ARTICLE IN PRESS

Biomaterials 26 (2005) 6296–6304

www.elsevier.com/locate/biomaterials

Nano-composite of poly(L-lactide) and surface grafted

hydroxyapatite: Mechanical properties and biocompatibility
Zhongkui Honga,b, Peibiao Zhanga,b, Chaoliang Hea,b, Xueyu Qiua,b, Aixue Liua,b,
Li Chena,b, Xuesi Chena,b,, Xiabin Jinga,b
a
State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences,
Changchun 130022, China
b
Graduate School of Chinese Academy of Sciences, Beijing 100039, China
Received 2 February 2005; accepted 7 April 2005
Available online 23 May 2005

Abstract

In order to improve the bonding between hydroxyapatite (HAP) particles and poly(L-lactide) (PLLA), and hence to increase
mechanical properties of the PLLA/HAP composite as potential bone substitute material, the HAP nano-particles were surface-grafted
with PLLA and further blended with PLLA. The structure and properties of the composites were subsequently investigated by the
mechanical property testing, the differential scanning calorimeter measurements (DSC), the scanning electron microscopy (SEM), the
polarized optical microscopy (POM), and the cell culture. The PLLA molecules grafted on the HAP surfaces, as inter-tying molecules,
played an important role in improving the adhesive strength between the particles and the polymer matrix. At a low content (
4 wt%) of
surface grafted-HAP (g-HAP), the PLLA/g-HAP nano-composites exhibited higher bending strength and impact energy than the pristine
PLLA, and at a higher g-HAP content (e.g., 20 wt%), the modulus was remarkably increased. It implied that PLLA could be
strengthened as well as toughened by g-HAP nano-particles. The results of biocompatibility test showed that the g-HAP existing in the
PLLA composite facilitated both adhesion and proliferation of chondrocytes on the PLLA/g-HAP composite film.
r 2005 Elsevier Ltd. All rights reserved.

Keywords: Polylactide; Hydroxyapatite; Composite; Mechanical properties; Biocompotibility; Cell culture

1. Introduction were lower than those of natural cortical bones. Thus, for
preparing a desired material that presents high mechanical
In order to avoid the second operation imposed on bone performance to match natural bones, inorganic fillers were
defect patient for taking out traditional metallic implants introduced into biodegradable polymers to fabricate filler/
made of stainless steel, titanium and its alloy, many polymer composites, such hydroxyapatite (HAP), b-
attempt have been made to obtain a novel material for tricalcium phosphate, or bio-ceramics [1–8]. Among them,
bone repairing in the last 20 years. Poly(lactic acid) (PLA), composites of HAP particles and biodegradable polymers
poly(glycolic acid), poly(
-caprolactone), and their copo- have been used clinically in various forms due to the good
lymers have attracted wide attention for their biodegrad- osteoconductivity and osteoinductivity of HAP and
ability in the human body. However, the mechanical biodegradability of poly(L-lactide) (PLLA) in the compo-
strength, toughness, and elastic modulus of these polymers sites. But in an ordinary PLA/HAP blending system, only
physical adsorption is achieved between HAP particles and
Corresponding author. State Key Laboratory of Polymer
PLA matrix, consequently, its mechanical properties are
Physics and Chemistry, Changchun Institute of Applied Chemistry, low and its load-bearing applications are limited.
Chinese Academy of Sciences, Changchun 130022, China. Tel.:
The interface adhesion of HAP particles and polymer
+86 431 5262112; fax: +86 431 5685653.
E-mail address: xschen@ciac.jl.cn (X. Chen). matrix plays a very important role among the major

0142-9612/$ - see front matter r 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biomaterials.2005.04.018
 ARTICLE IN PRESS
Z. Hong et al. / Biomaterials 26 (2005) 6296–6304 6297

factors affecting the properties of the PLA/HAP then tested. Normal tensile tests were conducted on an
composites. In order to increase the interfacial strength Instron 1121 machine at a crosshead speed of 1 mm/min.
between the two phases, various methods have been The tensile strength and modulus data were both
tried in the past [9–21]. obtained by averaging over five specimens.
The major objective of this paper is to explore a novel
approach to high mechanical properties of PLA/HAP 2.2.2. Bending strength and impact strength
composites, i.e. the hydroxyl groups on the surface of Rectangular bars having effective dimensions of
the HAP nano-particles are grafted with PLLA by 55  6  4 mm for bending and impact tests were cut
chemical bond and the g-HAP were further blended with from a 4-mm-thick plate which was compressed under
PLLA. In this way, the g-HAP particles can be easily 10 MPa at 175 1C and subsequently annealed at 115 1C
dispersed in the PLLA matrix and strongly tethered to for 1 h and naturally cooled to room temperature. A
the molecular chains of PLLA matrix. Thus, improved rectangle groove was made on one side of the specimens
mechanical properties would be expected. for notched Charpy impact strength measurement. The
3-point bending strength was measured by a universal
testing machine (Instron 1121, UK) at a crosshead speed
2. Materials and methods of 5 mm/min with a span of 40 mm, while measurement
of impact strength was performed on impact testing
2.1. Chemical reagents machine (JJ-20, China).

PLLA was prepared according to the literature [22]. 2.3. Differential scanning calorimeter measurements
Its molecular weight (Mw) was about 300,000. The HAP (DSC)
nano-particles with the atomic ratio Ca/PE1.67 were
acicular crystals of about 100 nm in length and 20–40 nm The thermal properties of PLLA and the PLLA/g-
in width. HAP nano-composites were measured by the differential
The preparation and the surface grafting of HAP scanning calorimetry (DSC-7, from Perkin–Elmer) at a
particles, and the preparation of the PLLA/g-HAP and heating rate of 10 1C/min from 20 to 200 1C. Before the
the PLLA/HAP composites have been described in our measurement, each sample was annealed at 115 1C for
previous paper [22] and are briefly depicted in Fig. 1. 1 h and about 10 mg sample was used.
The amount of grafted polymer determined by thermal Crystallinity of the PLLA in the composites was
gravimetric analysis was about 6 wt%. calculated from the following formula:
Crystallinity ð%Þ ¼ ðDH m =93:7Þ100%,
2.2. Measurements of mechanical properties where DHm indicates the melting enthalpy (J/g) that was
calculated from the fusion peak in DSC curve. And the
2.2.1. Tensile strength value 93.7 (J/g) is the theoretical enthalpy of completely
Dumbbell-shaped tensile test specimens with effective crystalline PLLA [23].
dimensions of 75 mm  5 mm  1 mm were prepared by
press-molding under 10 MPa pressure at 175 1C. One 2.4. Polarized optical microscopy (POM)
batch of these specimens was tested as molded; another
batch was subsequently annealed at 115 1C for 1 h and The crystal morphology of PLLA and the PLLA/g-
HAP composites were observed by POM (Linkam TM
Grafted PLLA 600). The samples used for POM observation were
prepared by casting 1 wt% chloroform solution on a
clean microscope cover-slide and drying at room
Surface grafting
temperature for 3 days to remove the residual solvent.
Mixed The dried samples were melted at 180 1C for 3 min and
HAP
g-HAP with then cooled to 115 1C at a rate of 40 1C/min. The POM
PLLA
observation was carried out as soon as the sample was
cooled to 115 1C.

2.5. Scanning electron microscopy (SEM)

PLLA matrix
SEM (XL30 ESEM FEG, PHILIPS) was used to
examine the selected impact fracture surface of PLLA
PLLA/g-HAP nano-composites
and the PLLA/g-HAP nano-composites, and to inves-
Fig. 1. Method for preparing of the PLLA/g-HAP nano-composites. tigate the dispersion of the fillers in the PLLA matrix.
 ARTICLE IN PRESS
6298 Z. Hong et al. / Biomaterials 26 (2005) 6296–6304

This microscope is a field emission SEM that allows low- 1.0  105 cells (in 1 ml of medium) were then placed, and
voltage operation and high-resolution observation. the plates were incubated at 37 1C and 5% CO2 for 24 h
and 1 week. The cover-slides were washed three times
2.6. Biocompatibility test with PBS, and fixed with 3% glutaraldehyde in PBS at
room temperature for 30 min. After thorough washing,
2.6.1. Specimen preparation for cell assay the cells were dyed with one drop of Giemsa stain
The PLLA/g-HAP, PLLA/HAP nano-composites (SIGMA) for 30 min, washed with distilled water, and
and the control PLLA were dissolved in chloroform to dried in air. Cell attachment and cell morphology were
form a 1 wt% homogeneous solution. The solution observed under the reverse microscope (TE2000U,
(50 ml) was coated onto a 24 mm  24 mm cover-slide NIKON). The pictures were taken by DIGITAL
(treated with 2% dimethyl dichlorosilane (Fluka)/ CAMERA DXM1200F (NIKON), and analyzed with
chloroform solution and dried at 180 1C for 4 h before NIH Image J, area fraction of cells in each cover-slide
use) and the solvent was removed by drying in the air for was obtained.
30 min and following dry in vacuum for 48 h at room
temperature. The cover slides were sterilized with UV
for 30 min.
3. Results and discussion
2.6.2. Cell culture
3.1. Mechanical properties
Full-thickness articular cartilage was obtained from
the femoral head and patellar femoral groove of aborted
Mechanical properties of the PLLA/g-HAP nano-
human fetuses (5 months) presented by a local hospital,
composites were evaluated by tensile, bending, and
and chipped into small pieces at the size of
impact strength measurements. In Fig. 2, the relation-
0.5  0.5  0.5 mm. Chondrocytes were isolated by
ship between the tensile strength and g-HAP content in
trypsin (2.5 mg/ml) (GIBCO) treatment of cartilage for
the composites was demonstrated. Obviously, for the
30 min, followed by type II collagenase (GIBCO) (2 mg/
annealed specimens, all the PLLA/g-HAP composites
ml) digestion for 3–4 h, and filtered through a nylon
gave higher tensile strengths than those of the PLLA/
sieve with a pore size of 88 mm. The cells were rinsed
HAP composites of corresponding compositions. The
three times with 0.1 M PBS by centrifugation at 1500 rpm
higher the g-HAP content, the larger the difference
for 5 min, and cultured in cell culture flasks in a density
between them because the tensile strength of the PLLA/
of 2.0  104/cm2 with Dulbecco’s modified Eagle’s
HAP composite decreased more rapidly with the filler
medium (GIBCO) supplemented with 10% FBS (GIB-
content than that of the PLLA/g-HAP composite. The
CO), 50 mg/l L-ascorbic acid (SIGMA), 10 mM HEPES
(SIGMA), 1.0  105/l penicillin (SIGMA) and 100 mg/l
Streptomycin (SIGMA), in a humidified incubator at
80
37 1C and 5% CO2. The medium was changed every 3 PLLA/HAP as moulded
days. After 7–10 days culture, the monolayer chondro- PLLA/HAP annealed
cytes were removed from the cell culture flasks by 75 PLLA/g-HAP as moulded
trypsin (2.5 mg/ml) and EDTA (0.2 mg/ml) (1:1,v/v) PLLA/g-HAP annealed
treatment, rinsed three times with 0.1 M PBS by 70
centrifugation at 1500 rpm for 5 min. The obtained
Tensile strength (MPa)

chondrocytes were re-suspended in the medium to adjust 65

cell density to 1.0  105/ml, and seeded on the cover-
slides coated with PLLA/g-HAP, PLLA/HAP and 60
PLLA, respectively, in 6-well cell culture plates at a
density of 1.0  105 cells/well, cultured in a humidified
55
incubator containing 5% CO2 at 37 1C. The medium
was replaced every 2 days.
50

2.6.3. Cell proliferation studies

Cell attachment and cell morphology on PLLA/g- 45
HAP, PLLA/HAP and PLLA at different time intervals
were studied. Cover-slides coated with PLLA/g-HAP, 40
PLLA/HAP and PLLA were placed into 6-well plates 0 5 10 15 20
(Costar), washed three times with PBS. Three milliliters Filler content (Wt%)
of medium was added to the wells to prevent the cover- Fig. 2. Effect of the filler content on the tensile strength of the nano-
slide from floating during cell seeding. Into each well, composites.
 ARTICLE IN PRESS
Z. Hong et al. / Biomaterials 26 (2005) 6296–6304 6299

un-annealed samples exhibited similar difference. It was Fig. 4 shows the dependency of the bending strength
noted that the tensile strength of the annealed PLLA/ and the bending modulus on the filler content. There
HAP samples displayed a decreasing tendency with the was a slight increase in the bending strength at low g-
HAP contents, while the annealed PLLA/g-HAP HAP contents up to 4 wt%. Beyond 6 wt%, however,
samples exhibited a maximum strength (about 75 MPa) the bending strength began to decrease almost in a linear
at g-HAP content of 4 wt%. All these experimental facts fashion, while the bending modulus was increased
indicated that the g-HAP play an important role in remarkably with the increasing g-HAP content. In
improving tensile strength of the composite, because the addition, the PLLA/g-HAP composite always showed
grafted PLLA molecules can enter the PLLA phase to higher bending strength and bending modulus than the
mix, crystallize and entangle with the molecular chains corresponding PLLA/HAP composite. These results
of PLLA matrix, thus the g-HAP particles are strongly were similar to those of the tensile test above.
tethered to PLLA matrix in the PLLA/g-HAP compo- The relationship between the inorganic particle
sites. The tensile properties of the composites were content and the notched Charpy impact energy for the
higher than those in our early reports [22], because the PLLA/HAP and the PLLA/g-HAP composites is illu-
crystallizable and relatively high Mw PLLA was used in strated in Fig. 5. Obviously, the PLLA/HAP and the
the present work. PLLA/g-HAP composites exhibited a maximum at 2
On the other hand, the tensile modulus of the and 4 wt% of the particle content, respectively. Beyond
composite increased with particle content, no matter these contents, the impact energy decreased with
the specimens were annealed or not, as shown in Fig. 3. increasing particle content rapidly. But the PLLA/g-
This is because the HAP or g-HAP particles reinforced HAP composite always showed higher impact energy
the composites successfully [23]. The g-HAP particles than the corresponding PLLA/HAP composite.
did not make the remarkable difference in tensile In summary, compared to the ordinary PLLA/HAP
modulus at a lower given particle content. This is composite, the PLLA/g-HAP composite showed im-
because the tensile modulus is measured at small proved tensile strength, bending strength, bending
deformation. At this stage, the PLLA tie-molecules do modulus and impact energy at the particle content of
not respond to the deformation. But at higher filler 4 wt%. However, the properties decreased with further
content, the PLLA/g-HAP composite displayed higher increasing filler content for both PLLA/g-HAP and
modulus than PLLA/HAP composite. Moreover, an- PLLA/HAP, but the decrease rate of the former was less
nealing at 115 1C for 1 h really improved the tensile than that of the latter. The tensile modulus and the
modulus, because the crystallinity was increased during
annealing, as revealed in the next section by DSC.
150 6
PLLA/g-HAP
PLLA/HAP
140
PLLA/g-HAP
4.0 PLLA/HAP
130

120
5

Bending modulus (GPa)

Bending strength (MPa)

3.5
Tensile modulus (GPa)

110

3.0 100

90
2.5 4

80
PLLA/HAP as moulded
2.0 PLLA/HAP annealed 70
PLLA/g-HAP as moulded
PLLA/g-HAP annealed
60
1.5 3
0 5 10 15 20 0 5 10 15 20
Filler content (Wt%) Filler content (Wt%)

Fig. 3. Plot of the tensile-modulus of nano-composite versus the filler Fig. 4. The dependency of bending strength and bending modulus on
contents. the filler content in the nano-composites.
 ARTICLE IN PRESS
6300 Z. Hong et al. / Biomaterials 26 (2005) 6296–6304

bending modulus increased with increasing filler content ical properties compared to the ordinary PLLA/HAP
for both PLLA/g-HAP and PLLA/HAP, but the composite in the filler content range examined. These
increase rate of the former was more than that of the improvements could be attributed to the presence of the
latter. Therefore, it could be concluded that the PLLA/ PLLA molecules grafted on the surface of HAP
g-HAP composite always exhibited improved mechan- particles. The grafted PLLA molecules played a role of
tie molecules between the reinforcing particles and
PLLA matrix.
5
PLLA/g-HAP
PLLA/HAP
3.2. DSC and POM observations
4
Impact strength / (Kj/m2)

As shown in previous section, annealing treatment

had a significant influence on the mechanical properties
3 of the PLLA/HAP and the PLLA/g-HAP composites.
In order to understand the reason, the composites were
investigated by DSC and POM.
2 The data calculated from DSC curves of pure PLLA
and the PLLA/g-HAP nano-composites in different
contents of g-HAP particles were listed in Table 1. It
1 was found that the crystallinity of the PLLA matrix
increased with the increasing g-HAP content from 1 to
10 wt% in the composite, beyond 10 wt%, however, the
0 5 10 15 20 crystallinity of the PLLA matrix began to decrease
Filler content (Wt%)
gradually, while the glass transition temperature (Tg)
Fig. 5. Notched Charpy impact energy as a function of the filler and the melting temperature (Tm) seemed to be
content in the nano-composites. independent of loading of g-HAP particles.

Fig. 6. POM micrographs of PLLA (A, B, C), PLLA/(8 Wt%)HAP (D, E, F), and PLLA/(8 Wt%) g-HAP (G, H, I).
 ARTICLE IN PRESS
Z. Hong et al. / Biomaterials 26 (2005) 6296–6304 6301

Fig. 6 showed the POM micrographs of pure PLLA, an effective heterogeneous nucleating agent. As a result,
the PLLA/HAP composite, and the PLLA/g-HAP the total crystallization speed of the PLLA/g-HAP
composite films. It was observed that at the initial composite was much higher than those of pure PLLA
crystallization stage, the PLLA/g-HAP composite dis- and the PLLA/HAP composite.
played the highest nucleation speed among the three
samples. It indicated that the g-HAP particles acted as
3.3. Micrographs of the fractured surfaces

Table 1 The SEM analysis has been performed in order to

Thermal properties of PLLA/g-HAP nano-composites determined by investigate the fracture mode of the three materials and
DSC the dispersion homogeneity of g-HAP particles in the
polymeric matrix. The micrographs have been taken on
Weight ratio of Tg(1C) Tm(1C) DHm(J/g)a Xc
PLLA/g-HAP (%)b
the impact fracture surface. For PLLA sample, there
were a lot of the parallel fracture lines in the direction of
100/0 60.3 158.2 34.5 36.8 stress (Fig. 7a), and upon high magnification (Fig. 7b)
98/2 61.4 158.3 35.8 38.2 some drawn deformations were seen at the edge of these
95/5 61.2 160.3 37.6 40.1
90/10 59.5 161.2 39.5 42.1
large fracture lines. Thus, the impact behavior of the
85/15 59.8 159.4 37.2 39.7 PLLA materials should be attributed to the semi-brittle
80/20 59.3 159.2 37.3 39.8 failure. But, for the PLLA/g-HAP composite containing
a
4 wt% g-HAP particles, the tough characteristics were
The DHm was calculated with respect to the weight of all PLLAs
demonstrated with a rough fracture surface due to the
present in the sample, including PLLA matrix and grafted PLLA.
b
Crystallinity of all PLLAs present. severely deformed PLLA matrix. The addition of g-HAP

Fig. 7. SEM micrographs of the impact fracture surface of PLLA (a)  130, (b)  2200; PLLA/g-HAP (4 wt%), (c)  130, (d)  8500; PLLA/g-
HAP (15 wt%), (e)  130, (f)  8500. The arrows show the agglomerate g-HAP particles in PLLA matrix.
 ARTICLE IN PRESS
6302 Z. Hong et al. / Biomaterials 26 (2005) 6296–6304

particles changed significantly the appearance of the PLLA/g-HAP composites. The composites prepared in
impact fracture surface to a pattern of multiple cracks. It this study and pure PLLA materials were subjected to
was also shown that few exposed particles were seen in the biocompatibility test using primary cultures of
the nano-composite containing 4 wt% g-HAP, since they human chondracytes cells as the model system (Fig. 8).
were completely embedded in the PLLA matrix. When It has been well known that cell adhesion is an
the g-HAP particle content increased to 15 wt% (Fig. 7e), important cellular process because it directly influences
a typical morphology of brittle failure with a smooth the following proliferation of cell and forming of bone
fracture surface and agglomerated g-HAP particles with tissue. In general, cell behavior and interaction with a
several micrometer in diameter (as indicated by the bioactive material surface are dependent on properties
arrows in Fig. 7f) were observed. such as topography, surface charge and chemistry
[24–27].
3.4. Biocompatibility test As shown in Fig. 9, the presence of bioactive g-HAP
particles in PLLA may have positive biological effects
In order to be used as the biomedical application, it is because the loss of the g-HAP particles that occur in
necessary to investigate the biological behaviors of the contact with the culture medium results in a coarse

Fig. 8. Morphology change of the human condracytes during the cell culture on PLLA (A, B), PLLA/HAP composite (C, D), and PLLA/g-HAP (E,
F) composite films.
 ARTICLE IN PRESS
Z. Hong et al. / Biomaterials 26 (2005) 6296–6304 6303

effects in the composites. These improvements could be

1(d) 7(d)
ascribed firstly to the grafted-PLLA molecules, which
40
played a role of tie molecules between the fillers and the
PLLA matrix, and secondly to the g-HAP particles
Area fractions (%)

30 which were uniformly distributed in the composites and

played the role of the heterogeneous nucleating agents in
the crystallization of the PLLA matrix. The PLLA/g-
20 HAP composites also demonstrated improved cell
compatibility due to the good biocompatibility of the
HAP nano-particles and the more uniform distribution
10
of the g-HAP nano-particles on the film surface. All of
these results indicated that the PLLA/g-HAP nano-
0 composites might have a promising medical application
PLLA PLLA/HAP PLLA/g-HAP in bone repair and in bone tissue engineering.
Materials
Fig. 9. Area fractions of the condracytes adhered and proliferated on
the surface of the PLLA, PLLA/HAP, and PLLA/g-HAP films.
Acknowledgements

This project is financially supported by the National

surface for the cell adhesion and proliferation. In Natural Science Foundation of China (No. 50273038,
addition, the g-HAP particles disengaged from compo- 20274048, 50373043), National Fund for Distinguished
site and exposed to body fluid might induce a micro- Young Scholar (No. 50425309) and the ‘‘863’’ Project
environment change, i.e., the alkalinization of the (2002AA326100) from the Ministry of Science and
medium, which has a positive influence on cell Technology of China, as well as by Chinese Academy
metabolism [28]. of Sciences (No. KJCX2-SW-H07).
When the PLLA/HAP composite was immerged in
body fluid, the water could easily permeate to the inner
side of the composite, resulting in a rapid degradation References
from the out side to the inner side and prompting the
lose of HAP filler, owing to the relatively low-interface [1] Kikuchi M, Suetsugu Y, Tanaka J, Akao M. Preparation and
adhesion between the particles and PLLA matrix, and mechanical properties of calcium phosphate/copoly L-lactide
several lacunas existed in the aggregated HAP particles composites. J Mater Sci Mater Med 1997;8:361–4.
[2] Shikinami Y, Okuno M. Bioresorbable devices made of forged
in composites. So, the PLLA/HAP composite seemed to composites of hydroxyapatite (HA) particles and poly L-lactide
be more effective for the cell adhesion in the initial stage (PLLA): Part I. Basic characteristics. Biomaterials 1999;20:
of cell culture. However, this advantage of the PLLA/ 859–77.
HAP composite was lost with the excessively rapid [3] Shikinami Y, Okuno M. Bioresorbable devices made of forged
degradation and destruction of the composite film on composites of hydroxyapatite (HA) particles and poly L-lactide
(PLLA). Part II: practical properties of miniscrews and mini-
which the cell adhered and proliferated. As shown in plates. Biomaterials 2001;22:3197–211.
Fig. 8D, after 7 days of incubation, a lot of white area [4] Wang M, Bonfield W. Chemically coupled hydroxyapatite-
appeared on the PLLA/HAP film surface because of the polyethylene composite: structure and properties. Biomaterials
degradation and damage of the material. In contrast, the 2001;22:1311–20.
[5] Choi D, Marra KG, Kumta PN. Chemical synthesis of hydro-
degradation behavior of the PLLA/g-HAP composite
xyapatite/Poly(
-caprolactone) composites. Mater Res Bull 2004;
seems to be more appropriate for the chondracytes 39:417–32.
growth on it. [6] Calandrelli L, Immirzi B, Malinconico M, Volpe MG, Oliva A,
Della Ragione F. Preparation and characterisation of composites
based on biodegradable polymers for ‘‘in vivo’’ application.
4. Conclusions Polymer 2000;41:8027–33.
[7] Boccaccini AR, Maquet V. Bioresorbable and bioactive polymer-
Bioglasss composites with tailored pore structure for tissue
Uniform g-HAP/PLLA nano-composites were suc- engineering applications. Compos Sci Technol 2003;63:2417–29.
cessfully prepared and exhibited improved mechanical [8] Verrier S, Blaker JJ, Maquet V, Hench LL, Boccaccini AR.
properties compared to corresponding PLLA/HAP PDLLA/Bioglasss composites for soft-tissue and hard -tissue
composites. Especially at g-HAP content of 4 wt%, the engineering: an in vitro cell biology assessment. Biomaterials 2004;
25:3013–21.
PLLA/g-HAP showed a maximum in tensile strength, [9] Borum-Nicholas L, Wilson Jr OC. Surface modification of
bending strength, and impact energy. It implied that the hydroxyapatite. Part I. Dodecyl alcohol. Biomaterials 2003;
g-HAP particles had both reinforcing and toughening 24:3671–9.
 ARTICLE IN PRESS
6304 Z. Hong et al. / Biomaterials 26 (2005) 6296–6304

[10] Nishizawa K, Toriyama M, Suzuki T, Kawamoto Y, Yokugawa [19] Liu Q, de Wijn J, van Blitterswijk CA. Covalent bonding of
Y, Nagata F. Surface modification of calcium phosphate ceramics PMMA, PBMA, and poly (HEMA) to hydroxyapatite particles.
with silane coupling agents. Chem Soc Japan 1995;1:63–7. J Biomed Mater Res 1998;40:257–63.
[11] Dupraz AMP, de Wijn JR, v.d. Deer S, de Groot K. [20] Dong GC, Sun JS, Yao CH, Jiang GJR, Huang CW, Lin FH. A
Characterization of silane-treated hydroxyapatite powders for study on grafting and characterization of HMDI-modified
use as filler in biodegradable composites. J Biomed Mater Res calcium hydrogen phosphate. Biomaterials 2001;22:3179–89.
1996;30:231–8. [21] Borum L, Wilson Jr OC. Surface modification of hydroxyapatite.
[12] Misra DN. Adsorption of zirconyl salts and their acids on Part II. Silica Biomater 2003;24:3681–8.
hydroxyapatite: Use of the salts as coupling agents to dental [22] Hong ZK, Qiu XY, Sun JR, Deng MX, Chen XS, Jing XB.
polymer composite. J Dent Res 1985;12:1405–8. Grafting polymerization of L-Lactide on the surface of hydro-
[13] Liu Q, de Wijn JR, Bakker D, van Blitterswijk CA. Surface xyapatite nano-crystals. Polymer 2004;45:6705–13.
modification of hydroxyapatite to introduce interfacial bonding [23] Xie XL, Liu XQ, Li Robert KY, Zhou XP, Zhang QX, Yu ZZ,
with polyactiveTM70/30 in a biodegradable composite. J Mater Mai YW. Rheological and mechanical properties of PVC/CaCO3
Sci Mater Med 1996;7:551–7. nanocomposites prepared by in situ polymerization. Polymer 2004;
[14] Liu Q, de Wijn JR, van Toledo M, Bakker D, van Blitterswijk 45:6665–73.
CA. Polyacids as bonding agents in hydroxyapatite/polyester- [24] Verrier S, Blaker JJ, Maquet V, Hench LL, Boccaccini AR.
ether (polyactiveTM70/30) composites. J Mater Sci Mater Med PDLLA/Bioglasss composites for soft-tissue and hard-tissue
1998;9:23–30. engineering: an in vitro cell biology assessment. Biomaterials 2004;
[15] Wang XJ, Li Y, Wei J, de Groot K. Development of biomimetic 25:3013–21.
nano-hydroxyapatite/poly (hexamethylene adipamide) compo- [25] Burridge K, Fath K. Focal contacts: transmembrane links
sites. Biomaterials 2002;23:4787–91. between the extracellular matrix and the cytoskeleton. Bioessays
[16] Liu Q, van Blitterswijk CA. A study on the grafting reaction of 1989;10(4):104–8.
isocyanates with hydroxyapatite particles. J Biomed Mater Res [26] Anselme K. Osteoblast adhesion on biomaterials. Biomaterials
1997;40:358–64. 2000;21(7):667–81.
[17] Liu Q, de Wijn JR, de Groot K, van Blitterswijk CA. Surface [27] Hott M, Noel B, Bernache-Assolant D, Rey C, Marie PJ.
modification of nano-apatite by grafting organic polymer. Proliferation and differentiation of human trabecular
Biomaterials 1998;19:1067–72. osteoblastic cells on hydroxyapatite. J Biomed Mater Res 1997;
[18] Liu Q, de Wijn JR, van Blitterswijk CA. Composite biomaterials 37(4):508–16.
with chemical bonding between hydroxyapatite filler particles and [28] Busa WB, Nuccitelli R. Metabolic regulation via intracellular pH.
PEG/PBT copolymer matrix. J Biomed Mater Res 1998;40:490–7. Am J Physiol: Regul Integr Comp Physiol 1984;246:R409–38.