Materials Research Express

ACCEPTED MANUSCRIPT

Bactericidal coating of paper towels via sustainable biosynthesis of silver

nanoparticles using Ocimum sanctum leaf extract
To cite this article before publication: Jaya Mary Jacob et al 2018 Mater. Res. Express in press https://doi.org/10.1088/2053-1591/aafaed

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Page 1 of 27 AUTHOR SUBMITTED MANUSCRIPT - MRX-111125.R1

1
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3 Bactericidal coating of paper towels via sustainable biosynthesis of silver nanoparticles
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6 using Ocimum sanctum leaf extract
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8 Jaya Mary Jacob 1, Merin Sara John 1, Aleena Jacob 1, Abitha P 1, Sruthy S Kumar 1,

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10 Reju Rajan 1, Suganthy Natarajan 2, Arivalagan Pugazhendhi 3*
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12 1
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Department of Biotechnology and Biochemical Engineering, Sree Buddha College of

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15 Engineering Pattoor, Kerala, India
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Department of Nanoscience and Technology, Alagappa University, Karaikudi, Tamil Nadu,
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India
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22 Innovative Green Product Synthesis and Renewable Environment Development Research
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24 Group, Faculty of Environment and Labour Safety, Ton Duc Thang University, Ho Chi Minh
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City, Vietnam
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31 *Corresponding Author Address:
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33 Dr. Arivalagan Pugazhendhi
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Innovative Green Product Synthesis and Renewable Environment Development Research
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38 Group
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40 Faculty of Environment and Labour Safety
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Ton Duc Thang University
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45 Ho Chi Minh City, Vietnam.
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47 Email: arivalagan.pugazhendhi@tdtu.edu.vn
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 AUTHOR SUBMITTED MANUSCRIPT - MRX-111125.R1 Page 2 of 27

1
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3 Abstract
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6 The present study reports the effective impregnation of silver nanoparticles (AgNPs)
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8 biosynthesized using Ocimum sanctum leaf extract onto conventional tissue paper for the

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10 inhibition of hospital borne pathogenic bacterial growth. The AgNPs biosynthesised using the
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leaf extract had face centered cubic lattice structure, nano scale dimensions, biocompatible

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15 coating of plant proteins and an absorbance maximum at 438 nm. Further, these nanoparticles
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17 were used to engineer tissue paper towels under conditions akin to room temperature. The
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presence of nanoparticles on the tissue paper filaments were confirmed using SEM analysis.
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22 Detailed analysis using XRD and FTIR indicated the presence of AgNPs on the paper fibres
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24 and the appreciable amounts of bio-active functional groups on the modified tissue paper when
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compared to the conventional ones. The bioactive layers aid the firm adherence of bacterial

cell wall proteins and there after the silver particles extend their bactericidal action on the
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31 attached bacteria. The antibacterial activity of the nano-engineered tissue paper towels were
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33 tested against common gram positive and gram negative pathogens. Further, cytotoxicity
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evaluation using MTT assay revealed that the AgNPs coated paper towels does not pose any
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38 unintended adverse effects on fibroblast cell lines. These results highlight that biogenic AgNPs
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40 coated paper towels are cost effective, eco-friendly and wholesome alternative for the
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conventional paper towels and can be an effective sanitizing product for decreasing the risk of
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45 health associated infections.
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47 Keywords: Silver Nanoparticle; Nanotechnology; Ocimum sanctum; Tissue paper;
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49 Antibacterial.
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52 1. Introduction
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55 Hospital acquired infections are becoming a serious issue worldwide, the transmission
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58 of which takes place majorly through medical devices, air, direct contact and utilization of
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60 unsterilized apparatus[1, 2]. Although efficient technologies and drugs are available, the

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Page 3 of 27 AUTHOR SUBMITTED MANUSCRIPT - MRX-111125.R1

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3 nanotechnological interventions that prevent the spread of bacterial infection via this mode are
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6 gaining momentum. Nowadays researchers are focusing on producing nanoparticle based auto
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8 sanitizing products for decreasing the risk of health associated infections [3, 4]. Even though

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10 conventional solvents and bleaching agents have been extensively used for sanitizing, the
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prospects of developing antimicrobial surfaces by impregnating or coating nanoparticles offer

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15 a long term, effective and non-toxic solution for the control of pathogens [5]. Majorly, hospital
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17 infections are spread through contaminated water, cloths, improper disposal of used tissue
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papers, cotton and other medical waste [6, 7]. Of particular concern are the paper products that
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22 are used widely in hospital environment. Due to their porous structure, fibres in paper are prone
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24 to bacterial colonization leading to continual contamination. The porous nature of paper fibres
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further facilitates biofilm formation thereby worsening the elimination prospects of these

bacterial colonies. In some circumstances, such as paper towels hanging in the splashing zone
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31 or those used for cleaning surfaces, they have been considered as potential sources of bacterial
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33 contaminations [8]. In this context intervention of nanotechnology to address this situation
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holds relevance.
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In this context, it is noteworthy that metallic nanoparticles like silver, gold, copper and
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41 other noble metals find immense application in healthcare, cosmetics and laundry products [9-
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11]. Advancement in nanotechnology research has cut down the production cost of
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nanomaterials to the extent that these smart materials have become an integral and affordable
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48 commodity for common man and commercial nanotechnology industry has demonstrated an
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50 exponential growth in recent times [12]. Among the different metallic nanoparticles used in
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52 consumer products, silver nanoparticles (AgNPs) have the highest consumer acceptability and
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55 potential for commercialization [13, 14]. The broad spectrum anti-microbial properties of nano
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57 silver has enabled its application in a wide variety of consumer products ranging from
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59 disinfecting medical products to water treatment systems [15, 16]. The release of free Ag+ ions
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1
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3 exerts antimicrobial effect by binding to the cell membrane proteins and inhibiting
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6 reproduction and cellular respiration[17]. The fast growing demand for AgNPs in health care
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8 application, the global market for AgNPs is expected to reach around $2.45 billion within 2022

pt
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10 [18]. However, the toxicity of AgNPs is still a major concern and often outweigh their
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usefulness in medical products[19, 20]. Considering these problems, nowadays green synthesis

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15 of AgNPs using plant extracts is widely practised. The biomolecules (alkaloids, phenolic
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17 compounds, terpenoids, co-enzymes) of plant and its extract act as reducing agents by
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converting the metals ions to nanoparticles rapidly and the biogenic AgNPs are known to be
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22 biocompatible, nontoxic and hence have better acceptability for human utility [21].
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25 The present study elaborates the development of antimicrobial tissue paper towels using
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silver nanoparticles biosynthesized using Ocimum sanctum (Tulsi) extracts. The bioactive

molecules present in Tulsi extract facilitate one-step, eco-friendly and economical synthesis of
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32 silver nanoparticle that are coated with a bio-derived layer that aids microbial disinfection. The
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34 results showed that the silver coatings on tissue paper towels significantly inhibit common
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36 bacterial microflora thereby decreasing the presence and spread of numerous human infections.
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39 2. Materials and methods
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42 2.1 Biosynthesis of AgNPs from Ocimum sanctum
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45 Fresh leaves of Ocimum sanctum were collected from Pattoor, Alappuzha, Kerala,
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47 India. The collected leaves were thoroughly washed thrice with distilled water and chopped
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49 into small pieces and 20 g of the leaves were boiled in 100 mL distilled water for 5 min. Then
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51
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52 the extract was filtered and collected. To 10 mL of the Tulsi leaf extract (TLE) 90 mL of silver
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54 nitrate (AgNO3) solution was added to make the final concentration to 10-3M and the solution
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56 was allowed to react at room temperature. Periodic sampling after 30 min was carried out to
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monitor the formation of AgNPs.
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3 2.2 Characterization methods
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6 The synthesized AgNPs were characterized using UV-vis spectroscopy (Systronics).
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8

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9 The absorbance spectra were recorded at wavelength of 300 to 800 nm. Further XRD
10
11 measurements were performed by CuK alpha radiation with wavelength of 1.5406 A⁰ in the
12
13 2θ range of 20⁰ to 80⁰ . The functional group of the synthesized AgNPs were determined

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using FTIR analysis. Further the morphological characterization of the AgNPs was performed
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18 by examining the surface morphology and size using Scanning electron microscopy (SEM) and
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20 Transmission electron microscopy (TEM) analysis.

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23 2.3 Coating of AgNPs over conventional tissue paper
24
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26 Conventional tissue paper (180 mm x 150 mm, 34-44 GSM) was used in this study.
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The tissue paper samples were dipped for 2 h in the aqueous solution of biogenic nano silver

30 particles and then air dried to fabricate the nanoparticle embedded tissue paper towel.
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33 2.4 Characterization of nanoengineered tissue paper towel
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36 The characterisation of both conventional tissue papers and nanoparticle embedded
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38 tissue paper were carried out using SEM, FTIR and XRD analyses. Further, the release studies
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40 for the conventional and antimicrobial tissue paper samples were carried out as per the protocol
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43 [22].
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46 2.5 Antibacterial activity of the nanoengineered tissue paper towel
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48 The antibacterial assay of nano-engineered tissue paper towel was assessed by using
49
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the Kirby-Bauer method against clinical pathogenic bacteria (Escherichia coli, Staphylococcus
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53 aureus and Klebsiella pneumoniae) on nutrient agar medium at 37 ⁰ C for 24 h. Freshly
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55 cultured bacterial colonies of the tested bacteria were taken and 100 µL of inoculum was spread
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57 on each agar plate. Nanoparticle embedded tissue paper were punched out into circles. Sterile
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60 Whattman filter paper loaded with plant extract, conventional tissue paper circles and

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3 chloramphenicol antibiotic discs (30 µg/disc each) were used in each plate and incubated at 37
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6 ⁰ C for 24 h. The plates were examined for the presence of zones of inhibition and the diameters
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8 of inhibition zones were measured.

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11 2.6 MTT cytotoxicity assay
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13

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14 In review of the recent studies on the risk of safety of AgNPs impregnated coatings to
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16 human beings, the present study examined the inhibitory effects of the AgNPs incorporated
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tissue paper on fibroblast cell lines using the calorimetric MTT assay. The assay is based on
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20

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21 the formation of purple formazan crystals by a reduction reaction initiated by the
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23 dehydrogenase enzymes secreted by metabolically active cells. The assay was carried out as
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25 per the protocols by Marziyeh et al. (2018). Cell viability was calculated by the following
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equation:
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𝑂𝐷𝑠𝑎𝑚𝑝𝑙𝑒
𝐶𝑒𝑙𝑙 𝑉𝑖𝑎𝑏𝑖𝑙𝑖𝑡𝑦 (%) = × 100
32 𝑂𝐷𝑐𝑜𝑛𝑡𝑟𝑜𝑙
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34
35 Where ODsample is the absorbance of the test sample and ODcontrol is the absorbance for untreated
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37 control cells.
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40 3. Results and discussion
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3.1 Biosynthesis of AgNPs from Ocimum sanctum

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Green synthesis of AgNPs using medicinal plants is one of the safest and non-toxic
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48 routes for the production of AgNPs. Among the different traditional medicinal plants, Ocimum
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50 tenuiflorum or Ocimum sanctum, has been a preferred source for researchers for initiating
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biosynthesis reactions and other extraction procedures for therapeutic process development
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55 [23-25]. This plant is commonly referred to as Tulsi, or Holy Basil, “Queen of plants” or
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57 “mother medicine of nature” and is abundantly available in India, Australia, Malaysia, West
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59 Africa and some parts of Arab countries [26, 27]. Plant extract based synthesis of AgNPs is
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1
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3 considered as most rapid and easier way [28]. The present study employed tulsi leaf extract
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6 (TLE) for AgNPs owing to the large amounts of bioactive components present in them.
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8 Literature reports that major bioactive in TLE like tannins, alkaloids, saponins, glycosides and

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10 aromatic compounds act as reducing agents and stabilizers aid redox reactions for the
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biosynthesis of AgNPs [new 1]. It has also been found that euginol is the major reducing agent

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15 in TLE that initiates the bioreduction of silver nitrate to AgNPs [29]. While the reducing
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17 potential of TLE bioactive play a major role in the formation of AgNPs, the size of the biogenic
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AgNPs also depend on other operating parameters like temperature and silver nitrate
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22 concentration [30]. Different enzymes, proteins and DNA has been used for nanoparticles
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24 synthesis, but they are expensive, unstable and temperature sensitive [31]. According to
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29
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AbdelRahim et al. high and low temperatures denature the enzymes and proteins and hence do

not aid AgNPs biosynthesis. Generally, temperatures ranging from 20-60 oC are conducive for
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31 the biosynthesis machinery [32]. Therefore, plant extract or biomolecules have been used
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33 effective for NPs synthesis [31]. They are stable and cheaper sources, which can with stand
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varied temperature, pH, salinity etc. So far numerous numbers of plants have been used as a
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38 reducing agent in metallic NPs synthesis [31, 33-35]. As for the precursor molar
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40 concentrations, it has been reported that increasing the silver nitrate concentration above 10-
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42 2
M results in the synthesis of larger particles [36] and usually a concentration of 1mM AgNO3
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45 is considered optimum [37]. The present study reports the use of 1mM AgNO3 for the
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47 biosynthesis reaction at room temperature. Taking into consideration, the debates regarding the
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49 toxicity and fate of nanoparticles and their synthesis routes, the utilization of the respective
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52 precursor at optimum concentration and room temperature holds relevance. The synthesis of
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54 AgNPs by the reduction of AgNO3 using TLE is monitored by the change in colour of the
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56 reaction mixture from colourless to yellow-brown. The visible colour change that indicates the
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formation of nano silver is depicted in Fig.1(a).
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1
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3 The UV-Vis spectra recorded from the reaction medium as a function of reaction time
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6 using O. sanctum leaves indicated an absorbance maximum at 428 nm [24]. Similar absorption
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8 bands owing to the electronic vibrations of AgNPs in resonance with the light waves have been

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10 reported earlier [38]. Further, the absorbance peak at 280 nm indicates the presence of
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biocompatible protein capping over the biogenic AgNPs [39]. Plants are known to contain

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15 bioactive molecules like polyphenols, carbohydrates, proteins, flavonoids, tanins etc., which
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17 induces the reduction and stabilization of metal ions [28, 40, 41]. This absorption peak has also
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been attributed to the electronic excitations in tyrosine and tryptophan residues of proteins
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22 acting as capping agents on the synthesized biogenic AgNPs in similar studies [24]. The protein
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24 capping acts as a biocompatible protective layer around the AgNPs and is also responsible for
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stabilizing the AgNPs [32]. In comparison with a previous using red cabbage extract, the

present result also clearly indicates that TLE has reduced Ag+ ions to Ag0 for the synthesis of
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31 AgNPs through its electron donation ability [40]. Some studies also suggest that higher
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33 concentration of plant extract produces smaller AgNPs having narrow size distribution of NPs
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compared to lower concentration of extract [42]. This may be the reason for getting narrower
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38 peak in UV Vis spectra, indicating smaller sized AgNPs due to higher concentration of TLE.
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40 3.2 Studies with XRD and FTIR analysis
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The crystalline nature of the biogenic AgNPs were analysed using X-ray diffraction
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45 that indicated prominent Bragg reflections at 2θ values of 37.7°, 43.8°, 64.2° and 77.28°
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47 corresponding to (111), (200), (220) and (311) respectively, typical of face centred cubic
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49 structure of silver (Fig. 1(b)). Our results are in concurrence with prior studies on
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52 biosynthesized silver particles using TLE [43]. FTIR analysis was carried out to detect the
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54 prominent functional groups on the biogenic Ag particles. Peaks at 1066.71 cm-1, 1117.42 cm-
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56 1
and 1616.24 cm-1, 1383.88 cm-1, and 3422.23 cm-1 corroborate to the C-O stretching, C-N
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stretch, C-H bending and O-H stretching respectively (Fig. 2(a)). The absorption peak at
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3 1616.24 cm-1 showed concurrence with the study by [13] which proved that proteins from TLE
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6 are associated with the AgNPs without altering the secondary structure of the protein. These
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8 prominent peaks confirm that proteins and other plant derived metabolites form a stabilizing

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10 coating over the biogenic Ag particles. The biolayer composed of amines, carboxylic and
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alcohol groups form biocompatible coatings that can extend the applicability of the

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15 biosynthesized AgNPs in medical and healthcare products [44].
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18 3.3 Studies with SEM and TEM analysis
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20 The particle size of the biogenic AgNPs was examined using the SEM analysis. The

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magnification at 25,000KX indicated uniform distribution of silver flakes in nanoscale (Fig.
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25 2(b)). The average diameter was measured using ImageJ software and was found to be around
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43 nm. Earlier, researchers had reported the biosynthesis of silver particles from the same

source in similar size ranges [44]. Previous reports using TLE showed spherical shaped AgNPs
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32 (30 nm) [24] and triangle shaped AgNPs (42 nm) synthesized using TLE [45]. Therefore, the
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34 results clearly proved that TLE potentially acted as reducing agent in the biological synthesis
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36 of AgNPs.
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39
The size and structure of the AgNPs were also analysed using TEM analysis. The TEM
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41 results proved that the AgNPs were almost in spherical shape. They were found as
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polydispersed form and the size ranged from 10 to 50 nm (Fig. 3 (a), (b)). The average size of
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the nanoparticles was found to be 30 nm. The TEM images also proved that the AgNPs were
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48 coated by organic layer over the nanoparticles surface. Although some of the AgNPs were
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50 found agglomerated due to the presence of the sticky organic layer over the nanoparticles.
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52 Majority of the particles appeared to be in physical contact but they were separated by the
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55 organic layer formed over the nanoparticles. In a recent work done by Shaik et al. suggested
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57 that the presence of thick organic layer over the AgNPs were from the several organic
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59 compounds of the plant extract and these bioorganic coating also reduces and stabilizes the
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3 nanoparticles[46]. The present results also produced smaller nanoparticles which are far better
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6 from the particles synthesized in previous studies using plant extracts [19, 47, 48].Therefore,
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8 the AgNPs synthesized using O. sanctum leaf extract were well characterized and further taken

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10 for antimicrobial studies.
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13 3.4 Nanoengineering of conventional paper towels using biogenic AgNPs

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16 The spread and control of nosocomial infections have of late been recognized as a
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serious challenge. The susceptibility of medical devices and basic amenities in a healthcare set
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21 up to serious infections is high and hence, the development of anti-microbial surfaces is of
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23 particular concern to address this scenario. The inherent advantages of coating Ag particles on
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25 leather and cotton fabrics to prevent bacterial colonies that cause skin infection has been
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reported earlier [49, 50]. In another study, the preparation of Se coated tissue paper towels

30 using chemically synthesized Se particles have been reported. The authors report Se particles
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32 of dimensions ranging from 50-100 nm having a surface coverage of 4.36% on the tissue paper
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34
35 [22]. Generally, Tulsi is an abundant medicinal plant in India and consumed by people of the
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37 country to prevent various nosocomial infections and disease [51]. Using them in synthesis of
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39 AgNPs will enhance the antibacterial property against infectious pathogen causing nosocomial
40
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infections [51]. In our study, Ag nanoparticles biosynthesized using tulsi leaf extract was
43
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44 impregnated into tissue paper towels to develop an antimicrobial surface. After impregnating
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46 the AgNPs, the tissue paper turned yellow in colour and it was stable for the period of 1 year
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48 (Fig. 4(a)). The impregnation of AgNPs was confirmed using SEM analysis revealed that the
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51 tissue paper fibres were deposited with nano sized silver particles while untreated tissue paper
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53 towels showed no such surface depositions (Fig. 4(b)). The nanoparticles formed agglomerates
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55 on few areas owing to the bioactive layer over the biosynthesized Ag particles [52]. The results
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58 of the present study are significant taking into account that the Ag particles used in the present
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60 study are derived from a medicinal plant extract. Using TLE extract will provide a safer and

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3 nontoxic route of using tissue paper coated with AgNPs. This type of tissue paper can be
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6 commercialized for using in general household purpose, hospitals and infant sanitary purposes.
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8 Further confirmatory studies were performed using XRD analyses using powdered

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9 a b
10 samples of the treated tissue paper that indicated prominent peaks at (2θ) values of 38.29 and
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44.16 corresponding to (111) and (200) of the FCC structure of Ag (JCPDS file number 89-

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15 3722) (Fig. 5(a)). The XRD spectral peaks confirm the presence of Ag particles on the tissue
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17 paper. Further, FTIR analysis of the treated and untreated samples clearly indicated that the
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19
AgNPs embedded tissue paper samples showed a larger number of prominent peaks in the
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22 FTIR spectra in comparison to the untreated sample (Fig. 5(b)). The IR peaks owe its presence
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24 to the biogenic AgNPs and the plant biomolecule coating over it [53]. The prominent peaks
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indicate the active bio molecules comprising of amines, carboxyl and alkene groups on the

treated sample. In this regard, it is noteworthy that the active functional groups on the surface
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31 of the treated sample can tether the microbial cell wall proteins thereby aiding the antimicrobial
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33 activity of the AgNPs [54].
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3.5 Antibacterial activity of Ag nanoengineered tissue paper
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38 Silver is known for its potential antibacterial activity[11]. In view of the potential anti-
39
40 bacterial properties of silver particles, the inhibitory role of the nanosilver embedded tissue
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42
paper towels against common clinical pathogens were analysed using the Kirby Bauer Disc
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45 diffusion assay. The bacteria under consideration were common hospital acquired human
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47 pathogens namely Staphylococcus aureus, Klebsiella pneumoniae and E.coli. The highest
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49 inhibitory activity was observed against E.coli followed by Staphylococcus aureus and
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52 Klebsiella pneumoniae. These findings affirm the microbicidal activity of the silver particles
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54 coated on the tissue paper surface. Our results also indicate that the untreated tissue paper
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56 samples and the leaf extract alone showed no significant inhibitory effect against the bacteria
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used in the study (Fig. 6). These observations confirm that silver particles when coated on paper
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3 towels enhance the effectiveness of the product in the control of nosocomial infections. In
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6 another relevant study, AgNPs coated cotton and leather surfaces have been reported to inhibit
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8 B. linens, P. acnes, B. cereus, and S. epidermidis [22]. The appreciable antimicrobial activity

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10 of the AgNPs coated tissue paper samples are noteworthy and may be attributed to the
11
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significant amount of bio active molecules present in bio-layer of O. sanctum. Synthesis of

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15 AgNPs using plant extract mainly relates with active molecules present in them involved in
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17 reduction of silver nitrate to elemental silver ions [40, 41, 55]. Studies have proved the use of
18
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plant rich in flavonoids for the reduction of AgNPs [41]. The natural antioxidant is plant extract
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22 help in the synthesis of narrow sized nanoparticles [40, 55]. Tulsi plants contains rich amount
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24 of antioxidant potentials which could also attribute the process of AgNPs synthesis [56]. The
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antibacterial property of TLE, may be due to the presence of essential oil components and

additionally coating of essential oils from TLE over AgNPs will increase the killing of bacterial
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31 pathogens [51]. These findings assert the need to utilize medicinal plant extracts for
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33 nanoparticle synthesis in the economical and effectiveness point of view.
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36 3.6 Ag release studies from the tissue paper
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39
Technological advances have accelerated the number of nanoparticle and associated
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41 products that are being used by human beings. Over the past few decades’ fabrication of AgNPs
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43
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over medical devices, cotton fabrics, leather food packaging etc. has been practiced [57, 58].
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However, the fate of nanoparticles in the environment is a much debated area. To ascertain that
46
47
48 the AgNPs are not released into the environment from the tissue paper, release studies were
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50 carried out using UV-vis absorption spectra. The time course spectral analysis is depicted in
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52 Fig. 7.
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55 It can be inferred that the samples did not show any significant peak in wavelength
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58 ranges from 400-450 nm. A peak in the afore-mentioned wavelength ranges is characteristic of
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60 Ag particles and absence of the same in the study indicates that the Ag particles are not released

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3 into the environment from the tissue paper [59]. These observations further ascertain the fact
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6 that the bio-capping over the AgNPs render long term stability to the nanoparticles and ensure
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8 their stable integration into the paper towels. The other peaks in the spectral analysis are

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10 characteristic of the biological functional groups on the AgNPs. On the basis of these
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observations, it can hence be concluded that the biomolecules on the biogenic Ag particles

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15 impregnated in the tissue paper samples bind to the cell wall proteins of bacteria and there after
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17 the silver particles extend its inhibitory action by contact inhibition [60].
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20 In addition to antibacterial activity, cytotoxicity is another important parameter

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influencing application of any material for industrial and medical purposes [61]. In the present
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25 study, cytotoxicity studies on fibroblast cell lines were carried out in order to analyze whether
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the tissue paper containing AgNPs pose any unintended adverse effects on cells. The in vitro

inhibitory effect on fibroblast cell line was determined by the MTT colorimetric assay. The
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32 time course cell viability studies depicted in fig. 8 indicate that the tissue paper towels treated
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34 with AgNPs indicate minimal cytotoxic effect which is almost comparable to that of the
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36 untreated tissue paper samples. It is noteworthy that conventional tissue paper samples also
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displayed cytotoxicity as evinced quantitatively by approximately 94% cell viability after a
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41 period of 72 h. The biogenic AgNPs treated fibroblast cell lines showed a marginal decrease in
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viability after 24 h of exposure. However, the percentage cell viability of AgNPs treated
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samples showed no significant difference with time after 24 h of study period. The inhibitory
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48 effects of the biogenic AgNPs coated paper towels were significantly less in comparison to that
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50 of cotton fabric containing polyaniline-AgNPs coatings [50]. The low toxicity of the biogenic
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52 AgNPs coated samples may be attributed to the biological properties of the plant peptide
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55 capping on the AgNPs [62].
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58 4. Conclusions
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3 In the present study, AgNPs were biosynthesized using Ocimum sanctum leaf extract
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6 in a simple one step process. The biosynthesized nanoparticles were found to have an average
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8 size of 40 nm, face centered cubic lattice structure, absorbance maxima at 428 nm and the

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10 presence of biocompatible capping over it. Further, these nanoparticles were used to engineer
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tissue paper towels under conditions akin to room temperature. The nanoengineered tissue

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15 paper towels were characterized using Scanning Electron Microscopic analysis that indicated
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17 the presence of nanoparticles on the tissue paper filaments. Further, antibacterial assays of the
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nanoengineered tissue paper towels against common gram positive and gram negative
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22 pathogens indicated effective antibacterial properties. In addition, there were larger amounts
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24 of proteins adsorbed on the surface of silver coated paper towels compared with uncoated paper
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29 growth on the surface of paper towels.
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towels, which might be an important reason why AgNPs coatings can inhibit diverse bacterial

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32 Acknowledgement
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35 The authors are thankful to the Department of Biotechnology and Biochemical
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37 Engineering, Sree Buddha College of Engineering, National Institution of Technology
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39 Karanataka, Maharaja`s College Cochin and Karunya University Coimbatore for extending the
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facilities for the conduct of the study.
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47 References
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50 Figure. 1(a) UV-visible spectra of AgNPs biosynthesized using Ocimum sanctum extract;
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16 samples treated with colloidal AgNPs (b) Conventional Tissue paper samples; inset
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35
36
37
38
39
40
41
42
43
pte

44
45
46
47
48
49
50
51
ce

52
53
54
55
56
Ac

57
58
59
60

27