Materials Science and Engineering C 32 (2012) 2508–2515

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Materials Science and Engineering C

journal homepage: www.elsevier.com/locate/msec

Synthesis and characterization of polyvinyl alcohol/cellulose cryogels and their

testing as carriers for a bioactive component
Oana Maria Păduraru, Diana Ciolacu, Raluca Nicoleta Darie, Cornelia Vasile ⁎
Romanian Academy, “Petru Poni” Institute of Macromolecular Chemistry, 41A, Grigore Ghica Voda Alley, 700487, Iasi, Romania

a r t i c l e i n f o a b s t r a c t

Article history: Novel physically cross-linked cryogels containing polyvinyl alcohol (PVA) and various amounts of microcrys-
Received 12 April 2012 talline cellulose were obtained by freezing/thawing technique. The main goal of this study was to improve
Received in revised form 11 June 2012 the properties and the performances of the pure PVA cryogels. The morphological aspects of the cryogels
Accepted 20 July 2012
were studied by scanning electron microscopy (SEM). The Fourier transform infrared spectroscopy (FT-IR)
Available online 27 July 2012
was used to reveal the presence of the interactions between the two polymers. Changes in crystallinity of
Keywords:
the samples were confirmed by X-ray diffraction (XRD) and by FT-IR spectroscopy. The modification of the
Polyvinyl alcohol thermal behavior induced by cellulose was studied by thermogravimetry. Rheological analysis revealed
Cellulose higher values of storage modulus (G′) for the cryogels containing higher amounts of cellulose. The degree
Cryogels and rate of swelling were controlled by the presence of the natural polymer in the network. The potential
Freezing-thawing application as bioactive compound carriers was tested, using vanillin as an active agent.
Bioactive compound © 2012 Elsevier B.V. All rights reserved.
Release

1. Introduction
Hydrogels are hydrophilic three-dimensional networks of poly-
mer chains which are capable of absorbing large amounts of water
or biological fluids [1]. Due to their properties, hydrogels have nu-
merous applications, including contact lenses, membranes for biosen-
sors, carriers for drug/aroma, in tissue engineering as scaffolds for
regenerating tissues and organs, as materials for artificial skin, as
cell carriers etc. [2–7]. Cast into films and dried, hydrogels are now
being used as biodegradable packaging materials for food, cosmetic
and pharmaceutical products [8]. Cryogels refer to hydrogels formed
at subzero temperature which provide special properties for different
bioengineering and biotechnological applications [9].
Polyvinyl alcohol (PVA) is a water-soluble, non-toxic, biodegrad-
able, biocompatible synthetic polymer with excellent film forming
properties [10]. Due to its hydroxyl groups present in each repeating
unit, PVA exhibits a strong hydrophilic and hydrogen bonding charac-
ter; thus it is able to form crosslinked hydrogels. Physical hydrogels
(cryogels) can be obtained by exposing PVA aqueous solutions to re-
peated cycles of freezing and thawing, which results in the formation
of crystallites. The main advantage of this technique is that no addi-
tional chemical crosslinker is being used. Comparing to chemically
⁎ Corresponding author at: Romanian Academy, “P. Poni” Institute of Macromolecu-
lar Chemistry, 41A Grigore Ghica Voda Alley, 700487, Iasi, Romania. Tel.: +40 0232
217454; fax: +40 0232 211299.
E-mail address: cvasile@icmpp.ro (C. Vasile).
crosslinked hydrogels, cryogels show higher elasticity and have in-
creased strength due to the crystalline regions which are capable of
better distributing a given mechanical load or stress [11,12].
PVA cryogels have been studied as controlled release carriers, as
barrier film for food packaging, as material for the preparation of
membranes used in chemical separation, as biomaterial in tissue en-
gineering, as wound dressing materials, food packaging etc. [13–16].
Wound dressings are used to promote the wound healing and to
create appropriate conditions for this process, while protecting the
wound from infections. It is widely accepted that a moist environ-
ment is desirable to enhance the wound healing so, some hydrogels
are being used with success as wound dressings, because they assure
an appropriate level of moisture at the interface between the dressing
and the wound, have the capacity to absorb exudates, prevent the
wound desiccation, are non-adhesive, have a cooling effect giving a
relief feeling to the patient, and in the same time they can deliver
an incorporated drug to the wound [17–19].
The use of natural polymers as components of polymeric materials
is of great interest, due to the diversity of their properties, but also
from economic and ecological reasons. PVA exhibiting high polarity
and water solubility is a good candidate for blends with natural poly-
mers [20]. Cryogels based on PVA/polysaccharides are suitable as
wound dressing materials because these hydrogels are flexible, me-
chanically strong, biocompatible, can be easily replaced, can assure
a moist wound environment and a barrier against microorganisms
[21–23].
Cellulose is the most abundant renewable resource on Earth. Sim-
ilar to PVA hydrogels, cellulose based hydrogels are also hydrophilic,
biodegradable, biocompatible, non-toxic [24]. In the human and

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 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515
animal body, cellulose exhibits a relatively low protein adsorption
and cell adhesion and a low immune response. In the same time,
this polysaccharide is not degradable in the body and is not digestible
due to the fact that human and animal cells do not synthesize cellu-
lases, the enzymes capable of degrading cellulose [25]. All these prop-
erties made the cellulose hydrogels suitable for various applications,
especially in the biomedical field [26–28].
Scientific literature presented different types of celluloses incor-
porated into PVA films and hydrogels, such as bacterial cellulose
[29], cellulose whiskers [30], cellulose nanofibres [31,32], cellulose
nanocrystals [33] and cellulose fibrous powder [34]. Various methods
of crosslinking, as well as different methods of cellulose dissolution
have been used in these kinds of studies. For example, Millon et al.
[29] used freezing/thawing method for obtaining PVA/bacterial cellu-
lose cryogels, while Abitbol et al. [33] used the same crosslinking
method in order to obtain PVA hydrogels reinforced with cellulose
nanocrystals from softwood Kraft pulp. Chang et al. [34] prepared
hydrogels using fibrous cellulose dissolved in NaOH/urea aqueous so-
lution and epichlorohydrin as a crosslinking agent.
The novelty of this study refers to the obtaining of PVA cryogels
containing microcrystalline cellulose by freezing-thawing method by
using a special dissolution method for cellulose in a water/NaOH mix-
ture, at low temperatures [35]. In our study we also tested the obtained
cryogels as carriers for the delivery of an antimicrobial and antioxidant
agent such as vanillin (4-hydroxy-3-methoxybenzaldehyde). Vanillin,
one of the most widely used flavoring agent, is produced naturally in
the specialized cells from the pods of the Vanilla sp. and synthetically
from eugenol, guaiacol or lignin [36]. It is being used as a flavoring
and antioxidant agent in food industry, but also in cosmetics and phar-
maceutical formulations [37,38]. It has been reported that vanillin ex-
hibits multifunctional effects such as antimutagenic, antiangiogenetic,
anti-colitis, anti-sickling, and analgesic effects [39]. The antimicrobial
properties of vanillin and its activity have been previously demonstrat-
ed against some bacteria (Escherichia coli, Pseudomonas aeruginosa, Lac-
tobacillus plantarum, Listeria innocua), but also against some yeast and
mold strains (Candida albicans, Penicillum expansum, Saccharomyces
cerevisiae) [40,41].
It was expected that the incorporation of microcrystalline cellu-
lose into PVA cryogels lead to an improvement of the properties and
an enhancement of the performances. These cryogels were character-
ized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffrac-
tion (XRD), thermal degradation, rheological measurements and “in
vitro” tested as drug carriers. Also the kinetics of swelling in phos-
phate buffer solution (PBS) pH 7.4 at 37 °C and of drug release was
studied.
2. Experimental
2.1. Materials
PVA with an average molecular weight of 146 000–186 000 and a
hydrolysis degree of 99% was purchased from Aldrich. Microcrystal-
line cellulose Avicell PH-101 was purchased from Fluka.
2.2. PVA/cellulose cryogels preparation
A PVA solution of 8 wt% concentration was prepared by dissolving
a certain amount of PVA into double distilled water, at 90 °C under
stirring for 8 hours. A clear solution was obtained.
Different amounts of microcrystalline cellulose were dispersed in
11/1 water/NaOH (w/w) mixture, stirred for 5 min and then frozen
at low temperature (− 30 °C). After defrosting, the obtained solutions
were stirred in order to obtain clear, transparent solutions [35].
Cellulose and PVA solutions were mixed in different ratios
(Table 1) and stirred for 5 min. The obtained mixtures were poured
into Petri dishes and frozen at − 20 °C for 12 hours, then thawed at
2509
Table 1
Cryogels composition.
Sample Cryogels composition (wt%)
PVA Cellulose
PVA 100 0
90/10 90 10
70/30 70 30
50/50 50 50
room temperature (+ 25 °C) for another 12 hours. The freezing/
thawing process was repeated three times.
The blank PVA cryogel was prepared pouring a certain amount of
PVA solution in a Petri dish and then exposed to three freezing/
thawing cycles (12 hours freezing/12 hours thawing).
The PVA and the PVA/cellulose cryogels were washed with
distilled water for 3 days and were dried by lyophilization for
24 hours, using a LABCONCO 117 freezing-dryer. Polymeric films
were obtained after lyophilization.
Due to the fact that cellulose cryogel could not be obtained by this
technique, the cellulosic powder was treated in the same conditions
as described above and used as blank sample.
2.3. Investigation methods
2.3.1. Scanning electron microscopy (SEM)
Scanning electron micrographs were taken on liquid nitrogen frac-
tured samples, with a Quanta 200 instrument. The fractured surface
was coated with a gold layer. Magnification is given on the images.
2.3.2. X‐ray diffraction measurements (XRD)
The measurements were carried out by means of a Bruker AXS D8
Advance X-ray diffractometer with a Cu Kα radiation source. The data
were collected in the 2θ = 2 ÷ 40° region. The crystallinity index (CrI)
was calculated by the area method, dividing the area of the crystalline
peak by the total area under diffractogram [42].
2.3.3. Fourier transform infrared spectroscopy (FT‐IR)
The FT-IR analysis was performed on a Vertex-70 (Bruker) appara-
tus, using KBr tablets containing a constant mass of 5 mg sample/
500 mg KBr. The spectra were recorded in the 4500–600 cm −1,
with 16 scans and a resolution of 4 cm −1. Three recordings were
done for each system and the average values were used for data inter-
pretation. The energy of the hydrogen bonds (EH) was calculated
using Eq. (1) [42].
EH ðkJÞ ¼ ð1=kÞ½ðυ0 −υÞ=υ0  ð1Þ
where k is a constant equal to 1.68 × 10 −2 kcal −1, υ0 is the standard
frequency corresponding to free ―OH groups (3650 cm −1), υ is the
frequency of the bonded ―OH groups (cm −1). The enthalpy of the
hydrogen-bond formation (ΔH) was evaluated using Eq. (2) [43]:
ΔH ðJ=gÞ ¼ 0:016ΔυOH þ 0:63 ð2Þ
where ΔυOH is the ―OH wavenumber shift (cm −1). The hydrogen-
bonding distance (R, A°) was obtained using Sederholm Eq. (3) [44].
 
−1 3
Δυ cm ¼ 4:43  10 ð2:84−RÞ ð3Þ
where Δυ = υ0 − υ (υ0 is the ―OH monomeric stretching frequen-
cy = 3600 cm −1; υ is the ―OH stretching frequency of the sample).
2.3.4. Thermogravimetry
The thermogravimetric analysis was performed on a Jupiter STA
449 F1—Netzsch instrument. Samples of 7 mg were heated up to
2510 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515
600 °C, with a heating rate of 10 °C/min, in open Al2O3 crucible, under
50 ml/min nitrogen flow.
2.3.5. Rheological measurements
Rheological measurements were performed with an Anton Paar
MCR301 rheometer, using a measurement system with a plate—
plate geometry of 25 mm diameter and 0.8 mm gap. The samples
were placed between the plates for 5 min to eliminate residual
shear history and then the experiments were carried out immediate-
ly. The linear viscoelastic range was determined performing an ampli-
tude sweep test at 23 °C, a shear strain of 0.1% being kept during the
further rheological tests. The measuring device was equipped with a
temperature unit that gave good temperature control (±0.05 °C). In
order to evaluate the rheological behavior with increasing tempera-
ture, the cryogels were heated from 20 to 80 °C with a heating rate
of 2 °C/min (temperature sweep tests). In the same time an oscillat-
ing stress was exerted to the samples with a constant frequency of
1 Hz.
2.3.6. Swelling behavior
Fluid absorbance studies are of great importance for biodegrad-
able materials and for those destined to be used as carriers for bioac-
tive agents, as the swelling behavior influences the released one. For
swelling measurements, the dried cryogels were weighed (Wd) be-
fore being immersed in PBS with a pH 7.4, at 37 °C. After immersion,
at different time periods, the samples were periodically removed
from the medium, the surface was gentle pressed with filter paper
and the weight of the sample (Wt) was gravimetrically determined.
The swelling degree (SD) was calculated according to Eq. (4).
SD ¼ ðW t −W d Þ=W d  100
where Wt is the weight of the sample after swelling in water at time t
and Wd is the weight of the dried cryogel (xerogel). Three measure-
ments were performed for each sample.
In order to determine the swelling kinetic of the water diffusion
into hydrogels, Eq. (5) was used:
nSW
F SW ¼ W t =W ∞ ¼ kSW t
where Fsw represents the swelling fraction, Wt and W∞ represent the
amount of water absorbed of the cryogel at time t, respectively at
equilibrium, ksw is the swelling constant related to the structure of
the cryogel and nsw is the swelling exponent, used to determine the
mode of transport of the solute [45]. This equation is applicable for
the initial stages of swelling.
2.3.6. Vanillin loading and release
The loading of the cryogels with vanillin was made using dry
cryogels in powder form which were mixed with vanillin. A specific
amount of PBS pH 7.4 was added over these mixtures (maximum
amount of liquid uptaken during swelling) and the mixtures were
left to swell at room temperature (25 °C) for at least 1 hour while
the entire amount of vanillin used, penetrates or attach to the matri-
ces. After that, the samples were freeze-dried using a LABCONCO
FreeZone, the ratio cryogel/vanillin remaining unchanged.
In vitro release studies were conducted in PBS solution of pH 7.4, at
37 °C. These conditions were selected in order to simulate the physi-
ological conditions from the wound, the wound dressing being a pos-
sible application for the new obtained cryogels.
Samples of dissolution medium were withdrawn periodically at
predetermined time intervals and the amount of released active
agent was determined spectrophotometrically at 228 nm, using a
UV–VIS Hewlett Packard 8540A spectrophotometer. The cumulative
amount of vanillin released was determined using the appropriate
calibration curve. The release measurements were made in triplicate.
In order to elucidate the kinetics of vanillin release from the
studied cryogels, the data were analyzed using the equation proposed
by Korsmeyer and Peppas—Eq. (6). This equation was applied only to
the first 60% of the total amount of vanillin released.
nr
Mt =M∞ ¼ kr t ð6Þ
where Mt/M∞ is the fractional release of the active agent (Mt and M∞
represents the amounts of active agent released at time t and at equi-
librium, respectively), kr is a constant characteristic of the polymer–
active agent interaction and nr is the diffusion exponent representing
the release mechanism.
For a value of nr = 0.5 the mechanism of the active agent release is
controlled by Fickian diffusion, while a value of nr = 1 indicates that
the release rate is independent of time and controlled by swelling
mechanism. Values of nr between 0.5 and 1 were regarded as an indi-
cator for the superposition of both phenomena. This type of release
mechanism was named anomalous (non-Fickian) transport. [46]
3. Results and discussions
3.1. SEM characterization
The freezing/thawing cycles of PVA/cellulose solutions determine
the formation of the ice crystals into the amorphous region, which
force the polymer chains to arrange themselves into small ordered re-
gions (crystallites) [47]. These crystallites act as physical crosslinks.
Upon thawing the ice crystals melt, leaving the porous structure of
the hydrogels unaltered. In this way these crystals act as porogen dur-
ing the hydrogel formation. The size and the number of crystallites
ð4Þ are increasing with repeated freezing/thawing cycles. Thus, the po-
rous walls consist of swollen amorphous PVA, while the crystallites
act as knots of the network. These crystalline knots assure high di-
mensional stability and elastic properties to the PVA cryogel [48]. In
the case of PVA/cellulose cryogels, cellulose chains are physically
entangled inside this 3D PVA network. The cryogels pores are appro-
priate sites for the encapsulation of various bioactive agents.
Fig. 1 shows the SEM cross-section images of the cryogels after
ð5Þ freeze-drying for 24 hours. The PVA cryogel has a structure with
pores of small dimensions which assure a higher surface for the con-
tact with water. The PVA/cellulose samples have a porous structure,
with pores which are interconnected, irregular and have walls
which become thicker with the increasing in the cellulose content.
Islands of cellulose appeared in the PVA matrix at a higher content
of cellulose in the cryogels because of partial compatibility of compo-
nents. The dimensions of the pores are in the range of 2 μm (for the
cryogel containing only PVA) and 7 μm (for the cryogels containing
cellulose), as determined from the SEM microphotographs. A porous
surface is beneficial for biomedical applications, in the transport of
gases through the wound dressing. For example a raised level of oxy-
gen in the tissue stimulates epithelization and fibroblasts prolifera-
tion [49].
3.2. X-ray diffraction measurements
Fig. 2 shows the XRD diffractograms of PVA, cellulose and the
studied cryogels. As it can be observed, all samples have a crystalline
structure and their crystallinity increases when cellulose is added.
The PVA diffractogram presents a single peak (strong) characteristic
to polymer crystallinity at about 2θ = 19.5°, while cellulose has the
structure of cellulose II, with the characteristic peaks of 2θ = 12°, a
well-defined peak at 2θ = 20° and the third peak at 21.7°.
In the diffractograms of the cryogels the intensities of the main
peaks of PVA and those of the cellulose are decreasing with the
decrease of the cellulose content. The same decrease is observed
for the Bragg peak from 12°, with a shift to lower values. These
 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515 2511

PVA 90/10

70/30 50/50
Fig. 1. SEM images of the cross-section of PVA and PVA/cellulose cryogels (magnification 2000×).

observations are a proof of cryogel formation between those two
polymers and their mutual influence.
The crystallinity index (CrI) for PVA was determined as 57%, while
the value for cellulose was found to be 66%. The values obtained for
the PVA/cellulose cryogels were intermediate between the values of
the blank samples. Thus, the sample containing 30 wt% cellulose has
60% crystallinity, while for the sample containing 50 wt% cellulose
the CrI increased to 63%.
3.3. FT‐IR measurements
In Fig. 3 the FT‐IR spectra of blank samples and of the PVA/
cellulose cryogels are shown. The absorption band which appears at
3436 cm −1 corresponds to the OH stretching vibration, including
the groups which are involved in intramolecular and intermolecular
hydrogen bonds. This band becomes broader with the addition of cel-
lulose into cryogels, due to the presence of an increased amount of
OH groups in the samples. The absorption band from 2925 cm −1,
which is assigned to the C–H stretching vibration from alkyl groups,
is shifted to lower wavenumber and its intensity increases with the
addition of cellulose. A strong band observed at 1631 cm −1 corre-
sponds to the absorbed water, while the band from 1384 cm −1 has
been assigned to the vibration of the CH2 groups.
The FT-IR spectra clearly indicate the presence of cellulose into
these cryogels. The band from 1431 cm −1, assigned to a symmetric
CH2 bending vibration is increasing in intensity with the increase of

140 1,2
PVA
1,0
120 90/10
0,8
70/30
0,6
100
50/50
0,4
Intensity, a.u.

80 0,2 Cellulose
%T

0,0
60 Cellulose
-0,2

40 -0,4
50/50
-0,6
70/30
20 -0,8
PVA
-1,0
0
5 10 15 20 25 30 35 40 4500 4000 3500 3000 2500 2000 1500 1000 500
2θ, degree Wavenumber (cm-1)

Fig. 2. X-ray diffractograms of the PVA, cellulose and their cryogels. Fig. 3. FT‐IR absorption band spectra of PVA, cellulose and PVA/cellulose cryogels.
2512 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515

Table 2 interactions between the components of the cryogels, through the

Spectral characteristics of PVA/cellulose cryogels. formation of hydrogen bonding.
Sample υOH H2O absorbed EH ΔH R (A°)
(cm−1) (cm−1) (kJ−1) (J/g)

PVA 3448 1631 13.8 55.8 2.80 3.4. Thermal behavior

90/10 3436.9 1631 14.6 55.6 2.80
70/30 3413.7 1631 16.2 55.2 2.79 The thermal properties of the systems were investigated by
50/50 3384 1637 18.2 54.8 2.79
thermogravimetry. The thermograms of PVA and of the studied sam-
Cellulose 3352 1645 20.4 54.3 2.78
ples obtained in nitrogen, with the corresponding first derivative
curves are shown in Fig. 4a and b.
The thermal decomposition of the cellulose has one step between
the cellulose content. This band is also known as the “crystallinity 30 and 124 °C, which corresponds to the water loss. The second step
band” for cellulose and an increase of its intensity reflects an en- of degradation, characterized by the highest weight loss (86.09%) is
hancement of the samples' degree of crystallinity [42]. The crystallin- represented by the unzipping of cellulose chains and the formation
ity of the cryogels is influenced by the components and is increasing of levoglucosan, followed by the decomposition to yield char and vol-
due to the higher amount of cellulose in the cryogels; data were atile products. The major degradation step for cellulose has the max-
also confirmed from the XRD measurements. imum temperature at 345 °C (Table 3) [50].
The band from 3448 cm −1 (OH stretching vibration) is shifted to The PVA thermogram shows four processes, easily observed from
lower wavenumbers, while the band from 1631 cm −1 (absorbed the DTG curves. The first process corresponds to the water loss,
water) is shifted to higher wavenumber for 50/50 ratio, as a conse- while the second to the partial dehydration of PVA together with
quence of the molecular interactions between PVA and cellulose. the polyene formation. The third step consists in polyene decomposi-
These FT‐IR data are correlated with rheological changes—see below. tion and the destruction of the macroradicals. The last step of thermal
Table 2 shows the spectral characteristics of the studied samples. degradation is represented by the advanced degradation of the poly-
It can be observed that the energy of the hydrogen bonds increases meric chain and by the scission reactions. The main degradation part
with the cellulose content in cryogels, which indicates a higher of PVA, where a high degree of mass loss occurs, appears between
number of hydrogen bonds probably because of the enhancement in 306 °C (Ton) and 419 °C (Tend) [51]. These temperatures are shifted
samples crystallinity. In the same time the values for the H bonding to lower values in case of PVA/cellulose cryogels. Also, the second
distances show a small decrease when cellulose is added into the step (217–306 °C) characteristic to PVA degradation disappears as a
cryogels. These data indicate the presence of some molecular consequence of the cellulose influence over the PVA cryogels.

Fig. 4. The TG (a) and DTG (b) curves of PVA, cellulose and PVA/cellulose cryogels.
 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515 2513

Table 3 of the cellulose content in the cryogels, improving the strength of

Thermogravimetric data of the studied samples. the material.
Sample Degradation Ton Tm Tend ΔW T50 wr Starting with 45 °C a decrease in the storage modulus appears
step (°C) (°C) (°C) (%) (°C) (wt%) which can be a consequence of the uncoiling of the crystalline regions
PVA I 31 132 217 6.5 381 3.4 and of the weakening and breaking of the hydrogen bonds. A gel–sol
II 217 287 306 4.9 transition takes place at 68 °C for PVA cryogel. This observation was
III 306 379 419 63.1 also confirmed by Hatakeyema et al., which found a temperature of
IV 419 448 513 21.5
70 °C for this transition [52]. This process is also observed for the hy-
90/10 I 34 136 207 2.6 370 8.5
II – – – – drogel containing 10% cellulose and is decreasing with the cellulose
III 207 369 417 74.7 content until it disappears for the sample containing the highest
IV 417 447 514 13.8 amount of cellulose (50%).
70/30 I 31 114 198 5.1 358 7.9
II – – – –
3.6. Swelling studies
III 198 358 412 73.0
IV 412 439 521 13.3
50/50 I 31 69 128 1.9 346 11.2 In vitro swelling and release studies were conducted at 37 °C (the
II – – – – human body temperature) in PBS solution having a pH 7.4 (the pH of
III 128 345 409 77.1
the wound fluids). The swelling degree was determined gravimetri-
IV 409 433 534 9.2
Cellulose I 30 71 124 2.1 345 11.2 cally. Adding a hydrophilic polymer into the PVA network, the in-
II 124 345 470 86.1 crease of the water sorption capacity of the hydrogel is expected.
Fig. 6 presents the swelling profiles of the PVA/cellulose cryogels as
a function of time. This figure shows that the swelling degree in-
The thermogravimetric parameters for each step of degradation creases with time, first rapidly and then slowly, reaching the equilib-
were evaluated from the above thermograms: the onset temperature rium swelling degree after ~ 200 min for all samples. It can be seen
(Ton), the temperature of maximum degradation rate (Tm), the tem- that the values for the swelling degree are increasing with the in-
perature corresponding to the end of each stage (Tend), the tempera- crease of cellulose content in the cryogels.
ture at which the mass loss is 50% (T50), the weight loss (ΔW%) and The overall crystallinity of the formed cryogels increases because
the residual mass (wr). These parameters are listed in Table 3. the freezing/thawing process leads to a more organized structure
It can be seen that the Ton and the Tm are decreasing with the in- of cellulose and PVA chains, and so to an increased strength. In the
crease of cellulose content, but the values are situated between the same time, the swelling behavior is strongly influenced by the pres-
temperatures corresponding to the pure components (blank sam- ence of hydrophilic polymer, cellulose, in the cryogels. By increasing
ples). The same trend is observed for the temperatures of the 50% the amount of cellulose, the hydrophilicity of the hydrogels increases,
mass loss from 381 °C characteristic to the PVA cryogel, to 345 °C inducing an increase of swelling.
for cellulose. The residual mass is increasing with the increase of the The equilibrium swelling degree (ESD) indicates the ability of a
cellulose content, from 3.40 wt% for the PVA to 11.18 wt% for the wound dressing to absorb wound fluids or exudates. The ideal dress-
highest content of cellulose in the cryogels. ing for a wound with a large amount of exudates is a dressing with a
high value of ESD. In the case of the PVA/cellulose cryogels the value
3.5. Rheological analysis for ESD (Fig. 7) is closely related to cryogels structure and composi-
tion, a high value of the ESD being associated with the presence of a
The use of oscillatory rheological analysis is a common method natural polymer.
used in cryogels characterization. Fig. 5 presents the evolution of In order to determine the kinetic parameters of swelling (nsw and
the storage modulus (G′) and of the loss modulus (G″) as a function ksw) graphs of ln Fsw as a function of ln t were plotted. The values for
of temperature, for the PVA cryogels with different amounts of the swelling exponent and for the swelling constant of the PVA/cellu-
cellulose. lose hydrogels, in PBS pH 7.4, at 37 °C are presented in Table 4. The
It can be seen that both modules are temperature dependent. As values of the swelling exponent for the studied cryogels indicate an
expected, the values of the G′ were higher than the values of G″ anomalous mechanism of swelling. In the case of anomalous swelling,
which indicates a predominant elastic response, typical for gels and the rate of polymeric chain relaxation is much lower than the rate of
solid-like materials. The values of the G′ increase with the increase

550
500
105
10 5 450
Swelling degree %

400
104 350
G' (Pa)

G'' (Pa)

10 4 300
250
103
200
10 3 150
PVA
102 PVA 100 90/10
90/10
50 70/30
70/30
50/50 10 2 50/50
1 0
10
20 30 40 50 60 70 80 0 100 200 300 400 1380 1400 1420 1440
Temperature ( 0C) Time (min)

Fig. 5. Temperature dependence of the storage modulus (filled symbols) and the loss Fig. 6. Swelling degree of the PVA/cellulose cryogels in phosphate buffer solution (pH
modulus (empty symbols) for the studied cryogels. 7.4) at 37 °C.
2514 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515

600

Cumulative amount of vanillin released (%)

50
550
40
500

450
30
ESD (%)

400

350 20

300
10
PVA
250 90/10
70/30
200 0
50/50

0 10 20 30 40 50 0 100 200 300 400 1360 1380 1400 1420 1440 1460
% Cellulose Time (min)
Fig. 7. Variation of the ESD with the cellulose content of the cryogels. Fig. 8. Vanillin release profiles from PVA/cellulose cryogels, in phosphate buffer solution
(pH 7.4) at 37 °C.

diffusion. This means that this case of swelling is a relaxation con-

trolled process [53].
The highest value for the swelling constant was obtained for the
cryogel containing 50 wt% cellulose. This suggests that the addition
of cellulose increases the rate of swelling.
3.7. In vitro release studies
The results obtained from vanillin release studies carried out on
PVA/cellulose cryogels containing 4.7% vanillin incorporated (Fig. 8)
show an initial burst release, after which the release rate slows as
the swelling equilibrium of the cryogels is reached. As it can be seen
in Fig. 8, the percentage of vanillin release from initial loaded amount
from PVA/cellulose cryogels is dependent of cellulose content in the
cryogels. An increase in cellulose content is equivalent with an in-
creased amount of the active agent released, as a consequence of
the cryogels morphology.
The release data were analyzed according to the model proposed
by Korsmeyer and Peppas (Eq. (6)). This model is used to analyze
the release of the active agent from matrices when the release mech-
anism is not well known or when more than one type of release phe-
nomenon is involved [46]. The kinetic parameters for vanillin release
from PVA/cellulose cryogels with various formulations are listed in
Table 5.
As observed, the release rate (kr) is increasing with the increase of
the cellulose content in cryogels. The values of the diffusion exponent
indicate an anomalous transport mechanism, which appear by cou-
pling Fickian diffusion with the relaxation of the cryogel network.
The addition of cellulose in the cryogels determines an increase in
the maximum amount of vanillin released (Qmax release) from
25.3% for the PVA cryogel without cellulose, to 43.4% for the cryogel
containing 50 wt% polysaccharide. In the same time, the period for
50% vanillin released (t1/2) is decreasing from 69 min for the PVA
cryogel to only 30 min for the 50 wt% cellulose in the sample. The
same trend is observed for the time of maximum amount of released
vanillin (tmax).
Both polymers used in this study are biocompatible and nontoxic,
being appropriate for use as wound dressing. In the same time these
Table 4
The swelling parameters of the PVA/cellulose cryogels.
two polymers are cheap and biodegradable; also the method used
for the cryogels obtaining is a technique which does not produce
any pollutant residues, so this is an environmentally friendly process.
PVA physical hydrogels present an increased strength among most
of the hydrogels due to their crystalline regions, but their main disad-
vantage is that swelling equilibrium occurs with a low rate [11]. The
scientific community tried to improve the properties of PVA cryogels
for biomedical applications, in general, and for wound dressing, in
particular. Kokabi et al. obtained PVA/clay cryogels and they discov-
ered that the presence of clay into the 3D networks of the cryogels
caused an increase in the crosslinking degree. The PVA/clay cryogels
exhibited great mechanical properties which made them appropriate
for wound dressing applications, although the swelling capacities of
these hydrogels decreased with increasing the clay percent [17].
Another attempt to obtain a wound dressing based on PVA cryogel
was the addition of sodium alginate (SA) to the PVA solution. Kim et
al. found that SA reduced the crosslinking process and weakened the
strength of the hydrogels, as well as the elongation at break. In the
same time, the swelling ability slowly increased with the amount of
SA up to a certain limit (20 wt%), then decreased. It was shown that
SA had insignificant effect on the drug release behavior [54].
In the case of PVA/dextran cryogels loaded with gentamicin, the
presence of the polysaccharide decreased the gel fraction, the
strength and thermal stability, but increased the swelling ability and
the porosity [55].
In the present study an improvement of the PVA cryogels proper-
ties in the presence of cellulose was obtained, especially the strength,
the swelling and the release capacity. These results demonstrate that
PVA/cellulose cryogels are appropriate candidates for wound dressing
materials.
4. Conclusions
Novel PVA/cellulose cryogels were obtained by freezing-thawing
technique, via physical crosslinking.
Table 5
The kinetic parameters of vanillin release from PVA/cellulose cryogels.
Sample PVA 90/10 70/30 50/50

nrel 0.82 ± 0.05 0.78 ± 0.04 0.70 ± 0.03 0.67 ± 0.03

Sample PVA 90/10 70/30 50/50
krel (min−1) * 10−3 3.7 ± 0. 1 7.3 ± 0. 2 10.1 ± 0. 2 19.7 ± 0. 5
nsw 0.143 ± 0.008 0.208 ± 0.005 0.179 ± 0.003 0.136 ± 0.009 Correlation factor (R) 0.98 0.98 0.98 0.98
ksw (min−0.1) 0.471 ± 0.007 0.460 ± 0.003 0.465 ± 0.002 0.842 ± 0.006 Qmax release (%) 25.3 33.4 32.2 43.4
Correlation 0.97 0.99 0.99 0.97 t1/2 (min) 69 47 40 30
factor (R) tmax (min) 390 390 270 210
 O.M. Păduraru et al. / Materials Science and Engineering C 32 (2012) 2508–2515
The SEM micrographs of cryogels indicated that the modifications
of the morphological aspects depend on the content of the natural
polymer. The crystallinity of the cryogels can be tailored by the cellu-
lose content. FT-IR spectra confirmed the presence of the interactions,
like hydrogen bonds, between the two components of the cryogels, as
cellulose and PVA. The rheological measurements showed that the
strength of the material can be enhanced by increasing the cellulose
content. The swelling capacities of the studied cryogels were im-
proved by the presence of cellulose in the network. The study regard-
ing the release of vanillin from PVA/cellulose matrices revealed an
increased percent of bioactive agent released with the increasing of
cellulose in the cryogels and also shortening of the half time and
maximum time of release.
The well-known biocompatibility of both polymers, together with
the absence of a chemical crosslinker, the increased cryogels' strength
and also the high degree of swelling and fast release of the bioactive
component from these cryogels make them promising matrices for
biomedical applications. Such hydrogels or films can be used as
potential materials both for wound dressing or as food packaging.
Acknowledgments
This research was financially supported by the grant of the
Romanian National Authority for Scientific Research, CNCS—UEFISCDI,
project number PN-II-ID-PCE-2011-3-0187.
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