journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

Available online at www.sciencedirect.com

www.elsevier.com/locate/jmbbm

Research Paper

Evaluation of polyvinyl alcohol composite

membranes containing collagen and bone particles
Nishar Hameeda,n, Veronica Glattauerb, John A.M. Ramshawb
a

Institute for Frontier Materials, Deakin University, Pigdons Road, Waurn Ponds, Geelong 3216, Australia
CSIRO Manufacturing Flagship, Bayview Avenue, Clayton 3169, Australia

ar t ic l e in f o

abs tra ct

Article history:

Composite biomaterials provide alternative materials that improve on the properties of the

Received 30 June 2014

individual components and can be used to replace or restore damaged or diseased tissues.

Received in revised form

Typically, a composite biomaterial consists of a matrix, often a polymer, with one or more

31 March 2015

llers that can be made up of particles, sheets or bres. The polymer matrix can be chosen

Accepted 1 April 2015

from a wide range of compositions and can be fabricated easily and rapidly into complex

Available online 9 April 2015

shapes and structures. In the present study we have examined three size fractions of

Keywords:

collagen-containing particles embedded at up to 60% w/w in a poly(vinyl alcohol) (PVA)

Polyvinyl alcohol

matrix. The particles used were bone particles, which are a mineralcollagen composite

Bone

and demineralised bone, which gives naturally cross-linked collagen particles. SEM

Demineralised bone

showed well dispersed particles in the PVA matrix for all concentrations and sizes of

Thermal properties

particles, with FTIR suggesting collagen to PVA hydrogen bonding. Tg of membranes shifted

Mechanical properties

to a slightly lower temperature with increasing collagen content, along with a minor

Cell culture

amount of melting point depression. The modulus and tensile strength of membranes
were improved with the addition of both particles up to 10 wt%, and were clearly
strengthened by the addition, although this effect decreased with higher collagen loadings.
Elongation at break decreased with collagen content. Cell adhesion to the membranes was
observed associated with the collagen particles, indicating a lack of cytotoxicity.
& 2015 Elsevier Ltd. All rights reserved.

1.

Introduction

In recent years, biomaterials have become widely used to

replace and restore the function of damaged or deteriorated
tissues, to support healing, to improve function and to correct
abnormalities (Nair and Laurencin, 2007). Metals, alloys and
ceramics have been used very successful as biomaterials;
however, in certain instances some shortcomings have
n

emerged, including corrosion, allergic tissue reactions of

metal ions, fabrication difculties and low mechanical reliability of some ceramics (Ramakrishna et al., 2001). Composite
biomaterials, based on polymers, provide an alternative that
may overcome certain limitations of metal and ceramic based
biomaterials. Composite biomaterials are multi-phased combinations of two or several biodegradable and biologically
derived components (Williams, 1999), which acquire new

Corresponding author. Tel.: 61 3 52271403.

E-mail addresses: nishar.hameed@deakin.edu.au (N. Hameed), veronica.glattauer@csiro.au (V. Glattauer),
john.ramshaw@csiro.au (J.A.M. Ramshaw).
http://dx.doi.org/10.1016/j.jmbbm.2015.04.005
1751-6161/& 2015 Elsevier Ltd. All rights reserved.

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

distinctive properties that the individual components cannot

achieve by themselves. Typically, a composite biomaterial
consists of a matrix having one or more llers which can be
made up of particles, sheets or bres. Polymers are widely
used as matrices for composites in various applications as
they are available in a range of compositions and forms
(solids, bres, fabrics, lms, and gels). Moreover, they can be
fabricated easily and rapidly into complex shapes and
structures.
In the present study we have examined collagencontaining particles embedded in a polymer, poly(vinyl alcohol), matrix. PVA has been shown to be suitable for various
biomedical applications (Kobayashi et al., 2001; Schmedlen
et al., 2002; Alexy et al., 2003; Weis et al., 2004; Yi et al., 2006;
Gupta et al., 2009; Sailaja et al., 2009; Jiang et al., 2011;
Sionkowska, 2011; Peng and Shen, 2011; Peng et al., 2012),
including as a base for composite biomaterials (Sionkowska,
2011). It can be readily modied to enhance the biological
properties (Schmedlen et al., 2002; Sailaja et al., 2009), and
soluble PVA arising from implants is readily cleared from the
body through the kidneys (Jiang et al., 2010). PVA is also well
suited for making composite materials, especially with biological polymers such as collagen and chitosan (Alexy et al.,
2003; Yi et al., 2006; Sionkowska, 2011; Peng and Shen, 2011;
Peng et al., 2012). Collagen for biomedical applications can be
prepared in a variety of formats (Ramshaw et al., 1996). In
some cases, the natural brous composition, such as found in
skin or tendon, is maintained when the tissue is comminuted
into a brous powder. In other cases, tissue is solubilised,
typically through an enzyme treatment to produce a puried
collagen solution (Miller and Rhodes, 1982). This solution can
be manipulated so as to reform a brous network, although
this network lacks the strength of the collagen bres that
have not been solubilised.
In the present study, we have examined whether a further,
different format collagen-based material is suitable as a
readily made, cost effective composite material for biomedical applications, particularly for hard tissue repair. Thus, we
have examined composites made with particles of naturally
cross-linked collagen, obtained through demineralisation of
bovine bone particles. For comparison we have used bone
particles in an un-modied, mineralised form. Bone particles
are readily available and can be readily demineralised if
required, derived, for example, from human, porcine and
bovine bone, and are in clinical use, for example in dental
applications (Ramrez-Fernndez et al., 2013; Figueiredo et al.,
2013; Ivanovic et al., 2014). Bone based material also represents a cost effective source of collagen, especially when
compared to soluble collagen. Three size fractions of each
type of particle were prepared and used to make composite
membranes with up to 60% w/w particles. These membranes
were examined by a number of methods. Scanning electron
microscopy (SEM) was used to study the morphology and
distributions of particles, while FTIR was used to look for
interactions between components. Differential scanning
calorimetry (DSC) was used to examine the glass transition,
Tg, and the melting temperature, Tc, while mechanical testing
looked at tensile strength, Young's modulus and axial strain.
A preliminary examination of cell compatibility was examined using L-929 mouse lung broblasts.

2.

Materials and methods

2.1.

Collagen-containing particles

39

Bovine bone particles were obtained from Waitaki Biosciences (New Zealand). Bone particles, for direct use (NB),
were separated into 2070 mm, 70100 mm and 100150 mm
fractions using a Fritsch Analysette 3PRO sieve system
(Oberstein, Germany). Production of demineralized bone
(DB) particles was as previously described (Glattauer et al.,
2010, 2011). Briey, unfractionated bone particles (NB) as
above were suspended in 0.6 M HCl and stirred intermittently
for 16 h at 4 1C. After settling, the acid was removed by
decanting and particles then further extracted each day for
6 days using the same conditions, before being washed in
several changes of H2O and freeze dried. The dried collagenbased particles were then fractionated into 2070 mm, 70
100 mm and 100150 mm fractions by sieving, as above. Bead
size distributions were veried by microscopy and image
analysis (Glattauer et al., 2010, 2011).

2.2.

Preparation of the composite lms

Poly(vinyl alcohol) (PVA), 99% hydrolyzed, with a molecular

weight, Mw 89,00098,000, was from SigmaAldrich. The PVA
samples were dried at 80 1C under vacuum for about 5 h to
remove moisture. The PVA composite membranes, containing NB or DB particles, were prepared using a solution of 1.0 g
PVA per 6.34 mL distilled water, stirred continuously at 90 1C
for 1 h. The PVA solution was cooled down to 20 1C and a preweighed amount of collagen particles was added. The mixture was stirred overnight to obtain a highly dispersed PVA/
collagen suspension. The suspension was evacuated to
remove any entrapped air bubbles and then a calculated
amount suspension was poured into rectangular glass plates
with edging (5 mm deep). In all samples, water was allowed
to evaporate at room temperature ( 20 1C). The nal thickness of the membranes was measured around 200720 mm.
Both types of particle, NB and DB in each of the 3 size ranges
(2070 mm, 70100 mm and 100150 mm) were used for composite membrane preparation.

2.3.

Scanning electron microscopy

The morphology of the composite sheets was examined by

scanning electron microscopy (SEM) using a Jeol Neoscope at
an activation voltage of 10 kV. The free surfaces were coated
with thin layers of gold before the observation.

2.4.

FTIR spectroscopy

The FTIR spectra of all the samples were measured on a

Bruker Vertex-70 FTIR spectrometer. The KBr disk method
was adopted to conduct the FTIR experiments. The sample
was mixed with KBr powder, grounded well and prepared to
KBr disks. The disks were dried under vacuum in an oven at
100 1C before the measurements. The spectra were recorded
at the average of 32 scans in the standard wavenumber range
of 4004000 cm  1 at a resolution of 4 cm  1.

40

2.5.

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

Differential scanning calorimetry (DSC)

DSC experiments were carried out using a TA-DSC model

Q200 instrument. The measurement was performed using
510 mg of the sample under an atmosphere of nitrogen gas.
The samples were rst heated to 100 1C and held at that
temperature for 5 minutes to remove the thermal history.
Then the samples were cooled to  50 1C at a rate of
20 1C/min, held for 5 minutes, and subsequently heated from
0 to 200 1C at 20 1C/min (second scan). Glass transition
temperature (Tg) values were taken as the midpoint of
transition in the second scan of DSC thermograms.

2.6.

Mechanical tests

The tensile behaviour of the materials was analysed using a

Lloyd LR 30K testing machine in tensile mode with a 100 N
capacity load cell and with a gauge length of 5 mm. The
specimen was a thin rectangular strip (25  6  0.2 mm3). The
loaddisplacement curves of the samples were obtained at a
strain rate of 1 mm/min at 75% relative humidity and 20 1C.
The stress and strain values were obtained using the standard equations. The stress, strain and modulus values were
evaluated from the elastic region and modulus values were
calculated at no more than 40 % strain to deformation.

2.7.

Cell culture

Cell adhesion on composite membranes was assessed using

L929 mouse broblasts (ATCC-CCL-1, Rockville, MD), a highly
adherent cell line. Prior to examination, the composite
membranes were sterilised by 15 kGy gamma irradiation.
Cells were grown in MEM containing GlutaMax (Life Technologies), plus 10% v/v foetal bovine serum, 1% NEAA (Gibco) and
1% Antibiotic-Antimycotic (Gibco-Invitrogen) at 37 1C in 5%
CO2. Cells at 80 % conuency were harvested using TrypLE
Express (Life Technologies) and washed twice with media.
Subsequently, cells were seeded onto membranes using
weighted stainless steel cell culture fences (Aix Scienticss,
Germany)(Fischer et al., 1990) at 25,000 cells/cm2 on various
composite materials in non-adherent, 24-well culture plates
(Costar, Corning, USA) and cultured in the same medium. Cell
adhesion was examined after 16 h using bright-eld optics
and after calcein-AM staining (Molecular Probes) following
the supplier's instructions, with uorescence optics, and
photographed using a Nikon TE2000-U microscope. The
calcein-AM staining detects live cells.

3.

3.1.

Microscopy

The composite membranes all had good exibility, and were

translucent, with the opacity increasing with the amount of
NB or DB particles that were incorporated as shown in Fig. 1.
The particles seem well dispersed (Fig. 1), with increasing
amounts leading to increased opacity. Those with a higher
opacity have the advantage of being easier to see in a surgical
application. The surface morphology of PVA/collagen composite membranes was observed using SEM (Fig. 2). This showed
the collagen particles, particularly at the surface of the PVA
matrix. The collagen particles were well dispersed in PVA at
all concentrations of particles and with all 3 particle size
fractions for both the bone particles (ND) and the collagenbased, demineralised bone particles (DB) (Fig. 1, Fig. 2). It is
possible that due to the weak interaction between collagen
and PVA at the interface, the collagen particles form into
small aggregates or individual particles and become well
dispersed in the polymer matrix.

3.2.

FTIR spectroscopy

FTIR spectroscopy was used to determine the inter- and

intra-molecular hydrogen bonding interactions in PVA/collagen composite membranes. The FTIR spectra of the composites are given in Fig. 3. The broad band in the 3000
3700 cm  1 region can be assigned to a wide distribution of
the stretching vibration of free, self-associated and interassociated hydroxyl groups of PVA and collagen. The selfassociated hydroxyl stretching band of PVA can be observed
at 3295 cm  1 (Ramakrishna et al., 2001). The characteristic
absorption bands of collagen were observed in composites
containing both bone and demineralised collagen particles.
The specic absorption peaks include 1639 cm  1 (CQO
stretching) for amide I; 1547 cm  1 (NH bending) for amide
II; 1239 cm  1 for amide III; 3302 cm  1 (OH stretching);
2951 cm  1 for aliphatic groups (  CH2 and CH3); and
1451 cm  1 for COO  . The intermolecular interactions between PVA and collagen may occur by the formation of hydrogen bonds at the interface. This may be enhanced for the DB
particles as the loss of mineral leads to a more porous
collagen particle. The hydroxyproline residues in collagen
and collagen peptides are involved in an extensive hydration
shell (Bella et al., 1994, 1995) and provide potential hydrogen
bonding sites, as well as the surface CQO groups that are

Results and discussion

Previously, composite materials including PVA based on

soluble and brous collagens have been described (Sarti and
Scandola, 1995; Sionkowska et al., 2009), but inclusion of
collagen-containing particles provides a different approach.
The new materials could be particularly useful for repair of
hard tissues, with the presence of residual growth factors in
the demineralised bone particles (Wildemann et al., 2007)
assisting in the tissue repair.

PVA

1%

10%

30%

50%

60%

Fig. 1 Photographs of PVA and PVA/collagen composite

membranes with increasing w/w concentrations of DB 20
70 lm collagen particles, as indicated.

41

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

500 m

500 m

500 m

Fig. 2 SEM surface images of PVA/collagen composite membranes containing 2070 lm DB particles; (a) DB at 10% w/w,
(b) DB at 30% w/w and (c) DB at 50% w/w.

3302cm

D60
DB
60
D50
DB
50
D30
DB
30

-1

1451cm

2951cm

-1

D10
DB
10

-1

D11
DB

-1

1547cm
-1
1637cm

1239cm

-1

1451cm

2951cm

-1

-1

-1

-1

3302cm

NB
30
W30

W10
NB 10
W1
NB 1
PVA
PVA

-1

1547cm

-1

1637cm

2951cm

-1

DB
30
D30
D10
DB
10
D11
DB

-1

-1

1547cm
-1
1637cm

-1

Wavenumber (cm )

3302cm

-1

NB 60
W60

Absorbance (a.u.)

Absorbance (a.u.)

W50
NB 50

1451cm

-1

W60
NB
60

-1

-1

1000 1500 2000 2500 3000 3500 4000

-1

Wavenumber (cm )

3302cm

2951cm

1239cm

PVA

1000 1500 2000 2500 3000 3500 4000

-1

Wavenumber (cm )

NB
50
W50
-1
2951cm
1239cm
-1
1451cm

1547cm

-1

W30
NB 30
NB 10
W10

-1

1637cm

W1
NB 1
PVA
PVA

-1

-1

DB
60
D60
D50
DB
50

DB 1
PVA

1000 1500 2000 2500 3000 3500 4000

1239cm
-1
1451cm

DB 10

1547cm
-1
1637cm

PVA

-1

3302cm
DB 60
DB 50
DB 30

Absorbance (a.u.)

1239cm

-1

Absorbance (a.u.)

-1

Absorbance (a.u.)

Absorbance (a.u.)

3302cm

W60
NB 60
NB 50
W50

1239cm

-1

1451cm

-1

2951cm

-1

W30
NB
30
-1

W10
NB 10

1547cm
-1
1637cm

NB
W11

PVA
PVA

1000 1500 2000 2500 3000 3500 4000

-1

Wavenumber (cm )

1000 1500 2000 2500 3000 3500 4000

-1

Wavenumber (cm )

1000 1500 2000 2500 3000 3500 4000

-1

Wavenumber (cm )

Fig. 3 FTIR spectra of PVA/collagen composites at room temperature with different weight percentages of collagen particles
(as indicated by the labels) and different collagen particle types and sizes, (a) DB 2070 lm, (b) DB 70100 lm, (c) DB 100
150 lm, (d) NB 2070 lm, (e) NB 70100 lm, and (f) NB 100150 lm.

also available for making hydrogen bonds with OH groups

in PVA.
To conrm the hydrogen-bonding interactions in the composites, the carbonyl stretching region, ranging from 1580 to
1700 cm-1, of the membranes was examined (Fig. 4). The FTIR
spectra of membranes containing DB2070 mm collagen particles
at various compositions are given in Fig. 4. The carbonyl bands
of collagen can be observed at 1637 cm-1 in the composites. A
new band is formed at 1618 cm-1 in composites containing
higher amounts of collagen (DB50 and DB60 wt%). This band is
due to the vibration of the hydrogen-bonded collagen carbonyl
groups at the PVAcollagen interface. It can be noticed that this
hydrogen bonded carbonyl band is not obvious in composites
containing low quantities of collagen. This can be attributed to

the limited hydrogen bonding interaction between collagen

carbonyl groups and PVA hydroxyl groups at these
compositions.

3.3.

Differential scanning calorimetry

The glass transition behaviour and melting temperatures of

the PVA/collagen membranes were obtained from the DSC
second heating scans (Fig. 5). PVA alone showed a glass
transition temperature (Tg) of 76 1C. In the composite membranes, a single Tg corresponding to the PVA rich phase was
observed. This transition broadens and shifts very slightly
towards lower temperatures as the concentration of collagen
increases (Fig. 5), perhaps due to the interfacial interactions

42

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

between the matrix and the ller. The size of this change was
small,o2.5 1C, even up to 60 wt% collagen addition. There
was no signicant difference seen between membranes prepared using DB or NB, or with different size particle preparations. There was also a minor of melting point depression
during second heating scan (Fig. 5). The melting point for
regenerated PVA is around 230 1C. This value dropped to
215 1C when the demineralised collagen content reaches
60 wt% of the whole composites. Again, no signicant difference was seen between the DB and NB samples, nor with
different size particle preparations. A melting temperature
depression is a characteristic feature of miscible polymer

3.4.

Mechanical tests

The tensile mechanical properties of the composite membranes were investigated (Fig. 6). Both the modulus and the
tensile strength of the PVA membrane were improved with
the addition of both DB and NB particles up to 10 wt%. At
higher loadings of particles, the modulus and the tensile
strength both decreased such that by 60 wt% particles the
tensile properties of the materials were poor. There was no
notable difference seen between DB and NB particles,
although the changes were somewhat larger at higher loadings with increased particle size. The un-modied PVA
membrane showed Young's modulus and tensile strength
of values of about 1430 MPa, 51.2 MPa respectively. There was
a maximum of 35 % improvement in modulus of PVA by the
addition of 10 wt% demineralised collagen particles, while the
tensile strength of PVA was improved by 30 wt% for the same
materials. Membranes including NB particles showed similar
properties. On the other hand, the elongation at break of the
membranes decreased with all increases in collagen loading,
with a slightly greater loss with increased particle size (Fig. 6).
Again there was little difference in the materials made with
either DB or NB particles. The increase in tensile strength and
modulus can be attributed to the higher interfacial adhesion
between PVA and collagen due to the hydrogen bonding
interaction between them. Thus, the PVA membrane was
noticeably strengthened by the addition of collagen-based
particles. The natural surface roughness of the particles may
be important in this strengthening.

100

100

80

80

Tg (C)

Tg (C)

Fig. 4 The carbonyl stretching region of PVA/collagen

composites, containing 2070 lm DB particles 160% w/w
particles, at room temperature.

materials having intermolecular interactions between the

components.

60
40
W20-70 m
NB 20 - 70 m
W70-100 m
NB 70 - 100 m
W100-1500
NB 100 - 150 m

20

60
40
DB
20 - 70m
m
D20-70
DB
70 - 100m
m
D70-100
D100-1500
m
DB
100 - 150 m

20
0

0
PVA

DB1 DB10 DB30 DB50 DB60

PVA

Wt % particles

300

300

250

250

200

200

Tc (C)

Tc (C)

Wt % particles

150
100

W20-70
m
NB
20 - 70
m
W70-100
NB
70 - 100m
m
W100-1500
m
NB
100 - 150
m

50

DB1 DB10 DB30 DB50 DB60

150
100
DB
20 - 70
m
D20-70
m
DB
70 - 100m
m
D70-100

50

DB
100 - 150
m
D100-1500
m

0
PVA

DB1 DB10 DB30 DB50 DB60

Wt % particles

PVA

DB1 DB10 DB30 DB50 DB60

Wt % particles

Fig. 5 The glass transition, Tg, and melting, Tc, temperatures of PVA/collagen membranes with NB and DB at various wt%
compositions.

43

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

1.0

60

40

20

2000

NB 20-70m
NB 70-100m
NB 100-1500m

0.8
0.6
0.4
0.2

0.0
W1

W10

W30

W50

W60

W1

Wt% of Collagen

W10

W30

1.0

W50

40

20

PVA

D30

D50

Wt% of Collagen

W1

D60

W10

W30

W50

W60

0.6
0.4

2000

0.0

0
D10

500

Wt% of Collagen

0.2

D1

1000

W60

DB 20-70m
DB 70-100m
DB 100-1500m

0.8

Axial Strain (%)

DB 20-70m
DB 70-100m
DB 100-1500m

PVA

1500

Wt% of Collagen

80

60

NB 20-70m
NB 70-100m
NB 100-1500m

PVA

Young's Modulus (MPa)

PVA

Tensile Stres (MPa)

Young's Modulus (MPa)

NB 20-70m
NB 70-100m
NB 100-1500m

Axial Strain (%)

Tensile Stres (MPa)

80

DB 20-70m
DB 70-100m
DB 100-1500m

1500

1000

500

PVA

D1

D10

D30

D50

D60

Wt% of Collagen

PVA

D1

D10

D30

D50

D60

Wt% of Collagen

Fig. 6 The mechanical properties of PVA/collagen membranes with NB and DB at various wt% compositions. (n Z 3 for each
data point).

Fig. 7 Growth of L929 broblasts on PVA/collagen membranes with NB (A) and DB (B) particles at 30 wt% compositions.
Particles were (A) 2070 lm and (B) 100150 lm. The images are shown as bright eld images with calcein-AM staining images
superimposed.

The mechanical data have been reported for the base

materials that have not been treated further to cross-link
the PVA. In some biomedical applications a rapid dispersion
of the PVA may be required and no cross-linking would be
required. However, in other applications a more durable
material may be preferred, which can be achieved by controlled cross linking of the PVA in the composite material.
PVA cross linking can be achieved by chemical methods, such
as with glutaraldehyde (Mansur et al., 2008), or by chemical
functionalisation of the PVA (Schmedlen et al., 2002). Alternatively, stabilisation of hydrated PVA can be achieved by a
series of freeze/thaw cycles (Wan et al., 2002), or by gamma
irradiation (Sharaf et al., 1999). Gamma irradiation has the
additional benet of sterilising the membranes.

3.5.

Cell culture

A preliminary examination of cell binding to the new composite materials was made. For these experiments only, the
cell culture samples were sterilised by gamma irradiation,
which also prevented the membranes from swelling and
dissolving. The L-929 broblasts did not bind to the PVA
(Fig. 7), consistent with the previous observation that it was
necessary to modify PVA, for example, with a cell binding
peptide, to enhance cell interactions (Schmedlen et al., 2002).
On the other hand, NB and DB particles have previously been
shown to be excellent substrates for cell adhesion and
culture, particularly when used in spinner culture (Glattauer
et al., 2010, 2011). In the present case, cell binding, shown by

44

journal of the mechanical behavior of biomedical materials 48 (2015) 38 45

calcein-AM staining, was observed at isolated locations

(Fig. 7), presumably when the collagen had penetrated the
PVA surface. The extent of the cell binding varied and was
moderate in some samples (eg: Fig. 7A), but was almost
absent in others (eg: Fig.7B). Most particles seem well buried
within the PVA and so are not available for cell attachment
and only those that are at, or have broken through to the
surface are available to bind cells. Examination of the composite material with ethidium homodimer-1, normally used
as part of a viability assay to identify dead cells, was not
readily possible as this reagent reacted readily with the
collagen particles in unseeded samples giving extensive red
staining (data not shown). However, visual inspection of
bright eld images did not show any adherent attached dead
cells. The live cells, as shown by the calcein-AM, were not
spreading after 16 h, possibly due to limited attachment sites,
and were not removed by further washing with PBS.
Thus, the present collagen-based particle composite materials with PVA give a new group of materials and seem
capable of being useful for certain biomedical applications.
The use of bone and demineralised bone particles introduces
a range of growth factors into the system (Zhang et al., 1997;
Blum et al., 2004; Wildemann et al., 2007). The growth factors
are more readily available from the demineralised bone
material (Zhang et al., 1997), although the amounts may
depend on the age of the bone material and on the demineralisations process (Peterson et al., 2004; Bae et al., 2006), such
that appropriate selection of particles may be necessary to
achieve the desired outcome. If additional strength or prolonged turnover are required, further modication of the
matrix through different cross-linking approaches is possible.
In the present study we have used bovine derived material,
but human bone and demineralised bone particles are readily
available as these materials have been approved for orthopaedic and dental applications. Also, other polymers such as
the readily resorbable polylactic/glycolic acid co-polymers
could be suitable. These new materials are easy to prepare
cost effectively and provide options for use in various
biomedical applications, particularly for hard tissue repairs.

Acknowledgement
This work was funded in part by an Alfred Deakin Research
Fellowship to NH.

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