Accepted Manuscript

Hydrothermal synthesis of Ag nanoparticles on the nanocellulose

and their antibacterial study

Xuewei Zhang, Haiyao Sun, Shengnan Tan, Jing Gao, Yujie Fu,
Zhiguo Liu

PII: S1387-7003(18)30998-5
DOI: https://doi.org/10.1016/j.inoche.2018.12.012
Reference: INOCHE 7200
To appear in: Inorganic Chemistry Communications
Received date: 13 November 2018
Revised date: 12 December 2018
Accepted date: 17 December 2018

Please cite this article as: Xuewei Zhang, Haiyao Sun, Shengnan Tan, Jing Gao, Yujie Fu,
Zhiguo Liu , Hydrothermal synthesis of Ag nanoparticles on the nanocellulose and their
antibacterial study. Inoche (2018), https://doi.org/10.1016/j.inoche.2018.12.012

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 ACCEPTED MANUSCRIPT

Hydrothermal synthesis of Ag nanoparticles on the nanocellulose and their

antibacterial study

Xuewei Zhang1,2,† , Haiyao Sun3, † , Shengnan Tan4 , Jing Gao1,2, , Yujie Fu1,2,, Zhiguo Liu1,2,*

1. Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry

University, Harbin 150040, People's Republic of China

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2. Engineering Research Center of Forest Bio-preparation, Ministry of Education, Northeast
Forestry University, Harbin 150040, People's Republic of China

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3. School of management and economics, Beijing Institute of Technology, Beijing 100081,
People's Republic of China

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4.Analysis and Test Center,Northeast Forestry University, Harbin 150040, People's Republic of
China

Abstract
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A facile hydrothermal method was developed to synthesize Ag nanoparticle on the
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nanocellulose in aqueous solutions in which the nanocellulose acts as the reducing

agent and stabilizer. The obtained Ag nanoparticles-nanocellulose nanocomposites

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were characterized by UV-Vis, FTIR, TEM, AFM and XRD measurements. The TEM
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and AFM observation results indicated that the synthesized silver nanoparticles are
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spherical and randomly distributed on the nanocellulose fibrils. The FTIR results
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confirmed that the nanocellulose is associated with the Ag nanoparticles. XRD

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measurements revealed that the silver nanoparticles are highly crystalline. The

antibacterial activity of the Ag nanoparticles-nanocellulose nanocomposites against

Gram-positive bacteria, Gram-negative bacteria and fungus was evaluated. The results

indicated that the silver nanoparticles-nanocellulose have high bactericidal efficiency

†
These Authors contributed equally to this work (co-first Author).
Corresponding author. E-mail: zguoliu@nefu.edu.cn; Tel.: +86-451-82192224; fax:
+86-451-82102082.
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against both bacteria and fungus.

Keywords: Ag nanoparticles; nanocellulose; antibacterial activity; Hydrothermal

synthesis

1. Introduction

Ag nanomaterials are not only widely used in the scientific research, but also

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applied in the daily life such as textiles, food storage containers and medical supplies

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due to their excellent comprehensive properties in mechanics, optics,

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superconductivity and antibacterial activity [1, 2]. Ag nanomaterials have also been

investigated for use in the biomedical devices and artificial implants to avoid bacterial
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infection [3-8].Because of their effective antibacterial activity, Ag nanoparticles (Ag
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NPs) have become as one of the most widely utilized nanomaterials in commercial

products such as home appliances, surface cleaners, room sprays, toys, textiles and
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medical devices [2, 9, 10]. The toxicity of the Ag nanoparticles and other
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nanomaterials on humans and the environment has been intensively investigated [2,
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11-13]. It is revealed that the stabilizing or capping agents of Ag nanoparticles play

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a very important role for their toxicity and antibacterial activity. Thus, it is very
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critical to develop different synthesis strategy and use novel stabilizing agents to

prepare Ag nanoparticles for further improving their antibacterial efficiency and

minimizing the possible adverse effects.

Nanocellulose, a sustainable and promising nanomaterial, has received

considerable attention in recent because of its unique structures, superb properties,

and natural abundance. Nanocellulose with various structures and surface chemistry

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properties can be produced from various sources of cellulose which is the most

abundant renewable biopolymer and almost inexhaustible raw material. Nanocellulose

consisted of β-1,4-linked D-anhydroglucose units has been explored in a wide variety

of applications[14]. Through different nanofibrillation and synthesis methods, uniform

nanofibrillated cellulose with a width ranging from 10 to100 nm and high aspect

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ratios can be fabricated [15]. Most importantly, abundant active hydroxyl groups on

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the nanocellulose surface are suitable for chemical modification. Consequently,

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nanocellulose with nanoscale features, high specific surface area, unique morphology,

low density, mechanical strength, renewability, biodegradability, and good

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biocompatibility can be considered as an ideal template to synthesis of noble metal
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nanomaterials.

There are some researches on the synthesis of cellulose/silver nanocomposite in

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recent [16, 17]. Since the common cellulose fibers are too large in comparison with
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Ag nanoparticles, the ratio of Ag nanoparticles in the composites is limited. In order

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to obtain more uniform and high ratio of Ag amount in the composites for biomedical
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application, it is desired to prepare Ag nanoparticles-nanocellulose nanocomposites.

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More recently, Tam et al. reported the preparation of silver nanoparticles/cellulose

nanocrystal hybrids with enhanced colloidal stability and antibacterial

performance[18]. The cellulose was modified with dopamine in their study. Toyoko

Imae et al. modified of cellulose with fourth generation poly(amido amine) dendrimer

to immobilize the Ag nanoparticle[19]. In these two studies, the chemical

modifications of cellulose with NH2-terminated molecules or polymer were required

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for Ag nanoparticle nucleation. The chemical modification process of cellulose could

induce the complexity of the products and alter the biocompatibilities of cellulose.

Consequently, development of facile and simple approaches to prepare Ag

nanomaterials-nanocellulose is still urgently needed owing to their potential

application in biomedical field. In this study, we developed a hydrothermal method

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for the synthesis of Ag nanoparticle on the nanocellulose in aqueous solutions in

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which the nanocellulose acts as the reducing agent and stabilizer. The nanocellulose

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was prepared by the TEMPO-mediated oxidization strategy [20].

2 Materials and methods

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2.1 Materials
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Microcrystalline cellulose and TEMPO were purchased from Sigma-Aldrich

(Shanghai China). AgNO3 (GA) was used without further purification. Other reagents
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including NaBr, NaClO, HCl, NaOH were analytical purity grade. Ultra-pure water
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was used throughout this study and its resistivity was >18 MΩ.cm.
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2.2 Preparation of nanocellulose by TEMPO-mediated oxidation method

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Microcrystalline cellulose (1g), TEMPO (0.016g) and NaBr (0.1g) were

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dissolved with 100 ml of pure water in a glass beaker. The obtained solution was then

heated in a water bath at 40 °C with continues stirring. 1.55 mL of NaClO solution

(20mmol, pH =10 (adjusted with 0.1M of HCl) ) was added in four batches with the

30 min interval. In the meantime, the solution pH was maintained at 10.0 by adding

appropriate volume of 0.5M NaOH solution. The oxidation product was collected by

centrifuging at 10000 rpm for 10 minutes. The obtained precipitate was washed with

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pure water until the supernatant became neutral. Recovery rate of the

TEMPO-oxidized celluloses after washing were over 90%.

2.3 Synthesis of Ag nanomaterials on the nanocellulose by hydrothermal method

0.1g of the dried TEMPO-oxidized nanocellulose was completely dissolved with

50 mL of pure water (assisted by sonication). The solution was then centrifuged at

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3000 rpm for 5 min to remove the large aggregate. The obtained supernatant was

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used for the synthesis. Nanocellulose solution (20 mL) and different volume of 1%

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AgNO3 （0.1 mL~0.5 mL）were mixed, respectively. The final solutions were

transferred into a Teflon-lined autoclave of 50 ml, and then sealed and maintained at
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140 °C for 4 h. Finally, the yellowish solutions were obtained.
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2.4 Characterization

2.4.1 UV–Vis absorption spectroscopy

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UV spectra were determined using a NanoPhotometer Model N60 ultramicro

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spectrophotometer (IMPLEM, Germany) .1.5 ul of the diluted solution was added to

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the spectrophotometer for measurement

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2.4.2 Fourier transform infrared spectroscopy (FTIR)

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FTIR spectra were measured by IR Affinity-1(Shimadzu) using KBr pellet method.

The sample was prepared by mixing the precipitate of the Ag nanoparticles (derived

from the centrifugation of the Ag colloidal solution) with a small amount of solid KBr.

FTIR spectra of pure microcrystalline cellulose and TEMPO oxidized nanocellulose

was also measured for comparison.

2.4.3 Transmission electron microscopy (TEM)

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The produced nanocomposites were imaged using JEM-2100 transmission electron

microscope (JEOL Ltd.) at 200 kV. The sample for TEM characterization was

prepared by placing 10 μl of the as-synthesized nanocomposites solution on a carbon

coated copper grid and dried at room temperature.

2.4.4 X-ray diffraction (XRD)

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Powder X-ray diffraction patterns were recorded with D / MAX 2200VPC (40 kV /

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40 mA). The XRD sample was centrifuged from the nanocomposites solution.

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2.4.5 Atomic force microscopy (AFM)

10 μl of the synthesized nanocomposites were dropped on the freshly cleaved mica

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surface. The samples were dried under a stream of pure Ar gas and stored in a
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desiccator prior to AFM imaging. The PicoPlus II AFM system from Molecular

Imaging Inc. was used. The resolution of the atomic force image is 512x512.
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2.5 Antibacterial tests

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The antibacterial tests of the nanocellulose nanocomposites were carried out with
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gram-negative bacteria E. coli, gram-positive bacteria S. aureus and the model fungus
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C. albicans. Penicillin was tested as the positive control. The lowest nanocomposites
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concentration that resulted in no microbial growth was determined as minimum

inhibitory concentration (MIC). 100 uL of freshly prepared microbial suspensions (1

× 108 CFU/ml) and 100uL of the tested samples with different concentrations were

mixed, and then transferred into 96-well plate. After 24 h of incubation at 37 °C for E.

coli and S. aureus, and 28 °C for C. albicans, the samples were determined by a

microplate reader (Biotek). Inhibition of zone of the synthesized nanocellulose

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nanocomposites was measured by the paper disk diffusion method. Nanocellulose

nanocomposites and control samples at 2MIC concentration were loaded on the filter

papers (6 mm diameter), and then gently placed onto the top of agar on which 108

cfu/ml bacterial suspensions including E. coli, S. aureus and C. albicans were

inoculated under sterile condition. After 24 h of incubation at 37 °C for E. coli and S.

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aureus, and 28°C for C. albicans, zone of growth inhibition around each sample was

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captured by digital camera.

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In order to further elucidate the antibacterial efficiency of the nanocomposites,

the inhibition rates of the nanocomposites synthesized with the different AgNO3
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quantity were compared. The bacterial density of activated E. coli, S. aureus and C.
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albicans was first diluted from 108 cfu / mL to 106 cfu / mL with broth. 100 uL of the

test samples (including positive control) at 2MIC concentration were mixed with 100
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uL of bacterial suspension, and incubated for 24 hours (E. coli, S. aureus at 37 ℃, C.

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albicans at 28 ℃). UV absorbance (OD) of the strain broth at 630 nm were

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measured and used for the calculation of inhibition rate. The inhibition rate is
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calculated as follows: A0 corresponds to the OD value of the culture broth before

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culture; At represents the OD value of the test sample. Acon is the OD value of the

mixed solution of broth and saline after incubation for 24 hours.

Inhibition rate (%) = 100 - 100 [(At - A0) / (Acon - A0)]

3. Results and discussion

3.1 Characterization of Ag nanoparticles- nanocellulose nanocomposites

Figure 1 shows the typical UV absorption spectra of the synthesized Ag

nanoparticles-nanocellulose solution. The obtained solution appeared as the light

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yellow color as shown in the inset in Figure 1. The strong absorption band peaked at

432 nm in Figure 1 can be attributed to the surface plasmon resonance band (PRB) of

the Ag colloid, which is in accordance with the previous studies [21-24]. Single PRB

also reveals that the obtained Ag nanomaterials are mainly spherical shape according

to the Mie theory, and does not form anisotropic particles. It has also been noticed that

the UV absorption band in present study located at relatively long-wavelength range

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in comparison with those of the Ag colloid in recent literatures [25, 26]. The position

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and intensity of the absorption peak of silver colloid is correlated with their shape,

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size and surface capping rate. Especially, the surface capping agents and capping rate

can significantly influence their UV absorption characteristics. Henglein et al. has

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demonstrated that the adsorption of nucleophilic species can modify the UV

absorption characteristics of the Ag nanoparticles [27]. In this study, nanocellulose is

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believed to be as both template and the capping agents of the Ag colloid.

Figure 2 shows the FTIR spectra of Ag nanoparticles-nanocellulose composites.

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The FTIR spectra of microcrystalline cellulose and TEMPO oxidized nanocellulose

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were also measured for comparison as shown in Figure 2. The peak at 3337 cm−1 in
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the composites spectra was ascribed to the hydrogen bond stretching vibration of
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−OH groups, and the absorption band peaked at 2916 cm−1 was ascribed to the

stretching vibration of C−H. The absorption at 1640 cm−1 was attributed to the
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bending vibration of −OH groups. The peak at 1061 cm−1 was attributed to the

stretching vibrations of C–O. In the FTIR spectra of the TEMPO oxidized

nanocellulose, the absorption at 1614 cm−1 was assigned to the sodium carboxylate

groups (−COONa) [28]. The disappearance of this absorption band in the Ag

nanoparticles-nanocellulose composites sample indicated the carboxylate groups may

involve the redox reaction. Furthermore, the absorption band attributed to the

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hydrogen bond stretching vibration in the nanocomposites sample was shifted to the

low wavenumber range in compared with the other two samples, which implied that

the −OH groups of nanocellulose may have the interaction with the surface of the Ag

nanoparticles.

Figure 3 (a) and (b) show the representative TEM image of the TEMPO oxidized

nanocellulose and Ag nanoparticles–nanocellulose nanocomposites. In Figure 3 (a),

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several hundred nanometers of the TEMPO oxidized nanocellulose can be identified.

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The nanocellulose tends to entangled with each other and form a cluster structure as

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the arrows shown in Figure 3 (a). In Figure 3 (b), a lot of the Ag nanoparticles can be

observed. It has been noted that most of Ag nanoparticles are attached on the
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nanocellulose template. These results confirmed that the nanoparticles are nucleated

and grown on the nanocellulose template. The measured size of the nanoparticle
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ranges from 20 to 180 nm. The average size of these nanoparticles is 86 nm. The size

distribution of the nanoparticles is showed in inset in Figure3 (b). Figure 3 (c) shows
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Energy dispersive X-ray (EDX) spectrum of the Ag nanoparticles–nanocellulose

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nanocomposites. This result confirmed the presence of silver metal in the

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nanocomposites . The Ag content is as much as 3.04 atom % in the average.

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The synthesized Ag nanoparticles–nanocellulose nanocomposites were further

characterized by atomic force microscope (AFM). Figure 4 (a) and (b) show the
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typical AFM image of the TEMPO oxidized nanocellulose and Ag nanoparticles–

nanocellulose nanocomposites，respectively. The nanocellulose appeared as many

rod-shaped whiskers, and these fibers bunched together into a nanosized tendon-like

structure. The section analysis of nanocellulose as indicated in the bottom of Figure 4

(a) indicates these rod-shaped whiskers have a width of 10-15 nm, a length of a few

hundred nanometers, and the height of 60 to 100 nm. These assembling structure of

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the nanocellulose and their stability are believed to be correlate with their surface

charge [29, 30]. In Figure 4 (b), many Ag nanoparticles can be observed on the

nanocellulose fibrils. During the AFM imaging, the position and height of these

nanoparticles was very stable which confirm these particles are firmly attached with

the nanocellulose fibrils and the AFM tips cannot disturb their structures. All these

AFM results are in accordance with those of TEM observation.

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In order to confirm the observed nanoparticles are made up of Ag atoms and has

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crystalline structures, XRD of the synthesized product was determined. Figure 5 (a)

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shows the XRD pattern of the TEMPO oxidized nanocellulose. There are no obvious

diffraction peaks from 30° to 90°. Figure 5 (b) shows the XRD pattern of the
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nanocomposites. The diffraction peaks (2θ values) at 37.9° 44.2 °, 64.1°,

77.2 °correspond to the (111), (200), (220) and (311) planes of the face centered cubic
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(FCC) Ag (JCPDS card no. 65-2871)[31].This result reveals the Ag+ ions have been

successful reduced under the hydrothermal conditions and formed highly crystalline
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structures.
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3.2 Formation mechanism of the Ag nanoparticles- nanocellulose nanocomposites

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It is believed that this is a redox reaction between nanocellulose and AgNO3 in

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which nanocellulose acts as the reducing agent to reduce Ag+. Nanocellulose is a

polysaccharide consisting of (1,4)-linked glucoses. There are a lot of OH group and

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end group on the nanocellulose fiber, which can be used to reduce Ag+ into Ag. After

mixing of AgNO3 solution and nanocellulose fibers, Ag+ can penetrate onto the of the

cellulose fibers. Due to the TEMPO oxidation in this study, the primary hydroxyl

group at the C6 position of the cellulose molecule is oxidized to the sodium

carboxylate group (COONa)[32]. The counter ion of sodium ion near oxidized

cellulose can be exchanged with the Ag+ to provide the cellulose-metal complex in

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the aqueous metal salt solution. Under the high temperature at 140℃, the adsorbed

Ag+ was reduced by OH groups of cellulose and formed as the initial nucleation site.

It should be pointed out that the hydrothermal process is necessary to accomplish

this reaction. The common heating method under standard atmospheric pressure has

been tested in the primary experiment, but did not produce a stable Ag colloidal

solution. As we demonstrated recently, the high temperature and high pressure derived

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from the hydro-thermal process are very useful to promote some redox reactions

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which cannot occur under standard atmospheric pressure[23].

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3.3 Antibacterial activities of the Ag nanoparticles- nanocellulose nanocomposites

The MIC of the Ag nanoparticles-nanocellulose nanocomposites was given in Table

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1. As shown in Table 1, the Ag nanoparticles-nanocellulose nanocomposites have a

good antibacterial activity against E.coli, C.albicans and S. aureus. Especially for
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E.coli, the Ag nanoparticles-nanocellulose nanocomposites indicated better

antibacterial efficiency. The bactericidal effect difference of the Ag

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nanoparticles-nanocellulose nanocomposites for different bacterial can be attributed to

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differences in cell wall structure and the abundance of functional groups on the cell

surface of different bacteria. The specificity of Ag nanoparticles for some bacteria has
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been recognized in several studies [33].

The zone of inhibition (ZOI) results of Ag nanoparticles-nanocellulose

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nanocomposites were shown in Fig. 6. The Ag nanoparticles-nanocellulose

nanocomposites synthesized with the different amount of AgNO3 and Penicillin were

investigated and compared. Penicillin has been frequently used as the positive control

to evaluate the antibacterial ability in the recent study[34]. It can be seen that Ag

nanoparticles-nanocellulose nanocomposites have good bactericidal effect against E.

coli. S. aureus and C.albicans. Especially for S. aureus, Ag

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nanoparticles-nanocellulose indicated a better antibacterial activity than the positive

control penicillin. Furthermore, its bactericide effects were increased with the

increasing the Ag used for the synthesis.

The inhibition ratio of the Ag nanoparticles-nanocellulose nanocomposites and the

control sample at 2MIC were further compared as shown in Figure 7. The Ag

nanoparticles-nanocellulose nanocomposites synthesized with the different amount of

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AgNO3 and penicillin were compared. It can be seen in Figure 7 that the Ag

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nanoparticles-nanocellulose nanocomposites have high bactericidal efficiency against

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E. coli. S. aureus and C.albicans. Furthermore, its bactericide effects were increased

with the increasing the amount of Ag used for the synthesis as shown in Figure 7. In a
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typical synthesizing condition (0.5 ml AgNO3 used in the synthesis), the obtained Ag

nanoparticles-nanocellulose nanocomposites have higher inhibition ratio than that of

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penicillin against E. coli..

In this study, the antibacterial activity of the nanocomposites is believed to be

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derived from the Ag nanoparticles. Some recent literatures have demonstrated the
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antibacterial activity about the different Ag nanomaterials [11, 35]. The release of
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silver atom or ions from Ag nanoparticles has been considered as one major route to
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produce bactericidal effects. Ag ions can interact with the cell membranes, nucleic

acids and proteins of the bacterial via the ligand exchange reactions to exert its
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activity. Additionally, the direct interaction of Ag nanoparticles with the surface of

bacterial cells also play important role for its antibacterial activity (particle-specific

effects). Ag atoms on the nanoparticles surface have a high affinity to the sulfur- and

phosphorus-containing compounds on the bacterial cell walls, which facilitate them

bind and enter into the bacterial cells to destroy it. In this study, the Ag nanoparticles

are randomly distributed on the nanocellulose template. Thus, the nanocellulose fibrils

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can also influence the nanocomposites antibacterial activity. It has been demonstrated

the nanocellulose fibrils have good biological compatibility, which facilitate the

interactions of the nanocomposites and bacterial. Therefore, the nanocellulose

templates are believed to enhance the antibacterial activity of the nanocomposites. In

addition, the Ag nanoparticles are firmly attached on the nanocellulose fibrils as

revealed by TEM and AFM observation, which facilitate them to be used for

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constructing more complex antibacterial products in biomedical field.

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4. Conclusion

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In this study, Ag nanoparticle on the nanocellulose has been synthesized by a

hydrothermal method. The obtained Ag nanoparticles-nanocellulose nanocomposites

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were characterized by UV-Vis, FTIR, TEM, AFM and XRD measurements. The FTIR

results verified that the nanocellulose is associated with Ag nanoparticle in the

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nanocomposites. The TEM and AFM observation results revealed that the synthesized

silver nanoparticles are spherical and firmly attached on the nanocellulose templates.
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XRD measurements revealed that the silver nanoparticles are highly crystalline. The
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antibacterial results indicated that the silver nanoparticles-nanocellulose

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nanocomposites have high bactericidal efficiency against both bacteria and fungus.
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Acknowledgements: The authors gratefully acknowledge the financial support by

National Key R&D Program of China (2016YFD0600805), the Fundamental

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Research Funds for the Central Universities (2572018CT01).

Author Contributions: X.Z. and H.S. synthesized and characterized the

nanocomposites, and analyzed the data. S.T. and J.G. contributed

reagents/materials/analysis tools. Y.F. and Z.L. designed the experiments and wrote

the paper.

Conflict of interest: The authors declare that they have no conflict of interest.

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References
[1] M.H. Chang, H.A. Cho, Y.S. Kim, E.J. Lee, J.Y. Kim, Thin and long silver nanowires self-assembled in
ionic liquids as a soft template: electrical and optical properties, Nanoscale Research Letters, 9 (2014)
330.
[2] P. Claudon, A. Violette, K. Lamour, M. Decossas, S. Fournel, B. Heurtault, G. Guichard,
Consequences of Isostructural Main‐Chain Modifications for the Design of Antimicrobial Foldamers:

PT
Helical Mimics of Host‐Defense Peptides Based on a Heterogeneous Amide/Urea Backbone.,
Angewandte Chemie International Edition, 49(2):333-336 (2010).

RI
[3] Y. Xiang, J. Lia, X. Liu, Z. Cui, X. Yang, K.W.K. Yeung, H. Pan, S. Wu, Construction of
poly(lactic-co-glycolic acid)/ZnO nanorods/Ag nanoparticles hybrid coating on Ti implants for

SC
enhanced antibacterial activity and biocompatibility, Materials Science and Engineering C, 79 (2017)
629-637.
[4] W.-Z. Yu, Y. Zhang, X. Liu, YimingXiang, ZhaoyangLi, S. Wu, Synergistic antibacterial activity of multi
NU
components in lysozyme/chitosan/silver/hydroxyapatite hybrid coating, Materials and Design, 139
(2018) 351-362.
[5] C. Mao, Y. Xiang, X. Liu, Z. Cui, X. Yang, K.W.K. Yeung, H. Pan, X. Wang, P.K. Chu, S. Wu,
MA

Photo-Inspired Antibacterial Activity and Wound Healing Acceleration by Hydrogel Embedded with
Ag/Ag@AgCl/ZnO Nanostructures, ACS Nano, 11 (2017) 9010-9021.
[6] C. Jin, X. Liu, L. Tan, Z. Cui, X. Yang, Y. Zheng, K.W.K. Yeung, P.K. Chu, S. Wu, Ag/AgBr loaded
Mesoporous Silica for Rapid Sterilization and Promotion of Wound Healing Biomaterials science, 6
D

(2018) 1735-1744.
[7] X. Xie, C. Mao, X. Liu, Y. Zhang, Z. Cui, X. Yang, K.W.K. Yeung, H. Pan, P.K. Chu, S. Wu, Synergistic
E

Bacteria Killing through Photodynamic and Physical Actions of Graphene Oxide/Ag/Collagen Coating,
PT

ACS Appl. Mater. Interfaces 9(2017) 26417-26428.

[8] X. Xie, C. Mao, X. Liu, L. Tan, Z. Cui, X. Yang, S. Zhu, Z. Li, X. Yuan, Y. Zheng, K.W.K. Yeung, P.K. Chu, S.
Wu, Tuning the Bandgap of Photo-Sensitive Polydopamine/Ag3PO4/Graphene Oxide Coating for Rapid,
CE

Noninvasive Disinfection of Implants, ACS Cent. Sci., 4 (2018) 724-738.

[9] R. Foldbjerg, X. Jiang, Teodora Miclăuş, C. Chen, H. Autrup, C. Beer, Silver nanoparticles – wolves in
sheep’s clothing?, Toxicology Research, 4 (2015) 563-575.
AC

[10] N. Durán, M. Durán, M.B.d. Jesus, A.B. Seabra, W.J. Fávaro, G. Nakazato, Silver nanoparticles: A
new view on mechanistic aspects on antimicrobial activity, Nanomed-Nanotechnol. , 12 (2016)
789-799.
[11] J. Liu, D.A. Sonshine, S. Shervani, R.H. Hurt, Controlled Release of Biologically Active Silver from
Nanosilver Surfaces, ACS nano, 4 (2010) 6903-6913.
[12] L. Tan, J. Li, X. Liu, Z. Cui, X. Yang, S. Zhu, Z. Li, X. Yuan, Y. Zheng, K.W.K. Yeung, H. Pan, Xianbao
Wang, S. Wu, Rapid Biofilm Eradication on Bone Implants Using Red Phosphorus and Near-Infrared
Light, Adv. Mater., (2018) 1801808.
[13] J. Li, L. Tan, X. Liu, Z. Cui, X. Yang, K.W.K. Yeung, P.K. Chu, S. Wu, Balancing Bacteria−Osteoblast
Competition through Selective Physical Puncture and Biofunctionalization of
ZnO/Polydopamine/Arginine-Glycine-Aspartic Acid-Cysteine Nanorods, ACS Nano 11 (2017)

14
 ACCEPTED MANUSCRIPT

11250-11263.
[14] M. Moniri, A.B. Moghaddam, S. Azizi, R.A. Rahim, A.B. Ariff, W.Z. Saad, M. Navaderi, R. Mohamad,
Production and Status of Bacterial Cellulose in Biomedical Engineering, Nanomaterials 7(2017) 257.
[15] W. Chen, H. Yu, S.-Y. Lee, T. Wei, J. Li, Z. Fan, Nanocellulose: a promising nanomaterial for
advanced electrochemical energy storage, Chem.Soc.Rev., 47 (2018) 2837-2872.
[16] R. Li, M. He, T. Li, L. Zhang, Preparation and properties of cellulose/silver nanocomposite fibers,
Carbohydrate Polymers, 115 (2015) 269-275.
[17] M.R.d. Moura, L.H.C. Mattoso, V. Zucolotto, Development of cellulose-based bactericidal
nanocomposites containing silver nanoparticles and their use as active food packaging, Journal of
Food Engineering, 109 (2012) 520-524.

PT
[18] Z. Shi, J. Tang, L. Chen, C. Yan, S. Tanvir, W.A. Anderson, R.M. Berry, K.C. Tam, Enhanced colloidal
stability and antibacterial performance of silver nanoparticles/cellulose nanocrystal hybrids, J. Mater.

RI
Chem. B, 3 (2015) 603-611.
[19] B. Ramaraju, T. Imae, A.G. Destaye, Ag nanoparticle-immobilized cellulose nanofibril films for

SC
environmental conservatio, Applied Catalysis A: General, 492 (2015) 184-189.
[20] A. Isogai, T. Saito, H. Fukuzumi, TEMPO-oxidized cellulose nanofibers, Nanoscale, 3 (2011) 71-85.
[21] Y. Zhang, H. Peng, W. Huang, Y. Zhou, X. Zhang, D. Yan, Hyperbranched Poly(amidoamine) as the
NU
Stabilizer and Reductant To Prepare Colloid Silver Nanoparticles in Situ and Their Antibacterial Activity,
J. Phys. Chem. C, 112 ( 2008) 2330-2336.
[22] Z. Liu, Z. Xing, Y. Zu, S. Tan, L. Zhao, Z. Zhou, T. Sun, Synthesis and characterization of L-histidine
MA

capped silver nanoparticles, Materials Science and Engineering C 32 (2012) 811-816.

[23] L. Biao, S. Tan, Y. Wang, X. Guo, Y. Fu, F. Xu, Y. Zu, ZhiguoLiu, Synthesis, characterization and
antibacterial study on the chitosan-functionalized Ag nanoparticles, Mat Sci Eng C-Mater, 76 (2017)
73-80.
D

[24] L. Biao, S. Tan, X. Zhang, J. Gao, Z. Liu, Y. Fu, Synthesis and characterization of
proanthocyanidins-functionalized Ag nanoparticles, Colloids and Surfaces B: Biointerfaces, 169 (2018)
E

438-443.
PT

[25] Z.G. Liu, Y.L. Wang, Y.G. Zu, Y.J. Fu, N. Li, N. Guo, R.S. Liu, Y.M. Zhang, Synthesis of
polyethylenimine (PEI) functionalized silver nanoparticles by a hydrothermal method and their
antibacterial activity study, Mat Sci Eng C-Mater, 42 (2014) 31-37.
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[26] B. Chudasama, A.K. Vala, N. Andhariya, R.V. Mehta, R.V. Upadhyay, Highly bacterial resistant silver
nanoparticles: synthesis and antibacterial activities, J Nanopart Res, 12 (2010) 1677-1685.
[27] A. Henglein, M. Giersig, Formation of Colloidal Silver Nanoparticles:  Capping Action of Citrate
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Journal of Physical Chemistry B, :, 103 (1999 ) 9533-9539.

[28] F. Cheng, C. Liu, X. Wei, T. Yan, H. Li, J. He, Y. Huang, Preparation and Characterization of
2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO)-Oxidized Cellulose Nanocrystal/Alginate Biodegradable
Composite Dressing for Hemostasis Applications, ACS Sustainable Chem. Eng. , 5 (2017) 3819-3828.
[29] Q. Xu, W. Li, Z. Cheng, G. Yang, M. Qin, TEMPO/NaBr/NaClO-Mediated Surface Oxidation of
Nanocrystalline Cellulose and Its Microparticulate Retention System with Cationic Polyacrylamide,
Bioresources, 9 (2013) 994-1006.
[30] W.R. Yao, Q.H. Xu, L.Q. Jin, Z.L. Cheng, Y. Gao, Effects of TEMPO/NaBr/NaClO-oxidation of
nanocrystalline cellulose on its properties, Chemistry & Industry of Forest Products, 35 (2015) 31-37.
[31] M.S.A.S. Shah, K. Zhang, A.R. Park, K.S. Kim, N.-G. Park, J.H. Park, P.J. Yoo, Single-step
solvothermal synthesis of mesoporousAg–TiO2–reduced graphene oxide ternary composites with

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enhanced photocatalytic activity, Nanoscale, 5 (2013) 5093-5101

[32] T. Saito, S. Kimura, Y. Nishiyama, A. Isogai, Cellulose Nanofibers Prepared by TEMPO-Mediated
Oxidation of Native Cellulose, Biomacromolecules, 8 (2007) 2485-2491.
[33] J.P. Ruparelia, A.K. Chatterjee, S.P. Duttagupta, S. Mukherji, Strain specificity in antimicrobial
activity of silver and copper nanoparticles, Acta Biomaterialia, 4 (2008) 707-716.
[34] T. Pandiyarajan, R. Udayabhaskar, S. Vignesh, R.A. James, B. Karthikeyan, Synthesis and
concentration dependent antibacterial activities of CuO nanoflakes, Materials Science and Engineering
C, 33 (2013) 2020-2024.
[35] S. Chernousova, M. Epple, Silver as Antibacterial Agent: Ion, Nanoparticle, and Metal, Angew.
Chem. Int. Ed., 52 (2013) 1636 - 1653.

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Figure legends

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Figure 1 UV absorption spectra of the diluted Ag colloidal solution (diluted 4 times of

the original colloid) in a typical experiment.

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Figure 2 The FTIR spectra of the Ag nanoparticles-nanocellulose nanocomposites,
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microcrystalline cellulose and TEMPO oxidized nanocellulose.

Figure 3 (a) and (b), The representative TEM image of TEMPO oxidized
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nanocellulose and the Ag nanoparticles-nanocellulose nanocomposites. 3 (c), Energy

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dispersive X-ray (EDX) spectrum of the nanocomposites.

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Figure 4 (c) and (d), The typical AFM image of TEMPO oxidized nanocellulose and
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the Ag nanoparticles-nanocellulose nanocomposites.

Figure 5 (a) and (b), XRD pattern of the TEMPO oxidized nanocellulose and Ag
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nanoparticles-nanocellulose nanocomposites.

Figure 6 The zone of inhibition (ZOI) results of Ag nanoparticles-nanocellulose

nanocomposites.

Figure 7 Inhibition ratios of the Ag nanoparticles-nanocellulose nanocomposites

synthesized from the different amount of AgNO3 at 2MIC against E. coli, S. aureus

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and C. albicans. Penicillin was also tested for comparison.

Figure 1

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Figure 3

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Figure4

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Figure 5
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Figure6

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Figure7
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Table 1 The MIC of the Ag nanoparticles- nanocellulose nanocomposites.

μg/mL

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E.coli S.aureus C.albicans
Ag NPs-nanocellulose 15.84 31.68 31.68

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Penicillin 3.90 7.80 7.80

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Hydrothermal synthesis of Ag nanoparticles on the nanocellulose and their

antibacterial study

Xuewei Zhang , Haiyao sun, Shengnan Tan , Jing Gao, , Yujie Fu,, Zhiguo Liu

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Graphical abstract

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Highlights

► Ag nanoparticles-nanocellulose was synthesized by a hydrothermal

method

►Ag nanoparticles-nanocellulose have been characterized by UV, FTIR,

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TEM, and XRD

► Ag nanocomposites have high bactericidal efficiency against both

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bacteria and fungus.

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