Cellulose (2014) 21:35733583
DOI 10.1007/s10570-014-0351-y
ORIGINAL PAPER
Cationic wood cellulose films with high strength
and bacterial anti-adhesive properties
Juho Antti Sirvio Anna-Kaisa Anttila Anna Maria Pirttila Henrikki Liimatainen
Ilkka Kilpelainen Jouko Niinimaki Osmo Hormi
Received: 2 April 2014 / Accepted: 3 July 2014 / Published online: 18 July 2014
! Springer Science+Business Media Dordrecht 2014
Abstract In this work, periodate oxidized birch
wood pulp and microfibrillated cellulose (MFC) were
cationized using Girards reagent T or aminoguani-
dine. Cationic celluloses were used to obtain films via
solvent-casting method, and the effects of the cation-
ization route and the cellulose fiber source on the
properties of the films were studied. Thermal and
optical properties of the films were measured using
differential scanning calorimetry and UVVis spec-
trometry, and the morphology of the films was
examined using an optical microscope and a field
emission scanning electron microscope. Bacterial
anti-adhesive properties of the films were also studied
Electronic supplementary material The online version of
this article (doi:10.1007/s10570-014-0351-y) contains supple-
mentary material, which is available to authorized users.
J. A. Sirvio (&) ! H. Liimatainen ! J. Niinimaki
Fiber and Particle Engineering Laboratory, University of
Oulu, P.O. Box 4300, 90014 Oulu, Finland
e-mail: juho.sirvio@oulu.fi
A.-K. Anttila ! A. M. Pirttila
Department of Biology, University of Oulu,
P.O. Box 3000, 90014 Oulu, Finland
I. Kilpelainen
Department of Chemistry, University of Helsinki,
P.O. Box 55, 00014 Helsinki, Finland
O. Hormi
Department of Chemistry, University of Oulu,
P.O. Box 3000, 90014 Oulu, Finland
using a modified leaf print method and against
Staphylococcus aureus and Escherichia coli. Both
cationizing agents exhibited similar reactivity with
periodate oxidized celluloses, however, MFC had
significantly higher reactivity compared to birch pulp.
The films with high tensile strength (39.145.3 MPa)
and modulus (3.57.3 GPa) were obtained from
cationized birch pulp, aminoguanidine modification
producing a film with slightly better mechanical
properties. Modulus of the films was significantly
increased (up to 14.0 GPa) when MFC was used as a
cellulose fiber source. Compared to the unmodified
MFC films, the cationic MFC films were less porous
and significantly more transparent; however, they had
slightly lower tensile strength values. It was found that
aminoguanidine modified celluloses had no culturable
bacteria on its surface and also exhibited resistance to
microbial degradation, whereas there were culturable
bacteria on the surface of Girards reagent modified
films and they were partially degraded by the bacteria.
Keywords Dialdehyde cellulose ! Cationic
cellulose ! Microfibrillated cellulose ! Mechanical
properties ! Bacterial anti-adhesive
Introduction
Natural resources are intensively studied for replacing
oil-based products. Due to their renewability, bio-
compatibility, biodegradability, wide availability and
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3574
non-toxicity, natural materials are considered as
sustainable alternatives to obtain chemicals and
materials compared to nonrenewable sources. One
important group of natural materials is biopolymers,
such as polysaccharides, among which cellulose,
hemicellulose and starch are extensively investigated,
in particular.
Cellulose is the most abundant natural polymer and
thus one of the most potential resources to replace oil-
based products (Kamide 2005). Cellulose is a linear
polysaccharide consisting of b-1,4-glycosidic linked
D-glucose [the repeating unit is called the anhydro-
glucose unit (AGU)]. Every AGU has three reactive
hydroxyl groups which allow variety modification of
cellulose. However, the hydroxyl groups of cellulose
are connected via tight hydrogen bond network due to
which cellulose has a limited reactivity and solubility
compared to other similar polymers such as starch.
Despite the low reactivity, there is great diversity of
the chemical modifications that can be conducted on
cellulose to improve or change its properties. Perio-
date oxidation is a way to introduce highly reactive
aldehyde groups on cellulose backbone (Jackson and
Hudson 1937). These aldehyde groups can be further
derivatized using various reaction routes (Crescenzi
et al. 1984; Maekawa and Koshijima 1991; Hou et al.
2007). Even though periodate is toxic and expensive
chemical, it can be recycled after oxidation (Hearon
et al. 1978; Liimatainen et al. 2013). Furthermore,
periodate oxidation and following modifications can
be conducted in a mild aqueous environment, making
it an environmentally feasible cellulose modification
route.
So far, periodate oxidized cellulose has been
utilized to obtain materials to be used in applications
such as water treatment (Liimatainen et al. 2011a,
2012a), chromatography (Kim and Kuga 2001a), a
temporary paper wet strength agent in a paper making
(van Brussel-Verraest et al. 2003) and a production of
nanocellulose (Liimatainen et al. 2012b; Tejado et al.
2012). Previously, we have also shown that sustain-
able packaging materials can be obtained from anionic
wood cellulose pulp (Sirvio et al. 2013a). However,
there is no reports of the preparation of cellulose-based
film materials through cationization of different per-
iodate oxidized cellulose materials. In this work, two
wood cellulose materials, birch pulp and commercial
microfibrillated cellulose (MFC), were cationized by
using Girards reagent T or aminoguanidine after
123
Cellulose (2014) 21:35733583
periodate oxidation of cellulose fibers. The cellulose
films were made by solvent-casting method and the
effects of the cationization agent and fiber source on
the film morphology and optical, thermal, bacterial
anti-adhesive properties, and mechanical properties
were studied.
Materials and methods
Raw material and chemicals
Bleached birch cellulose pulp (Betula pendula) was
obtained as dry sheets and used as a fiber material after
disintegration in deionized water. Properties of cellu-
lose are presented elsewhere (Liimatainen et al.
2011b). The commercial grade microfibrillated wood
cellulose (MFC, KY-100G) was obtained from Daicel
Industries Ltd. (Japan).
All of the chemicals used in the periodate oxidation
and in the further derivatizations (NaIO4, [(CH3)3-
N?CH2CONHNH2]Cl-, NH2NHC(NH)NH2!HCl), and
in the analysis of the aldehyde content (NH2OH!HCl,
CH3COONa!2H2O) were obtained as p.a. grade from
SigmaAldrich (Germany) and were used without
further purification. Deionized water was used through-
out the experiments.
Chemical modification of birch cellulose
Dialdehyde celluloses were obtained from birch
cellulose (DAC1) and MFC (DAC2) by the method
described earlier (Sirvio et al. 2011). In brief, 9 g of
cellulose was oxidized using 7.38 g of sodium perio-
date at 55 "C for 3 h in the absence of light. After
filtration and washing with deionized water, product
was stored at 4 "C in a non-dried state. Yields of the
DAC1 and DAC2 were 82 and 78 %, respectively.
Cationizations were conducted by similar method
described previously (Sirvio et al. 2011a). Girards
reagent T or aminoguanidine hydrochloride was dis-
solved to 200 ml of deionized water and the pH of the
solution was adjusted to 4.5 using a dilute HCl solution.
2 g of DAC1 or DAC2 was added and reaction was
allowed to proceed for 3 days under a magnetic stirring
at room temperature. Products from DAC1 was filtrated
and washed with deionized water. Products from DAC2
were purified using centrifugation at 7,500 g. After
removal of decant, product was washed with deionized
Cellulose (2014) 21:35733583
water and centrifuged again. Washing was repeated two
times. All of the products were stored at 4 "C in a non-
dried state. Products from Girards reagent T modifi-
cation of DAC1 and DAC2 were designed as GC1 and
GC2, respectively, whereas products from aminogua-
nidine modification of DAC1 and DAC2 were designed
as AGC1 and AGC2, respectively.
The aldehyde contents of the oxidized celluloses
were determined by an oxime reaction as reported
previously (Sirvio et al. 2011) and cationic group
content of modified celluloses were determined using an
elemental analyzer (PerkinElmer CHNS/O 2400 Series
II) by analyzing nitrogen content of the dried sample.
Preparation of wood cellulose films
The cellulose-based films were prepared by a solvent-
casting method. Cationic celluloses were dispersed into
water using a magnetic stirrer and 10 % of glycerol
(based on cellulose mass) was added as a plasticizer. The
dispersions were stirred using an UltraTurrax (Ger-
many) disperser (11,000 rpm) for 15 min to obtain
homogeneous dispersions, degassed, cast onto the
polystyrene tray and allowed to dry for 3 days at room
temperature to obtain films with a grammage of 30
g/m2. Films of GC1, GC2, AGC1 and AGC2 were
designated as GC1F, GC2F, AGC1F and AGC2F,
respectively. An unmodified MFC film, which was
fabricated in similar way, was used as a reference. The
surfaces of the films faced toward the tray and toward
the air during the drying process were designated as the
bottom and top sides, respectively. The visual appear-
ances of the films were studied using an optical
microscope.
Characterization of wood cellulose films
Field emission scanning electron microscope
A FESEM (Zeiss Ultra Plus, Carl Zeiss SMT AG,
Germany) was used to examine the morphologies of
the film surfaces. The films were sputter-coated with
carbon, and an accelerating voltage of 15 kV was used
during imaging.
Fourier transfer infrared spectrum
FTIR spectra of films were recorded using a Bruker
FT-IR spectrometer (USA) using spectral width
3575
ranging from 4,000 to 400 cm-1 with 2 cm-1 resolu-
tion and an accumulation of 32 scans. Prior the FTIR
measurement, films were compressed using mechan-
ical press to obtain sufficient film thickness.
Transparency of the anionic cellulose films
The transparency of the cellulose films was deter-
mined using a Shimadzu UVVis spectrometer
(Japan). A slide of each film was placed in the cuvette
stand so that the beam was oriented 90" with respect to
the slide.
X-ray diffraction
For X-ray diffraction (XRD) analyses, the samples
were analyzed between 2h = 5" and 114" with an
angle step size of 2h = 0.02" on a D8 Discover X-ray
diffractometer (Bruker AXS Inc., Madison, MI, USA)
equipped with a Co Ka (40 kV/35 mA) source. The
degree of crystallinity of the cellulose films in terms of
the crystallinity index (CrI) was calculated from the
peak intensity of the main crystalline plane (200)
diffraction (I200) at 22.8" and from the peak intensity at
18.0" associated with the amorphous fraction of
cellulose (Iam) according to following equation:
I200 # Iam
CrI & 100 %
I200
Differential scanning calorimetry
Differential scanning calorimetry (DSC) was used to
study the thermal properties of the cellulose films
(DSC823 Mettler Toledo Schwerzenbach, Switzer-
land). 2.2 g of the sample contained in aluminum pan
was heated from 25 to 400 "C with a heating rate of
10 "C/min under N2 gas at a flow rate of 50 ml/min.
Mechanical properties
The films were cut into 15 mm 9 150 mm stripes and
were conditioned at 23 "C and 50 % humidity for 48 h
before being tested. The specific tensile strength (r/q),
strain and specific modulus (E/q) were determined
according to standard ISO 527-3 using a Lorentzen
and Wettre tensile strength tester (Sweden). A test
speed of 50 mm/min was used. The initial distance
between grips was set to 100 mm. Each test was
replicated five times and the results are presented as
123
3576
averages of these measurements. The tensile strength
(r) and modulus (E) were calculated from r/q and E/
q, respectively, where q represents the density.
The film thicknesses were measured at five random
positions on the film using a Lorentzen and Wettre
thickness meter (Sweden); the reported results repre-
sent the average of the five measurements.
Bacterial anti-adhesive properties and resistance
to degradation
The films ability to support microbial growth on the
surface was tested using a modified leaf print method
(Hirano and Upper 1983). Briefly, a 2.5 cm2 piece of
film was pressed for 1015 s on LuriaBertani (LB)
agar medium, removed, and placed to another position
on the agar medium. The cultures were grown
overnight at 22 and 37 "C, and bacterial colonies
were counted on the culture plates.
Bacterial anti-adhesive properties and resistance
against bacterial degradation were tested on cultures
of Staphylococcus aureus and Escherichia coli.
Overnight cultures of S. aureus and E. coli were
grown in 5 ml of liquid LB at 37 "C and 100 ll of the
overnight culture was spread on LB agar medium. The
film was placed over the bacterial cells on the LB
medium plate and the culture was grown overnight at
37 "C. The presence of bacterial growth around, under
and on top of the film was analyzed visually and
microscopically.
Results and discussion
Preparation of wood cellulose films
To obtain the cationic celluloses, birch pulp and
commercial MFC were both periodate oxidized in
identical conditions. The aldehyde content of DAC1
and DAC2 were calculated to be 1.68 and 1.97 mmol/
g, respectively. We have previously shown that one
way to improve the efficiency of the periodate
oxidation is to reduce the particles size of cellulose
(Sirvio et al. 2011b). Compared to birch pulp cellu-
lose, MFC consisted of smaller fibers (FESEM images
of both fibers is presented elsewhere (Sirvio et al.
2013b) and thus had a higher reactive surface area.
Consequently, MFC was oxidized to higher aldehyde
content than birch pulp.
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Cellulose (2014) 21:35733583
Both DACs were further derivatized with amino-
guanidine (AGC1 and AGC2) or Girards reagent T
(GC1 and GC2), resulting in formation of stable imine
structures due to the delocalization of the imine double
bond making the carbon atom of the imine double
bond less susceptible to nucleophilic attack (Clayden
et al. 2001). Reaction routes are presented in the
Scheme 1. Both reaction routes leads to the formation
of cellulose with a cationic charge as Girards reagent
T bears a tertiary amine group and aminoguanidine a
highly basic guanidine group. As it can be seen in
Table 1, both reagents showed similar reactively with
DACs, however, there is a significant difference
between the reactivity of DAC1 and DAC2. When
DAC1 was used as a starting material, approximately
half of the aldehyde groups were reacted with
cationization agents, whereas about 70 % of aldehyde
groups in DAC2 were converted to corresponding
imine groups during the cationizations. Again, the
better reactively of MFC based DAC was most likely
due to the smaller particle size and thus due to more
accessible reaction sites. Yields of the cationic cellu-
loses varied from 73 to 99 % (Table 1). No clear
connection between obtained yields and cationization
agent or cellulose source used was observed. Yield of
the Girards reagent T modified cellulose was same
regardless of the used cellulose source, whereas
significantly higher yield was obtained using MFC
instead of birch pulp when aminoguanidine was used
as a cationization agent.
All of the cationic celluloses formed strong self-
standing films via solvent-casting method. On the
other hand, no intact films were obtained from
unmodified celluloses as they were uneven and got
stuck on the tray. However, small slides were obtained
from the MFC films and were used as a reference.
Figure 1 presents the optical microscope images of
the cellulose films. It can be seen that there is no
significant differences between aminoguanidine and
Girards reagent T modified cellulose films. However,
the films produced from MFC had clearly less intact
fibers on their surfaces. Also, the lengths of the visible
fibers were significantly shorter in AGC2 and GC2
compared to AGC1 and GC1. This agrees well with
the fiber size of the starting materials.
Optical properties of the cellulose films were
studied using a UVVis spectrometer and spectra are
shown in Fig. 2. There were slight differences in the
optical properties of the films produced by different
Cellulose (2014) 21:35733583 3577

OH
-
Cl
* O O
*
+ N
H N N
N
O NHHN
H N O
2 H + O -
- Cl
H Cl + +
OH t. N N
OH
a) ca
NaIO4
* O O * O O H
HO * * H
2 H
OH O O N N
+
b) H N OH
H
H2N -
* O O
ca Cl *
t. +
H N
N H
H NH HN
+ + -
- N N Cl
Cl H
H NH2 NH2

Scheme 1 Periodate oxidation of cellulose and synthesis of cationic celluloses using a Girards reagent T and b aminoguanidine
hydrochloride

Table 1 The charge Sample DAC Aldehyde content Yield Charge density Conversion
densities of cationized (mmol/g) (%) (mmol/g) (%)
celluloses, aldehyde group
content of starting DAC, AGC1 DAC1 1.68 73 0.76 49
yields, and the conversion
GC1 DAC1 1.68 80 0.76 51
of aldehydes to
corresponding imine groups AGC2 DAC2 1.97 99 1.19 68
GC2 DAC2 1.97 80 1.16 71

Fig. 1 Optical microscope images of cellulose films: a AGC1F, b GC1F, c AGC2F, d GC2F and e unmodified MFC

123
3578 Cellulose (2014) 21:35733583

AGC2F 1,567.32 cm-1 (Sirvio et al. 2011a). Additionally,

40
sharp bands of carbonyl and nitrogennitrogen bonds
35 GC1F were observed at 1,693.49 and 924.81 cm-1, respec-
30 GC2F tively. In the spectrum of AGC1F, the characteristic
Transmittance (%)

25
band of carbonnitrogen double bond vibration and
AGC1F nitrogenhydrogen bond bending were observed at
20
1,679.55 and 1,643.59 cm-1, respectively (Zhang
15 et al. 1999). Also the carbonnitrogen single bond
10 vibration band was observed at 1,318.13 cm-1.
5
Same bands were observed in spectra of the films
MFC prepared from MFC (spectra not shown), with a
0
300 400 500 600 700 800 slightly higher intensity due to the better reactivity of
Wavelength (nm) DAC2. FTIR spectra confirmed a successfully chem-
ical modification of DAC with both of the reagents.
Fig. 2 UVVis spectra of the cellulose films
Field emission scanning electron microscope

reagent as the AGC1F and GC1F had transmittances
of 21 and 32 % at 600 nm. When MFC was used as a
starting material, the transmittance of aminoguanidine
modified film increased to 40 %. However, the
transmittance of GC2F was slightly lower than that
of GC1F. Compared to the cationic cellulose films, the
unmodified MFC film was opaque. Thus, the cationic
modification significantly improved optical properties
of the cellulose films.
Fourier transfer infrared spectrum
FTIR spectra of the cellulose films prepared from
birch cellulose are presented in Fig. 3. FTIR spectra of
unmodified cellulose and DAC are presented in our
previous study (Sirvio et al. 2011a). In the spectrum of
GC1F, the characteristic band of imine bond between
Girards reagent T and DAC was observed at
Effect of the chemical modifications and the cellulose
sources on the morphology of the cellulose films was
studied using FESEM (Fig. 4). Images with higher
magnifications can be seen Online Resource 1. When
birch cellulose was used as a starting material, the top
side of the film produced via aminoguanidine modi-
fication was rougher and higher amount of intact
cellulose fibers were visible compared to film obtained
via Girards reagent T modification. In addition, some
pores were observed at the bottom side of AGC1F,
whereas the bottom side of GC1F was very even and
significantly less porous, indicating that Girards
reagent T modification of birch cellulose results in
formation more compact film.
When MFC was used as a starting material,
opposite effect was observed. The top side of the
AGC2F had a fewer amount of cellulose fibers visible
on its surface than GC2F. On the surface of the GC2F,

Fig. 3 FTIR spectra of GC1F (a) and AGC1F (b)

123
Cellulose (2014) 21:35733583
Fig. 4 FESEM images of the top side of a AGC1F, b GC1F, e AGC2F, f GC2F and g MFC, and the bottom side of c AGC1F, d GC1F
and h MFC
few very long cellulose fibers were observed as seen in
the middle of the Fig. 4f. AGC2F had very smooth
bottom side, whereas there were few small pores on
the bottom side of GC2F (Online Resource 1).
As it was already observed with optical microscope,
there is a large difference on the upper side of the films
produced from birch cellulose and MFC. Whereas the
upper side of the AGC1F consisted of web-like
structure of intact cellulose fibers, with fines filling
the voids between fibers, the upper side of the AGC2F
was mainly flat and only few intact fibers are observed
on the surface.
It was observed that the intact cellulose fibers are
more visible on the films produced by cationization of
MFC compared to the unmodified MFC film. How-
ever, the surfaces of the AGC2F and GC2F were more
even around the fibers than that of the MFC film. Also,
there were visible pores on the bottom side of the MFC
film, whereas especially the bottom side of the AGC2F
was very smooth and even. Possible reason for these
3579
observations is that cationization of MFC leads to
spontaneous fibrillation of the smaller fibers of MFC
in some extent, thus producing lots of small cellulose
particles. These small fibers produce the films with the
smooth and even surface. However, the bigger parti-
cles may not be fibrillated as effectively as small fibers
and are thus easily visible on the surface of the cationic
MFC films.
Wide-angle X-ray diffraction
X-ray diffractograms of the cellulose films are
presented in Fig. 5. All of the films showed charac-
teristic peaks of cellulose I with the main 2h diffrac-
tion angles close to 14.8", 16.8", and 22.8" associated
with the 110, 110 and 200 crystalline planes,
respectively (French 2014). The chemical treatment
wasnt observed to have high impact on the crystal-
linity of cellulose as AGC1F and GC1F had CrI of 56.6
and 58.9 %, respectively, being close to the initial CrI
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3580
MFC
GC2F
Intensity
AGC2F
GC1F
AGC1F
BRICH
20 40
2
Fig. 5 X-ray diffractograms of wood cellulose films
of the birch cellulose (56.6 %). Slight increase in the
crystallinity of GC1F might be due to the dissolution
of the amorphous parts of cellulose during modifica-
tion. The MFC film had the crystallinity of 74.1 %
which was slightly decreased after cationization with
Girards reagent T to 69.0 %. On the other hand, CrI of
the MFC was unchanged after aminoguanidine mod-
ification. It can be concluded that the both reaction
routes keeps the crystalline structure of the starting
cellulose well intact.
Differential scanning calorimetry
The results of the DSC analysis of the cellulose films
are presented in Fig. 6. In the curve of the birch pulp
and MFC films, broad endothermic peaks appeared at
354 and 351 "C, respectively, followed by the exo-
thermic peaks at 370 "C. Endothermic peaks are
mainly due to the formation and evaporation of
levoglucosan, formed as a depolymerization product
of cellulose (Soares et al. 1995). Exothermic peaks are
due to the char formation. After derivatizations, both
endothermic and exothermic peaks appeared at lower
temperature, e.g. endothermic peaks of the Girards
reagent T modified birch pulp and MFC films were
observed at 259 and 254 "C, respectively. Compared
to Girards reagent T modification, aminoguanidine
modified showed endothermic peaks at about 20 "C
lower temperature, indicating slightly lower thermal
stability of AGC1F and AGC2F than GC1F and GC1F.
Chemical modification of cellulose is known to
modify thermal properties of cellulose and thermal
123
Cellulose (2014) 21:35733583
Fig. 6 DSC curves of the cellulose films
degradation temperature is usually observed to
decrease (Kim and Kuga 2001b). Same phenomenon
was also observed in the present study. Decrease of the
crystallinity and consequently the increase of the
amorphous region of the cellulose are known to favor
the dehydration decomposition pathway of the cellu-
lose which take place by the chain scission and char
formation (Han et al. 2010). The dehydration decom-
position pathway is known to take place at lower
temperature than formation of levoglucosan. How-
ever, as it was observed in the present XRD study, all
the modified celluloses showed similar crystallinity as
their starting materials and thus the lower degradation
temperature is most unlikely due to the dehydration. It
is possible that the cationization of cellulose decreases
the polymerization degree, lowering the thermal
energy request to product the levoglucosan, which
was observed as a decreased degradation temperature
(Vicini et al. 2004).
Mechanical properties of wood cellulose films
Thicknesses and the mechanical properties of the
cellulose films are presented in Table 2. As already
observed in FESEM images, the cationization of birch
cellulose with Girards reagent T results in the
formation of the more dense film compared to
aminoguanidine. Therefore, AGC1F was 34 % thicker
than GC1F. The opposite effect of the chemical
treatment on MFC can also be seen, AGC2F being
slightly thinner compared to GC2F. The MFC film was
observed to be thickest of the films, indicating that
cationization of celluloses results in formation of more
compact films. This is consistent with FESEM studies
Cellulose (2014) 21:35733583 3581

Table 2 Thicknesses and the mechanical properties of the cellulose films

Film Thickness (lm) r/q (kNm/kg) r (MPa) Strain (%) E/q (MNm/kg) E (GPa) Density (g/cm3)

AGC1F 29.0 0.0 43.8 3.0 45.3 3.1 5.3 0.5 7.0 2.9 7.3 3.0 1.0 0.0
GC1F 21.6 2.7 28.7 2.7 39.1 3.6 4.4 0.4 2.6 0.2 3.5 0.2 1.4 0.1
MFC 32.6 1.0 87.1 3.9 79.2 3.5 4.4 0.5 13.2 0.6 12.0 0.5 0.9 0.1
AGC2F 22.2 0.7 40.6 3.3 55.4 4.5 3.5 0.6 10.2 0.4 14.0 0.5 1.4 0.0
GC1F 25.4 1.4 44.2 4.6 53.1 5.5 3.3 1.1 10.1 0.3 12.1 0.4 1.2 0.1

Fig. 7 The test of anti-adhesive properties of AGC2F a and lost strength after overnight culture as the film became torn
GC2F b on cultures of Escherichia coli. The bacterial culture when lifted from the agar with forceps
was growing around, but not under or on top of the films. GC2F

in section Field emission scanning electron
microscope.
Results also show that the two chemical modifica-
tion routes used in this study had different effect on the
birch pulp as AGC1F has 53, 169 and 109 % higher r/
q, E/q and E values than to GC1F, r values of being
statistically same. However, when MFC was used as a
starting material, both reaction routes resulted in the
formation of films with statistically same r/q, r and
E/q. Interestingly, AGC2F had 16 % higher E value
than GC2F.
Compared to birch pulp, MFC produced films with
higher mechanical strength and modulus. Especially,
moduli of MFC based films were significantly higher,
e.g. GC2F had 246 % higher E value than GC1F.
However, it was observed that cationization of MFC
decreased the mechanical properties of films, e.g. r
value of the unmodified MFC films was 43 and 49 %
higher than those of AGC2F and GC2F, respectively.
Only the E value of AGC2F was increased 16 % during
the cationization. It is known that periodate oxidation
can lead to degradation of cellulose fibers (Liu et al.
2012), which might result in decrease of mechanical
properties of cationized MFC films compared to
unmodified MFC film. It is also possible that high
cationic group content (over 1 mmol/g) of AGC2F and
GC2F creates electric repulsion between cellulose
fibers, leading to decreased mechanical properties. All
of the films exhibited similar elongation at break and
strain values of the films varied from 3.3 to 5.3 %,
AGC1F having the highest and GC1F having the lowest
strain. Strain values of the cationic MFC films were
slightly lower compared to the cationic birch pulp films,
e.g. AGC2F has 66 % lower strain than that of AGC2F.
We have previously shown that anionic birch pulp
can be used to obtain sustainable film materials with r
and E values comparable with those of synthetic
polymers such as polyethylene and poly(vinyl chloride)
(Sirvio et al. 2013a). The strength values obtained with
cationized birch pulp films were similar, thus materials
in this study are also comparable with materials
obtained from oil-based resources. Additionally, the

123
3582
cationic MFC films had even higher tensile strength and
significantly higher moduli than those of produced from
birch pulp.
Bacterial anti-adhesive properties of wood
cellulose films
The leaf print test on SOC agar medium demonstrated
that the GC1F and GC2F films can support bacterial
growth of 8 and 12 CFU/mm2 on the surface, respec-
tively. The AGC1F and AGC2F films produced no
bacterial colonies, i.e. the surface of these films
contained no culturable bacteria. When the films were
subjected to cultures of E. coli and S. aureus, bacteria
were growing around the films on the LB agar
medium, but not on top or under the films (AGC2F
is shown in Fig. 7a). The AGC1F and AGC2F retained
the structure when subjected to microbial degradation
on LB agar medium, but GC1F and GC2F films
became partially degraded by the bacteria, as the film
material lost strength after overnight culture (GC2F is
shown in Fig. 7b). This was seen as loss of the film
structure when the film was lifted from the medium
with forceps. These results suggest that the amino-
guanidine treatment increases the durability of the
wood cellulose film to bacterial degradation.
Conclusions
The cationic cellulose films were produced from birch
pulp and commercial MFC using sequential periodate
oxidation and cationization by Girards reagent T or
aminoguanidine. Both cationization routes produced
smooth self-standing films with good mechanical prop-
erties. In addition, aminoguanidine treatment resulted in
films possessing bacterial anti-adhesive properties and
resistance to microbial degradation. The films produced
from MFC were significantly stronger than those
produced from birch pulp. Even though, the unmodified
MFC film had a slightly higher tensile strength than
cationized films, the cationic films exhibited signifi-
cantly higher transparency. This work demonstrated that
Girards reagent T and aminoguanidine can be used to
obtain films with good mechanical properties, moderate
transparency and bacterial anti-adhesive properties.
Acknowledgments This work was carried out as part of the
Future Biorefinery Program of Finnish Bioeconomy Cluster
123
Cellulose (2014) 21:35733583
FIBIC Ltd. We would like to thank VTT Espoo for their pulp
analysis. Dr. Juha Heiskanen is gratefully acknowledged for his
kind assistance in the DSC analyses.
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