Carbohydrate Polymers 124 (2015) 3542

Contents lists available at ScienceDirect

Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Antimicrobial and antioxidant surface modication of cellulose bers

using layer-by-layer deposition of chitosan and lignosulfonates
Hui Li a,b,c, , Lincai Peng b,d
a

Food Safety Research Institute, Kunming University of Science and Technology, Kunming 650500, China
State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510640, China
Key Laboratory of Pulp and Paper Science & Technology of Ministry of Education of China, Qilu University of Technology, Jinan 250353, China
d
Faculty of Chemical Engineering, Kunming University of Science and Technology, Kunming 650500, China
b
c

a r t i c l e

i n f o

Article history:
Received 26 June 2014
Received in revised form 18 January 2015
Accepted 26 January 2015
Available online 12 February 2015
Keywords:
Layer-by-layer
Chitosan
Lignosulfonates
Antimicrobial activity
Antioxidant activity
Cellulose bers

a b s t r a c t
To confer cellulose bers antimicrobial and antioxidant activities, chitosan (CS)/lignosulfonates (LS) multilayers were constructed on bers surfaces through layer-by-layer deposition technique. The formation
of CS/LS multilayers on cellulose bers surfaces was veried by X-ray photoelectron spectroscopy (XPS)
and zeta potential measurement. The surface morphologies of CS/LS multilayers on bers surfaces were
observed by atomic force microscopy (AFM). The results showed that characteristic element (i.e. N and
S element) content increased with increasing bilayers number, the surface LS content increased linearly
as a function of bilayers. Zeta potential of modied bers was inversed after deposition of each layer.
AFM phase images indicated that the cellulose microbrils on bers surfaces were gradually covered
by granular LS aggregate. The antimicrobial testing results demonstrated that CS/LS multilayers modied bers with CS in the outermost layer exhibited higher antimicrobial activity against Escherichia coli.
The antioxidant testing results showed that antioxidant activity of CS/LS multilayers modied bers was
better than that of original bers under the same oxidation conditions.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
Cellulose bers are one of the most abundant, renewable,
biodegradable and biocompatible natural polymers. Cellulose
bers and their derivatives have been used in a variety of applications in several areas, such as textile industry, paper industry,
packaging industry and medical eld (Kalia, Thakur, Celli, Kiechel,
& Schauer, 2013; Vuoti et al., 2013). However, the cellulose bersbased materials are particularly easy to be attacked by fungi and
bacterial during use and storage (Vartiainen et al., 2004). Microbial growth on the bers-based materials leads to irreversible
changes of a destructive character, which are the results of oxidation, hydrolysis and ssion of cellulose chains (Silva et al.,
2011). These chemical changes ultimately give rise to the materials degradation, strength loss and even increasing the risk of
infection in the medical eld (Szostak-Kotowa, 2004; Dong et al.,
2014). These detrimental effects can be avoided or controlled by

Corresponding author at: Food Safety Research Institute, Kunming University of

Science and Technology, Jingming South Road 727, Kunming 650500, China.
Tel.: +86 087165920293.
E-mail address: lihuiscut@126.com (H. Li).
http://dx.doi.org/10.1016/j.carbpol.2015.01.071
0144-8617/ 2015 Elsevier Ltd. All rights reserved.

antimicrobial and antioxidant modication of cellulose bers using

specic agents (Martins et al., 2012).
The selection of antimicrobial and antioxidant agent depended
on the mechanism of antimicrobial and antioxidant activities, toxicity and cost (Ammayappan & Moses, 2009). In recent years,
natural and eco-friendly antimicrobial and antioxidant agents have
attracted considerable attentions with increasing awareness of
environment protection and concern for infectious diseases control
(Kenny et al., 2014; Brewer, 2011). Chitosan (CS) is a natural cationic
polysaccharide. Owing to its antimicrobial, nontoxic, hemostatic,
biocompatible and biodegradable properties, chitosan has been
widely used in various scientic elds, including biotechnology
(Suginta, Khunkaewla, & Schulte, 2013), agriculture (Coqueiro,
Maraschin, & Piero, 2011), food-preservation (Aider, 2010), medical and pharmaceutical areas (Cline et al., 2013). Lignins are one of
most useful natural resources and millions tons of technical lignins
are produced globally each year, mainly as a by-product of the
pulping industry (Calvo-Flores & Dobado, 2010). Lignins are natural polyphenolic compounds that contain phenolic groups, which
possess antioxidant characteristics (Dizhbite, Telysheva, Jurkjane,
& Viesturs, 2004; Garca, Toledano, Andrs, & Labidi, 2010). Most
studies have revealed the efcacy of different source technical
lignins (such as kraft lignin, lignosulfonates, ethanol lignin) as

36

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

antioxidants in different composite materials (rubber, plastic, thermomechanical pulp) (Agarwal, Prasad, Sharma, & Setua, 2014;
Pouteau, Dole, Cathala, Averous, & Boquillon, 2003; Schmidt, Rye, &
Gurnagul, 1995; Pan, Kadla, Ehara, Gilkes, & Saddler, 2006), as well
as cosmetics and pharmaceutical products (Vinardell, Ugartondo,
& Mitjans, 2008; Ugartondo, Mitjans, & Vinardell, 2008; Ugartondo,
Mitjans, & Vinardell, 2009). Recent industrial applications of technical lignins are mainly based on lignosulfonates (LS). Compared
with other technical lignins, LS recovered from the spent pulping liquids from sulte pulping exhibit excellent water-solubility
and anionic character due to their functional sulfonate groups (Kim
et al., 2013). LS have been employed in a wide range of specialty
products, such as dispersant (Yang, Qiu, Zhou, & Lou, 2007), surfactant (Trufanova, Parfenova, & Yarygina, 2010), resin adsorbent
(Liang, Song, Huang, Zhang, & Chen, 2013). Moreover, the antioxidant and antimicrobial activities have extended their potential
applications (Lora & Glasser, 2002; Faustino, Gil, Baptista, & Duarte,
2010; Kang, Li, Chang, & Fan, 2011).
The electrostatic layer-by-layer (LBL) deposition technique, as
reported rst by Decher (1997), has proved to be a simple and
versatile surface modication method via constructing nanostructured multilayer with tailored composition and properties on solid
substrate surface (Zhang et al., 2004; Li, Biagioni, Finazzi, Tavazzi,
& Piergiovanni, 2013a). The basic ideal of LBL method is the
alternate deposition of polycations and polyanions on the solid surfaces through electrostatic interactions (Elosua, Torres, Hernaez,
Matias, & Arregui, 2013). A great variety of substances including polyelectrolytes (Joseph, Ahmadiannamini, Hoogenboom, &
Vankelecom, 2014), inorganic nanoparticles (Li, Fu, & Peng, 2013b)
and enzyme (Xing, Eadula, & Lvov, 2007) have been employed
to fabricate the LBL multilayers. Recent studies showed that
polyelectrolyte/polyelectrolyte or polyelectrolyte/nanoparticles
multilayers created by LBL deposition technique have been widely
utilized to modify the surfaces of cellulose bers (Wgberg, 2012).
Some improved physical properties and special functionalities can
be given for the modied bers, such as high scalability (Marais,
Utsel, Gustafsson, & Wgberg, 2014), high porosity (Lu et al., 2007),
electroconductibility (Agarwal et al., 2009; Hu et al., 2013), magnetism (Cranston & Gray, 2006), ame resistance (Laufer, Kirkland,
Morgan, & Crunlan, 2012), superhydrophobicity (Zhao, Tang, Wang,
& Lin, 2010; Zhang, Wang, Wang, & Li, 2012; Gustafsson, Larsson,
& Wgberg, 2012), etc. Additionally, antimicrobial modication of
cellulose bers-based materials mentioned using LBL deposition
technique has attracted a lot of interest, and several related studies have been reported (Gomes, Mano, Queiroz, & Gouveia, 2013).
However, numerous related studies involving using synthetic polyelectrolytes (like polystyrolsulfon acid, polyving akohol, polyacrylic
acid) or photocatalytic nanoparticles (like Ag, ZnO, TiO2 ) as LBL
building blocks (Joshi, Khanna, Shekhar, & Jha, 2011; Iiiergrd,
Rmling, Wgberg, & Ek, 2013; Zahran, Ahmed, & EI-Rae, 2014;
Iamphaojeen & Siriphannon, 2012; Li et al., 2013b), and so far
there are only few reports concerning the alternate deposition of
natural polyelectrolytes with opposite charges on cellulose bers
surfaces (Deng et al., 2011; Gomes, Mano, Queiroz, & Gouveia,
2012).
The aim of this study was to prepare bio-compatible cellulose bers with antimicrobial and antioxidant activities, which
will have a broad range of potential application in medical,
packaging and other health-related areas. For this purpose, natural antimicrobial and antioxidant agents (CS and LS) were
deposited on cellulose bers surfaces via electrostatic layer-bylayer deposition technique. The chemical composition, growth
manner and surface morphologies of CS/LS multilayers were measured, respectively. Furthermore, the antimicrobial and antioxidant
activities of CS/LS multilayers modied cellulose bers were also
evaluated.

Scheme 1. Schematic representation of CS/LS multilayers deposition process on

bers surfaces.

2. Experimental
2.1. Materials
Fully bleached eucalyptus kraft pulp bers were kindly provided by Guangxi Hektar Paper Industry Co., Ltd (Guangxi, China).
Before use, the bers were washed and the carboxyl groups
of the bers were converted to their sodium forms according
to Marais et al. (2014). The chitosan (CS) with 95% degree of
deacetylation and radical inducer 2,2 -azobis(2-amidinopropane)
dihydrochloride (ABAP) were purchased from Aladdin Reagent Co.
Ltd. (Shanghai, China). The CS aqueous solution was prepared by
dissolving CS in 0.1 M CH3 COOH, adjusting pH value to 4 using
HCl and NaOH. The lignosulfonates (LS) were kindly provided by
Jiangmen Sugarcane Chemical Factory Co. Ltd. (Guangdong, China).
Before use, the LS were prepared as described in our previous work
(Li et al., 2013b). The ultrapure water (Millipore, MA, USA) with a
resistivity of 18 M/cm was used in all experiments, and all other
chemicals were analytical grade and used without further purication.
2.2. Preparation of CS/LS multilayers on cellulose bers surfaces
Multilayers composed of CS and LS on cellulose bers surfaces have been fabricated by an alternating immersion process
(Scheme 1). The bers were rst immersed into CS aqueous solution (0.5 g/L, pH 4) for 15 min (Step 1), then rinsed thoroughly three
times using ultrapure water to remove excess CS and drying in air
(Step 2). Subsequently, the bers were immersed into LS aqueous
solution (0.5 g/L, pH 4) for 15 min (Step 3), which was followed by
the same rinsing and drying procedure (Step 4). Repeating Steps
14 n times fabricates a multilayers containing alternating n layers
of CS and n layers of LS, which was dened as (CS/LS)n .
2.3. Characterization
2.3.1. X-ray photoelectron spectroscopy (XPS) measurements
The XPS measurements were carried out using a Kratos
Axis Ultra spectrometer (Kratos Analytical, UK) equipped with a
monochromated A1 K X-ray source (150 W). Prior to XPS measurement, the cellulose bers were made into pulp sheets by vacuum
ltration. The test samples in circular form (about 0.5 cm2 ) were
cut from the sheet and mounted with screw-on clips to the sample holder. The samples were degassed overnight under vacuum
in a turbo-pumped chamber to stabilize the moisture content in
the sample matrix. At least 3 different spots were analyzed on
each sample. The chemical elements present on the samples were
indentied from survey spectra.
2.3.2. Zeta potential measurement
A Mtek SZP-06 zeta potential analyzer (BTG Group,
Germany) based on the streaming potential method was
used to measure the zeta potential of modied bers. Before
test, the modied cellulose bers were dispersed in water

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

37

Fig. 1. XPS spectra of original and modied cellulose bers: (a) original cellulose bers, (b)(d) (CS/LS)1 , (CS/LS)3 and (CS/LS)5 multilayers modied cellulose bers,
respectively.

to form 3% consistency ber suspension, the pH of ber

suspension was adjusted to 4 by using HCl and NaOH. Each
sample was tested in triplicate and the average value was reported
in this work.
2.3.3. Atomic force microscopy (AFM) analysis
The bers for AFM analysis were taken from the same pulp
sheets for XPS analysis, and the sample preparation method was
as described by Liu, Fu, Zhu, Li, and Zhan (2009). A commercial
Multimode Nanoscope IIIa AFM system (Veeco, Santa Barbara, CA,
US) was used to observe the surfaces characteristics of modied
cellulose bers. The AFM system was equipped with a J-type scanner and a standard silicon cantilever with a resonance frequency
of 290320 kHz. The scan was operated in a tapping mode in air at
room temperature with a relative humidity of 65%. Several scans
were performed from different parts of the samples and representative images were chosen for presentation. The scanning size was
1.5 1.5 m. Software Version 5.12r3 (Veeco Co., USA) was used
for online data recording and software WSxM (Nnaotec Electronica, Spain) was used for ofine data analysis. No image processing
except attening was made.
2.4. Antimicrobial activity of CS/LS multilayers modied cellulose
bers
The method used for testing the antimicrobial activity of modied cellulose bers was as described by Qian et al. (2009).
Gram-negative Escherichia coli was selected as representative
microorganism. The procedure is as follows: The pulp sheet (0.1 g)
cut from XPS analysis sheet sample sterilized by autoclaving and
5 mL E. coli suspension (ca. 106 CFU/mL) were mixed and shaken
at 200 rpm at 37 C for 1 h. After shaking, a series of dilutions were
made and then 100 L of dilution was spread on LuriaBertani agar
in a Petri dish. The plates were incubated at 37 C for 24 h and the
number of colonies was counted. At least three repeated tests were
carried out for each sample. The growth inhibition degree of E. coli
can be quantied by the following equation:
Degree of growth inhibition for E. coli =

AB
100%
A

(1)

where A and B are the number of colonies of the control and tested
samples, respectively.

2.5. Antioxidant activity of CS/LS multilayers modied cellulose

bers
2.5.1. ABAP-initiated oxidation treatment
The oxidation treatment was carried out in a reactor kettle
equipped with an automatic temperature control system, pressure
control and mechanical stirring. ABAP-initiated oxidation treatment conditions were as follows: 10% ABAP (relative oven-dried
bers), O2 pressure 140 kPa, temperature 60 C, reaction time (0, 2,
6, 12, 24, 48 h), stirring speed 120 rpm, 10% consistency in 50 mM
phosphate buffer (pH 7.0). After oxidation treatment, the bers
were washed thoroughly with ultrapure water and ltered.
2.5.2. Physical properties testing
The cellulose bers treated by ABAP-initiated oxidation were
made into handsheets with a grammage of 80 g/m2 using a semiautomatic sheet former equipped with circulation water. Prior to
test, the handsheets were kept at constant temperature and humidity (23 C, 50% relative humidity) for at least 24 h. The zero-span
tensile strength was determined using a Pulmac zero-span tensile tester (Pulmac International Inc, Middlesex, USA) according to
ISO 15361:2000 standard. The intrinsic viscosity values of cellulose bers were determined using a cupri-ethylenediamine (CED)
solution according to the standard method SCAN-CM 15:88. The
degree of polymerization (DP) was calculated using the following
MarkHouwinkSakurada equation (Sihtola, Kyrklund, Laamanen,
& Palenius, 1963).
DP0.905 = 0.75 []
where [] is the intrinsic viscosity value of cellulose bers.
3. Results and discussion
3.1. Formation of CS/LS multilayers on cellulose bers surfaces
XPS was used to examine the surface chemical composition of
original and modied cellulose bers during the LBL deposition
process. The survey XPS spectra of original and modied bers
are shown in Fig. 1. The original bers almost consisted of cellulose, distinctive peaks at 284.6 eV and 532.6 eV indicated the
presence of carbon and oxygen, respectively (Fig. 1a). For the modied bers, distinctive peaks at 167.7 eV, 284.6 eV, 399.2 eV, and

38

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

Fig. 2. Chemical structure of chitosan.

532 eV indicated the existence of sulfur, carbon, nitrogen and oxygen (Fig. 1bd). Here, the S 2p peak observed was attributed to
sulfonic groups in LS macromolecule, whereas N 1s peak observed
belonged to amino groups in CS macromolecule (Fig. 2). Besides,
there were obvious increase in peaks intensities of sulfur and
nitrogen with the increasing bilayers number, demonstrating the
growth of CS/LS multilayers on cellulose bers surfaces.
XPS has been shown to be a useful tool to determine lignin content of bers surfaces (Johansson, Campbell, Koljonen, & Stenius,
1999). The surface lignin content of cellulose bers can be calculated from the components relative amount of C 1s peak after
deconvolution (Gustafsson, Ciovica, & Peltonen, 2003). A high resolution C1s spectrum can be deconvoluted into different carbon
components, i.e. carbon with different chemical environments
according to Dorris and Gray (1978). The types of chemical bonds of
carbon in pure LS can be categorized into ve groups: C1 (C C), C2
(C O), C3 (O C O, C O), C4 (O C O) and C5 (C S). On the other
hand, pure cellulose contains only C2 and C3 carbons. Therefore, the
surface LS content can be evaluated by determining the percentage
of C1, C4 and C5 if the sample only consisted of cellulose and LS. As
reported by our previous work (Li, Liu, Fu, & Zhan, 2011), the C4 and
C5 cannot be used for calculation the LS content due their low content in LS and poor tting accuracy (because of a small difference
between C2 and C5, they were tted in one peak), respectively. The
ber surface LS content (LS ) can by quantied by determining the
relative amount of C1 carbon, as shown by the following equation:
LS =

C1
100%

(2)

where C1 represents the area of C1 peak divided by the total area

of the C 1s peak, is a correction factor for the presence of contaminants, and is the area of the C1 peak divided by the total are
of C1s peak for the puried LS, here is 47% as reported before (Li
et al., 2011). It is worth noting that the CS also presented on the
modied bers surfaces, which contains C2, C3 and C bonded to N
(287.3 eV). These three carbons in CS molecule also have contribution to division of C 1s peak. However, they affect C2 and C3 carbons
more than C1 carbon, so the calculation error for Eq. (2) is minor.
XPS analysis results are illustrated in Table 1. For original cellulose bers, a small C1 peak was detected. This is due to presentation
of a very small amount of residual lignin on fully bleached pulp surface. Hence, 3.7% was employed for value in Eq. (2) to eliminate
the C1 contaminants from sample. With the LBL process proceeded, the peaks intensities of C1 and C2 increased, demonstrating
increased amount and coverage of building blocks on original cellulose ber surface since both CS and LS have C1 and C2 carbons.
Furthermore, the calculated surface LS content were plotted as a
function of bilayers number in Fig. 3. There is an almost linear
relationship between surface LS content and number of bilayers
(R2 = 0.972), suggesting that the consecutive deposition of cationic
CS and anionic LS on ber surface is stepwise and the LBL deposition process is very consistent from layer to layer and highly
reproducible.

Fig. 3. Surface content of LS on cellulose bers as a function of bilayers number.

The surface charge of modied cellulose bers was monitored

with zeta potential measurements. The zeta potential of modied
bers during LBL deposition process is presented in Fig. 4. The original cellulose bers had a negative potential of 29.7 mV. When
the CS was deposited on the ber surface, the zeta potential of
ber surface reversed to positively charge of 13.6 mV, subsequent
deposition of LS onto cellulose bers reversed the zeta potential
to 10.9 mV. Alternating and regular zeta potential reversals were
observed with the further deposition of each oppositely charged CS
and LS, further demonstrating that the deposition process of CS and
LS on ber surface could be achieved in a reproducible way, which
is consistent with the results from XPS analysis.
3.2. Surface morphologies of CS/LS multilayers on cellulose bers
surfaces
The surface morphologies of cellulose bers modied with CS/LS
multilayers investigated by AFM are shown in Fig. 5. The original
cellulose ber surface composed of regular ordered microbrils
exhibited a uniform topography with a root-mean-square (RMS)
roughness of about 13.57 nm (Fig. 5a). As the number of bilayers
increased, some granular substances gradually appeared on the
brillar surface (Fig. 5b and d). When (CS/LS)5 multilayers were
formed on cellulose ber surface, the brillar surface was almost
completely covered with granular substance. The amount of these
granular substances increased with the increasing number of bilayers, resulting in the increase in surface RMS roughness of modied
cellulose bers. The cellulose bers surfaces modied with (CS/LS)1 ,
(CS/LS)3 and (CS/LS)5 multilayers have RMS roughness of 21.51 nm,
25.83 nm and 32.68 nm, respectively. These granular substances

Fig. 4. Zeta potential changes of modied cellulose bers during the LBL deposition
process.

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

39

Table 1
XPS results from cellulose bers modied by CS/LS multilayers.
Number of
bilayers
0
1
2
3
4
5

LS (%)

C 1s total = 100%
C1 (%)

C2, C5 (%)

C3 (%)

C4 (%)

3.7
10.3
18.1
24.2
29.5
31.7

80.3
68.2
61.5
62.6
65.4
65.7

15.1
19.4
18.0
10.7
3.4
1.5

0.9
2.1
2.4
2.5
1.7
1.1

0
14.1
30.6
43.6
54.9
59.6

Fig. 5. Surface morphologies of original and modied cellulose bers: (a) original ber, (b)(d) (CS/LS)1 , (CS/LS)3 and (CS/LS)5 multilayers modied bers, respectively, (e)
CS alone-modied bers.

were regarded as LS granules aggregate. This is attributed to three

reasons: (i) the morphological character of ber surface modied
with CS alone was very similar to that of original ber surface and
the CS cannot be distinguished from the ber surface as presented
in Fig. 5e; (ii) LS granules tend to aggregate in acid solution due
to hydrogen-bonding interactions (Nyman, Rose, & Ralston, 1986);
(iii) the amount of granular substances increased with the increase
in the bilayers number, which is corresponding to increased surface
LS content.

outermost layer was better than that of the samples with

LS in the outermost layer. Furthermore, the growth inhibition
degree increased with the increasing bilayers number when the
CS in the outermost layer. These results may be caused by

3.3. Assessment of antimicrobial activity of cellulose bers

modied by CS/LS multilayers
The degree of growth inhibition for E. coli was examined in
CS/LS multilayers modied bers with different surface compositions and deposited bilayers. It was found from Fig. 6 that
all the modied cellulose bers had E. coli inhibition activity
because of the presence of CS. The growth inhibition degree
of (CS/LS)4.5 multilayer modied cellulose bers reached to
97%, while the growth inhibition degree of (CS/LS)5 multilayer modied cellulose bers was less than 63%, indicating
that the antimicrobial activity of the samples with CS in the

Fig. 6. Growth inhibition degree for E. coli by cellulose bers modied with (CS/LS)n
multilayers having different bilayers number.

40

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

Fig. 7. DP (a) and zero-span tensile strength (b) of original and modied cellulose bers versus oxidation reaction time.

The presence of ROO and Cell-OO will cause both depolymerization and propagation of the radical chain. LS are a phenolic polymers
which inhibit ABAP-initiated oxidation reactions by trapping the
chain-propagating ROO. The LS phenolic polymers gives up their
phenolic hydrogen atom to ROO, producing phenoxyl radical
PhO, terminating chain reaction between ROO and cellulose.
Simultaneously, colored non-radical products such as quinones
were formed by PhO coupling reaction.
4. Conclusions

Scheme 2. Schematic diagram illustrating ABAP radical induced oxidation mechanism.

three reasons: (1) CS has antimicrobial activity; (2) the E. coli

absorption and immobilization capacities of CS were increased
because of its hydrophilicity and higher positive charge; (3) the
increase in surface RMS roughness of modied cellulose bers led
to higher specic area, which may cause efcient contact and interaction between CS and E. coli.
3.4. Assessment of antioxidant activity of cellulose bers modied
by CS/LS multilayers
Degree of polymerization reects the average length of cellulose
chains. The zero-span tensile strength is a widely used index for
evaluating the average strength of individual ber. Fig. 7 shows
plots of DP and zero-span tensile strength of original and modied
cellulose bers against oxidation reaction times. It is clearly seen
that DP and zero-span tensile strength of original cellulose bers
dramatically decreased as the oxidation reaction time increased,
which is due to attack of radical to cellulose chains, thereby causing
depolymerization and strength loss of cellulose bers. For the CS/LS
multilayers modied cellulose bers, the DP and zero-span tensile
strength gently decreased with increasing oxidation reaction time.
As the number of bilayers increased, the changes in the DP and
zero-span tensile strength of modied bers became level off. This
can be explained that LS layers on modied bers surfaces acted
as natural radical scavenger, which protected cellulose ber from
radicals attacking.
In this study, a probable radical oxidation mechanism is shown
in Scheme 2. Radical initiator ABAP is a water-soluble azo, which
can rapidly decomposed at 60 C, producing two carbon-centerd
radicals (R). R reacted with oxygen to generate peroxyl radical
(ROO), which attracted the cellulose, resulting in cellulose chain
fragmentation and generating cellulose radical (Cell). The Cell
reacted with oxygen to produce Cell-OO and attracted cellulose.

The antimicrobial and antioxidant surfaces modication of cellulose bers were achieved through constructing CS/LS multilayers
on bers surfaces via layer-by-layer deposition technique. The
increase in characteristic elements (N and S) of CS/LS multilayers with the number of bilayers was observed, and the surface LS
content of modied cellulose ber linearly increased as a function
of bilayers. The surface zeta potential of modied cellulose bers
was inversed after each deposition step. The AFM phase images
showed that the granular LS aggregate gradually covered the cellulose bers surfaces as the LBL deposition proceeded, resulting in
the increase in bers surfaces roughness. The antimicrobial testing
results demonstrated that cellulose bers modied by CS/LS multilayers exhibited higher antimicrobial activity against E. coli, the
degree of E. coli growth inhibition for a (CS/LS)4.5 multilayer modied cellulose bers reached up to 97%. The E. coli growth inhibition
degree increased with the increasing bilayers number, especially
E. coli growth inhibition degree of modied cellulose bers with
CS in the outmost layer was better than that of the cellulose bers
with LS in the outmost layer. The antioxidant testing results showed
that there was almost no change in the DP and zero-span tensile
strength of modied bers after ABAP-initiated oxidations due to
the presence of radical scavenger LS. Moreover, the antioxidant
activity of CS/LS multilayers modied cellulose bers increased
with the bilayers number increased.
Acknowledgements
This work was supported by the Applied Basic Research Program
of Yunnan Province (no. 2014FD008), Talent Training Program of
Yunnan Province (no. KKSY201305002), State Key Laboratory Open
Foundation of Pulp and Paper Engineering of China (no. 201323)
and Open Foundation of Key Lab of Pulp and Paper Science &Technology of Ministry of Education (no. 08031349).
References
Agarwal, K., Prasad, M., Sharma, R. B., & Setua, D. K. (2014). Novel bio-degradable
lignin reinforced NBR composite. International Journal of Energy Engineering, 4,
4762.

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

Agarwal, M., Xing, Q., Shim, B. S., Kotov, N., Varahramyan, K., & Lvov, Y. (2009).
Conductive paper from lignocellulose wood microbers coated with a nanocomposite of carbon nanotubes and conductive polymers. Nanotechnology, 20, 18.
Aider, M. (2010). Chitosan application for active bio-based lms production and
potential in food industry: Review. LWTFood Science and Technology, 43,
837842.
Ammayappan, L., & Moses, J. J. (2009). Study of antimicrobial activity of aloevera,
chitosan, and curcumin on cotton, wool, and rabbit hair. Fibers and Polymers, 10,
161166.
Brewer, M. S. (2011). Natural antioxidants: Sources, compounds, mechanisms of
action, and potential applications. Comprehensive Review in Food Science and
Food Safety, 10, 221247.
Calvo-Flores, F. G., & Dobado, J. A. (2010). Lignin as renewable raw material. ChemSusChem, 3, 12271235.
Cline, P. B., Antoine, V., Denis, B., Laurent, V., Laurent, D., & Catherine, F. (2013).
Development and characterization of composite chitosan/active carbon hydrogels for a medical application. Journal of Applied Polymer Science, 128, 29452953.
Coqueiro, D. S. O., Maraschin, M., & Piero, R. M. D. (2011). Chitosan reduces bacterial
spot severity and acts in phenylpropanoid metabolism in tomato plants. Journal
of Phytopathology, 159, 488494.
Cranston, E. D., & Gray, D. G. (2006). Formation of cellulose-based electrostatic layerby-layer lms in a magnetic eld. Science and Technology of Advanced Materials,
7, 319321.
Decher, G. (1997). Fuzzy nanoassembilies: Toward layered polymeric multicomposites. Science, 277, 12321237.
Deng, H. B., Wang, X. Y., Liu, P., Ding, B., Du, Y. M., Li, G. X., et al. (2011). Enhanced
bacterial inhibition activity of layer-by-layer structured polysaccharide lmcoated cellulose nanobrous mats via addition of layered silicate. Carbohydrate
Polymers, 83, 239245.
Dizhbite, T., Telysheva, G., Jurkjane, V., & Viesturs, U. (2004). Characterization of the
radical scavenging activity of ligninsNatural antioxidants. Bioresource Technology, 95, 309317.
Dong, C., Ye, Y., Qian, L. Y., Zhao, G. L., He, B. H., & Xiao, H. N. (2014). Antibacterial
modication of cellulose bers by grafting -cyclodextrin and inclusion with
ciprooxacin. Cellulose, 21, 19211932.
Dorris, G. M., & Gray, D. G. (1978). The surface analysis of paper and wood bres by
ESCA. Cellulose Chemistry and Technology, 12, 721734.
Elosua, C., Torres, D. L., Hernaez, M., Matias, I. R., & Arregui, F. J. (2013). Comparative
study of layer-by-layer deposition technique for poly(sodium phosphate) and
poly(allylamine hydrochloride). Nanoscale Research Letters, 8, 539545.
Faustino, H., Gil, N., Baptista, C., & Duarte, A. P. (2010). Antioxidant activity of lignin
phenolic compounds extracted from kraft and sulphite black liquors. Molecules,
15, 93089322.
Garca, A., Toledano, A., Andrs, M. A., & Labidi, J. (2010). Study of the antioxidant
capacity of Miscanthus sinensis lignins. Process Biochemistry, 45, 935940.
Gomes, A. P., Mano, J. F., Queiroz, J. A., & Gouveia, I. C. (2012). Layer-by-layer deposition of antibacterial polyelectrolytes on cotton bres. Journal of Polymer and the
Environment, 20, 10841094.
Gomes, A. P., Mano, J. F., Queiroz, J. A., & Gouveia, I. C. (2013). Layer-by-layer deposition of antimicrobial polymers on cellulosic bers: A new strategy to develop
bioactive textiles. Polymers for Advanced Technologies, 24, 10051010.
Gustafsson, E., Larsson, P. A., & Wgberg, L. (2012). Treatment of cellulose bres with
polyelectrolytes and wax colloids to create tailored highly hydrophobic brous
networks. Colloids and Surfaces, A: Physicochemical and Engineering Aspects, 414,
415421.
Gustafsson, J., Ciovica, L., & Peltonen, J. (2003). The ultrastructure of spruce kraft
pulps studied by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). Polymer, 44, 661670.
Hu, L. B., Zheng, G. Y., Yao, J., Liu, N., Weil, B., Eskilsson, M., et al. (2013). Transparent
and conductive paper from nanocellulose bers. Energy & Environmental Science,
6, 513518.
Iamphaojeen, Y., & Siriphannon, P. (2012). Immobilization of zinc oxide nanoparticles on cotton fabrics using poly 4-styrenesulfonic acid polyelectrolyte.
International Journal of Materials Research, 103, 643647.
Iiiergrd, J., Rmling, U., Wgberg, L., & Ek, M. (2013). Tailoring the effect of
antibacterial polyelectrolyte multilayers by choice of cellulosic ber substrate.
Holzforschung, 67, 573578.
Johansson, L. S., Campbell, J. M., Koljonen, K., & Stenius, P. (1999). Evaluation of
surface lignin on cellulose bers with XPS. Applied Surface Science, 144145,
9295.
Joseph, N., Ahmadiannamini, P., Hoogenboom, R., & Vankelecom, I. F. J. (2014). Layerby-layer preparation of polyelectrolyte multilayer membranes for separation.
Polymer Chemistry, 5, 18171831.
Joshi, M., Khanna, R., Shekhar, R., & Jha, K. (2011). Chitosan nanocoating on cotton
textile substrate using layer-by-layer self-assembly technique. Journal of Applied
Polymer Science, 119, 27932799.
Kalia, S., Thakur, K., Celli, A., Kiechel, M. A., & Schauer, C. L. (2013). Surface modication of plant bers using environment friendly methods for their application
in polymer composites, textile industry and antimicrobial activities: A review.
Journal of Environmental Chemical Engineering, 1, 97112.
Kang, S. M., Li, B., Chang, J., & Fan, J. (2011). Antioxidant abilities comparison of lignins
with their hydrothermal liquefaction products. BioResources, 6, 243252.
Kenny, O., Smyth, T. J., Walsh, D., Kelleher, C. T., Hewage, C. M., & Brunton, N. P. (2014).
Investigating the potential of under-utilised plants from the asteraceae family
as a source of natural antimicrobial and antioxidant extracts. Food Chemistry,
161, 7986.

41

Kim, S., Fernandes, M. M., Matam, T., Loureiro, A., Gomes, A. C., & Cavaco-Paulo,
A. (2013). Chitosan-lignosulfonates sono-chemically prepared nanoparticles:
Characterization and potential applications. Colloids and Surface, B: Biointerfaces,
103, 18.
Laufer, G., Kirkland, C., Morgan, A. B., & Crunlan, C. (2012). Intumescent multilayer
nanocoating, made with renewable polyelectrolytes, for ame-retardant cotton.
Biomacromolecules, 13, 28432848.
Li, F., Biagioni, P., Finazzi, M., Tavazzi, S., & Piergiovanni, L. (2013). Tunable green
oxygen barrier through layer-by-layer self-assembly of chitosan and cellulose
nanocrystals. Carbohydrate Polymers, 92, 21282134.
Li, H., Fu, S. Y., & Peng, L. C. (2013). Surface modication of cellulose bers
by layer-by-layer self-assembly of lignosulfonates and TiO2 nanoparticles:
Effect on photocatalytic abilities and paper properties. Fibers and Polymers, 14,
17941802.
Li, H., Li, H., Fu, S. Y., & Zhan, H. Y. (2011). Surface hydrophobicity modication of
cellulose bers by layer-by-layer self-assembly of lignosulfonates. BioResources,
6, 16811695.
Liang, F. B., Song, Y. L., Huang, C. P., Zhang, J., & Chen, B. H. (2013). Adsorption of hexavalent chromium on a lignin-based res equilibrium thermodynamics kinetics.
Journal of Environmental Chemical Engineering, 1, 13011308.
Liu, H., Fu, S. Y., Zhu, J. Y., Li, H., & Zhan, H. Y. (2009). Visualization of enzymatic
hydrolysis of cellulose using AFM phase imaging. Enzyme and Microbial Technology, 45, 274281.
Lora, J. H., & Glasser, W. G. (2002). Recent industrial applications of lignin: A sustainable alternative to nonrenewable materials. Journal of Polymers and Environment,
10, 3948.
Lu, Z. H., Eadula, S., Zheng, Z. G., Xu, K. R., Grozdits, G., & Lvov, Y. (2007). Layer-bylayer nanoparticle coatings on lignocelluloses wood microbers. Colloids and
Surfaces A: Physicochemical and Engineering Aspects, 292, 5662.
Martins, N. C. T., Freire, C. S. R., Pinto, R. J. B., Fernandes, S. C. M., Pascoal, N. C., Silvestre,
A. J. D., et al. (2012). Electrostatic assembly of Ag nanoparticles onto nanobrillated cellulose for antibacterial paper products. Cellulose, 19, 14251436.
Marais, A., Utsel, S., Gustafsson, E., & Wgberg, L. (2014). Towards a superstrainable paper using the layer-by-layer technique. Carbohydrate Polymers, 100,
218224.
Nyman, V., Rose, G., & Ralston, J. (1986). The colloidal behaviour of kraft lignin and
lignosulfonates. Colloids and Surfaces, 21, 125147.
Pan, X., Kadla, J. F., Ehara, K., Gilkes, N., & Saddler, J. N. (2006). Organosolv ethanol
lignin from hybrid poplar as a radical scavenger: Relationship between lignin
structure, extraction conditions, and antioxidant activity. Journal of Agricultural
and Food Chemistry, 54, 58065813.
Pouteau, C., Dole, P., Cathala, B., Averous, L., & Boquillon, N. (2003). Antioxidant
properties of lignin in polypropylene. Polymer Degradation and Stability, 81,
918.
Qian, L., Guan, Y., Ziaee, Z., He, B. H., Zheng, A., & Xiao, H. N. (2009). Rendering cellulose bers antimicrobial using cationic -cyclodextrin-based polymers included
with antibiotics. Cellulose, 16, 309317.
Schmidt, J. A., Rye, C. S., & Gurnagul, N. (1995). Lignin inhibits autoxidative degradation of cellulose. Polymer Degradation and Stability, 49, 291297.
Sihtola, H., Kyrklund, B., Laamanen, L., & Palenius, I. (1963). Comparison and conversion of viscosity and DP-value determined by different method. Paperi Ja Puu,
45, 225232.
Silva, C., Matam, T., Kim, S., Padro, J., Prasetyo, E. N., Kudanga, T., et al. (2011).
Antimicrobial and antioxidant linen via laccase-assisted grafting. Reactive and
Functional Polymers, 71, 713720.
Suginta, W., Khunkaewla, P., & Schulte, A. (2013). Electrochemical biosensor application of polysaccharides chitin and chitosan. Chemical Reviews, 113, 54585479.
Szostak-Kotowa, J. (2004). Biodeterioration of textiles. International Biodeterioration
& Biodegradation, 53, 165170.
Trufanova, M. V., Parfenova, L. N., & Yarygina, O. N. (2010). Surfactant properties of
lignosulfonates. Russian Journal of Applied Chemistry, 83, 10961098.
Ugartondo, V., Mitjans, M., & Vinardell, M. P. (2008). Comparative antioxidant and
cytotoxic effects of lignins from different sources. Bioresource Technology, 99,
66836687.
Ugartondo, V., Mitjans, M., & Vinardell, M. P. (2009). Applicability of lignins from
different sources as antioxidants based on the protective effects on lipid
peroxidation induced by oxygen radicals. Industrial Crops and Products, 30,
184187.
Vartiainen, J., Motion, R., Kulonen, H., Rtt, M., Skytt, E., & Ahvenainen, R. (2004).
Chitosan-coated paper: Effects of nisin and different acids on the antimicrobial
activity. Journal of Applied Polymer Science, 94, 986993.
Vinardell, M. P., Ugartondo, V., & Mitjans, M. (2008). Potential application of
antioxidant lignins from different sources. Industrial Crops and Products, 27,
220223.
Vuoti, S., Laatikainen, E., Heikkinen, H., Johansson, L. S., Saharinen, E., & Retulainen,
E. (2013). Chemical modication of cellulosic bers for better convertibility in
packaging application. Carbohydrate Polymers, 96, 549559.
Wgberg, L. (2012). Cellulose bers and brils as templates for the layer-bylayer (LBL) technology. In G. Decher, & J. B. Schlenoff (Eds.), Multilayer thin
lms: Sequential assembly of nanocomposite materials (pp. 171187). Weinheim:
Wiley-VCH Verlag GmbH & Co. KGaA.
Xing, Q., Eadula, S., & Lvov, Y. (2007). Cellulose berenzyme composites fabricated
through layer-by-layer nanoassembly. Biomacromolecules, 8, 19871991.
Yang, D. J., Qiu, X. Q., Zhou, M. S., & Lou, H. M. (2007). Properties of sodium lignosulfonates as dispersant of coal water slurry. Energy Conversion and Management,
48, 24332438.

42

H. Li, L. Peng / Carbohydrate Polymers 124 (2015) 3542

Zahran, M. K., Ahmed, H. B., & EI-Rae, M. H. (2014). Surface modication of cotton
fabrics for antibacterial application by coating with AgNPsalginate composite.
Carbohydrate Polymers, 108, 145152.
Zhang, M., Wang, S. L., Wang, C. Y., & Li, J. (2012). A facile method to fabricate
superhydrophobic cotton fabrics. Applied Surface Science, 261, 561566.
Zhang, X., Shi, F., Yu, X., Liu, H., Fu, Y., Wang, Z. Q., et al. (2004). Polyelectrolyte multilayer as matrix for electrochemical desposition of gold clusters:

Toward super-hydrophobic surface. Journal of the American Chemical Society, 126,

30643065.
Zhao, Y., Tang, Y. W., Wang, X. G., & Lin, T. (2010). Superhydrophobic cotton fabric
fabricated by electrostatic assembly of silica nanoparticles and its remarkable
buoyancy. Applied Surface Science, 256, 67366742.