Psidium guajava Linn.; HT-29 cells; antioxidant;

antibacterial.

Introduction

Nanotechnology is one of the active research areas in the modern material science. Nanomaterials research has received the considerable attention in worldwide due to its unique properties. A size of the nanoparticle usually ranges from one dimension less than 100nm (Lakhan et al., 2020).

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Nanoparticles, nano films, nano rods and fullerenes are different variety of nanomaterials.

Among these variety nanoparticles have the ability to expose zero dimension with biochemical applications. Past 10 years, the uniform size and shape dependent nanoparticles synthesized increased because which is frequently used in advanced technology (Ebrahimzadeh et al., 2020). Recently, nano-biotechnology has acquired significance in the field of biology, material science and biomedicine. An authentic and non- harmful combination of nanoparticles has a great advance in the nanoparticle’s development.

There are various techniques accessible in nanoparticles synthesis process such as micro wave assist process, sono-chemical, electrochemical, radiation assist and photochemical method. The critical part of these techniques was very dangerous and utilize harmful synthetic compounds which may prompt potential health effects and environmental effects. Plants sources used nanoparticles synthesizing methods have a lot of focus due to its eco-friendly and utilization of nontoxic chemicals (Bharathi & Bhuvaneshwari, 2019).

Among the various inorganic metal nanoparticles, silver (Ag) nanoparticles have received considerable attention in the field of biomedical research. AgNPs gained more attention due to its unique physical, biological and chemical properties (Roy et al., 2019). Recently synthesizing silver nanoparticles through biological method using microorganisms, plant extract and marine algae. Among these the plant mediated AgNPs has high biological properties (Shah et al., 2020).

The plant associated AgNPs has excellent absorption capacity and utilizes the chemical component present in the plant materials and is not toxic to environment (Vijaya anand et al., 2019). Plants contain a wide variety of secondary metabolite including tannins, terpenoids, alkaloids, and flavonoids which have been proved to have anti-microbial, antitumor antioxidant, and other biological activities can be of great attention in therapeutic treatments (Lakshmanan et al.,2018). Plants have the capability to reduce metal ion into metal nanoparticles. Silver has been widely used as healing property and antibacterial agent for many years due to its antiseptic nature and huge therapeutic values. Furthermore, few salts and derivatives of silver economically fabricated as antimicrobial agents. At low concentration of silver-based drugs are effectively affect bacteria and viruses but not for healthy human cells (Mollick et al., 2019). Cancer kills the lives of millions of people worldwide every year. The development of anticancer drug has a great challenge in damaging cancer affected cells without unnecessary toxicity. Hence, over the past few years, the focus of researchers on the development of cancer medications from natural ingredients has increased significantly (Khandelwal et al., 2020). Malignancy is an obstruction in the mechanism of controlling of typical development, multiplication and cell death. Silver nanoparticles act as antitumor by slowing progressive growth of tumor cells (Gomathi et al., 2020).

Psidium guajava Linn. usually known as guava, of the family Myrtaceae is a native plant of tropical America, however presently developed all over the world (Perez et al.,2008). The guava leaves have a long history of medicinal uses that are still used today. Guava leaves, roots and fruits have been used for the prevention and treatment of diarrhea, gastroenteritis and dysentery.

A liquid leaf extract is used to reduce blood glucose level in diabetics (Olatunde yet al., 2018).

High level of antibacterial activity was detected in P. guajava Linn. leaves (Geetha et al., 2017).

Moreover, the various bioactive compounds, such as polyphenol, flavonoids, polysaccharides, enzymes, proteins and ascorbic acid in the guava leaf was responsible for the reduction of silver ions and antioxidant activity (Wang et al., 2018). Guava leaf extract reported to have chemotherapeutic effects and suppress the proliferation of various cancer cells including colon, prostate, oral epithelium, adenocarcinoma and murine leukemia (Ryu et al., 2012).

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Phytochemical screening of leaf extract of guava leaf was carried out according to the methods for identification of carbohydrates, proteins, phenols, terpenoids, alkaloids, coumarins, steroids, paleobotanies, quinones, saponins, flavonoids, and tannins using standard phytochemical method.

Among all the variety of medicinal species studied, we can highlight P. guajava Linn. that are highly implicated in therapeutic medicine against bacterial infection, inflammation, pain and cancer, (Mahfuzul et al., 2007). The aim of the present study is to synthesis a rapid and eco- friendly AgNPs using fresh leaves of P. guajava Linn. and its antibacterial, antioxidant and anticancer activities.

Materials and methods Materials used:

Silver nitrate (AgNO3), Sodium hypo chloride, 2,2- Diphenyl-1-picrylhydrayl (DPPH), Dimethyl sulfoxide (DMSO), Luria Bertani broth and Muller Hinton Agar- HiMedia, India. Dulbecco’s modified eagle medium (DMEM), Fetal Bovine Serum (FBS), Trypsin-EDTA, Antimycotic and antibiotic solution were purchased from Gibco, Thermo fishers scientific, India. 3-(4,5-Dimethyl- 2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) - Sigma Aldrich.

Collection and preparation of Guava leaf extract:

Psidium guajava Linn. used for the preparation of leaf extract were collected from a local region, Chennai, Tamilnadu washed and sterilized with sodium hypo chloride. 25gm of guava leaves were accurately weighed and crushed using a blender and finely macerated. After homogenization 100 ml double deionized water was added and heated over water bath maintained at 80ºC for 15 minutes; Now the leaf extract was cooled at room temperature and then filtered through Whatmann No.1 filter paper (pore size 25µm) and used immediately for the AgNPs synthesis process. (Sun et al., 2014).

Phytochemical evaluation of guava leaf extract:

Fresh extract of guava leaves were used for the phytochemical analysis, such as test for saponins, phenols, tannins, terpenoids and flavonoids for screening the bioactive chemical compounds in the guava leaf extract were carried out using the standard procedure as described by Bisht et al., 2016.

Biosynthesis of silver nanoparticles:

50ml of aqueous guava leaf extract was mixed with 50ml of 1mM AgNO3 solution. Then the reaction mixture stirred using magnetic stirrer for 24h (Ragunandhan et al., 2011).

Characterization of silver nanoparticles:

The synthesized silver nanoparticles were preliminarily confirmed by UV-visible Spectroscopy (Perkin-Elmer MA, USA), range between 350-750 nm. The possible functional groups in the guava leave extract reduced AgNPs were analyzed using Fourier Transform Infrared (FTIR Perkin-Elmer, MA, USA) spectra in the range of 4000-400 cm-1. The crystalline nature of silver nanoparticles was confirmed by X-Ray Diffraction pattern (XRD-Bruker AXS, Inc., Madison, USA). The size and morphology of the nanoparticles were identified by using Scanning Electron Microscope (SEM). Energy Dispersive X-ray (EDX) used for the identification of existence elements of silver (Kumar et al., 2017).

Antibacterial activity by agar well diffusion method:

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Antibacterial activity of guava leaf extract and AgNPs were tested against Escherichia coli, Klebsiella sp., Pseudomonas sp. Those bacterial stains were maintained in pre-cultured in nutrient broth overnight in a rotary shaker at 37°C. The cultured bacteria were spread on the Muller Hinton agar using sterile cotton swab. Wells (6mm) were created with the help of sterilized micropipette tips and different concentration (15 µl, 25 µl, 50 µl, 100 µl) of prepared AgNPs and guava leaf extract were loaded into wells on plate. The plates were then incubated for 18 h at 370C, and zone of inhibition were recorded (Asha et al. 2017).

Determination of free radical scavenging activity by DPPH assay:

0.1 mM DPPH solution was prepared with ethanol. Different concentration of guava leaf extract, synthesized silver nanoparticles and ascorbic acid (e.g., 50, 100, 150, 200 and 250µg/mL) added into prepared DPPH solution. Ascorbic acid was considered as a standard and a blank as DPPH solution without sample was used. The mixture of solution was vortexed and then incubated at 37℃for 30 minutes. Finally, the absorbance of the solution mixture was measured spectrophotometrically at 517nm (Keshari et al. 2020).

Determination of invitro cytotoxic activity by MTT assay:

The present work was carried out in human colon cancer cells (HT-29). The cell lines were purchased from National Centre for Cell Science (NCCS), Pune, India. They were grown in DMEM (Dulbecco’s Modified Eagle’s Medium) medium with 10% Fetal Bovine Serum, 1%

antibiotic and antimitotic solution and maintained in 5% CO2 incubator at 37C. The HT-29 cells (1×104cells/well) were seeded into 96 well plats and treated with AgNPs in various concentration (15, 25, 50, 75 and 100 μg/mL) for 24 h with serum free media. The spent medium was discarded from cells at the end of the treatment period. 0.5mg/mL MTT prepared in 1 X PBS was added and incubated at 37˚C for 4 h using CO2 incubator. The medium containing MTT was aspirated from the cells after incubation period and washed using 200μL of PBS. The developed crystals were dissolved using 100μL of DMSO and the formazan dye turns to purple blue color.

The absorbance was measured at 570nm using microplate reader (Khorrami et al. 2019).

Result

Phytochemical analysis:

The guava leaf extract has been analyzed for their phytochemical constituents. The test results confirmed that the leaves extract contains secondary metabolites like carbohydrates, alkaloids, flavonoids, phenolic compounds, saponins, sugars, etc. The results were qualitatively expressed by positive sign and show the observed color changes in various phytochemical tests (Table 1).

Table 1. Phytochemical analysis

S. No Phytochemical components Guava leaf

1 Carbohydrates ̅

2 Tannins +

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3 Saponins +

4 Alkaloids ̅

5 Flavonoids ̅

6 Quinones ̅

7 Glycosides +

8 Terpenoids +

9 Phenols +

10 Steroids +

11 Anthraquinones +

Characterization of synthesized AgNPs UV-visible spectroscopy analysis:

The formation of silver nanoparticles from guava leaf extract observed using UV-visible spectroscopy. When the leaf extract was mixed with silver nitrite solution, slowly it started to change colour from greenish brown to dark brown color due to the reduction of Ag+ to Ag0 (Fig 1). The surface plasmon resonance peak was raised after 24 h of incubation. A broad peak was observed at 418nm which further confirmed the synthesis of AgNPs as shown in (Fig 2).

Figure 1 - A) Pidium guajava leaf B) leaf extract of Pisidium guajava Linn. and C) synthesized silver nanoparticles after 24 h of incubation

A B C

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Figure 2- UV – VIS spectra of Biosynthesis of silver Nanoparticles

Fourier transform infra-red analysis:

FTIR analysis was to identified the functional groups associated with AgNPs, biosynthesized from P. guajava Linn. leaf extract, which is responsible for capping, reduction and efficient stabilization of the AgNPs. (Fig 3) shows some pronounced absorbance bands observed at 3739 cm-1 [N-H stretching (amine group)], 2918 cm-1 [C-H (sym/Asym) aliphatic], 2360 cm-1 [aromatic rings], 2029 cm-1 [aliphatic nitro compounds NO2 symmetric], 1975 cm-1 [ Ether linkage], 1053 cm-1 [C-C (aromatic benzene)]. There is a significant change in the vibrational spectra of stabilized nanoparticles when compared with increase in the concentration of leaf extract of P. guajava Linn. The absorption peak at 3739 cm-1 gets narrower and broader due to proteins, which are formed between the amine groups.

Figure 3. Fourier transform infrared spectrum of silver nanoparticles

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X-ray diffraction analysis:

Analysis of AgNPs using X-ray diffraction confirmed the crystalline nature of particles (Fig 4).

The XRD spectrum shows four main predominant Braggs diffraction peaks which are positioned at 2θ values of 38.0°, 44.1°, 64.4°, and 77.4° which corresponds to 111, 200, 220, and 311 facets of the face-centered cubic structure of AgNPs. These diffraction peaks are consistent with the standard Ag crystals, indicating that the synthesized AgNPs are of crystalline in nature. The unassigned peaks (*) observed in the XRD spectrum indicates the crystallization of plant phytochemicals phase presence in guava leaf extract occurred on the surface of the AgNPs.

Figure 4- X-ray diffraction spectrum of silver nanoparticles

Energy dispersive X-ray spectrometer analysis:

Energy dispersive X-ray (EDX) spectrometer set up the presence of natural identification of the silver and homogenous assignment of silver nanoparticle. Assessment of AgNPs by Energy Dispersive X-ray (EDX) spectrometer showed the existence of elemental indication of the Ag and homogenous distribution of synthesized AgNPs. The sharpe sign pinnacle of a capably settled the decrease of AgNO3 to AgNPs. The upstanding pivot communicates the quantity of X-ray tallies while the equal hub shows energy in KeV. Detection lines for the main releasing energy for Ag were clarified and these communicate with peaks in the spectrum, thus giving affirmation that Ag has been properly recognized and present in the solution (Fig 5).

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Figure 5- Energy dispersive X-ray spectrum of silver nanoparticles

Scanning electron microscopy (SEM) analysis:

The Scanning electron microscopy (SEM) has been used to identify the size, shape and morphology of silver nanoparticles. Size and shape of the silver nanoparticles depend on the precursor, reducing agents used to synthesize nanoparticles. The size of silver nanoparticles were found at 20 nm and spherical and some particles were round in shape (Fig 6).

Figure 6- Scanning electron microscope image of silver nanoparticles

Antibacterial activity by agar well diffusion assay:

The agar well diffusion method was employed for screening the antibacterial activity of green synthesized silver nanoparticles and aqueous extract of P. guajava Linn against Gram negative bacteria. The aqueous leaf extract and green synthesized AgNPs showed antibacterial activity in all concentrations (15, 25, 50 and 100 µL) against test pathogens such as E. coli, Pseudomonas sp. and Klebsiella sp. and the results are shown in table 2. The highest zone of inhibition was found against Pseudomonas sp. and minimum level antibacterial activity were observed in E. coli for aqueous extract of P. guajava Linn. The synthesized AgNPs showed highest zone of inhibition was found in Klebsiella sp. and lowest inhibition was found in E. coli (Fig 7). The present study clearly indicates the antibacterial activity was dose-dependent manner and it was

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revealed synthesized silver nanoparticles showed potential antibacterial activity than aqueous extract of P. guajava Linn.

Table 2: Antibacterial activity of silver nanoparticles

Figure 7- A) Silver nanoparticles treated bacterial pathogen and B) Guava leaf extract treated bacterial pathogens.

Antioxidant activity by DPPH assay:

The antioxidant properties of guava leaf extract and synthesized AgNPs were examined by a DPPH radical scavenging assay. In this study the antioxidant activity was found in dose- dependent way. The synthesized AgNPs displayed the highest free radical inhibition (80.2%) at 250 µg/ml. Likewise, the guava leaf extract showed the highest scavenging effect (62.21 %) respectively (Fig 8). The DPPH scavenging activity of synthesized nanoparticles exhibited significant inhibition when compared with standard ascorbic acid. Hence the biosynthesized silver nanoparticles showed higher antioxidant activity than guava leaf extract.

Bacterial species

Zone of inhibition

15 μl 25 μl 50 μl 100 μl

Leaf extract

AgNPs Leaf extract

AgNPs Leaf

extract AgNPs Leaf

extract AgNPs

E. coli 4 mm 4 mm 5 mm 7 mm 7 mm 6 mm 8 mm 10 mm

Klebsiella sp. 5 mm 5 mm 6 mm 6 mm 7 mm 21 mm 10 mm 25 mm Pseudomonas

sp. 5 mm 5 mm 7 mm 6 mm 8 mm 9 mm 14 mm 10 mm

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Figure 8: DPPH radical scavenging activity

Invitro cytotoxic activity by MTT assay:

Invitro cytotoxic activity of guava leaf mediated silver nanoparticles was analyzed by MTT assay against HT-29 colon cancer cells. The cells were treated with different concentration of AgNPs, the untreated cells considered as control. The cell viability was assessed following the treatment of cells with different concentrations of AgNPs (15, 25, 50, 100 and 500 μg/mL). The results of the MTT assay revealed that the cell viability decreased with increase in the concentration of the synthesised AgNPs Fig C. The IC50 value was found around 68.26 µg/ml. The synthesized silver nanoparticles treated colon cancer cells changed its morphology and the cell count was reduced when compared to control well (Fig 9). It proved the AgNPs from guava leaf extract has a tendency to kill cancer cells and it acted as anti-cancer activity for colon cancer.

Figure 9: A) HT-29 control cells, B) AgNPs treated cells and C) Cytotoxicity of AgNPs

Discussion

Green synthesis of silver nanoparticles was done by guava leaf extract. Guava leaf extract has important biological components like Tannins, Saponins, Glycosides, Phenols, Anthraquinones detected by phytochemical analysis and alcoholic, protein and nitrogen groups were found by FTIR analysis acts as potential reducing and capping agent to synthesize AgNPs (Nasar et al., 2019; Roy et al., 2019). The synthesized silver nanoparticles were confirmed by visual observation of color changes in solution. The dark brown colored colloidal solution indicating the

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silver ions reduction reaction due to the response of surface plasmon resonance (Pirtarighat et al., 2019). UV spectroscopy analysis used to assess the color change of reaction mixture. The increasing color intensity of colloidal solution increases adsorption rate due to the bulk amount of silver ions reduced and more nanoparticles integrated (kaur et al., 2019). Our report ultimately similar to previous studies. (Parvataneni et al., 2020).

The XRD analysis used to determine the crystalline nature of silver nanoparticles. The crystalline size of the synthesized AgNPs was calculated by the Debye-Scherrer equation, D = Kλ/β cos θ.

Where D = average crystallite size, K - shape factor (0.9), λ - wavelength of Cu Kα radiation (1.541 Å), β - FWHM of refection (in radians) located at 2θ and θ - angle of reflection (in degrees) was used to relate the crystallite size to the line broadening. The average crystallite size of prepared AgNPs was found to be about 25.8 nm (Saxena et al., 2012). Babak sadhegi et al., 2015 reported that green synthesized silver nanoparticles using seed aqueous extract of pistacia atlantica showed similar results. The average size, distribution and surface morphology of synthesized silver nanoparticles have been analyzed using SEM coupled with EDX respectively.

Integrated silver nanoparticles have been found to be crystalline aggregated and spherical in size and different size ranges. Such differentiations in size and shape are common when using various biological compounds using synthesis process (Kedi et al., 2018). Previously similar results have been found reported for silver nanoparticles from plant extract as reducing agents (logeshwari et al., 2015; Awwad eta l., 2009).

The antibacterial mechanism of silver nanoparticles which releases silver ion into the bacterial cells and inactivated the bacterial enzymes. It inhibits DNA replication, cause cytoplasm damage and reduced adenosine triphosphate level and destroy the bacterial cells (Hajmohamadi et al., 2019). Because nanoparticles are smaller in size and have a larger surface area gives more effective in antibacterial activity (Bilal et al., 2017). The smaller nanoparticles have a tendency to penetrate into the bacterial cell membrane for the destruction of DNA and cause cell death (Bhakya et al., 2015). Table-2 represented the biosynthesized silver nanoparticles showed potential antibacterial activity than guava leaf extract. Radhika Parvataneni, 2018 reported that aqueous leaf extract of Scoparia dulcis L. synthesized AgNPs showed maximum antibacterial activity against E. coli (13 mm) and Pseudomonas sp. (11 mm) at 1mg/mL of concentration.

Ephedra procera C. A. Mey. mediated silver nanoparticles showed good antibacterial activity against E. coli and Klebsiella sp. (Muhammad et al., 2019). There are many studies and reports that plant mediated silver nanoparticles has strong antimicrobial activity. These AgNPs were effective in killing a wide variety of bacterial pathogens involved in various infectious diseases (Guerra et al., 2020).

The increased formation of reactive oxygen species (ROS) in the living cells as a results of oxidation process, and it is extremely unhealthy due to vigorous oxidation-reduction potential.

Under normal biological condition, the generation of ROS and antioxidant capacity of cells ought to be balance, whereas beneath imbalance condition, the cells start the oxidation process to develop oxidative stress. (Roy et al., 2019) Free radicals can cause cellular damages in the human body, which is generally neutralized by antioxidants substances in the plant materials. Most of the antioxidants are known to slow down the progression of chronic diseases (Ansar et al., 2020).

The antioxidant capacity of guava leaf extract and synthesized silver nanoparticles were found that that AgNPs have more radical scavenging activity than leaf extract. The antioxidant capacity was observed due to the presence of phytochemicals like phenolic functional groups in the plant extract as capping agents of the nanoparticles (Kumar et al., 2016).

In the current research, the impact of guava leaf extract synthesized silver nanoparticles on the cytotoxicity of HT-29 cells was analyzed. Colon cancer is the third most dangerous cancer in

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world wide. The silver nanoparticles were able to inhibit the HT-29 cells in a dose dependent manner (Muthu Irulappan et al., 2010). Silver nanoparticles have pro-apoptotic and antioxidant potential which acted as therapeutic agent for cancer cells. When silver ions enter into the cancer affected cells which blocks the DNA repair enzymes it can suppress DNA multiplication (Biresaw et al., 2020). This might be attributed to two effects, intracellular Reactive Oxygen Species (ROS) generation by AgNPs and its activity on the key cellular components, leading to cell death (Samari, F et al., 2019). Factors such as particles size and surface charges have greater dissemination and high penetration in the tumor cells. Cancerous cells having a different pore size compared to other cells, the size-controlled silver nanoparticles can be exhibited effectively in the therapy of cancer (Amendola et al., 2010). Previous study reported that a Zingiber officinale synthesized AgNPs inhibits the HT-29 cells at 150.8 µg/ml (Venkatadri et al., 2020). In our study reported that the cytotoxic effect of guava leaf synthesized silver nanoparticles that can halt the cell proliferation at lower concentration in colon cancer.

Conclusion

In conclusion, the silver nanoparticles developed using P. guajava Linn leaf aqueous extract by rapid, non-toxic and environment friendly green approach method. The synthesized silver nanoparticles were characterized by UV-spectroscopy, SEM, XRD and FTIR. The biosynthesized AgNPs exhibited DPPH scavenging property and excellent antibacterial activity against E. coli, Pseudomonas sp. and Klebsiella sp. and, the excellent anticancer activity against HT-29 colon cancer cells. Our finding confirmed that the green synthesized silver nanoparticles showed potent pharmacological activity.

Conflict of interest

The authors have no conflicts of interest regarding this investigation.

Acknowledgments

The authors would like to thank Er. A.C.S. Arun Kumar, President, Dr. M.G.R Educational and Research Institute University for providing the necessary facilities. We would like to thank the Department of Biotechnology, Dwaraka Doss Goverdhan Doss Vaishnav College for providing the necessary facilities. We acknowledge the Nanotechnology Research Centre (NRC), SRMIST for providing the research facilities.

References

[1] Lakhan, M. N., Chen, R., Shar, A. H., Chand, K., Shah, A. H., Ahmed, M., ... & Wang, J.

(2020). Eco-friendly green synthesis of clove buds extracts functionalized silver nanoparticles and evaluation of antibacterial and antidiatom activity. Journal of microbiological methods, 173, 105934.Blunkett, D. (1998, July 24). Cash for competence.

Times Educational Supplement, p. 15.

[2] Ebrahimzadeh, M. A., Naghizadeh, A., Amiri, O., Shirzadi-Ahodashti, M., & Mortazavi- Derazkola, S. (2020). Green and facile synthesis of Ag nanoparticles using Crataegus pentagyna fruit extract (CP-AgNPs) for organic pollution dyes degradation and antibacterial application. Bioorganic chemistry, 94, 103425.

(13)

[3] Bharathi, D., & Bhuvaneshwari, V. (2019). Evaluation of the cytotoxic and antioxidant activity of phyto-synthesized silver nanoparticles using Cassia angustifolia flowers. BioNanoScience, 9(1), 155-163.

[4] Roy, A., Bulut, O., Some, S., Mandal, A. K., & Yilmaz, M. D. (2019). Green synthesis of silver nanoparticles: biomolecule-nanoparticle organizations targeting antimicrobial activity. RSC advances, 9(5), 2673-2702.

[5] Shah, Z., Hassan, S., Shaheen, K., Khan, S. A., Gul, T., Anwar, Y., ... & Suo, H. (2020).

Synthesis of AgNPs coated with secondary metabolites of Acacia nilotica: An efficient antimicrobial and detoxification agent for environmental toxic organic pollutants. Materials Science and Engineering: C, 111, 110829.

[6] Mariadoss, A. V. A., Ramachandran, V., Shalini, V., Agilan, B., Franklin, J. H., Sanjay, K., ... & Ernest, D. (2019). Green synthesis, characterization and antibacterial activity of silver nanoparticles by Malus domestica and its cytotoxic effect on (MCF-7) cell line. Microbial pathogenesis, 135, 103609.

[7] Lakshmanan, G., Sathiyaseelan, A., Kalaichelvan, P. T., & Murugesan, K. (2018). Plant- mediated synthesis of silver nanoparticles using fruit extract of Cleome viscosa L.:

assessment of their antibacterial and anticancer activity. Karbala International Journal of Modern Science, 4(1), 61-68.

[8] Mollick, M. M. R., Rana, D., Dash, S. K., Chattopadhyay, S., Bhowmick, B., Maity, D., ... & Chattopadhyay, D. (2019). Studies on green synthesized silver nanoparticles using Abelmoschus esculentus (L.) pulp extract having anticancer (in vitro) and antimicrobial applications. Arabian journal of chemistry, 12(8), 2572-2584.

[9] Khandelwal, R., Arora, S. K., Phase, D. M., Pareek, A., & Ravikant. (2020, May). Anti- cancer potential of green synthesized silver nanoparticles. In AIP Conference Proceedings (Vol. 2220, No. 1, p. 020046). AIP Publishing LLC.

[10] Gomathi, A. C., Rajarathinam, S. X., Sadiq, A. M., & Rajeshkumar, S. (2020). Anticancer activity of silver nanoparticles synthesized using aqueous fruit shell extract of Tamarindus indica on MCF-7 human breast cancer cell line. Journal of Drug Delivery Science and Technology, 55, 101376.

[11] Gutiérrez, R. M. P., Mitchell, S., & Solis, R. V. (2008). Psidium guajava: a review of its traditional uses, phytochemistry and pharmacology. Journal of ethnopharmacology, 117(1), 1-27.

[12] Olatunde, O. O., Benjakul, S., & Vongkamjan, K. (2018). Antioxidant and antibacterial properties of guava leaf extracts as affected by solvents used for prior dechlorophyllization. Journal of food biochemistry, 42(5), e12600.

[13] Wang, L., Wu, Y., Xie, J., Wu, S., & Wu, Z. (2018). Characterization, antioxidant and antimicrobial activities of green synthesized silver nanoparticles from Psidium guajava L.

leaf aqueous extracts. Materials Science and Engineering: C, 86, 1-8.

[14] Ryu, N. H., Park, K. R., Kim, S. M., Yun, H. M., Nam, D., Lee, S. G., ... & Ahn, K. S.

(2012). Induces Anticancer Activity by Suppressing AKT/Mammalian Target of Rapamycin/Ribosomal p70 S6 Kinase in Human Prostate Cancer Cells. J Med Food, 15(3), 231-241.

(14)

[15] Mahfuzul Hoque, M. D., Bari, M. L., Inatsu, Y., Juneja, V. K., & Kawamoto, S. (2007).

Antibacterial activity of guava (Psidium guajava L.) and neem (Azadirachta indica A.

Juss.) extracts against foodborne pathogens and spoilage bacteria. Foodborne pathogens and disease, 4(4), 481-488.

[16] Raghunandan, D., Mahesh, B. D., Basavaraja, S., Balaji, S. D., Manjunath, S. Y., &

Venkataraman, A. (2011). Microwave-assisted rapid extracellular synthesis of stable bio- functionalized silver nanoparticles from guava (Psidium guajava) leaf extract. Journal of Nanoparticle Research, 13(5), 2021-2028.

[17] Sun, Q., Cai, X., Li, J., Zheng, M., Chen, Z., & Yu, C. P. (2014). Green synthesis of silver nanoparticles using tea leaf extract and evaluation of their stability and antibacterial activity. Colloids and surfaces A: Physicochemical and Engineering aspects, 444, 226- 231.

[18] Bisht, R., Chanyal, S., & Agrawal, P. K. (2016). Antimicrobial and phytochemical analysis of leaf extract of medicinal fruit plants. Asian J. Pharm. Clin. Res, 9(4), 131-136.

[19] Kumar, S. V., Karpagambigai, S., Rosy, P. J., & Rajeshkumar, S. (2017). Phyto-assisted synthesis of silver nanoparticles using solanum nigrum and antibacterial activity against salmonella typhi and Staphylococcus aureus. Mechanics, Materials Science &

Engineering Journal, 9(1).

[20] Keshari, A. K., Srivastava, R., Singh, P., Yadav, V. B., & Nath, G. (2020). Antioxidant and antibacterial activity of silver nanoparticles synthesized by Cestrum nocturnum. Journal of Ayurveda and integrative medicine, 11(1), 37-44.

[21] Asha, S., Thirunavukkarasu, P., & Rajeshkumar, S. (2017). Green synthesis of silver nanoparticles using mirabilis jalapa aqueous extract and their antibacterial activity against respective microorganisms. Research Journal of Pharmacy and Technology, 10(3), 811- 817.

[22] Khorrami, S., Zarepour, A., & Zarrabi, A. (2019). Green synthesis of silver nanoparticles at low temperature in a fast pace with unique DPPH radical scavenging and selective cytotoxicity against MCF-7 and BT-20 tumor cell lines. Biotechnology Reports, 24, e00393.

[23] Nasar, M. Q., Khalil, A. T., Ali, M., Shah, M., Ayaz, M., & Shinwari, Z. K. (2019).

Phytochemical analysis, Ephedra Procera CA Mey. Mediated green synthesis of silver nanoparticles, their cytotoxic and antimicrobial potentials. Medicina, 55(7), 369.

[24] Pirtarighat, S., Ghannadnia, M., & Baghshahi, S. (2019). Green synthesis of silver nanoparticles using the plant extract of Salvia spinosa grown in vitro and their antibacterial activity assessment. Journal of Nanostructure in Chemistry, 9(1), 1-9.

[25] Kaur, A., Preet, S., Kumar, V., Kumar, R., & Kumar, R. (2019). Synergetic effect of vancomycin loaded silver nanoparticles for enhanced antibacterial activity. Colloids and Surfaces B: Biointerfaces, 176, 62-69.

[26] Parvataneni, R. (2020). Biogenic synthesis and characterization of silver nanoparticles using aqueous leaf extract of Scoparia dulcis L. and assessment of their antimicrobial property. Drug and chemical toxicology, 43(3), 307-321.

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[27] Saxena, A., Tripathi, R. M., Zafar, F., & Singh, P. (2012). Green synthesis of silver nanoparticles using aqueous solution of Ficus benghalensis leaf extract and characterization of their antibacterial activity. Materials letters, 67(1), 91-94.

[28] Aygün, A., Özdemir, S., Gülcan, M., Cellat, K., & Şen, F. (2020). Synthesis and characterization of Reishi mushroom-mediated green synthesis of silver nanoparticles for the biochemical applications. Journal of pharmaceutical and biomedical analysis, 178, 112970.

[29] Kedi, P. B. E., Meva, F. E. A., Kotsedi, L., Nguemfo, E. L., Zangueu, C. B., Ntoumba, A.

A., ... & Maaza, M. (2018). Eco-friendly synthesis, characterization, in vitro and in vivo anti-inflammatory activity of silver nanoparticle-mediated Selaginella myosurus aqueous extract. International journal of nanomedicine, 13, 8537.

[30] Logeswari, P., Silambarasan, S., & Abraham, J. (2015). Synthesis of silver nanoparticles using plants extract and analysis of their antimicrobial property. Journal of Saudi Chemical Society, 19(3), 311-317.

[31] Awwad, A. M., & Salem, N. M. (2012). Green synthesis of silver nanoparticles byMulberry LeavesExtract. Nanoscience and Nanotechnology, 2(4), 125-128.

[32] Khodadadi, S., Mahdinezhad, N., Fazeli-Nasab, B., Heidari, M. J., Fakheri, B., & Miri, A.

(2021). Investigating the Possibility of Green Synthesis of Silver Nanoparticles Using Vaccinium arctostaphlyos Extract and Evaluating Its Antibacterial Properties. BioMed Research International, 2021.

[33] Bilal, M., Rasheed, T., Iqbal, H. M. N., Hu, H., & Zhang, X. (2017). Silver nanoparticles:

biosynthesis and antimicrobial potentialities. International Journal of Pharmacology, 13(7), 832-845.

[34] Bhakya, S., Muthukrishnan, S., Sukumaran, M., Muthukumar, M., Kumar, S. T., & Rao, M. V. (2015). Catalytic degradation of organic dyes using synthesized silver nanoparticles: a green approach. Journal of Bioremediation & Biodegredation, 6(5), 1.

[35] Guerra, J. D., Sandoval, G., Avalos-Borja, M., Pestryakov, A., Garibo, D., Susarrey-Arce, A., & Bogdanchikova, N. (2020). Selective antifungal activity of silver nanoparticles: A comparative study between Candida tropicalis and Saccharomyces boulardii. Colloid and Interface Science Communications, 37, 100280.

[36] Ansar, S., Tabassum, H., Aladwan, N. S., Ali, M. N., Almaarik, B., AlMahrouqi, S., ... &

Alsubki, R. (2020). Eco friendly silver nanoparticles synthesis by Brassica oleracea and its antibacterial, anticancer and antioxidant properties. Scientific Reports, 10(1), 1-12.

[37] Kumar, V., Bano, D., Mohan, S., Singh, D. K., & Hasan, S. H. (2016). Sunlight-induced green synthesis of silver nanoparticles using aqueous leaf extract of Polyalthia longifolia and its antioxidant activity. Materials Letters, 181, 371-377.

[38] Sriram, M. I., Kanth, S. B. M., Kalishwaralal, K., & Gurunathan, S. (2010). Antitumor activity of silver nanoparticles in Dalton’s lymphoma ascites tumor model. International journal of nanomedicine, 5, 753.

(16)

[39] Biresaw, S. S., Damte, S. M., & Taneja, P. (2020). Green Synthesized Silver Nanoparticles: A Promising Anticancer Agent. International Journal of Nanoscience, 19(04), 1950027.

[40] Samari, F., Baluchi, L., Salehipoor, H., & Yousefinejad, S. (2019). Controllable phyto- synthesis of cupric oxide nanoparticles by aqueous extract of Capparis spinosa (caper) leaves and application in iron sensing. Microchemical Journal, 150, 104158.

[41] Parvataneni, R. (2020). Biogenic synthesis and characterization of silver nanoparticles using aqueous leaf extract of Scoparia dulcis L. and assessment of their antimicrobial property. Drug and chemical toxicology, 43(3), 307-321.

[42] Amendola, V., Bakr, O. M., & Stellacci, F. (2010). A study of the surface plasmon resonance of silver nanoparticles by the discrete dipole approximation method: effect of shape, size, structure, and assembly. Plasmonics, 5(1), 85-97.

[43] Venkatadri, B., Shanparvish, E., Rameshkumar, M. R., Arasu, M. V., Al-Dhabi, N. A., Ponnusamy, V. K., & Agastian, P. (2020). Green synthesis of silver nanoparticles using aqueous rhizome extract of Zingiber officinale and Curcuma longa: In-vitro anti-cancer potential on human colon carcinoma HT-29 cells. Saudi Journal of Biological Sciences, 27(11), 2980-2986.

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