Acta Biomaterialia 7 (2011) 2769–2781

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Acta Biomaterialia
journal homepage: www.elsevier.com/locate/actabiomat

Review

Nanoscale hydroxyapatite particles for bone tissue engineering

Hongjian Zhou, Jaebeom Lee ⇑
Department of Nanomedical Engineering, College of Nanoscience and Nanotechnology, Pusan National University, Miryang 627-706, Republic of Korea

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

Article history: Hydroxyapatite (HAp) exhibits excellent biocompatibility with soft tissues such as skin, muscle and
Received 7 December 2010 gums, making it an ideal candidate for orthopedic and dental implants or components of implants. Syn-
Received in revised form 11 March 2011 thetic HAp has been widely used in repair of hard tissues, and common uses include bone repair, bone
Accepted 16 March 2011
augmentation, as well as coating of implants or acting as fillers in bone or teeth. However, the low
Available online 1 April 2011
mechanical strength of normal HAp ceramics generally restricts its use to low load-bearing applications.
Recent advancements in nanoscience and nanotechnology have reignited investigation of nanoscale HAp
Keywords:
formation in order to clearly define the small-scale properties of HAp. It has been suggested that nano-
Biomaterials
Bioceramics
HAp may be an ideal biomaterial due to its good biocompatibility and bone integration ability. HAp bio-
Nanoscale particles medical material development has benefited significantly from advancements in nanotechnology. This
Hydroxyapatite feature article looks afresh at nano-HAp particles, highlighting the importance of size, crystal morphology
Bone tissue engineering control, and composites with other inorganic particles for biomedical material development.
Ó 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

1. Introduction
Bone is a natural organic–inorganic ceramic composite consist-
ing of collagen fibrils containing embedded, well-arrayed, nano-
crystalline, rod-like inorganic materials 25–50 nm in length [1–
3]. Structural order in bone occurs at several hierarchical levels
and reflects the materials and mechanical properties of its compo-
nents (Fig. 1). Hydroxyapatite (HAp) is chemically similar to the
inorganic component of bone matrix – a very complex tissue with
general formula Ca10(OH)2(PO4)6. The close chemical similarity of
HAp to natural bone has led to extensive research efforts to use
synthetic HAp as a bone substitute and/or replacement in biomed-
ical applications [4,5].
Tissue engineering is intensively researching solutions that
have the potential to reduce the complications related to current
treatment methods. Tissue engineering can be defined as an inter-
disciplinary field that applies the principles of engineering and life
sciences to develop biological substitutes that restore, maintain or
improve tissue function [6]. This concept involves three main strat-
egies: the use of isolated cells or cell substitutes to replace limited
functions of the tissue; utilization of tissue-inducing substances
such as growth factors; and scaffolds to direct tissue development.
An ideal scaffold for bone tissue engineering is a matrix that acts as
a temporary substrate allowing cell growth and tissue develop-
ment. This occurs initially in vitro and eventually in vivo. The scaf-
fold should be able to mimic the structure and biological function
of the native extracellular matrix (ECM) in terms of both chemical
⇑ Corresponding author.
E-mail address: jaebeom@pusan.ac.kr (J. Lee).
composition and physical structure. Scaffolds used for tissue engi-
neering applications should also be biocompatible; able to provide
appropriate mechanical support; exhibit favorable surface proper-
ties such as promoting adhesion, proliferation and differentiation
of cells; and provide an environment in which cells can maintain
their phenotypes.
Recently, HAp has been used for a variety of biomedical applica-
tions, including matrices for drug release control and bone tissue
engineering materials [8,9]. Since HAp has chemical similarity to
the inorganic component of bone matrix, synthetic HAp exhibits
strong affinity to host hard tissues. Chemical bonding with the host
tissue offers HAp a greater advantage in clinical applications com-
pared to most other bone substitutes such as allografts or metallic
implants [10]. The main advantages of synthetic HAp are its biocom-
patibility, slow biodegradability in situ, and good osteoconductive
and osteoinductive capabilities [1,11]. A study by Taniguchi et al.
showed that sintered HAp exhibits excellent biocompatibility with
soft tissues such as skin, muscle and gums. Such capabilities have
made HAp an ideal candidate for orthopedic and dental implants
or components of implants. Synthetic HAp has been widely used
to repair hard tissues. Common uses include bone repair, bone aug-
mentation, as well as coating of implants or acting as fillers in bone
or teeth [12–18]. However, the low mechanical strength of normal
HAp ceramics restricts its use mainly to low load-bearing applica-
tions. Recent advances in nanoscience and nanotechnology have
reignited interest in the formation of nanosized HAp and the study
of its properties on the nanoscale.
Nanocrystalline HAp powders exhibit improved sinterability
and enhanced densification due to greater surface area, which
may improve fracture toughness, as well as other mechanical

1742-7061/$ - see front matter Ó 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.actbio.2011.03.019
2770 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
Fig. 1. The hierarchical structure of bone at its various length scales. The microstructure of cortical bone consists of osteons with Haversian canals and lamellae, and at the
nanoscale, the structural units are collagen fibers composed of bundles of mineralized collagen fibrils. Copyright Elsevier and reproduced with permission [7].
properties [11]. Moreover, nano-HAp, compared to coarser crystals,
is expected to have better bioactivity [19]. Thus, nano-HAp parti-
cles can be utilized for engineered tissue implants with improved
biocompatibility over other implants. Nanotechnology has the po-
tential to significantly benefit development of HAp biomedical
materials. To our knowledge, several reviews of nanocrystalline
calcium orthophosphates have been published in recent years.
For example, Dorozhkin et al. [20,21] reviewed the current state
of technology and recent developments of various nanosized and
nanocrystalline calcium orthophosphates, involved in synthesis
and characterization as well as biomedical and clinical applica-
tions. Moseke et al. [22] reviewed the synthesis and properties of
tetracalcium phosphate (TTCP) in biomaterial applications such
as cements, sintered ceramics and coatings on implant metals;
Johnson et al. [18] reviewed the compression, flexural and tensile
properties of calcium phosphate (CaP) and CaP–polymer compos-
ites for applications in bone replacement and repair; Tran et al.
[23] summarized studies that have demonstrated enhanced
in vitro and in vivo osteoblast functions (e.g. adhesion, prolifera-
tion, synthesis of bone-related proteins and deposition of cal-
cium-containing mineral) on nanostructured metals, ceramics,
polymers, and composites. After reviewing these feature articles
to avoid any redundancy, we focus on calcium orthophosphate,
and characterize its properties in the condition of nano-HAp with
different morphologies and porous structures-materials that offer
great promise as bone substitutes and/or replacements in biomed-
ical applications. Moreover, we summarize how composites of HAp
and other inorganic nanomaterials can enhance the bioactivity and
biocompatibility of HAp – an area that has become the focus of re-
cent research. The remainder of this feature article is organized
into five sections. In the Section 2, the synthesis of morphologically
different nano-HAps is introduced. Section 3 discusses the fabrica-
tion of the porous structure of nano-HAp. Section 4 reviews the
bio-orthopedic properties of nanoscale HAp for application in bone
tissue engineering. Section 5 introduces composites of HAp and
other inorganic nanomaterials for enhancing the bioactivity and
biocompatibility of HAp. Finally, in Section 6, we provide a sum-
mary and our own perspectives on this active area of research.
2. Synthesis of nanoscale HAp
HAp (Ca10(OH)2(PO4)6) nano- and microcrystals with multiform
morphologies (separated nanowires, nanorods, microspheres,
microflowers and microsheets) have been successfully synthesized
by many powder processing techniques, including sol–gel synthe-
sis [24–28], solid state reactions [29], co-precipitation [30], hydro-
thermal reactions [31], microemulsion syntheses [32] and
mechanochemical synthesis [33].
2.1. HAp nanoparticles
From a practical application perspective, suitable nano- or
micromaterials with specific morphologies not only need to be
capable of being synthesized in large quantities with a desired
composition, reproducible size and structure, but also of being
prepared and assembled using environmentally responsible tech-
niques. Recently, environmentally friendly synthetic methodolo-
gies, including molten-salt synthesis, hydrothermal processing,
biomimic synthesis and template synthesis, have been imple-
mented as viable techniques for the synthesis of a range of materi-
als [34,35]. Nanosized HAp particles can be prepared by a variety of
techniques such as mechanochemical synthesis [36], combustion
preparation [37] and various wet chemistry techniques [38,39].
Among the most reported precipitation processes, chemical agents
such as citric acid [40,41], amino acids [42] and ethylenediamine-
tetraacetic acid (EDTA) [43,44] have been used to mediate HAp
nucleation and crystal growth. These modifiers exert significant
control over crystal morphology due to affinity between the mod-
ifying agent and the HAp crystals. However, there has been less fo-
cus on the precipitation kinetics of nucleation and growth, which
are related to the degree of supersaturation, SHAp. This value can
be calculated as follows:
IAPHAp
S¼ ; ð1Þ
K sp
where IAPHAp is the ionic activity product expressed as:
IAPHAp ¼ ½Ca2þ 5 ½PO3 3  5 3
4  ½OH cCa2þ cPO3 cOH : ð2Þ
4
The brackets represent ion concentrations of the respective spe-
cies and c values are the activity coefficients of the ions. Ksp is the
solubility product of HAp. With a higher degree of supersaturation,
a greater driving force for precipitation, i.e. a faster precipitation
rate, was expected with increasing IAPHAp [45].
Biological mineralization (or biomineralization) is the process
of in vivo inorganic material formation. The new theory of ‘‘aggre-
 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
gation-based crystal growth’’ [46] and the new concept of the
‘‘mesocrystal’’ [47] highlight the roles of nanoparticles in biological
crystal engineering. Mimicking the formation of natural CaP, hard
tissues contribute significantly to the biological function of engi-
neered materials. Many advances have been made in biomaterial
research with the rapid growth of nanotechnology. The study of
CaPs is a specific area in nanotechnology, which may be applied
readily in the repair of hard and soft skeletal tissues [48–50]. Moll-
azadeh et al. [51] prepared HAp crystals using an in situ biomi-
metic process in the presence of polyvinyl alcohol (PVA). They
systematically investigated the effects of polymer amount and
molecular weight on the physical properties of HAp crystals. The
results indicated that the development (size and shape) of HAp
nanocrystals precipitated in an aqueous solution of PVA was inver-
sely related to the polymer molecular weight (i.e. the smallest
crystallite size was observed with the highest PVA molecular
weight). It is thought that HAp formation is initiated through the
interaction of Ca2+ ions with the negative side groups on the poly-
mer surface. The larger number of reaction sites in the higher
molecular weight PVA polymer led to a higher number of HAp nu-
clei, and therefore a smaller crystallite size.
The hydrothermal method, a typical solution-based approach,
has proven to be an effective and convenient process to prepare
various inorganic materials with diverse, controllable morpholo-
gies and architectures [52–55]. The environmentally acceptable
advantages of this method include easily controllable reaction con-
ditions, relatively large scale and high yield in terms of quantity of
the desired products, and frequent use of water as the reaction
medium. Zhang et al. [56] developed a general strategy for the syn-
thesis of HAp nano- and microstructures using water as a reaction
medium through a simple hydrothermal process. Several dominant
morphologies were achieved (nanorods, nanowires, microsheets,
burr-like microspheres and microflowers). The pH value plays a
crucial role in obtaining Ca10(OH)2(PO4)6 samples with various
morphologies. The use of trisodium citrate also has an important
Fig. 2. Schematic for the formation and morphology evolution mechanism of Ca5(PO4)3OH samples with various morphologies based upon different pH values [56].
2771
influence on product shape. Possible formation mechanisms for
Ca10(OH)2(PO4)6 nano- and microcrystals with diverse morphology
are presented in Fig. 2. A strong blue emission peak at approxi-
mately 428 nm was observed at room temperature, and the photo-
luminescence (PL) intensity of this emission varied with
Ca10(OH)2(PO4)6 morphology over a range of pH values. CO 2 radi-
cals in the HAp lattice interstitials may be responsible for self-acti-
vated luminescence. These types of phosphors do not contain
metal ions as activators and contain no toxic elements; thus, they
are considered environmentally friendly luminescent materials.
The microsized Ca10(OH)2(PO4)6 sample prepared at pH 5.0 with
strong blue emission, spherical morphology, non-aggregation and
high crystallinity can potentially be used as a new, efficient and
environmentally friendly blue luminescent material.
Qiu et al. [57] prepared nanocrystalline HAp by precipitation
with the aid of ultrasonic atomization using Ca(NO3)24H2O and
(NH4)2HPO4 as raw materials. The results showed that the synthe-
sis method used in this study can effectively shorten reaction time
while improving powder homogeneity compared to other pub-
lished methods. It was also found that addition of a small amount
of the surfactant glycine during precipitation synthesis can reduce
HAp nanoparticle agglomeration. However, Poinern et al. [1] and
LeGeros [11] developed a chemical route to synthesize HAp using
calcium nitrate and potassium hydrogen phosphate as the main
raw materials. This hydrothermal method also used ultrasonic irra-
diation followed by heat treatment to manufacture nanosized HAp.
This study has shown that nano-HAp particles with a spherical
morphology in the nanometer range (approximately 30 nm ± 5%)
can be synthesized using a hydrothermal chemical precipitation
method incorporating low-power sonic irradiation. The crystallin-
ity and morphology of HAp depends on the Ca/P ratio, ultrasonic
irradiation and temperature. An ultrasonic power of 50 W and
temperatures in the range of 400 °C are sufficient to produce the
nano-HAp. This offer an economic route for the synthesis of
nano-HAp with a strong scale-up capability.
2772 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
Li et al. [58] studied the crystalline behaviors of HAp influenced
by different forms of citrates and explained this behavior using the
classical theory of crystal nucleation and growth in solutions. They
found that the supersaturation value decreased, i.e. the speed of
crystal nucleation decreased, with increasing citric acid monohy-
drate (C6H8O7H2O, CiA) concentration or with decreasing triso-
dium citrate (Na3C6H5O72H2O, NaCit) concentration. The
presence of CiA inhibited the fast-growing of (0 0 l) planes and in-
creased the crystallinity of (h k 0) planes. This resulted in homoge-
neously grown particles rather than the prismatic particles
obtained when synthesis is performed without CiA. By raising
the synthesis temperature, finer and more homogenously grown
particles could be prepared in the presence of CiA. With increased
ripening time, a higher crystallinity of HAp with lower amounts of
CaO phase was obtained. This could be explained by crystal growth
in solutions, i.e. prolonged maturation time resulting in a higher
amount of unreacted Ca(OH)2 that can be involved in the crystal
growth of CaP powders.
Utilizing nanotechnology, calcium and phosphate can be
manipulated at the molecular level and assembled to produce
materials with unique structural and functional properties. The
preparation of CaP powders with a particular morphology, stoichi-
ometry, crystallinity and crystal size distribution is important in
biomedicine and materials science. Various processes have been
employed to prepare CaP powders, including co-precipitation
[59], sol–gel process [60,61], spray pyrolysis [62], hydrothermal
synthesis [63,64], emulsion processing [65,66] and mechanochem-
ical method [67]. Among these methods, the sol–gel method has
received the most attention because of its well-known inherent
advantages, namely homogeneous molecular mixing, low process-
ing temperature, and ability to generate nanocrystalline powders,
bulk amorphous monolithic solids and thin films [68]. Natarajan
et al. [69] prepared nanosized HAp particles using the sol–gel
method from the water-based solution of calcium and phosphorus
precursors. In that study, two calcium precursors, i.e. calcium ni-
trate tetrahydrate and calcium acetate, were chosen. The influence
of aging period, pH, viscosity and sintering temperature on the
crystallinity and morphology of HAp particles were investigated
for the two calcium precursors and a triethyl phosphate precursor.
The morphology of nano-HAp when the phosphorous precursor
was used was dependent on the type of calcium precursor used.
Gopi et al. [70] reported the synthesis and characterization of
nano-HAp powders by a novel ultrasonic coupled sol–gel synthe-
sis. The resulting powders were sintered by conventional means
at different temperatures. These results show that nano-HAp pow-
ders synthesized by ultrasonic coupled sol–gel synthesis showed a
remarkable particle size reduction compared with the conven-
tional sol–gel method, hence; these powders could be used as a
coating material in biomedical applications.
With the increasing need to develop clean, non-toxic and envi-
ronmentally friendly techniques, HAp powders have been ex-
tracted using bioproducts such as corals, cuttlefish shells, natural
gypsum, natural calcite and bovine bone [71,72]. Chemical analysis
has shown that these products, typically considered as biowaste,
are rich sources of calcium in the form of carbonates and oxide.
One such biowaste is chicken eggshells. Every day, a million tonnes
of eggshells are generated as biowaste around the globe. Eggshell
represents 11% of the total egg weight and is primarily composed
of calcium carbonate (94%), calcium phosphate (1%) and organic
matter (4%) [37]. Besides being economically cheap and plentiful
in nature, eggshells have been shown to be biocompatible with the
human body during implantation but not osteoconductive. Hence,
converting these powders into HAp prior to implantation is advan-
tageous. Sanosh et al. [73] reported a simple sol–gel precipitation
technique to synthesize nano-HAp powders using CaO derived
from chicken eggshells. Their study shows that biowaste egg shells
can effectively be utilized for synthesizing pure nano-HAp
powders.
Particle formation by sol–gel is a very complex process. It in-
volves nucleation, growth, aggregation and agglomeration [74,75].
Lee et al. [76] also synthesized highly sinterable, nanosized HAp
powders using a wet chemical route with recycled eggshell and
phosphoric acid as calcium and phosphorous sources, respectively.
Raw eggshell was easily converted to CaO by the calcining process,
and phosphoric acid was mixed with the calcined eggshell by a wet
ball-milling method. The observed phases of the powder synthesis
process were dependent on the mixing ratio (wt.%) of the calcined
eggshell to phosphoric acid and the heating temperature. The
ball-milled, nanosized HAp powder, which has an average particle
size of 70 nm, is fully densified at 1300 °C for 1 h. The Ca/P ratio
for the stoichiometric composition of HAp was controlled by adjust-
ing the mixing ratio. In addition, HAp can be successfully produced
from recycled eggshells, seashells and phosphoric acid. The phases
obtained depend on the ratio of calcined eggshells/seashells to
phosphoric acid, the calcination temperature and the mechano-
chemical activation method (ball milling or attrition milling) [77].
Mostaghaci et al. [78] synthesized nanosized HAp using an Iranian
strain of Serratia. Results showed that the optimum powder pro-
duction was achieved at approximately pH 8 and a temperature
of 37 °C. The powder particles were single crystals and ranged in
size from 25 to 30 nm. Moreover, particle shapes and the sizes were
relatively uniform and exhibited lower agglomeration relative to
conventional methods. This powder could be used in the regenera-
tion of bone defects, fabrication of medical implants, and as a vector
for pharmaceutical and biological materials such as genes.
2.2. HAp nanorods and nanoflakes
Bone is a composite, consisting of HAp nanorods embedded in a
collagen matrix. In the biomineralization process of vertebrate
hard tissues, some specific molecules control the nucleation and
growth of inorganic crystals (HAp), resulting in the formation of
hierarchical structure of teeth and bones with superior mechanical
properties [79,80]. Thus, controlled syntheses of apatitic crystals
with various morphologies have been the focus of intensive re-
search in order to understand more completely their biominerali-
zation and utility in industrial and biomedical applications [81].
Inspired by biomineralization, molecular manipulation of inor-
ganic crystals with organic growth modifiers gradually developed
into a powerful tool for the design of novel tissue engineering
materials [82,83].
The molecule–template combination exerts significant control
over crystal morphology and has been discussed in many articles
[84–87]. A variety of organic molecules such as hexadecyltrimeth-
yl-ammonium bromide (CTAB), sodium dodecyl sulfate (SDS), ami-
no acids, proteins and monosaccharides have been used to
synthesize HAp with fibrous- and flake-like morphologies [86,88–
92]. Kalita et al. [93] synthesized bioactive HAp (Ca10(PO4)6(OH)2)
ceramic powder in the lower end of the nanoregime using micro-
wave radiation, which offers several advantages. The applied
microwave power of 600 W, pH of the suspension, mole ratio of
Ca/P in the starting chemicals, and the chelating effect of EDTA
served as important factors in the synthesis of nanocrystalline
HAp powder. Results confirmed a highly crystalline nanopowder
(5–30 nm) with elemental composition of Ca and P in the HAp
phase and possessing mixed (elliptical and rod-shaped) morphol-
ogy. Tari et al. [94] reported that using poly(sodium 4-styrene sul-
fonate) (PSSS) as a nucleation- and growth-controlling agent
resulted in the precipitation of well-crystallized HAp nanoparticles
through microemulsion. During PSSS mixing with a calcium precur-
sor, rod-like micelles were formed, which control the morphology
and crystallization of nano-HAp. The investigations showed that
 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
the obtained HAp nanorods have an average width and length of
about 30 and 200 nm, respectively. Shanthi et al. [95] reported that
CTAB is used as the capturing material for successfully prepared
nano-HAp rods at ambient temperature and normal atmospheric
pressure. A cytotoxicity test using a normal cell line indicated the
biocompatible range of the nanocrystalline HAp and its fairly
non-toxic nature.
Calcium phosphates comprise a large family of compounds with
important biological applications-including osteologic implant for
coatings, grafts, scaffolds and bone cavity fillings, and vehicles for
drug, protein and gene delivery-due to their similarity with the
mineral constituents of human bones and teeth. The properties
of calcium phosphate, including bioactivity, biocompatibility, solu-
bility, mechanical properties and absorption, can be tailored over
wide ranges by controlling particle composition, size, morphology
and assembly. For these reasons, it is of great importance to devel-
op synthesis methods focused on the precise control of particle
size, morphology and chemical composition. Jiang et al. [96] pre-
sented an approach using CaP that offered careful size and struc-
tural control via a template-guided process. In their study, HAp
nanorods and well-aligned hybrid HAp composites were prepared
via a template-guided synthesis procedure (a schematic illustra-
tion of the formation of HAp nanorods is shown in Fig. 3). First,
phosphate ester was used as a structure-directing agent for prepar-
ing HAp nanorods. After hydrothermal treatment, CaP nanorods
with well-controlled particle size and porosity can be obtained.
Second, carboxymethyl cellulose (CMC) molecules were found to
be effective in controlling particle size and the subsequent align-
ment of HAp. HAp composites with well-organized microstruc-
tures were prepared using CMC. CMC has carboxyl groups that
can attract Ca2+ ions, thereby guiding the growth of HAp grains.
These proposed processes can also be applied to the preparation
of CaP materials with well-controlled microstructures using other
similar templates for a wide range of applications.
HAp is the basic component of natural bone. In the bone forma-
tion process, HAp mineralization is controlled by collagen, which is
a special protein containing an ionic group that can interact with
HAp, and a dispersive group, which can stabilize HAp in the phys-
iological environment. However, dental structures are also com-
posed of HAp, which is needle-like and forms under the control
of proteins. Nearly the entire biological mineralization process is
a crystallization process controlled by organic components.
Fig. 3. Schematic illustration of the formation of HAp nanorods. Copyright Elsevier and reproduced with permission [97].
2773
Recently, considerable attention has been paid to the biomimetic
mineralization process because this process may lead to the fabri-
cation of novel materials that cannot be produced by conventional
methods [97–101]. Zhang et al. [102] reported the mineralization
of nano-HAp on self-assembled collagen; the nanocomposite
formed has a similar structure and biocompatibility to natural
bone. Tang et al. [103] reported the fabrication of nano-HAp under
the control of amino acids and found that both bone mimetic plate-
like and dental-mimetic rod-like nano-HAp can be produced under
the control of different amino acids. This method makes it possible
to control nano-HAp mineralization by purely synthetic polymers
rather than natural biomacromolecules. Since the synthetic poly-
mer has a relatively low cost and can be more easily designed,
nano-HAp synthesized by this method has many potential
applications. Yao et al. [104] employed double-hydrophilic block
copolymer (DHBC) poly-(vinylpyrrolidone)-b-poly(vinylpyrroli-
done-alt-maleicanhydride)-b-poly-(vinylpyrrolidone) (PVP-b-
P(NVP-alt-MAn)-b-PVP) to synthesize a biomimetic template for
HAp nanocrystal synthesis. Needle-like HAp nanocrystals can be
formed in the presence of PVP108–P(NVP-MAn)28–PVP108, as shown
in Fig. 4. Compared to the HAp nanocrystals formed in the presence
of poly-(vinylpyrrolidone) (PVP) homopolymer, those formed with
DHBC are more stable and do not precipitate in water after prepa-
ration. The crystallization process and the morphology of the final
nano-HAp crystals can be controlled by adjusting the DHBC molec-
ular structure. Cho et al. [105] prepared nanosized HAp powders
with high crystallinity and appropriate stoichiometry by a high-
temperature flame spray pyrolysis process from spray solutions
containing PEG. The mean sizes of HAp powders obtained from
PEG spray solutions were changed from several tens to several
hundred of nanometers according to the PEG concentrations in
the spray solutions at a post-treatment temperature of 800 °C.
HAp powders post-treated at a low temperature of 400 °C had a fi-
ber-like morphology. On the other hand, post-treated HAp pow-
ders at temperatures of 600 and 1000 °C had a rod-like
morphology with a low aspect ratio and spherical-like morphology,
respectively. The mean sizes of HAp powders post-treated at tem-
peratures of 600 and 1000 °C were 32 and 213 nm, respectively.
The application of nanostructured HAp with different morpholo-
gies for bone tissue engineering has been introduced above. How-
ever, several other applications of nano-HAp are also in progress.
Surface modifications of HAp nanoparticles have been performed
2774 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
Fig. 4. TEM images of HAp nanocrystals templated by PVP108–P(NVP-MAn)28–PVP108 after the 1 day (a), 8 day (b) and 13 day (c) preparations. Copyright ACS and reproduced
with permission [104].
in order to modulate their colloid stability, prevent dissolution in the
case of low pH, prevent inflammation, serve as an intermediate layer
to allow strong bond formation between HAp–polymer matrices,
and potentially enhance its bioactivity or improve its conjugation
ability with special functional groups [106–108]. HAp nanoparticles
have also served as non-viral carriers for drug delivery and gene
therapy because of their established biocompatibility, ease of han-
dling and well-known adsorption affinity [109–112]. Furthermore,
HAp nanoparticles can be stably loaded with radioisotopes [111].
After loading with genes or drugs by adsorption, HAp nanoparticles
provide a protective environment that shields them from degrada-
tion while providing a convenient pathway for cell membrane pen-
etration and the controlled release of the genes/drugs [112]. The
research results indicate the potential of nano-HAp in gene delivery
and as drug carriers [112,113]. Readers who are interested in learn-
ing more may refer to several elegant and more comprehensive re-
views, e.g. those by Dorozhkin [20,21,114].
3. Porous structure of nanoscale HAp
HAp ceramics have been widely used as artificial bone substi-
tutes because of their high biocompatibility, bioaffinity and osteo-
conductibility. However, induction of bone growth into HAp blocks
is unsatisfactory, because it is very slowly replaced by host bone
after implantation. For this reason, porous bodies and granules of
HAp ceramics have been developed and have been widely used
in clinical settings. However, due to the closed structure of conven-
tional porous HAp, which has non-uniform pore geometry and low
interpore connections, it is very difficult for implant pores to be-
come completely filled with newly formed host bone [115]. How-
ever, porous HAp ceramics with highly interconnecting structures
have been developed, and osteoconduction can occur deep inside
such ceramics. Wang et al. [116] systematically studied the effects
of electrical polarization of porous HAp ceramics using two types
of cylindrical porous HAp ceramics with high and low interpore
connections (HAp-H and HAp-L, respectively) on bone ingrowth.
Electrical polarization was effective in enhancing bone ingrowth
through all the pores of HAp-H implants; however, this advantage
was not apparent in the HAp-L implants. This suggested that en-
hanced bone ingrowth into HAp porous bodies due to electrical
polarization may be a co-operative interaction between the osteo-
conductivity of HAp porous bodies and the enhanced osteogenic
cell activity induced by large charges stored on pore surfaces.
The size and density of interpore connections, as well as those
of pores, are important factors for osteoconduction into the central
area of porous HAp. Diverse characteristics, such as pore size, pore
shape, pore interconnectivity and total porosity of the scaffold, are
considered important factors for successful tissue regeneration
[117,118]. Microwave heating has also been applied to fabricate
interconnective porous structured bodies by foaming as-synthe-
sized, calcium-deficient HAp (Ca-deficient HAp) precipitate con-
taining H2O2. The porous bodies were sintered by a microwave
process with activated carbon as the embedding material to pre-
pare nano- and submicron-structured ceramics. The study results
suggest that porous carbonated biphasic CaP ceramics with a nano-
structure promote osteoblast adhesion, proliferation and differen-
tiation [119]. In conclusion, porous carbonated biphasic calcium
phosphate (BCP) ceramics with a nanostructure are simple and
quick to prepare using microwaves. Compared to those produced
by conventional sintering, carbonated BCP ceramics may be better
bone graft materials.
Since synthesized HAp is very brittle, it cannot be used for load-
bearing bone replacements. Hence, implant materials composed of
hard and soft phases (composite materials) are used for total bone
replacement [120]. Composites of HAp with polymers such as poly-
methyl methacrylate, poly(3-hydroxybutyrate-co-3-hydroxyval-
eate) and polyacrylic acid show improved mechanical properties,
as well as good biocompatibility and bioactivity [121,122]. Degrad-
able and non-degradable polymers are used in controlled drug
delivery, scaffolds for tissue engineering, wound dressing, cosmetic
skin masks and protective clothing. Imai, Furuichi and co-workers
[122,123] reported hierarchical laminated architecture and porous
structures after calcinations of the polymer composite (poly(-
acrylic acid)) at 700 °C. Polyacrylamide hydrogel is a biomaterial
and non-degradable water-based polymer used as tissue filler.
Joshy et al. [124] investigated mineralization of HAp in a UV-poly-
merized acrylamide gel matrix by varying precursor concentration
and pH (pH 8–10). During polymerization, diammonium hydrogen
phosphate ions were implanted in the gel matrix and subsequently
immersed in calcium nitrate solution. Thin, laminated, macropo-
rous structures, embedded with nanospheres and ribbons of HAp
were mineralized. The HAp was found to be oriented along the
c-axis, which could lead to preferential binding of acidic proteins
on its surface. The laminated structures displayed a resorbable nat-
ure, whereas flake-like structures obtained at higher concentra-
tions were found to be bioactive. This composite could be an
alternative to the use of silicone gel to avoid the long-term risk
of fibrosis and migration when implanted. Muthutantri et al.
[125] developed a novel fabrication technique, a combination of
slurry dipping and electrospraying, to produce HAp foams as po-
tential matrices for bone tissue engineering applications. Slurry-
dipped and electrosprayed scaffolds for different time intervals
were compared with foams prepared by the individual methods
of dipping and electrospraying. Significant differences in strut
crack distribution and strut thickness and porosity were observed
on sintered foams prepared under various conditions. All sintered
structures had average porosities in the range of 84–94% and desir-
able pore interconnections, whereas the combined method pro-
duced foams of uniform pore distribution, thicker struts and
improved mechanical properties.
 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
A new direct rapid prototyping process called low-temperature
deposition manufacturing (LDM) was proposed to fabricate scaf-
folds [126]. This process integrated extrusion/jetting and phase
separation and can therefore fabricate scaffolds with hierarchical
porous structures, creating an ideal environment for new tissue
growth. Scanning electron microscopy (SEM) images of fabricated
scaffold structures with different polymer matrices are shown in
Fig. 5. The interconnected computer-designed macropores allow
cells in new tissues to grow throughout the scaffold. Moreover,
the parameter-controlled micropores allow nutritional compo-
nents in, and metabolic wastes out. The macrocellular morphology,
microcellular morphology, porosity and mechanical properties of
poly(a-hydroxy acid)-tricalcium phosphate (TCP) composite scaf-
folds prepared by the proposed method were investigated. These
highly controllable scaffolds may play an important role in tissue
engineering. LDM could also be combined with multinozzle depo-
sition or cell deposition to accurately control materials or cells
point-by-point. Sinha et al. [127] report a novel method of produc-
ing HAp–PVA microspheres suitable for biomedical application.
Spray drying is a well-established industrial process that produces
fine ceramic powders. Integrating this with a method akin to bio-
mineralization provides a direct route to produce HAp micro-
spheres with highly controlled morphological features.
4. Bio-orthopedic properties of nanoscale HAp
Bone substitutes are required to repair segmental defects
caused by the removal of infected tissue or bone tumors. The most
desirable form of bone substitutes, in such cases, is autologous
bone. However, autografts are not always available and may result
in morbidity at the donor site. An allograft is preferred in some
cases, but the possible immune response and disease (i.e. human
immunodeficiency virus (HIV) or hepatitis B) transmission are det-
rimental to the recipient [128]. Bone graft substitutes have at-
tracted much attention because of their advantages over both
autografts and allografts [129]. In the case of a bone, an optimized
biomaterial should be as biomimetic as possible, i.e. it should con-
sist of poorly crystalline, carbonate-substituted apatite with suffi-
cient mechanical properties [130]. HAp artificial bones are now
widely used in clinical practice and show satisfying repair function
in a series of studies [131–133]. Nonetheless, there are some weak-
nesses, such as weak intensity and slow degradation. To strengthen
HAp, researchers have carried out omnidirectional investigations
and the challenge in HAp development is whether biological prop-
erties can match mechanical properties [134].
Recently, our group reported HAp coating on scratched areas of
a human tooth and HAp disks by the immersion method in a HAp
colloidal solution (620 lm average diameter dispersed in deion-
ized water) [135]. The surface morphologies of the scratched area
Fig. 5. SEM micrographs of scaffolds fabricated with different polymer matrices. Copyright ACS and reproduced with permission [126].
2775
after immersion for 1–3 months were investigated and showed
that the damaged surfaces were remarkably recovered. The
mechanical property and chemical stability of the HAp coating lay-
ers on both specimens were then determined via the Vickers hard-
ness test and concentration measurement of extracted Ca2+ ions,
respectively, after strong acidic treatment. The cellular behavior
of mouse calvaria-derived pre-osteoblastic cells (MC3T3-E1) was
also examined on the HAp layers regenerated on micro-scratched
HAp disks for the purpose of their potential applications on maxil-
lofacial bone conservation and reconstruction for prosthetic den-
tistry, and artificial disk preparation of a vertebral column. These
results of HAp coating on the scratched areas of the tooth suggest
that this technique could be suitable for the development of long-
term prevention of micro-cleavage and tooth health supporters to
reduce discoloration, and for other maxillofacial and orthopedic
applications. Another interesting result published by our group
concerned the regeneration of a micro-scratched tooth enamel
layer by nanoscale HAp solution [136]. Nanoscale HAp powders
with a mean particle size of 200 nm were used to regenerate the
enamel layers of damaged teeth. An artificially scratched tooth
was immersed in a nanoscale HAp powder suspension in deionized
water (70 wt.% HAp) at 37 °C for a period of 1–3 months. After
3 months, the scratched surface was finally inlaid with HAp and
the roughness increased from 2.80 to 5.51. Moreover, the hardness
of the newly generated HAp layer on the crown was similar to that
of the innate layer. Ca2+ and PO3
4 ions from the HAp powders dis-
solved in deionized water were precipitated on the tooth to pro-
duce cement pastes on the enamel surface due to its high degree
of recrystallization, resulting in a hard, newly regenerated HAp
layer on the enamel layer. This nanoscale HAp powder solution
might be used to heal decayed teeth as well as to develop tooth-
whitening materials.
In addition, our group reported the clinical evaluation of SB-1™
as a synthetic bone in sinus bone grafting. The SB-1™ used consists
of synthetic HAp with 500–1400 lm particle size and has the same
chemical composition as the inorganic part of human bones. Rab-
bits with damaged bones were treated surgically with SB-1™ and
the progress of bone recovery was monitored using X-ray tech-
niques. The result showed that the SB-1™ was well integrated with
the surrounding host bone and promoted induction of new bone,
leading to bonding with newly formed bone and recovery of dam-
aged bone tissue. In particular, it was found through an in vivo
study with rabbit that SB-1™ was superior in terms of bonding
with host bones as well as induction and integration with new
bone compared to the allografts and bone grafts with low HAp
contents.
Zhu et al. [128] evaluated the osteoconductive properties of
nano-HAp material and its potential application as artificial bone
in repairing bone defects, and attempted to analyze the scientific
2776 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
basis of these properties. Their animal model of bone defects was
based on the bilateral radius of 39 New Zealand white rabbits,
which were randomly divided into an experimental group (bone
defect repaired with nano-HAp artificial bone), a control group
(bone defect repaired with HAp artificial bone) and a blank group
(defect left empty). The experimental group was found to stimulate
a callus bonier than the control and blank groups. These differences
in bone conduction are statistically significant (P < 0.05). Therefore,
nano-HAp artificial bone can potentially be used for bone defect
treatment. Osteosarcoma is a primary malignant bone tumor, most
prevalent in children and adolescents, and is usually highly aggres-
sive and eventually lethal. Despite multimodal therapies, there is
no effective approach to treat this malignant disease. Shi et al.
[137] observed the biological response of osteosarcoma cells to
two kinds of nano-Hap: nano-HAp-S and nano-HAp-L. These nano-
spheres have the same crystallinity (phase) and morphology but
differ in size. Cells treated with both forms of nano-HAp were
inhibited and mainly led to apoptotic cell death. The caspase-9-
dependent intrinsic apoptotic pathway plays a role in this. Inter-
estingly, the suppression and apoptosis of osteosarcoma cells were
directly related to the size of nanoparticles, and the larger-sized
nano-HAp was more effective than the smaller particles. In the col-
lagen matrix, tens to hundreds of these nano-blocks combine into
self-assembled biomaterials that have remarkable physical and
chemical features such as unique mechanical strength, insensitiv-
ity to growth/dissolution and flexible structures [138,139]. Thus,
features of smaller HAp nanoparticles may more closely resemble
features of HAp during biomineralization than features of the lar-
ger HAp particles that are conventionally used. Therefore, nano-
HAp may promote osteoblast adhesion, proliferation and synthesis
of alkaline phosphatase and lead to rapid repair of hard tissue in-
jury [140,141].
Nano-HAp has been reported to be a better candidate in bio-
medical applications. Cai et al. [142] prepared nano-HAp, typically
Fig. 6. Bone tissue cross-section of N-HA scaffold under phase contrast (a) 3 and (c) 12 weeks post-surgery with corresponding cross-polarized light micrographs shown in
(b,d) representing birefringence of collagen strands. 200 original magnification. S, scaffold; M, mineralized bone; C, collagen birefringence; V, vessel. Copyright ACS and
reproduced with permission [144].
20 ± 5, 40 ± 10 and 80 ± 12 nm in diameter, and studied their ef-
fects on proliferation of two bone-related cells: bone marrow mes-
enchymal stem cells (MSCs) and osteosarcoma cells (U2OS). Cell
culture experiments showed improved cytophilicity of the nano-
phase mineral relative to conventional HAp. Greater cell viability
and proliferation of MSCs were measured for nano-HAp, remark-
ably so for the 20 nm particles. Interestingly, the growth of osteo-
sarcoma cells was inhibited by nano-Hap, and 20 nm particles
were the best retardant. Nano-HAp has been suggested to exhibit
favorable cell proliferation to optimize biological functionality,
for which particle size is believed to play a key role. These
in vitro findings are of great significance for understanding the
cytophilicity and biological activity of nanoparticles during bio-
mineralization. Li et al. [143] investigated the effects on highly
malignant melanoma cells of nano-HAp particles with different
morphologies. Three types of HAp particles with different mor-
phologies were synthesized and co-cultured with highly malignant
melanoma cells using phosphate-buffered saline (PBS) as a control.
A precipitation method with or without citric acid addition as a
surfactant was used to produce rod-like nano- and micron-sized
HAp particles, respectively, and a novel oil-in-water emulsion
method was employed to prepare ellipse-like nano-HAp particles.
Experiment results indicated that the particle nanoscale effect,
rather than particle morphology of HAp, was more effective in
inhibiting highly malignant melanoma cell proliferation. Appleford
et al. [144] found that the bone formation and angioconductive po-
tential of HAp scaffolds closely matched to trabecular bone in a ca-
nine segmental defect after 3 and 12 weeks of implantation.
Histomorphometric comparisons were made between naturally
forming trabecular bone (control) and defects implanted with scaf-
folds fabricated with micro-size HAp (M-HAp) and nanosized HAp
(N-HAp) ceramic surfaces. As shown in Fig. 6, no significant differ-
ences were identified between the two HAp scaffolds; however,
significant bone in-growth was observed after 12 weeks with
 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
43.9 ± 4.1% and 50.4 ± 8.8% of the cross-sectional area filled with
mineralized bone in M-HAp and N-HAp scaffolds, respectively. This
study showed the potential of trabecular bone modeled, highly
porous and interconnected HAp scaffolds for regenerative
orthopedics.
5. Composites of HAp and inorganic nanomaterials
The low fracture toughness and poor wear resistance of HAp can
be improved by adding second-phase reinforcement. Carbon nano-
tubes (CNTs) have already shown their potential as effective rein-
forcements for HAp and other ceramics [145–148] to improve
fracture toughness. Reports are available on the processing of
HAp–CNT composite coatings for orthopedic implants through
plasma spraying [149–152], laser surface alloying [153,154], elec-
trophoretic deposition [155,156] and aerosol deposition [157]. In
addition to conventional sintering [158,159] and hot isostatic
pressing [160,161], spark plasma sintering (SPS) [161–163] has
also been employed to fabricate free-standing HAp–CNT compos-
ites. Omori et al. [161] reported consolidation of multiwalled
(MW) CNTs by SPS followed by dip-coating with HAp and a second
round of SPS consolidation of the coated preform. They observed a
consolidated coating of HAp on CNTs without crack formation. An-
other study on SPS of HAp–CNT composites used nano-HAp pow-
ders and CNTs, mixed by ball milling, as the starting material.
Sarkar et al. [162] found the fracture toughness of HAp–CNT com-
posite to be as high as 1.27 MPa m0.5, with 2.5 vol.% CNT addition,
which was a 30% increase over HAp. Xu et al. [163] performed
extensive mechanical stirring to homogeneously mix CNTs with
spray-dried HAp powder to prepare feedstock for SPS processing.
The Young’s modulus has been reported to be 131 GPa with
2 vol.% CNT addition, although no direct comparison with HAp
has been reported. The biocompatibility of CNT-reinforced HAp
composite has also been studied by Xu et al. This group determined
the beneficial effect of CNT on osteoblast cell proliferation. Lahiri
et al. [164] investigated CNT-reinforced HAp composite synthe-
sized using SPS. Quantitative microstructural analysis suggests
that CNTs play a role in grain boundary pinning and are responsible
for improved densification and retention of nanostructure
throughout the thickness of the sintered pellet. HAp crystal forms
a coherent interface with CNTs, resulting in a strong interfacial
bond. The uniform distribution of 4 wt.% CNTs in the HAp matrix,
good interfacial bonding and fine HAp grain size help to improve
fracture toughness by 92% and elastic modulus by 25% compared
to a HAp matrix without CNT.
Xiao et al. [165] introduced a simple and effective approach to
functionalize MWCNTs by in situ deposition of HAp to improve
hydrophilicity and biocompatibility. The scheme of biomineraliza-
Fig. 7. Scheme of biomineralization mechanism of the preparation of HAp–PEG–MWCNTs. Copyright Elsevier and reproduced with permission [165].
2777
tion mechanism of HAp–PEG–MWCNT preparation is shown in
Fig. 7. First, two types of pre-functionalized MWCNTs were pre-
pared: acid-oxidized MWCNTs, and MWCNTs covalently modified
by PEG. The influence of acid-oxidation time, pre-phosphorylation
and PEGylation of MWCNTs on in situ growth of HAp was further
investigated in simulated body fluid (SBF) with ionic concentra-
tions of 2, 5 and 10 times, respectively, at 37 °C for 24 h. The results
showed that these factors have positive effects on HAp crystal
growth, with PEGylation of MWCNTs playing a key role during
deposition. Finally, the methyl thiazolyl tetrazolium (MU) assay
was performed to evaluate cytotoxicity, which showed that PEGy-
lated MWCNTs wrapped by HAp crystals have the best
biocompatibility.
Inspired by self-assembly of nano-HAp association with the
67 nm periodic microstructure of collagen, Liao et al. [166] used
MWCNTs with an approximately 40 nm bamboo periodic micro-
structure as a template for nano-HAp deposition to form a nano-
HAp–MWCNT composite. Spindle-shaped units consisting of an
assembly of near-parallel, fibril-like nano-HAp polycrystals were
formed and oriented at a specific angle to the long axis of CNTs.
Spindle-shaped units detached from the MWCNT template are able
to maintain the ordered parallel structure of the nano-HAp poly-
crystal fibril. Tang et al. [167] found that biomimetic synthesis of
HAp on SDS functionalized MWCNTs by using an alternate soaking
process (ASP) in Ca/P solutions. The results showed that nano-HAp
crystals were formed on SDS functionalized MWCNTs, and miner-
alized MWCNTs remained in a dispersed state. HAp–MWCNT
nanohybrids, combining the osteconductive properties of HAp
and the excellent mechanical properties of MWCNTs, will provide
a promising material for bone tissue engineering.
Balani et al. [151] synthesized HAp-reinforced with sub-
micrometer Al2O3 and CNTs as a coating on a Ti–6Al–4 V substrate
via plasma spraying. The addition of Al2O3 and CNTs to HAp shows
improvements in the hardness and elastic modulus by 65% and
50%, respectively, compared to HAp. Consequently, HAp–Al2O3–
CNT coatings have been nano-scratched to determine their wear
performance. Reinforcement of HAp by Al2O3 decreases wear vol-
ume by more than 13 times, whereas HAp–Al2O3–CNT coating
shows an even further decreased wear volume of 5 times com-
pared to that of HAp–Al2O3 coating. Ajeesh et al. [168] prepared
sintered iron oxide–HAp nanocomposite ceramics from powders
produced through a co-precipitation process. The phase purity
and bioactivity of the composites were analyzed as a function of
percentage of iron oxide in the composite. In all the prepared com-
posites, HAp retains the same phase identity and high X-ray opac-
ity as a composition containing 40 wt.% iron oxide. Increased cell
viability and cell adhesion showed that the prepared composite of-
fers considerable potential for bone tissue engineering
2778 H. Zhou, J. Lee / Acta Biomaterialia 7 (2011) 2769–2781
applications. Jiang et al. [169] designed and synthesized nano-HAp
crystals carrying incorporated Fe ions by a co-precipitation meth-
od. The results showed that lattice substitution or incorporation
occurs between Fe ions and nano-HAp, which is quite different
from the mechanical mixture of Fe3O4 nanoparticles and nano-
HAp crystals. The nano-Fe–HAp with a synthetic molar ratio of
1:2 (calculated by Fe3O4:HAp) exhibited a needle-like crystal shape
and relatively good crystallinity. However, when the molar ratio
was increased to 1:1, some Fe3O4 nanoparticles were separated
from the nano-Fe–HAp crystals. Magnetization measurements
showed that nano-Fe–HAp crystals were in a superparamagnetic
state, proving that the existence of Fe ions in nano-HAp crystals
contributed to the magnetic properties of nano-Fe–HAp crystals.
As one component of biomedical composites, nano-Fe–HAp has
the potential to promote bone regeneration.
Bioactive nano-titania ceramics with excellent biomechanical
compatibility and bioactivity were prepared by using HAp additive
as a grain growth inhibitor. The mechanical properties of nano-
titania ceramics were analogous to those of human bone. The
amount of HAp additive played an important role in determining
the grain/particle size of nano-titania ceramics, which had a great
effect on osteoblast proliferation in cell culture experiments. Cell
culture experiments also showed that the bioactive HAp additive
itself also significantly affected the cytocompatibility of nano-tita-
nia ceramics. These results indicated that the content of HAp addi-
tive not only had an effect on the bioactivity of nano-titania
ceramics due to the bioactivity of the additive itself, but also had
an effect both on the biomechanical compatibility and bioactivity
of nano-titania ceramics by adjusting the grain/particle size of
the ceramics. The HAp and ZrO2 nanopowders were prepared by
chemical reactions and alcohol–aqueous solution heating, respec-
tively. The nanosized HAp-ZrO2 powders with a homogeneous dis-
tribution could then be synthesized by ball-milling. HAp-ZrO2
bioceramics with small grain sizes could be obtained by using
hot-press sintering technology [170]. The in vitro biocompatibility
of these bioceramics was studied, and the results showed that
there was no reaction between HAp and ZrO2 powders-which
may be attributed to the very short sintering time of the hot-press
sintering- and that HAp-ZrO2 bioceramics possess non-toxic and
non-allergenic properties.
Biomaterials science has found many ways to enhance different
properties of synthetic HAp by introducing various ionic substitu-
tions into the apatite structure [171]. One of the most interesting
substitutions is the incorporation of Si, which considerably im-
proves the bioactivity of synthetic apatite [172]. The role of Si in
bioactive processes involved in new bone tissue formation is well
known [173]. Carlisle first reported the presence Si traces during
bone mineralization at early stages of calcification, and this has
proven the influence of an Si-deficient diet in diseases such as oste-
oporosis [174]. Therefore, Si-substituted HAp (Si–HA) was chosen
as an appropriate candidate for the preparation of an improved
CaP material and was subsequently produced by chemical synthe-
sis [175,176]. Solla et al. [177] chemically synthesized Si–HA and
presented it as a new material with enhanced bioactivity. Si–HAp
films were deposited by pulsed laser deposition (PLD), using tar-
gets composed of HAp mixtures with different Si-containing
sources such as SiO2 and diatomaceous earth. Analysis revealed
that Si is successfully incorporated into the HAp structure, as well
as traces of other elements such as Na, Fe or K.
6. Summary and perspective
Nanophase HAp bioceramics have gained importance in the bio-
medical field due to their superior biological and biomechanical
properties. Development of HAp biomedical materials will benefit
from advances in nanotechnology. Several methods for synthesiz-
ing HAp on the nanoscale have evolved in the past few decades.
Due to the chemical similarity between HAp and mineralized bone
of human tissue, synthetic HAp exhibits a strong affinity to host
hard tissues. A significant amount of research in this area is ex-
pected to be focused on the nanoscale for enhanced applications
as resorbable scaffolds that can be replaced by endogenous hard
tissues over time. In the future, the ability to functionalize surfaces
with different molecules of varying natures and dimensions based
on their means of attachment to cells, as well as the potential to
nanostructure the surface physically, chemically and topographi-
cally, will enable selective targeting within biological systems to
show specificity towards individual proteins and peptides. Under-
standing the influence of nano-HAp particle size, crystal morphol-
ogy controls and interfaces with cells from a higher level is
essential for the future development of nanotechnology and bio-
technology. Such an interdisciplinary approach is very compli-
cated, and the effective collaboration of scientists from different
disciplines is necessary.
Acknowledgments
This research was supported by the International Research and
Development Program of the National Research Foundation of Kor-
ea (NRF) funded by the Ministry of Education, Science and Technol-
ogy (MEST) of Korea, and National Fisheries Research and
Development Institute (Grant Nos. K20091003000, FY2009,
20100434961-00).
Appendix A. Figures with essential colour discrimination
Certain figures in this article, particularly Figures 4, 6, 7, are dif-
ficult to interpret in black and white. The full colour images can be
found in the on-line version, at doi:10.1016/j.actbio.2011.03.019.
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