Acta BIOMATERIALIA

Acta Biomaterialia 1 (2005) 411419

www.actamat-journals.com

In vitro degradation behavior of a novel bioresorbable

composite material based on PLA and a soluble CaP glass
M. Navarro a, M.P. Ginebra a,*
, J.A. Planell a, C.C. Barrias b,c
, M.A. Barbosa b,c

a
Reference Centre for Bioengineering of Catalonia (CREBEC), Biomaterials, Biomechanics and Tissue Engineering Division,
Department of Materials Science, Universitat Polite`cnica de Catalunya, Av. Diagonal 647, 08028 Barcelona, Spain
b
INEBInstituto de Engenharia Biomedica, Laboratorio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal
c
Universidade do Porto, Faculdade de Engenharia, Departamento de Engenharia Metalurgica e Materiais,
Rua Dr Roberto Frias, 4200-465 Porto, Portugal

Received 27 October 2004; accepted 29 March 2005

Abstract

Poly(a-hydroxy acids), and in particular polylactic acid (PLA), are nowadays amongst the most used bioabsorbable materials.
However, this polymer may not meet some application requirements due to inadequate mechanical properties and or its degradation
characteristics. A possible strategy to tackle this problem is the incorporation of an inorganic phase into the polymeric matrix. In
this work a new fully biodegradable composite material made with PLA and calcium phosphate soluble glass particles has been
developed. The behaviour of the PLA/glass composite has been analysed during its degradation in simulated physiological condi-
tions by means of weight loss, molecular weight and thermal properties analysis and electron microscopy observation. The results
showed that the incorporation of phosphate glass particles into the polymer signicantly accelerated the degradation of the PLA and
induced the formation of calcium phosphate precipitates at the composite surface.

2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Keywords: Biodegradable composite; Polylactic acid; Phosphate glass; In vitro degradation; Bone regeneration

1. Introduction have been followed to overcome these limitations are

In the last few decades, polymers, and particularly
bioabsorbable polymers such as aliphatic polyesters,
have attracted great interest for several biomedical
applications. Poly(a-hydroxy acids), and specically
polylactic acid (PLA), are amongst the most well known
bioabsorbable materials and have been used in numer-
ous applications such as sutures, pins, screws and drug
delivery systems [1,2]. However, PLA may not com-
pletely meet the requirements for some applications
due to its insucient mechanical properties or its degra-
dation characteristics. Amongst the approaches that
*
Corresponding author. Tel.: +34 934011089.
E-mail address: maria.pau.ginebra@upc.es (M.P. Ginebra).
the use of dierent copolymers and the incorporation
of an inorganic phase into the polymeric matrix. In
the rst approach, copolymers of the PLA stereoisomers
have been combined as well as dierent polymers, in
order to obtain a wide range of properties. In the second
approach, the inorganic phase can be either bres [3] or
particles of dierent ceramics or glasses [46]. Ideally,
the incorporation of calcium phosphate ceramics and
glasses into the polymeric matrix would improve not
only the mechanical properties of the material, but also
its bioactivity and biological behaviour.
Several studies incorporating calcium phosphate
ceramics or bioactive glasses into a biodegradable
polymeric matrix have been carried out for the develop-
ment of composites for facial bone augmentation,
drug delivery systems, tissue engineering scaolds and

1742-7061/$ - see front matter
2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.actbio.2005.03.004
412 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419
orthopaedic applications [79]. In general, the addition
of these inorganic phases resulted in an increase of the
bioactivity of the material. However, hydroxyapatite
and other calcium phosphate ceramics hardly (or very
slowly) dissolve under physiological conditions [10,11].
The same goes for the bioactive silicate based glasses.
Therefore, it is of interest to develop a composite where
both phases are completely degradable, in order to en-
sure the complete erosion of the material.
From this perspective, phosphate glasses have been
selected as the inorganic phase of the composite materi-
als developed. Specically, G5 is a soluble phosphate
glass in the system P2O5CaONa2OTiO2, which has
been extensively studied in terms of its mechanical pro-
perties and degradation [12]. Besides, it has been shown
to be non-cytotoxic in previous biocompatibility studies
[13]. In this work the eect of the incorporation of G5
soluble glass particles on the degradation process of a
PLA matrix will be evaluated.
PLA degradation is a complex process that has been
extensively investigated [14,15]. Indeed, it is known that
there are numerous variables, such as the materials
chemical structure, crystallinity, molecular weight, pro-
cessing conditions, shape and size, aecting the polymer
degradation mechanisms. The addition of an inorganic
phase into the polymeric matrix increases the complexity
of the degradation pattern of the material. Some studies
have reported dramatic changes in polymer degradation
with the incorporation of some calcium phosphate com-
pounds [16,17].
The scope of this study is to perform a chemical and
morphological analysis of the behaviour of a PLA/G5
composite material during its degradation in simulated
physiological conditions.
2. Materials and methods
2.1. Materials
Polylactic acid with a 95L/5DL ratio (PURAC, Neth-
erlands) and a molecular weight (Mw) of 3.34 105 was
used as the polymeric matrix. For the reinforcement, a
soluble phosphate glass coded G5, with the following
molar composition 44.5P2O544.5CaO6Na2O5TiO2,
was used. The making of this glass is explained in detail
elsewhere [12]. Briey, the reagents were mixed homoge-
neously in a platinum crucible and heated at 1350 C for
3 h. After that, the mixture was cast and annealed on a
metallic preheated plate. Finally, the glass was milled in
an agate ball mill and glass particles were sieved at 80 lm.
2.2. Composite preparation
Composite lms were obtained using the solvent cast-
ing method. First, a 5% (w/v) solution was prepared by
dissolving the PLA in chloroform for 48 h. Once dis-
solved, the glass particles were introduced (0 and 50%
by weight) and mixed into the PLA + chloroform solu-
tion until the glass particles were well distributed in
the matrix. The mixture was cast in a Teon mould
and the chloroform was allowed to evaporate for 48 h.
After that, thin lms of composite and PLA were ob-
tained and cut into small pieces. The PLA and PLA/
glass pieces (hereafter PLA/G5) were compression moul-
ded using a hydraulic press (IQAP LAP, Spain) at
160 C. The pressure was increased progressively to
100 bars. After pressing, the moulded plates with a
thickness of 1.5 mm were cut into rectangular slabs
(50.8 12.7 mm) using a rotator diamond saw (Struers,
Minitom, StruersTech, Denmark). For the study of the
degradation of the materials, PLA and PLA/G5 speci-
mens were immersed in a simulated body uid (SBF) re-
ported by Kokubo et al. [18] at 37 C for 6 weeks.
2.3. Weight loss
The degradation of the materials was evaluated by
means of weight loss measurements. The surface area
and initial weight (W0) of three specimens of each mate-
rial (PLA and PLA/G5) were measured before being
immersed in polystyrene containers with the SBF. A sur-
face/volume ratio of 1 g/10 ml was kept constant during
the test. Each week, the specimens were recovered from
the uid and dried at 37 C until the weight stabilized.
After that, the weight was recorded (Wt) and the
uid was changed. The percentage of weight loss was
determined using the following equation: %W =
(W0
Wt) 100/W0, where W0 is the initial weight of
the specimen and Wt is the weight of the dried specimen
at time t.
2.4. Dierential scanning calorimetry (DSC)
Small pieces from the PLA and PLA/G5 specimens
after the dierent stages of their preparation and after
0, 2, 4 and 6 weeks of degradation were weighed for
the analysis. Samples were analyzed on a TA Instru-
ments model 2029 dierential scanning calorimeter.
Approximately 10 mg of each material were placed in
aluminium pans. Samples were heated from 20 to
200 C at 10 C/min using nitrogen as the purge gas.
Thermograms were analyzed to determine the thermal
transitions and percentage of crystallinity (%Xc) of the
samples. The crystallinity percentage was calculated
using the reference heat of enthalpy (93.1 J/g) for theo-
retically 100% crystalline PLA [19]. In the case of the
PLA/G5 material, the heat of fusion (DHf) and the heat
of crystallization (DHc) of the samples were recalculated
according to the amount of inorganic phase present at
each time. The new heats of fusion and crystallization
(DH 0f and DH 0c ) were calculated as: DH 0 = DH/Xp, where
 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419 413

Xp is the fraction of polymer matrix present at each The radiation was supplied by an Ar+ laser tuned to
time. This fraction was determined by means of a stan- the green line at 516.5 nm. Data were taken at frequen-
dard ash method. The specimens were heated in an elec- cies between 300 and 1400 cm
1 using a Jobin Yvon
tric furnace at 600 C for 3 h to burn out the organic T64000 spectroscope.
matrix and then reweighed in order to calculate the ller
fraction.
3. Results
2.5. Size exclusion chromatography (SEC)
3.1. Weight loss
Weight average molecular weight (Mw) of PLA and
PLA/G5 composite samples was determined by size
Fig. 1 displays the weight variation of the PLA and
exclusion chromatography (SEC) using a modular sys-
PLA/G5 glass plates during the degradation period.
tem, composed of an isocratic pump, a vacuum degas-
For comparison, the weight loss for the glass G5 has
ser, a refractive index detector (all from Knaeur), and
also been included as measured in a previous study
a viscometer/right angle laser light scattering (RALLS)
[12]. However, it has to be kept in mind that this curve
dual detector (T60, Viscotek). Separations were per-
corresponds to the degradation of a block of the glass,
formed in a set of three PLgel 10 lm MIXED-B col-
and not to particles smaller than 80 lm, which is the
umns (Polymer Laboratories). Narrow polystyrene
particle size incorporated in the G5/PLA composite
standards (Polymer Laboratories) were used for calibra-
material. The PLA weight loss increased slowly during
tion. Analysis were performed at 40 C, using THF as
the whole period. However, for the PLA/G5 composite
the eluent at a ow-rate of 1.0 ml/min. PLA and PLA/
a more complex behaviour was observed. After 3 weeks
G5 samples were dissolved in chloroform (2 mg/ml), di-
of degradation, the composite material experienced a
luted in THF and ltered (0.22 lm) before injection.
weight loss of 25%. After this time there was a decrease
PLA/G5 samples were previously centrifuged at
in the weight loss, which means that the material experi-
1000 rpm for 5 min in order to eliminate the glass parti-
enced a weight gain. This increase was attributed to the
cles. The Mw loss percentage was calculated using an
formation of a new calcium phosphate phase that pre-
equation similar to the one presented for the mass loss.
cipitated on the material surface.
2.6. Surface characterization
3.2. Evolution of the molecular weight of the PLA
The evolution of the materials surface was evaluated
by means of scanning electron microscopy (SEM), X-ray Mw values obtained during processing and degrada-
diraction (XRD), Raman spectroscopy and Fourier tion of the PLA and PLA/G5 samples, as well as the
transform infrared (FTIR) spectroscopy. evolution of the molecular weight loss percentage are
At dierent degradation times, the materials were re-
moved from the SBF and washed with deionised water.
The materials were coated with gold and their surface
was observed by SEM (coupled with energy dispersive 25
X-ray (SEM-EDX), Jeol JSM 6400) to evaluate the evo-
lution of the material morphology during degradation. 20
The precipitates formed on the surface of the com-
PLA
posite were scratched and analysed by XRD in a
Weight loss (%)

15 PLAG5
BraggBrentano h/2h Siemens D-500 diractometer G5
with CuKa radiation selected by means of a secondary
graphite monochromator. The starting and the nal 2h 10

angles were 4 and 70, respectively. The step size was

0.05 2h and the measuring time 3 s per step. 5
For FTIR spectroscopy, the scratched precipitates
were mixed with potassium bromide (KBr) and very thin 0
tablets were obtained for the analysis. The FTIR spectra
of the samples were obtained using a FTIR spectropho- 0 1 2 3 4 5 6
tometer (Bomem MB-120) at frequencies in the 400 Time (weeks)
4000 cm
1 range.
Fig. 1. Weight loss versus degradation time for the two materials
For FTRaman spectroscopy, the analysis was car- studied (error bars not shown if smaller than symbols). The weight loss
ried out directly on the surface of the precipitates lo- curve of the glass (G5) has also been included to illustrate its
cated on the composite surface without scratching. degradation behaviour (see Ref. [12]).
414 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419

Table 1 of molecular weight was slightly higher for the PLA

Molecular weight loss values of PLA after dierent stages of the than for the PLA/G5 composite.
preparation process and dierent degradation times (mean SD of
three injections)
3.3. DSC
Material Mw (105)
PLAraw 3.34 0.03 Table 2 shows a summary of the characteristic
PLAlm 3.39 0.02
PLApressed (PLA, 0 weeks) 2.10 0.05
parameters obtained by DSC both for the plain PLA
PLA, 2 weeks 1.72 0.02 and for the PLA matrix of the composite, throughout
PLA, 4 weeks 0.92 0.04 the degradation period. A decrease in the glass transi-
PLA, 6 weeks 0.68 0.04 tion temperature (Tg) was observed for both materials
PLA/G5pressed 1.95 0.03 after dierent periods of immersion in the uid. Simulta-
PLA/G5, 2 weeks 1.27 0.04
PLA/G5, 4 weeks 1.07 0.04
neously, the percentage of crystallinity of the PLA sam-
PLA/G5, 6 weeks 0.76 0.03 ples increased in both materials, gradually in the
PLAraw, as-received; PLAlm, after solvent casting; PLApressed, after
polymeric materials and very steeply in the composite,
hot pressing. especially during the rst 2 weeks of degradation.

3.4. SEM analysis

80
SEM analysis showed remarkable changes in the
70 morphology of the composite surface during the degra-
dation period in SBF as shown in Fig. 3. At the initial
Molecular weight loss (%)

60
time (Fig. 3a), a at polymeric surface interrupted by
50 some protuberances due to the glass particles was ob-
served. However, after 2 weeks of immersion in SBF
40
at 37 C, the surface of the glass particles began to
30 emerge to the composite surface because of their reacti-
PLA vity with the aqueous medium. Some of the most pro-
20 PLA/G5
truding particles lead to the formation of a CaP
10 compound on the composite surface, and others origi-
nated numerous cracks in the material (see Fig. 3b).
0 After 4 weeks the particles continued to break the poly-
0 1 2 3 4 5 6
meric surface and some of the cracks became larger. The
Time (weeks)
CaP precipitated was spread in dierent zones of the
Fig. 2. Molecular weight (Mw) loss versus degradation time for the composite surface (Fig. 3c). After 6 weeks of degrada-
two dierent materials (PLA, PLA/G5) (error bars not shown if tion (Fig. 3d), a wide part of the material surface was
smaller than symbols). covered with the precipitates. The morphology of this
precipitate can be observed in Fig. 4. A globular or
spherical and interconnected porous structure was
formed, and tiny crystals could also be observed on its
shown in Table 1 and Fig. 2, respectively. Results shown
surface (Fig. 4b). Precipitates were formed on both sides
in Table 1 indicate that the initial dissolution of the PLA
of the samples, on the top and also on the side lying on
in chloroform did not aect the Mw. However, the com-
the bottom of the container.
pression-moulding process signicantly aected the
molecular structure of the polymeric matrix, since the
PLA lost 37% of its initial Mw. The incorporation of
the glass particles into the polymer (PLA/G5) did not Table 2
Thermal properties of the two dierent materials along the degrada-
cause signicant dierences in comparison with the
tion process in SBF at 37 C (value SD)
Mw value of the pressed PLA (PLApressed). Concerning
Material Tg (C) %Xc
the degradation behaviour, both materials presented a
similar behaviour during the immersion period studied PLA 58.61 0.58 2.41 1.63
PLA, 2 weeks 56.53 1.86 6.88 1.42
(6 weeks), showing a continuous decrease of the molec-
PLA, 4 weeks 56.43 1.42 8.95 1.58
ular weight. Nevertheless, the results obtained by SEC PLA, 6 weeks 54.03 2.40 9.68 2.93
indicated that the reduction of the molecular weight of PLA/G5 57.95 0.09 3.71 1.32
the PLA in the PLA/G5 composite within the rst 2 PLA/G5, 2 weeks 57.89 0.05 33.56 3.45
weeks of immersion was double that observed for the PLA/G5, 4 weeks 57.85 0.12 38.1 3.01
PLA/G5, 6 weeks 56.82 0.01 38.56 2.62
plain PLA. After 4 and 6 weeks of degradation, the loss
 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419 415

Fig. 3. Composite surface and microstructure of the calcium phosphate layer formed at the material surface after dierent times of immersion in SBF
at 37 C: (a) t = 0 weeks (arrows show some of the G5 particles); (b) t = 2 weeks; (c) t = 4 weeks; and (d) t = 6 weeks.

3.5. Precipitates characterisation

The Ca/P molar ratios of the precipitates formed

after dierent periods of the composite degradation in
SBF are shown in Table 3. EDX results indicated that
the molar ratio increased from approximately 0.54
(which is the Ca/P ratio of the G5 glass) to 1.39 after
4 weeks of degradation, and nally increased to 1.5 after
6 weeks of immersion in SBF.
Fig. 5 displays the X-ray diractogram of the calcium
phosphate compound precipitated on the composite sur-
face. The presence of only one wide peak between 19.72
and 38.5 indicates the formation of an amorphous
phase.
Fig. 6 shows the FTIR spectrum of the CaP precipi-
tated on the composite surface after 6 weeks of degrada-
tion. The spectrum presented the main bands between
400 and 1250 cm
1 (Fig. 6a) followed by a band close
to 1650 cm
1 and a broad band at 30003500 cm
1.
The last two bands are attributed to the presence of

Table 3
Ca/P molar ratio of the G5 glass and the precipitate formed at the
material surface after dierent periods of time (value SD)
Time (weeks) Ca/P ratio
0 0.53 0.04
2 0.91 0.09
4 1.39 0.12
Fig. 4. Morphology of the CaP precipitate formed on the PLA/G5
6 1.5 0.15
composite material.
416 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419

200 slightly acidic due to PLA degradation): octacalcium

180 phosphate (OCP), dicalcium phosphate dihydrate
(DCPD) and an amorphous calcium phosphate (ACP)
160
[20].
140 The bands located at 1133 and 1001 cm
1 could be
Intensity (a.u.)

120 attributed to the m3 and m1 vibrational modes of the

100 PO4 group and that at 737 cm
1 could be due to the
dPOH mode of the DCPD [21]. The bands found at about
80
1041, 560 and 924 cm
1 could be attributed to the m3 and
60 m4 modes of the PO3
4 group and the dPOH mode of the
40 HPO2
4 group of the OCP, respectively [21,22]. How-
20 ever, none of them corresponds to the most characteris-
tic bands of these compounds.
0
10 20 30 40 50 Nonetheless, these last three bands could also be as-
Diffraction angle (2) signed to ACP [23], since they are very close to the main
bands of this CaP according to Stutman et al. [24] (1050,
Fig. 5. Diractogram of the calcium phosphate compound precipi-
940, and 560 cm
1). On the other hand, the bands found
tated at the composite surface.
at 1041 and 737 cm
1 could also be attributed to the
presence of the a and b pyrophosphates (a,b-Ca2P2O7)
[25].
90 The structural changes of the phosphate glass as well
as the structure of the new precipitates formed were ana-
80
lysed by Raman spectroscopy. Raman spectra shown in
Transmitance (a.u.)

70 Fig. 7 correspond to the structural evolution of the glass

particles during the complete period of degradation. The
60 Raman spectrum of the G5 glass presented the charac-
teristic shifts of this metaphosphate glass: the (PO2)asym
50 stretch at approximately 1260 cm
1, the (PO2)sym stretch
at approximately 1170 cm
1 and the (POP)sym stretch
40
near 690 cm
1, which correspond to Q2 or metaphos-
30 phate groups, and the (PO3)sym stretch at 1040 cm
1 that
4000 3500 3000 2500 2000 1500 1000 500 belongs to Q1 units or pyrophosphate groups, the TiO4
(a) Wavenumbers (cm1) stretch at 900 cm
1 and the TiO6 units stretch at
630 cm
1 [2528].
However, the spectrum of the glass surface (precipi-
90
tates) after 4 and 6 weeks of immersion in SBF has
80
undergone a dramatic change. Most of the shifts corre-
Transmitance (a.u.)

70
50
60 45

50 40 CaP precipitate
35
Intensity (a.u.)

40
30
30 25
1250 1000 750 500
20
(b) Wavenumbers (cm1) G5
15
Fig. 6. FTIR spectra of the CaP precipitate after 6 weeks of immersion
in SBF at 37 C. 10

5
400 600 800 1000 1200 1400
water molecules. Fig. 6b shows in more detail the bands
due to the dierent PO vibrations. Wavenumber (cm1)
There are three possible calcium orthophosphates Fig. 7. Raman spectra of the G5 glass and the CaP precipitate after 6
able to develop under these test conditions (37 C, pH weeks of immersion in SBF at 37 C.
 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419
sponding to the Q2 metaphosphate groups have notably
decreased in intensity or disappeared, while new bands
corresponding to Q1, pyrophosphate groups close to
1040 and 745 cm
1 and Q0 units or orthophosphate
groups close to 960 cm
1 were formed due to the degra-
dation process of the glass. The shifts close to 1040 and
745 cm
1 correspond to the presence of the (PO3)sym
and (POP)sym assignments of the b-Ca2P2O7 [25,27]
while the shift located about 960 cm
1 can be attributed
to ACP.
4. Discussion
In this work the evolution of a composite material
consisting of a PLA matrix reinforced with a CaP solu-
ble glass under in vitro simulated physiological condi-
tions was studied. The material was analysed in terms
of its weight loss percentage, morphological changes at
the surface and the variations of the physico-chemical
properties in the polymeric and vitreous phases. The
PLA degradation pattern has been widely studied
[14,15] and, therefore, PLA samples were used as a con-
trol in order to evaluate the eect of the glass particles in
the polymeric matrix during the material degradation.
Some authors have reported studies regarding the
hydrolytic degradation of poly(a-hydroxy) acids blends
with several calcium compounds, such as CaCO3, HA
or b-TCP [16,29]. In general, there is a lack of consensus
in the literature with respect to the eect of dierent
additives on the hydrolytic degradation of polymers.
Not only the basicity of the inorganic phase but also
its solubility play major roles in the modication of
the polymer degradation process, since the availability
of the basic ions or groups able to neutralize the poly-
mer carboxyl end groups depends on the solubility of
the inorganic ller.
In this study, a soluble CaP glass has been incorpo-
rated in the PLA matrix in order to obtain a fully
degradable blend. This fact presents an advantage in
comparison to those polymers blended with poorly sol-
uble inorganic phases, in the case of applications requir-
ing temporary devices.
In general, the results obtained in this study indicated
that the incorporation of the glass particles in the poly-
mer modied the degradation of PLA and induced inter-
esting changes in the material surface morphology.
The weight loss studies showed signicant dierences
between the PLA and the PLA/G5 composite. PLA dis-
played a slow but continuous weight loss, while the com-
posite experienced a more complex behaviour. During
the rst 3 weeks the weight loss was signicantly higher
for the PLA/G5 than for the PLA; however, afterwards
the trend was inverted and an increase in the weight
of the composite was observed. The dierences between
the two materials during the rst 3 weeks of degradation
417
can be attributed mainly to two dierent events occur-
ring in the composite material. The rst event is related
to the presence of the PLA/glass interfaces and surface
cracks (see Fig. 3b) that facilitate both uid penetration
and leakage of the degradation products, accelerating
the degradation of the material. The second event is re-
lated to the solubility and mass loss of the glass particles
themselves. Indeed, G5 is a soluble glass, which dis-
solves by a supercial hydrolytic degradation mecha-
nism [12]. This dissolution mechanism is highly
dependent on the surface area and size as well as on
the pH of the surrounding medium, the solubility being
higher in acidic media. PLA degradation generates an
acidic environment, which thus enhances the degrada-
tion of the small glass particles.
In relation to the weight increase that takes place
after 3 weeks of degradation (see Fig. 1), it could be
attributed to two dierent phenomena: the formation
of a hydrated calcium phosphate precipitate on the
material surface, and the buering eect of the degrada-
tion by-products of the G5 glass on the autocatalytic
degradation process of the polymer. Both points will
be discussed in more detail later.
SEC results showed that during hot pressing a signi-
cant loss of molecular weight was produced both in the
polymer and in the composite material (Table 1). Be-
sides, SEC analysis showed that during immersion in
SBF both materials underwent a progressive decrease
of the Mw, indicating that the degradation process took
place from the beginning of the immersion period, in
agreement with previous studies [14].
The fact that the composite material lost twice the
Mw value of the PLA after 2 weeks of degradation sug-
gested an acceleration of the degradation process of the
composite as compared to the plain PLA. This could be
explained again by the enhancement of the water pene-
tration through the cracks and discontinuities in the
polymer matrix produced by the glass particles. After
4 weeks, the Mw loss rate slightly diminished in the case
of the composite material and became similar to the Mw
loss of the PLA. This behaviour could be attributed to
the buering eect caused by the degradation products
of the phosphate glass, which even if not basic, are less
acidic than the degradation products of the PLA [12]. It
is well known that the degradation mechanism for the
PLA is a bulk mechanism autocatalysed by carboxyl
end groups formed by chain cleavage [30]. Therefore,
the degradation products released from the glass parti-
cles could act as a buering agent, hindering the auto-
catalytic process and hence, levelling o the weight
loss and the Mw loss of the composite.
Thermograms from the DSC analysis of the two
materials, displayed their thermal transitions along the
degradation time (Table 2). The glass transition temper-
ature values for both materials decreased during the
degradation period. This decrease is consistent with
418 M. Navarro et al. / Acta Biomaterialia 1 (2005) 411419
the decrease of the molecular weight during the immer-
sion period reported previously.
Simultaneously, the percentage of crystallinity of the
PLA increased gradually with increasing time, while the
crystallinity of the polymeric matrix of the PLA/G5
composite experienced a strong increase after the rst
2 weeks. This behaviour can be related both to the high
Mw loss underwent for the PLA/G5 material after 2
weeks of degradation and with the weight loss shown
in Fig. 1. Due to enhanced uid penetration in the
PLAglass interface, the hydrolysis process that leads
to the chain cleavage was accelerated, the amorphous re-
gions being the most degraded. As a consequence, the
relative percentage of the crystalline phase increased
[15]. After the initial degradation, the increase in the per-
centage of crystallinity levelled o since the percentage
of amorphous phase, more susceptible of degradation,
was progressively reduced.
In relation to the morphological changes, SEM
observations revealed the formation of calcium phos-
phate precipitates in the composite surface. Fig. 3 shows
the evolution of the material surface after dierent peri-
ods of time. The G5 particles incorporated in the PLA
matrix reacted with the uid, originating a globular
structure that emerged in the material surface. After 6
weeks, the material surface was partially covered by
the CaP formation. The calcium phosphate compound
presented a porous structure whose topography could
enhance the biological response of the composite
material.
G5 degradation yielded to the formation of Q0 units
(orthophosphate groups) and Q1 units (pyrophosphate
groups) according to Raman and FTIR spectra. These
units are able to form bonds with the Ca2+ ions dis-
persed in the SBF, generating the observed precipitates.
XRD and FTIR analysis of the CaP formation
showed that, in general, the spectra were constituted
by broad bands (Figs. 5 and 6) which suggest the pres-
ence of an amorphous phase. Given the resemblance
of the FTIR bands and the Raman shifts to the most
characteristic bands and shifts of an amorphous calcium
phosphate phase and the b-Ca2P2O7, it is possible that
the calcium phosphate compound formed at the com-
posite surface is a combination of these CaP phases,
where the b-Ca2P2O7 is present in a quasi-amorphous
state.
On the other hand, the Ca/P ratio obtained for the
precipitate after 4 and 6 weeks of immersion in the uid,
matched that corresponding to ACP, which can vary be-
tween 1.18 and 1.5, depending on the environmental
conditions, according to Termine et al. [31]. Moreover,
Zahidi et al. [32] reported an ACP corresponding to
an amorphous OCP with a ratio of 1.33, while dierent
studies of the Ca/P molar ratio of ACPs reported values
close to 1.5 [33]. ACP represents a 3040% of the min-
eral phase of bone [31] and it is usually a transient phase
to the formation of OCP, DCPD or even HA [34,35].
However, the fact that the ACP formed did not evolve
to any crystalline phase during the period studied could
be related to the presence of pyrophosphate ions as de-
tected in FTIR and Raman analysis, since their presence
can increase the stability of ACP [36].
The incorporation of glass particles into the poly-
meric matrix not only reinforced the material mechani-
cally [37], but also induced the formation of a CaP
precipitate that can facilitate the interaction between
the material and the bone tissue, which is an important
feature for bone regeneration applications. In addition,
the presence of G5 particles induced other morphologi-
cal changes such as the formation of cracks in the mate-
rial surface due to the reactivity of the glass particles as
well as the separations at the PLA/glass interface. These
cracks could facilitate the release of dierent degrada-
tion products to the surrounding medium, accelerating
the degradation of the material.
Finally, given the dierences between in vitro and in
vivo systems, it will be necessary to analyse the in vivo
behaviour of this novel composite material. Indeed, in
vivo the local chemical changes are smoothed by the
dynamic ow of the physiological uids, which is in con-
trast with the semi-static conditions of in vitro testing.
In addition, it should be assumed that other mechanisms
of degradation would take place when the material is
implanted, such as enzymatic degradation, cell-mediated
degradation and phagocytosis, which indeed could aect
the nal degradation process of the material.
5. Conclusions
A PLA/G5 biodegradable composite material was
developed and its behaviour during a 6 week period of
in vitro degradation was analysed. The calcium phos-
phate glass incorporated in the polymeric matrix in-
duced morphological changes such as the formation of
cracks in the material, which accelerated the degrada-
tion of the material. Additionally, it reacted with the
aqueous medium inducing the formation of a CaP pre-
cipitate, which could enhance the interaction between
the material and the bone tissue.
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