Available online at www.sciencedirect.

com

Surface & Coatings Technology 202 (2008) 3142 3147

www.elsevier.com/locate/surfcoat

Electrochemical deposition of octacalcium phosphate micro-fiber/chitosan

composite coatings on titanium substrates
Xiong Lu a,b,, Yang Leng b , Qiyi Zhang b,c
a

Key Lab of Advanced Technologies of Materials, Ministry of Education, School of Materials Science and Engineering,
Southwest Jiaotong University, Chengdu 610031, China
Department of Mechanical Engineering, Hong Kong University of Science and Technology, Kowloon, Hong Kong, China
c
The College of Chemical Engineering, Sichuan University, Chengdu 610065, China
Received 13 July 2007; accepted in revised form 19 November 2007
Available online 4 December 2007

Abstract
Calcium phosphate/chitosan composites have been extensively studied as bone substitutes, tissue engineering scaffolds, or bone cements. In
the present study, we have prepared octacalcium phosphate (OCP) micro-fiber/chitosan composite coatings through an electrochemical deposition
method. OCP coatings with microporous structure, which are woven by micro-fibers with 2030 m in length and 0.11 m in width, have a
good ability to incorporate chitosan. This novel OCP micro-fiber/chitosan composite coating could have broad applications in the biomedical
engineering.
2007 Elsevier B.V. All rights reserved.
PACS: 81.15.Pq; 87.68.+z
Keywords: Electrochemical deposition; Composite coatings; Octacalcium phosphate; Chitosan

1. Introduction
Titanium is the most widely used metallic biomedical
materials used in orthopedics due to its good mechanical
properties. Various techniques have been developed to deposit
bioceramic coatings on titanium substrates to improve their
surface bioactivity as reviewed by de Groot et al. [1]. Among
them, electrochemical deposition (ED) is one of the most
commonly used methods. Ease of processing control, variability
of the coating composition, the possibility of protein delivery
and the suitability for complex implant geometries have made
the ED method increasingly popular [2]. Different types of
calcium phosphate (Ca-P) bioceramic coatings have been
achieved with the ED method, such as dicalcium phosphate
dehydrate (DCPD, CaHPO42H2O) [3], dicalcium phosphate
Corresponding author. Key Lab of Advanced Technologies of Materials,
Ministry of Education, School of Materials Science and Engineering, Southwest
Jiaotong University, Chengdu 610031, China. Tel.: +86 28 87634023; fax: +86
28 87601371.
E-mail addresses: luxiong_2004@163.com (X. Lu), meleng@ust.hk
(Y. Leng), qyzhang-scu@163.com (Q. Zhang).
0257-8972/$ - see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.surfcoat.2007.11.024

anhydrous (DCPA, CaHPO4) [4], octacalcium phosphate (OCP,

Ca8(HPO4)2(PO4)45H2O) [5], and hydroxyapatite (HA, Ca10
(OH)2(PO4)6) [6,7].
In recent years, there is a tendency to develop organic
materials/Ca-P composite coatings, such as protein/Ca-P,
collagen/Ca-P, on Ti substrates in order to obtain new types
of coatings that are more biocompatible and bioactive [8,9].
Chitosan/Ca-P coatings also attract much research interesting
[1015]. Chitosan is the copolymer of glucosamine and Nacetyl glucosamine that derives from partially deacetylated
chitin and the idealized chemical structure of chitosan is shown
in Fig. 1 [16]. In the past twenty years, chitosan has drawn
considerable attention in the biomedical areas because it has
many important properties such as good biocompatibility,
minimal foreign body reaction, excellent antimicrobial activity,
the ability to be molded in various geometries, and the
suitability for cell ingrowth and osteoconduction [17]. Nowadays chitosan become one of the most promising biopolymers
for tissue engineering and orthopedic applications and considerable attention has been given to chitosan-based materials.
Chitosan/CaP composites have also been widely studied as
bone substitute, tissue engineering scaffolds, and bone cements,

X. Lu et al. / Surface & Coatings Technology 202 (2008) 31423147

Fig. 1. The chemical structure of idealized chitosan.

which demonstrates increased bioactivity and biodegradation

together with sufficient mechanical strength [1822].
Heretofore there are several reports about Ca-P/Chitosan
composite coatings prepared by ED method. Redepenning et al.
prepared composite coatings containing DCPD/chitosan by ED
and the composites were converted to HA/chitosan composites
through bathing the samples in 0.1 M NaOH for 7 days at room
temperature [12]. Wang et al. developed a hybrid Ca-P/chitosan
coating through ED and this hybrid coating showed an
improved bone marrow stromal cell attachment [10,11]. Note
that the Ca-P phase in the hybrid coatings was a mixture of OCP
and HA. Pang et al. employed electrophoretic approach to
deposit chemically precipitated HA nanoparticles and chitosan
together to obtain composite coatings with pure HA phase [14].
OCP is one important type of Ca-P bioceramics that has been
regarded as the precursor of biological apatite and recent studies
has described the positive role of OCP in osteoconduction and
osteoinduction because it is more resorbable and enhances more
bone formation than HA does [23,24]. To the author's
knowledge, although there were several reports about controlling OCP growth under various condition [25,26], the study of
pure OCP micro-fiber/chitosan composite coatings on titanium
substrates is still missing. The objective of this study is to
prepare pure OCP micro-fiber/chitosan coatings on Ti substrate
using ED method. In this paper, experimental results on the
fabrication and characterization of OCP/chitosan composite
coatings were reported and the electrochemical deposition
mechanism was also discussed.
2. Experimental
Titanium specimens used in this work were commercial pure
titanium (Grade 4, CP Ti) obtained from Baoji Special Iron and
Steel Co. LTD., China. Before ED processing, the titanium was
cut to make plate specimens of 10 mm 10 mm 1 mm in
dimensions. The specimens were etched in an acidic mixture to
remove the natural oxide layer and increase the surface
roughness. The acidic etching was conducted in a mixed
solution of 98% H2SO4 and 36% HCl and deionized water
(volume ratio of H2SO4: HCl: H2O = 1:1:1) at 60 C for 1 h.
After etching, the specimens were cleaned by ultrasound in
deionized water for 15 min.
ED was conducted in the cell that had three electrodes: a
titanium plate as the working electrode, a platinum plate as the
counter electrode and a saturated calomel electrode (SCE) as the
reference electrode. The titanium plate was welded to connect
copper wire by an electric resistance welder. The weld spot and
the copper wire were sealed with silicone gel. Ca-P/chitosan
coating were prepared on cathodic Ti plates in the electrolyte of

3143

an aqueous solution of 12.5 mM Ca(NO3)2 and 5 mM

NH4H2PO4 and 4 wt.% chitosan. According to theoretical
analysis, high Ca-P concentrations help OCP crystal growth,
which guide us to use high concentration Ca-P solution [27].
The pH value of the electrolyte was buffered by HClTris at
pH = 5. Pure Ca-P coating without chitosan also was prepared as
a comparison. The ED was conducted one hour for each
specimen at room temperature using a potentiostat/galvanostat
(PGP201 Radiometer, Denmark) with the current maintained at
1.0 mA/cm2. During the process, the electrolyte was kept static
without any stirring so as to obtain well crystalline OCP. The
titanium plates with coatings were then rinsed with deionized
water and dried at 50 C overnight.
Scanning electron microscopy (SEM) (JSM 6300, JEOL,
Japan), transmission electron microscopy (TEM) (Philips
CM20, Philips Analytic, The Netherlands) and High Resolution
TEM (HRTEM) (JSM 2010, JEOL, Japan) were used to examine
the morphology and crystal structures of the calcium phosphate
deposits. The TEM samples were extracted from the substrate by
an ultrasonic vibration method [28]. The coatings on the
titanium surfaces were also examined using a thin film X-ray
diffractometer (TF-XRD) (X'pert pro-MPD, PANalytical, The
Netherlands). The TF-XRD measurements were performed on a
stage using a Cu-K (wavelength = 1.54056 ) X-ray source
with a step rate of 0.01 per second. Fourier transform-infrared
spectroscopy (FTIR) (FTS 6000, Bio-Rad, USA) was used to
determine the chemical composition of the deposited coatings.
The tensile adhesion strength of the as-prepared coating was
evaluated using a universal mechanical testing machine (Model
5567, Instron, USA). The sample was fixed on the stage of the
testing machine and the coating was glued a titanium stub
(diameter 10 mm) that was connected to the crosshead through a
grip. The glue was cured for 48 h at room temperature before
tensile testing. A tensile load at the crosshead speed of 1 mm/
min was applied to the coating/substrate interface until failure
occurred. The adhesion strength was calculated as the load at
failure divided by the coated area bonded to the stub. The
reported adhesion strength of the coatings was calculated by
averaging measurements of five specimens.
In order to evaluate bioactivity of the coatings, the coatings
were immersed in 200 ml of an acellular simulated body fluid
(SBF) with ion concentrations close to that of human blood
plasma at 36.5 C for 7 days. The SBF recipe followed that of
Kokubo [29] and was prepared by dissolving reagent grade
NaCl (8.035 g), NaHCO 3 (0.355 g), KCl (0.225 g),
K2HPO43H2O(0.230 g), MgCl26H2O (0.311 g), CaCl2
(0.293 g), and Na2SO4 (0.072 g) into 1 l double-distilled
water and buffering at pH 7.4 with tris-hydroxymethylaminomethane [(CH2OH)3CNH2] and 1 M hydrochloric acid (HCl).
The SBF was refreshed every two days in order to keep the ion
concentration stable.
3. Results
Both the OCP/Chitosan and pure OCP coatings are
uniformly deposited across the surfaces of titanium substrate.
The morphology of coatings under different conditions is totally

3144

X. Lu et al. / Surface & Coatings Technology 202 (2008) 31423147

different. In OCP/chitosan coatings formed at pH = 5, crystalline

OCP micro-fibers were observed (Fig. 2a, b) while large flaky
OCP crystals were found in pure OCP coatings (Fig. 2c).
However, the OCP/chitosan coatings formed at pH = 4.5 have
morphologies similar to those of the pure OCP coatings. SEM
observations (Fig. 2a, b) revealed that the OCP fibers are around
2030 m in length while the widths range from 0.1 to 1 m.
The aspect ratio of as-synthesized fibers could be as large as
100. The variety of length and width of fibers may be due to the
difference of the rate of crystal growth of fibers during synthesis
process. Note that the morphology of OCP fibers are similar to
Fig. 3. TF-XRD spectra of different samples: (a) Ti substrate; (b) Pure OCP
coatings; (c) fiber-like OCP/chitosan coatings.

those obtained by Iijima et al., which might suggest that

chitosan has similar function to amelogenin that controls the
oriented and elongated growth of OCP crystals [26]. Fig. 1a and
b demonstrated that OCP micro-fibers were woven together to
form porous structures. Chitosan was entrapped in the recesses
and covered on the fibers as observed by SEM. On the other
hand, the pure OCP coatings were mainly composed of large
flake-like crystals with the sized around 500 nm in width as
revealed by SEM (Fig. 2c) and TEM micrographs (Fig. 5c).
The TF-XRD spectra showed that the deposits crystals were
OCP. in spite of flake-like or fiber-like crystals. The pure OCP
coatings exhibited distinct diffraction peaks at 2 = 4.7, 26.0,
31.6 corresponding to the (100), (002), (402) crystal plane of
OCP (see Fig. 3). These peaks were the characteristic peaks of
well-crystallized OCP as listed in JCPDS card #79-0423. Fiberlike OCP/chitosan coatings also had all the typical OCP
diffraction peaks which demonstrated that the micro-fibers were
OCP. Besides, the XRD pattern of fiber-like OCP/chitosan
coatings also revealed a weak and broad band at 2 ranged from
10 to 20 that was the contribution from the chitosan content in
the composite coatings.
The clear evidence of chitosan existence in the fiber-like
OCP/chitosan coatings was from FT-IR investigation (Fig. 4).
All the characteristic peaks of OCP were observed in the

Fig. 2. SEM micrographs of OCP/chitosan coatings and pure OCP coatings: (a)
Fiber-like OCP reinforced chitosan coatings; (b) Amplified image of (a); (c)
Flake-like pure OCP coatings.

Fig. 4. FTIR spectra of different samples: (a) Pure OCP coatings; (b) Flake-like
OCP/chitosan coatings.

X. Lu et al. / Surface & Coatings Technology 202 (2008) 31423147

3145

Fig. 5. TEM micrographs of individual OCP fiber and flake: (a) bright field image of OCP fiber; (b) SAD of (a); (c) Bright field image of OCP flake; (d) SAD of (c).

spectra. A broad band related to the OH stretching vibration of

the hydrate water in OCP was noticed at 3400 cm 1. The
spectral range 9501200 cm 1 contains the symmetric and the
asymmetric PO stretching modes of the phosphate groups.
The spectral range 550700 cm 1 contains the bending mode of
the phosphate group [30,31]. In addition, the unique peaks of
OCP are the stretching mode of HPP42 (867 and 917 cm 1) and
the hydrogen bending modes of waters of crystallization in OCP
crystal (~ 1640 cm 1) [2]. Additional distinct peaks due to the
chemical functional groups of chitosan appeared only in the
spectrum of fiber-like OCP/chitosan coatings, which supported
that chitosan well incorporated in the composite coatings. The
peaks at 2966, 2932, 2869 cm 1 caused by aliphatic CH
stretching band of CH2 and CH3 [32] and 1458 cm 1 were
assigned to CH2 [33]. Another major peak at wave number
1730 cm 1 was assign to carbonyl group due to the presence of
acetyl group which confirms that the used chitosan is a partially
deacetylated product [34]. Besides these, the weak band at
1380 cm 1 was assigned to CH bending and CH stretching
of CH3 [35]; the weak band at 1243 cm 1 represented the free
primary amino group (NH2) at C2 position of glucosamine,
which is the major functional group of chitosan. A peak at
1400 cm 1 represented CO stretching mode of primary
alcoholic group (CH2OH) [36]. Note that there were several
regions that the bands of OCP overlapped with that of chitosan:
the significant band of amide group at 1655 cm 1 due to the

C_O stretching (amide I) overlapped with that of OCP

1640 cm 1; the broad band in the region 35503350 cm 1
due to the OH stretching vibration of hydrate water in the OCP

Fig. 6. High resolution TEM micrograph of the interface between OCP and
chitosan.

3146

X. Lu et al. / Surface & Coatings Technology 202 (2008) 31423147

crystal overlapped with the broad band caused by the NH

stretching vibration [37]. Apart from above mentioned, no
extraneous functional group was detected for the composite
coatings. The FTIR results confirmed that chitosan macromolecules were intact well in the composites and did not
stimulate any unwanted impurity phases.
The orientation and crystal structure of an individual OCP
fiber and flake, the interface of OCP/chitosan were further
examined by TEM, selected area diffraction (SAD) and
HRTEM. Fig. 5 presented the typical bright field image and
SAD results of single crystal OCP fibers and flakes. Both the
diffraction patterns of the flake-like and fiber-like crystals were
0)
indexed as that of OCP with B = [110], in which the (11
spacing of 0.938 nm revealed the unique OCP structure [2].
HRTEM showed the lattice fringe of crystalline OCP gradually
transited to amorphous chitosan, which proved that OCP and
chitosan had a smooth interface (Fig. 6). The corresponding
Fast-Fourier Transformation (FFT) pattern (Fig. 6, inset) of the
HRTEM fringes indicated the diffraction pattern of the OCP
structure with the [110] zone axis, which was in excellent
agreement with the SAD patterns in Fig. 5.
The tensile test showed that the adhesion strength of the pure
OCP coatings was 1.31 0.33 MPa while the OCP/Chitosan

composite coatings showed the adhesion strength of 7.13

1.99 MPa. It is well known that the adhesion strength for
coatings formed by deposition is usually rather low, which is
also proved by our results. The adhesion strength was improved
by adding the chitosan into the composite coatings. This
increase in adhesion strength could be attributed to the crosslink
effects of chitosan formed during the electrodeposition.
Bioactivity is defined as the property of the material to develop
a direct, adherent, and strong bonding with the bone tissue [38].
It has been already confirmed that the bioactivity of orthopedic
materials can be examined by their capability of forming a
bone-like apatite on their surfaces in SBF with ion concentrations nearly equal to those of human blood plasma [3941].
Currently, SBF has been widely used for the assessment of the
bioactivity of various materials and for the formation of bone
apatite on various implants in vitro. In the present study, SEM
observation showed that a layer of calcium phosphate (Ca-P)
deposited on both the pure OCP and OCP/chitosan coatings
after 7 days SBF immersion, which indicated the good
bioactivity of these coatings (see Fig. 7).
4. Discussion
The results of this work demonstrated the possibility of the
fabrication of OCP/chitosan composite coatings in acidic
chitosan/supersaturated Ca-P mixing solution through an ED
method. Generally chitosan is insoluble in alkaline conditions.
However, it can dissolve easily in acid solution and takes a
positive charge and becomes a cationic polyelectrolyte through
the following equation [14]:
ChitNH2 H2 O ChitNH
3 H2 O

Then, in the electrochemical cell, electric field enables the

motion of the charged chitosan macromolecules towards the
cathode surface. On the cathode, the following reaction happens
which generate hydroxyl ion (OH), resulting in an increasing
pH at the electrode surface:
2H2 O 2e H2 2OH

Then OCP precipitates are formed on the cathodic titanium

surface with all the reactants needed for Ca-P formation [27]:
OH H2 PO4 HPO2
4 H2 O

3
OH HPO2
4 PO4 H2 O

3
8Ca2 2HPO2
4 4PO4 5H2 O
3
Ca8 HPO2
4 2 PO4 4 5H2 O

At the same time, the chitosan loses its charge and forms an
insoluble deposit on the cathode surface:

ChitNH
3 OH ChitNH2 H2 O

Fig. 7. SEM micrographs show the Ca-P formation on the as-prepared coatings
immersed in SBF after 7 days: (a) Pure OCP coatings; (b) OCP/chitosan
coatings.

Therefore, it can be speculated that two events will take place

at the cathode surface: OCP will precipitate on the substrate due
to locally increased pH; and positively charged chitosan will

X. Lu et al. / Surface & Coatings Technology 202 (2008) 31423147

also move to the cathode by electric attraction [11]. Finally, a

Ca-P/chitosan composite coating will be formed on the cathodic
substrate through ED process from the theoretical point of view.
The most interesting finding of this research is that chitosan
may have the ability to modulate the morphology of OCP
crystals. SEM and TEM micrographs revealed that chitosan was
tightly entrapped within OCP micro-fibers. These results were
further verified by XRD and FTIR analysis. However, it is still
not clear how chitosan interact with OCP during the crystallization process. One explanation might be that chitosan prefers
to be deposited on the microporous coatings woven by OCP
fibers with the high aspect ratio due to the large specific surface
area of the micro-fibers. In the presence of an electric field, the
ionized chitosan is more easily deposited at the cathode surface
and then chitosan itself may also modulate Ca-P precipitation
and help to form OCP micro-fibers. It was hypothesized that the
modulation action of chitosan to some extent is comparable
with that of some proteins, such as amelogenin proteins [25,26].
The modulating action leading to more fibers formation is a
positive feedback and helps to attract more chitosan deposition.
Considering the two independent processes, i.e. OCP and
chitosan deposition, both of them contribute to the formation of
OCP micro-fiber reinforced chitosan composite coatings
simultaneously and synergistically.
5. Conclusions
Composites of chitosan and calcium phosphate have been
studied widely as bone substitute, tissue engineering scaffolds,
or bone cements. This report presented the first stage
experiment results of the OCP micro-fiber reinforced chitosan
composite coatings on titanium substrates synthesized through
an electrodeposition method. OCP has good osteoconductivity
and even can enhance more bone formation than HA can.
Chitosan helps to enhance osteoblast attachment and proliferation and inhibit fibroblast proliferation. The as-prepared OCP/
chitosan coatings combine both the advantages of OCP and
chitosan and the microporous structures of the coatings woven
by OCP fibers is in favor of osteoblast adhesion and tissue
ingrowth. This novel OCP micro-fiber/chitosan composite
coating could be used to increase the bioactivity of titanium
and have broad applications in the biomedical engineering.
Acknowledgement
This project was financially supported by the National
Natural Science Foundation of China (no. 30700172).
References
[1] K. de Groot, J.G.C. Wolke, J.A. Jansen, Proc. Inst. Mech. Eng. 212 (1998)
137.

3147

[2] X. Lu, Z.F. Zhao, Y. Leng, J. Cryst. Growth 284 (2005) 506.
[3] M. Kumar, H. Dasarathy, C. Riley, J. Biomed. Mater. Res. 45 (1999) 302.
[4] M.H. Prado Da Silva, J.H.C. Lima, G.A. Soares, C.N. Elias, M.C.
de Andrade, S.M. Best, I.R. Gibson, Surf. Coat. Technol. 137 (2001) 270.
[5] S. Lin, R.Z. LeGeros, J.P. LeGeros, J. Biomed. Mater. Res. 66A (2003)
819.
[6] M. Shirkhanzadeh, J. Mater. Sci., Mater. Med. 9 (1998) 67.
[7] H.B. Hu, C.J. Lin, P.P.Y. Lui, Y. Leng, J. Biomed. Mater. Res. 65A (2003)
24.
[8] X.L. Cheng, M. Filiaggi, S.G. Roscoe, Biomaterials. 25 (2004) 5395.
[9] Y. Liu, E.B. Hunzikerc, N.X. Randalld, K. de Groot, P. Layrolle,
Biomaterials. 24 (2003) 65.
[10] J. Wang, A. van Apeldoorn, K. de Groot, J. Biomed. Mater. Res. 76A
(2006) 503.
[11] J. Wang, J. de Boer, K. de Groot, J. Dent. Res. 83 (2004) 296.
[12] J. Redepenning, G. Venkataraman, J. Chen, N. Stafford, J. Biomed. Mater.
Res. 66 (2003) 411.
[13] J. Pena, I. Izquierdo-Barba, M. Garca, M. Vallet-Reg, J. Eur. Ceram. Soc.
26 (2006) 3631.
[14] X. Pang, I. Zhitomirsky, Mater. Chem. Phys. 94 (2005) 245.
[15] R.A.A. Muzzarelli, G. Biagini, A. DeBenedittis, P. Mengucci, G. Majni, G.
Tosi, Carbohydr. Polym. 45 (2001) 35.
[16] K. Okuyama, K. Noguchi, M. Kanenari, T. Egawa, K. Osawa, K. Ogawa,
Carbohydr. Polym. 41 (2000) 237.
[17] A.D. Martino, M. Sittinger, M.V. Risbud, Biomaterials 26 (2005) 5983.
[18] Y. Yokogawa, K. Nishizawa, F. Nagata, T. Kameyama, J. Sol-Gel. Sci.
Technol. 21 (2001) 105.
[19] Y. Zhang, M. Zhang, J. Biomed. Mater. Res. 62 (2002) 378.
[20] H.K. Xua, C.G. Simon, Biomaterials 26 (2005) 1337.
[21] Y. Zhang, M. Zhang, J. Biomed. Mater. Res. 55 (2001) 304.
[22] S. Takagia, L. Chow, S. Hirayama, F. Eichmiller, Dent. Mater. 19 (2003)
797.
[23] S. Kamakura, Y. Sasano, T. Shimizu, K. Hatori, O. Suzuki, M. Kagayama,
K. Motegi, J. Biomed. Mater. Res. 59 (2002) 29.
[24] P. Habibovic, C.M. van der Valk, C.A. van Blitterswijk, K. de Groot,
G. Meijer, J. Mater. Sci. Mater. Med. 15 (2004) 373.
[25] M. Iijima, J. Moradian-Oldak, J. Mater. Chem. 14 (2004) 2189.
[26] M. Iijima, Y. Moriwaki, H.B. Wen, A.G. Fincham, J. Moradian-Oldak,
J. Dent. Res. 81 (2002) 69.
[27] X. Lu, Y. Leng, Biomaterials 26 (2005) 1097.
[28] X. Lu, Y. Leng, Biomaterials 25 (2004) 1779.
[29] A. Oyane, H. Kim, T. Furuya, T. Kokubo, T. Miyazaki, T. Nakamura,
J Biomed Mater Res. 65A (2003) 188.
[30] A. Stoch, A. Brozek, S. Bazewicz, W. Jastrzebski, J. Stoch, A. Adamczyk,
I. Roj, J. Mol. Struct. 651653 (2003) 389.
[31] B.O. Fowler, E.C. Moreno, W.E. Brown, Arch. Oral Biol. 11 (1966)
477.
[32] N. Shanmugasundaram, P. Ravichandran, P. Reddy, N. Ramamurty, S. Pal,
K. Panduranga Rao, Biomaterials 22 (2001) 1943.
[33] Y.X. Xu, K.M. Kim, M.A. Hanna, D. Nag, Ind. Crops Prod. 21 (2005)
185.
[34] A. Pawlak, M. Mucha, Thermochim. Acta 396 (2003) 153.
[35] S. Wang, Q. Huang, Q. Wang, Carbohydr. Res. 340 (2005) 1143.
[36] R. Murugan, S. Ramakrishna, Biomaterials 25 (2004) 3829.
[37] K. van de Velde, P. Kiekens, Carbohydr. Polym. 58 (2004) 409.
[38] L.L. Hench, J. Am. Ceram. Soc. 74 (1991) 1487.
[39] T. Kokubo, H.M. Kim, M. Kawashita, Biomaterials 24 (2003) 2161.
[40] T. Kokubo, H.M. Kim, M. Kawashita, T. Nakamura, J. Mater. Sci., Mater.
Med. 12 (2004) 99.
[41] T. Kokubo, H. Takadama, Biomaterials 27 (2006) 2907.