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Acta Biomaterialia 6 (2010) 4457–4475

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Amorphous calcium (ortho)phosphates

Sergey V. Dorozhkin *
Kudrinskaja sq. 1-155, Moscow 123242, Russian Federation

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

Article history: Amorphous calcium phosphates (ACPs) represent a unique class of biomedically relevant calcium
Received 4 May 2010 orthophosphate salts, having variable chemical but essentially identical glass-like physical properties,
Received in revised form 22 June 2010 in which there is neither translational nor orientational long-range ordering of the atomic positions. Nor-
Accepted 23 June 2010
mally, ACPs are the first solid phases, precipitated after a rapid mixing of aqueous solutions containing
Available online 4 July 2010
ions of Ca2+ and PO34 ; however, other production techniques are known. Interestingly, ACPs prepared
by wet-chemical techniques were found to have a relatively constant chemical composition over a rela-
tively wide range of preparation conditions, which suggests the presence of a well-defined local struc-
Calcium orthophosphate
tural unit, presumably with the structure of Ca9(PO4)6 – so-called Posner cluster. However, the
Apatite presence of similar clusters in ACPs produced by other techniques remains uncertain. All ACPs are ther-
modynamically unstable compounds and, unless stored in dry conditions or doped by stabilizers, spon-
taneously tend to transform to crystalline calcium orthophosphates, mainly to calcium apatites. This
solution instability of ACPs and their easy transformation to crystalline phases are of a great biological
relevance. Specifically, the initiating role ACPs play in matrix vesicle biomineralization raises the impor-
tance of ACPs from a mere laboratory curiosity to that of a key intermediate in skeletal calcification. In
addition, due to significant chemical and structural similarities with calcified mammalian tissues, as well
as excellent biocompatibility and bioresorbability, all types of ACPs are very promising candidates for the
manufacture of artificial bone grafts. This review summarizes the current knowledge on the occurrence,
preparation, composition, structure, major properties and biomedical applications of ACPs. To assist read-
ers in looking for the specific details on ACPs, a great number of references have been collected and
Ó 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

1. Introduction enamel [9], as well as in the mitochondria [1] and sarcoplasmic

reticulum [10] of some cells. Despite intensive efforts, the accumu-
In nature, amorphous phases exist extensively with readily lated evidence for ACP as an integral mineral component in major
moldable isotropic properties, and serve as structural materials. hard tissues, such as bones and teeth, is equivocal and has for
For example, amorphous structures represent about 20% of approx- many years been the subject of considerable debate [11–24]. Fur-
imately 60 different inorganic compounds or minerals formed by thermore, recent studies on bone and teeth formation have sug-
living organisms. These biologically formed minerals are often gested the presence of transient amorphous mineral precursors
called biominerals, while the process of their formation is called and a universal strategy for calcium carbonate-based [21,25] and
biomineralization [1]. A recent review on the subject indicates that calcium orthophosphate-based [21,22,25–28] biomineralization
many biominerals are formed by amorphous precursors and, fur- in both vertebrates and invertebrates.
thermore, the amorphous phases may possess fluidic properties An interesting study on a potential role of ACP in facilitating
that impart new processing capabilities to the system [2]. Among assembly of hydroxyapatite (HA) nanoparticles into highly ordered
existing biogenic amorphous minerals, those composed of calcium structures has been published recently [29]. Higher-order HA
orthophosphates are most abundant in the teeth and exoskeletal architectures were detected only when the starting particles were
structures of marine invertebrates [2–6]. On the other hand, the aggregates of nanospheres with HA cores and ACP shells. Surface
existence of similar amorphous calcium phosphate (ACP) minerals ACP initially linked HA nanoparticles in a way that allowed a par-
in vertebrate organisms has not been well established experimen- allel orientation of the HA nanoparticles and then was incorpo-
tally except in highly specialized locations such as the inner ear rated into HA by phase transformation to produce more ordered
structures of embryonic sharks [1], mammalian milk [7,8], dental architectures with the characteristic features of apatites in biolog-
ical structures. Further, it was demonstrated that enamel- and
* Tel.: +7 499 255 4460. bone-like apatites could be prepared by using nanodimensional
E-mail address: HA and ACP under the control of various biological additives

1742-7061/$ - see front matter Ó 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
4458 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

[29]. This study points to an important role ACP might play in bone as amorphous or crystalline materials based on the macroscopic
formation. However, due to a lack of absolute proof, the question as properties such as their external shapes, fracture mechanisms
to the occurrence of ACP phases in newly mineralized tissues of and optical properties long before X-ray diffraction techniques
vertebrates remains unanswered [30]. and other methods became available to reveal the atomic struc-
This review is devoted to amorphous calcium orthophosphates, tures of these materials. Only in the past century has an under-
which are of a great biomedical importance due to their chemical standing of the microscopic nature of amorphous materials
and structural similarities to calcified mammalian tissues. A com- become possible [79]. However, there is still much debate concern-
plete list of available calcium orthophosphates, together with their ing the exact nature of these materials. For example, in a recent
standard abbreviations, chemical formulae and major properties, is article, Sheng et al. [81] state: ‘‘the atomic arrangements in amor-
given in Table 1 [31,32]. Since all of these compounds are calcium phous alloys remain mysterious at present”.
orthophosphates, strictly speaking, all abbreviations in Table 1 are An amorphous structure is distinctly different from a densely
incorrect; however, they are extensively used in the literature. As packed assembly of microcrystals and is closely related to the struc-
the majority of calcium orthophosphates are crystalline, this re- ture of a liquid phase. Ideally, an amorphous solid should be de-
view is devoted to the detailed description of ACP (emphasized scribed by the model of a perfectly random structure [82];
by bold font in Table 1). Furthermore, with a few important excep- however, this is the boundary condition. As such, the structure of
tions, neither ion-substituted ACPs [33–45], nor ACP-containing amorphous solids is normally described in terms of statistical distri-
composites [46–72] are considered and discussed. The readers butions. Nevertheless, prior to a further description, one must spec-
interested in either these topics and/or other types of amorphous ify the existing atomic length scales. The shortest length scale
calcium phosphates (e.g. amorphous calcium polyphosphate [73– usually used to describe the structure of a material consists of an
77] and amorphous calcium metaphosphate [78]) are referred to atom and its nearest neighbors, out to perhaps a distance of two or
the original publications. three atoms. All solids and liquids have some structure on this scale,
which is called a short-range order (SRO). For crystalline solids,
2. Basic definitions and knowledge on amorphous solids structural order persists over much longer distances (at least tens
or hundreds of atomic distances), such that the atoms occupy sites
According to the laws of thermodynamic, a perfect infinite crys- in a periodic three-dimensional array. Such materials are said to
tal cannot exist in the real world. Various disorders in the forms of have a LRO and include most metals and many covalently bonded
vacancies, interstitial atoms, impurities, dislocations, grain bound- solids. Non-crystalline solids, including glasses, lack a LRO and are
aries, surfaces and other interfaces disrupt the periodicity of other- said to be amorphous even though they can have a SRO that is quite
wise ‘‘perfect” crystals and in many cases determine their physical well defined [83].
properties. By contrast, highly disordered solids are those solids According to the available literature, for each particular atom of
that are so irregular that the concept of a reference crystal lattice any solid there exists a SRO of 2–5 Å, a medium-range order
must be abandoned. Such highly disordered materials are called (MRO) of 5–20 Å and a LRO at distances exceeding 20 Å [84,85].
amorphous materials [79]. As wikipedia, the free encyclopedia, In the case of covalent materials, in which a directed chemical bond-
has it: an amorphous (from the Greek term aloquo1, which ing is dominant, a SRO can be characterized in terms of the well-de-
means ‘‘shapeless” or ‘‘without form”) solid is a solid, in which fined coordination polyhedra, which, in many cases, appear to
there is no translational and orientational long-range order (LRO) concur with the unit cells. The definition of a MRO is more conten-
of the atomic positions [80]. Early researchers categorized solids tious and it is helpful to subdivide MRO into three subcategories.

Table 1
Existing calcium orthophosphates and their major properties [31,32].

Ca/P molar Compound Formula Solubility at 25 °C, Solubility at pH stability range in aqueous
ratio log (Ks) 25 °C, g l1 solutions at 25 °C
0.5 Monocalcium phosphate monohydrate Ca(H2PO4)2H2O 1.14 18 0.0–2.0
0.5 Monocalcium phosphate anhydrous Ca(H2PO4)2 1.14 17
1.0 Dicalcium phosphate dihydrate (DCPD), CaHPO42H2O 6.59 0.088 2.0–6.0
mineral brushite
1.0 Dicalcium phosphate anhydrous (DCPA), CaHPO4 6.90 0.048
mineral monetite
1.33 Octacalcium phosphate (OCP) Ca8(HPO4)2(PO4)45H2O 96.6 0.0081 5.5–7.0
1.5 a-Tricalcium phosphate (a-TCP) a-Ca3(PO4)2 25.5 0.0025
1.5 b-Tricalcium phosphate (b-TCP) b-Ca3(PO4)2 28.9 0.0005
b b d
1.0–2.2 Amorphous calcium phosphate (ACP) CaxHy(PO4)znH2O, n = 3–4.5; 5–12
1.5–1.67 Calcium-deficient hydroxyapatite (CDHA)e Ca10x(HPO4)x(PO4)6x(OH)2xf 85.1 0.0094 6.5–9.5
(0 < x < 1)
1.67 Hydroxyapatite (HA, HAp or OHAp) Ca10(PO4)6(OH)2 116.8 0.0003 9.5–12
1.67 Fluorapatite (FA or FAp) Ca10(PO4)6F2 120.0 0.0002 7–12
1.67 Oxyapatite (OA or OAp) Ca10(PO4)6O 69 0.087
2.0 Tetracalcium phosphate (TTCP or TetCP), Ca4(PO4)2O 38–44 0.0007
mineral hilgenstockite
These compounds cannot be precipitated from aqueous solutions.
Cannot be measured precisely. However, the following values were found: 25.7 ± 0.1 (pH 7.40), 29.9 ± 0.1 (pH 6.00), 32.7 ± 0.1 (pH 5.28). The comparative extent of
dissolution in acidic buffer is: ACP  a-TCP  b-TCP > CDHA  HA > FA.
Stable at temperatures above 100 °C.
Always metastable.
Occasionally, CDHA is named precipitated HA.
In the case x = 1 (the boundary condition with Ca/P = 1.5), the chemical formula of CDHA looks as follows: Ca9(HPO4)(PO4)5(OH).
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4459

At the shortest length scale (5 Å), a near-MRO describes the material changes from a supercooled liquid, with properties one
connections among the coordination polyhedra. At the next length would expect from a liquid state material, to a solid. The tempera-
scale (5–8 Å), an intermediate MRO can be associated with corre- ture at which this transition occurs is called the glass transition
lations between pairs of preferred dihedral angles for neighboring temperature. If a cooling rate is faster than the rate at which atoms
bonds. Finally, on a yet larger length scale (8–20 Å), a far-MRO and/or molecules can be organized into a more thermodynamically
can be associated with the total dimensionality of the covalently favorable crystalline state, then an amorphous solid will be formed.
bonded amorphous network. Fig. 1 [86] represents an excellent In contrast, if atoms and/or molecules have a sufficient time to be
visual demonstration of the differences between a SRO and a MRO. organized into structures with two- or three-dimensional order,
Further details on this topic are available in the literature [84,85]. then a crystalline (at least, a semicrystalline) solid will be formed.
In covalent solids, bond angles and bond lengths, as well as a Furthermore, in many cases amorphous materials can be produced
number of the nearest neighbors, are all part of the appropriate by additives, which interfere with the ability of the primary con-
bonding scheme. Thus, due the nature of chemical bonding, even stituent to crystallize. For example, addition of soda to melted sil-
truly amorphous materials have some structural SRO and, perhaps, icon dioxide results in amorphous window glass, and addition of
some MRO. For example, MRO regions of 15 Å in dimensions and glycols to water results in a vitrified solid [80]. More to the point,
comprising about 100 atoms have been directly observed in amor- amorphization of many solids can achieved by applying mechani-
phous carbon [87]. Some order in two-dimensional projections of cal forces, e.g. by intensive milling [90].
thin amorphous three-dimensional structures was found [88]. In From a thermodynamic point of view, amorphous materials are
addition, covalent amorphous solids were found to exhibit a at best metastable. Given sufficient time, they tend to transform to
MRO at length scales up to 20 Å or so [85]. Such MRO clusters crystalline phases that are thermodynamically more stable. Inter-
are called paracrystals [83]. These paracrystals have a crystalline estingly, when an unstable crystalline solid is transformed to an
topology but the atomic positions are highly distorted from those amorphous phase, this transformation frequently exhibits features
of a perfect crystal. However, in solids there is a serious problem that are associated with ordinary melting, i.e. amorphization fre-
of very small particles. Specifically, if the crystal sizes are extre- quently begins at grain boundaries, surfaces or other defect sites,
mely small, it is difficult to make a distinction between the truly as does ordinary melting. Further, as the transformation proceeds,
amorphous and crystalline solids. Namely, if a powder consists of a sharp interface that separates the amorphous materials from the
tiny perfect crystals with dimensions of 2  2  2 nm (8 nm3) or untransformed crystalline material is always observed [79].
less, both this powder and any bulk materials prepared from this
powder (e.g. by compaction) will be amorphous, simply due to 3. Amorphous calcium phosphates (ACPs)
the fact their sizes are below the minimum value of LRO. Addition-
ally, in very small crystals a large fraction of the atoms are located 3.1. History
at/or near surface. Relaxation of the surface and various interfacial
effects distort the atomic positions, decreasing the structural order. According to Eanes [20], the history of ACPs looks as this: ‘‘In
Thus, even the most advanced structural characterization tech- 1955, Robinson and Watson [91] were the first to suggest that a
niques, such as X-ray, neutron and electron diffraction, as well as substantial portion of newly formed mineral in young bone was
transmission electron microscopy (TEM), have difficulties in distin- not crystalline. Instead, they described this early mineral as being
guishing between the amorphous and crystalline structures on more similar in character to an amorphous-like precipitate they
these length scales [80,89]. had prepared in a study on synthetic HA [92]. This precipitate,
Many studies have revealed that the majority of solids can be which appeared initially in their synthesis when sufficiently con-
found or prepared in an amorphous state. For example, cooling centrated solutions of CaCl2 and Na2HPO4 were mixed at room
strongly reduces atomic and/or molecular mobility. Thus, in princi- temperature and neutral pH, had as its most distinctive features
ple, given a sufficiently high cooling rate, any liquid can be trans- an extremely fine, non-crystalline texture when examined by
formed into an amorphous solid. As cooling is performed, the TEM and no discernible electron diffraction pattern. This latter fea-
ture led them to infer that the considerably more diffuse electron
diffraction pattern of newly formed bone mineral as compared to
more mature bone mineral, although still apatitic in character,
indicated the presence also of an amorphous component” [20].
However, Boskey [19] has reported another story: ‘‘In 1964, Dr.
Paul Tannenbaum, a graduate student in periodontics at Columbia,
and a research assistant in Dr. Posner’s laboratory at hospital for
special surgery, was studying the effect of fluoride on apatite crys-
tal size. He prepared a synthetic apatite by mixing high concentra-
tions (30 mM) of calcium chloride and (20 mM) sodium acid
phosphate in buffer, and, being anxious to confirm that the precip-
itate which formed was apatite, pelleted it by centrifugation, dried
it with acetone and placed it on a holder for analysis by wide-angle
X-ray diffraction (XRD). The pattern obtained (Fig. 2.1, bottom) was
broad and diffuse, with a maximum at 30° 2h had no features,
and was clearly not apatite. Dr. Posner suggested that Dr. Tannen-
baum did not have the settings correct on the X-ray diffractometer,
but since it was late on Friday, decided to correct the settings on
Monday. On Monday, the sample, which had been left on the
diffractometer over the weekend, was again subjected to XRD
analysis, but now the pattern had the appearance of a poorly
Fig. 1. A formation of skydivers illustrates disorder on an intermediate length scale.
Each skydiver has a simple set of rules for bonding to the next skydiver (SRO) but
crystalline apatite (Fig. 2.1, middle). Dr. Tannenbaum was certain
there is a sufficient flexibility for different patterns of ordering to be created on the that the settings on the diffractometer were not different from
scale of a few body lengths (MRO). Reprinted from Ref. [86] with permission. those he had used previously. Instructed by Dr. Posner to repeat
4460 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

Fig. 2. Fragments of: (2.1) XRD patterns (Cu Ka radiation, 0.154 nm) of synthetic ACP (bottom), poorly crystalline CDHA (middle) and highly crystalline HA (top); (2.2)
infrared spectra of ACP (upper), CDHA (middle) and HA (lower). Fig 2.2 is reprinted from Ref. [123] with permission.

the experiment, he observed the same phenomenon. Immediately 3.2.1. Wet chemistry
after being mixed, the precipitate formed was amorphous, while By the early 1970s researchers had already established that the
after several hours, it converted to poorly crystalline apatite. It final and stable product of a reaction between calcium and ortho-
seemed plausible to Dr. Posner that were such an ‘‘amorphous” phosphate salts in neutral or basic aqueous solutions was crystalline
material (i.e. one that did not give a crystalline diffraction pattern) stoichiometric HA. However, stoichiometric, highly crystalline HA
present in bone, along with the apatite, it might account for the could only be prepared at elevated temperatures; thus, in aqueous
broad diffraction pattern of bone mineral” [19] (It should be solutions CDHA is formed instead [31,32]. During CDHA precipita-
emphasized that both Chow et al. [93] and Eanes [94] published tion, over a broad range of solution conditions, an ACP precursor
corrigenda to this story by Boskey). phase is formed [107–117], in some cases, via a short intermediate
In the 1960s, both XRD and infrared spectroscopic techniques stage of octacalcium phosphate (OCP) formation [118,119]. Data
were used to obtain a quantitative estimate of the amorphous con- are available that CDHA crystallization from ACP simply involves
tent of bone mineral, and then, based on the methods used in poly- an increase in LRO in the structure [110]. It should be emphasized
mer chemistry, an algorithm to estimate the amount of ACP in that by the mid-1970s ACP had been found not to be a mandatory
bones was developed [95–97]. Early XRD estimates indicated the precursor to CDHA: in dilute aqueous solutions CDHA was found
presence of 30% or more of a non-crystalline mineral in bones to precipitate without going through an ACP precursor [120]. A mod-
of several animal species. Later estimates by X-ray radial distribu- el was subsequently developed to illustrate factors influencing the
tion analysis placed the upper limit of 10% ACP in bones and nature of non-stoichiometric amorphous precursor phases precipi-
brought into question whether all X-ray amorphous bone minerals tating in highly supersaturated solutions [121].
were truly non-crystalline [17,98–101]. However, further studies The basic approach for synthesizing ACP still consists of a spon-
by higher-resolution techniques have shown that 99% of the min- taneous precipitation by mixing concentrated aqueous solutions of
eral in bone is a poorly crystalline ion-substituted calcium-defi- calcium and orthophosphate ions, first developed in 1953 by Wat-
cient hydroxyapatite (CDHA) of biological origin [19]. son and Robinson [92]. Another commonly used method is to pre-
Morphological evidence establishing the extent of ACP in skeletal pare an acidic (pH 4–5) subsaturated aqueous solution of a calcium
tissues is equally ambiguous. Although some studies [91,102–106] orthophosphate salt (e.g. dicalcium phosphate dihydrate (DCPD))
report a presence of small spheroidal particles atypical of crystalline and afterwards to induce precipitation by rapid addition of a strong
material, primarily in actively metabolizing regions, most TEM stud- base (e.g. NaOH, KOH, NH4OH) to reach the desired solution pH
ies of bones do not even mention finding such possibly amorphous [122]. Vigorous mixing is highly desirable. By means of both ap-
structures. Furthermore, during aging, the amount of ACP in bones proaches, various types of ACPs have been prepared from solutions
and teeth decreases while the crystalline forms of biologically encompassing a wide range of pH (from 6.5 to 13), Ca/P ionic ra-
formed ion-substituted CDHA increase during early stages of forma- tios (from 0.1 to 10), calcium and orthophosphate concentrations
tion [3,11]. Since physical and morphological evidences for ACP in (from 0.002 to 1 mol l1), as well as at temperatures from 0 to
skeletal tissue have been difficult to establish directly, much of our 50 °C. However, the Ca/P ratio of the mixing reagents (classically,
progress in clarifying the possible roles of ACP in biogenic calcifica- Ca(NO3)24H2O and (NH4)2HPO4) is typically kept within 1.50–
tion has come from both synthetic and in vitro studies [20]. 1.67, while the mixing solutions are frequently rendered basic by
addition of NH4OH [46,110,122–126]. At acidic pH, crystalline
3.2. Preparation calcium orthophosphates are normally precipitated. However, in
the presence of stabilizers (Mg and/or citrates), ACP could be
There are various methods for producing ACPs. We will look precipitated at solution pH of 6.0–6.5 [127]. No information on
first at wet chemistry methods, then at mechanical and thermal ACP precipitation from even more acidic aqueous solutions has
techniques. been found in the literature. The obtained precipitates should be
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4461

Fig. 3. XRD patterns of: (3.1) freeze-dried precipitates prepared from aqueous solutions containing different amounts of polyethylene glycol (note a shift of the center of the
broadened peak from 32° toward 31° with increasing polyethylene glycol/Ca molar ratio, which implies some structural differences in the resulting ACPs); (3.2) heat-
treated (800 °C) ACPs prepared from aqueous solutions containing different amounts of polyethylene glycol. Reprinted from Ref. [142] with permission.

collected shortly after the preparation (the sooner, the better), be- example, an ACP was prepared using a mechanochemical method
cause in aqueous media ACP is spontaneously converted to other involving a dry mixture of DCPD and Ca(OH)2 reactants with a
calcium orthophosphates, mainly to CDHA [110,128]. Furthermore, Ca/P ratio of 1.67 [147]. Other authors have shown that a pro-
it was shown that the final calcium orthophosphate (a dry powder) longed high-energy ball milling of either a-TCP, b-TCP powder in
would be amorphous if, beside the appropriate key factors of the ethanol or a dry mixture of ACP and DCPD powders lead to ACP for-
synthesis (a high concentration of reagents, a basic solution pH mation after 24 h [148–150]. However, there is a non-negligible
and a low temperature), both a high addition rate and a mandatory risk of powder contamination (ball wear) when using this process-
freeze-drying of the precipitates were employed [39,110,126,129]. ing route over extended periods to obtain an ACP [30]. Further-
The precipitated ACP phases appear to be spherical (Fig. 4a and b) more, a crystalline to amorphous transition has been detected for
in an electron microscope (diameter 30–100 nm), unlike the nee- various calcium orthophosphates at very high (up to 10 GPa) pres-
dle-like crystals of CDHA (Fig. 4c and d). The solution pH, the con- sures (Fig. 5) [166,167].
centration of the mixing reagents and the preparation temperature
all affect the ACP particle sizes: a higher supersaturation produces 3.2.4. Thermal
smaller ACP particles [123]. Although ACP can be prepared with a Finally, ACPs might be prepared using high-energy processing
Ca/P molar ratio as low as 1.2 (at low pH: see Fig. 6) or as high as at elevated temperatures. This method is based on a rapid quench-
1.7 (at high supersaturation), departure from a Ca/P of 1.5 has ing of melted calcium orthophosphates occurring, for example,
been shown to be due to surface-adsorbed soluble phases, which during plasma spraying of HA [151–163]. A plasma jet, which pos-
can be washed away, or to occluded Ca, respectively [98]. sesses very high temperatures (5000–20000 °C), partly decom-
poses HA. Furthermore, it has been suggested that thermal
3.2.2. Non-aqueous solutions and solvents (sol–gel) spraying produces the amorphous phase, not only due to the high
In addition, ACPs can be easily prepared in either non-aqueous cooling rate, but also because removal of hydroxyl ions makes it
or solvent + water media [125,130–144]. The presence of organic more difficult for the crystalline phase to form [156]. This generally
compounds and/or solvents results in a decrease in the dielectric leads to calcium orthophosphate phases with variable composi-
constant. Therefore, all ions in solutions appear to be less solvated tions, often containing impurities, which is not convenient for
than those in water. The consequence of this is a strong decrease of preparation of pure ACPs. Interestingly, the amorphization degree
solubility and an increase in precipitation kinetics, which simpli- of plasma-sprayed HA coatings appeared to correlate with the
fies amorphization [145]. Furthermore, in such systems, complexes presence of vacancies of hydroxyl ions in the structure of HA: the
of calcium with organic agents can be formed. This favors ACP for- more vacancies there were in the apatite structure due to missing
mation, which is attributed to the coordinated complexing agents hydroxyl sites, the more ACP was present in the resultant coatings
remaining in the structure of ACP [136]. The influence of the pres- [155]. This conclusion was drawn based on the fact that particles
ence of organic solvents to an amorphization degree of precipitated resident in plasma for a longer period of time will lose more struc-
calcium orthophosphates is well illustrated in Fig. 3.1. In some tural water. Other studies have shown more amorphous phase lo-
cases, incorporation of organic compounds into ACPs has been de- cated adjacent to the substrate and a gradient tending towards a
tected [125,142]. Interestingly, replacement of a freeze-drying lower ACP content at the top of the coating [30]. Therefore, in plas-
stage of a wet ACP precipitate by oven drying at 80 °C resulted in ma-sprayed HA coatings, amorphous phases are in intimate mix-
its transformation to CDHA [131]. This process was ascribed to tures with both crystalline calcium orthophosphates and other
an internal hydrolysis of a part of orthophosphate ions of ACP to compounds, such as CaO [9], and so far nobody has succeeded in
those of CDHA according to the schemes [131,146]: separating ACPs from the coatings. However, the amorphous re-
PO3 2  gions in plasma-sprayed HA coatings might be mapped using a
4 þ H2 O ! HPO4 þ OH ð1Þ
scanning cathodoluminescence microscopy technique [159]. Fur-
Ca9 ðPO4 Þ6 þ H2 O ! Ca9 ðHPO4 ÞðPO4 Þ5 OH ð2Þ thermore, due to a number of uncertainties, the reproducibility
of such experiments is poor; thus, the plasma-spaying technique
is not considered as a valuable method to produce ACPs. A flame
3.2.3. Mechanical spray synthesis, in which a liquid precursor solution is fed through
In addition to the aforementioned solution-based methods, var- a capillary into a burning methane/oxygen supporting flame,
ious types of ACPs can be prepared by dry chemical techniques. For seems to be more preferable to produce ACPs at high temperatures
4462 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

Fig. 4. Bright-field transmission electron micrographs of ACP ? CDHA transformation at reaction times of (a) 5 min, (b) 3 h, (c) 9 h, (d) 48 h. Reprinted from Ref. [168] with

Fig. 6. Ca/P molar ratios of washed (dashed line) and unwashed (solid line) ACP
Fig. 5. Effect of pressure on XRD patterns of HA and hydrated TCP at 1 bar and high precipitates as a function of their formation pH. Reprinted from Ref. [169] with
pressures. Reprinted from Ref. [167] with permission. permission.
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4463

[164,165]. Unfortunately, it remains unclear to what extent the remain unknown and, in the available literature, the various ACPs
high-temperature ACPs are similar to the dry-heated dehydrated are distinguished only by Ca/P ratio. Since most ACPs have a Ca/P
ACPs prepared by wet-chemistry. Further details on and additional ratio close to 1.5 (see below), a term ‘‘amorphous TCP” (ATCP) is
examples of ACP preparation can be found in a recent review [30]. standard in the literature [30,132,146,150,164,175–181]. Other
To conclude this part, one should mention an interesting at- terms such as ‘‘amorphous carbonated apatite” [39,182,183],
tempt to precipitate separately hydroxyapatites of Mg, Ca, Sr and ‘‘amorphous CaHPO4” [132], which is equal to ‘‘amorphous DCPA”
Ba from basic supersaturated orthophosphate solutions containing (Table 1), ‘‘amorphous Ca8(HPO4)2(PO4)4” [132], which is equal to
a 10:6 divalent cation/PO4 molar ratio [170]. The first phase which ‘‘amorphous OCP” (Table 1), ‘‘amorphous OCP” [134], ‘‘amorphous
precipitated in the case of Mg, Ca and Sr had a 3:2 ratio (i.e. that of Ca10(PO4)6(OH)2” [184], which is equal to ‘‘amorphous HA” (Table
a TCP), while only Ba went directly to the stable 10:6 HA phase. 1) and ‘‘amorphous dicalcium phosphate” [185] are rare but have
Precipitated magnesium orthophosphate was amorphous and did been already mentioned. It should be noted that ACPs with the
not convert to a magnesium-deficient HA. Precipitated calcium Ca/P ratio <1.0 currently remain unknown.
orthophosphate formed ACP, which converted to CDHA by a solu-
tion-mediated autocatalytic mechanism [124]. Precipitated stron- 3.4.1. Precipitated ACPs
tium orthophosphate was not amorphous but poorly crystallized Although first described in 1953 [92], quantitative chemical
and readily converted in solution to a strontium HA. Thus, the studies on precipitated ACPs were not reported until 1965, when
smaller alkaline earth cation systems tended to form the more sta- methods were devised to isolate large amounts of unstable solids
ble amorphous 3:2 compounds [170]. for analysis. To minimize changing during sample drying, those
methods utilized filtration and/or centrifugation to wash excess ions
3.3. Morphology of precipitated ACPs from ACP slurries, then freezing wet ACPs under high vacuum to re-
move any remaining entrapped solvent by sublimation [122]. Early
When viewed by TEM, ACP solids precipitated from aqueous chemical studies [128,186] on ACPs prepared at pH 10.5, filtered,
solutions usually have a curvilinear appearance rather than the washed and lyophilized, showed that the Ca/PO4 molar ratio was
faceted and angular shape of crystalline calcium orthophosphates very close to 1.5, suggesting a TCP composition (as Ca3(PO4)2nH2O
[171,172]. However, this curvilinear aspect has only been clearly [97,187]). No OH ions were found in this compound. Furthermore,
established for dried ACP. The morphological form of highly hy- the electron spin resonance spectra of vanadyl (VO2+) ions adsorbed
drated flocculent solids that appear initially in freshly precipitated on ACP formed under varying conditions also revealed that ACP is
ACP suspensions is not known. What is observed when drops of either a non-crystalline form of hydrated TCP or a solid solution with
these suspensions are placed on carbon-coated grids, excess solu- the composition Ca3(PO4)l.87(HPO4)0.2 [188]. Other researchers re-
tion removed and air-dried are irregularly shaped, anastomosing ported a stoichiometry of the precipitated ACP more akin to OCP
aggregates of low-contrast, disk-shaped particles varying widely [130,132,134,189–194], DCPA [132] and DCPD [195]; however, with
in lateral dimensions (from 0.01 lm to >5 lm) [172,173]. These a few exceptions [132,134] the terms ‘‘amorphous OCP” and ‘‘amor-
highly flattened particles represent collapsed, desolvated residues phous DCPD” were not introduced: although the Ca/P ratio of early
of the initial wet ACP flocculates. formed ACP phases was varied between 1.35 and 1.38, which was
As ACP suspensions age, high-contrast particles with a more close to that of OCP (Table 1), the authors of Refs. [192–194] used
spherical aspect begin to appear, initially evolving as bud-like exten- the term ACP2 to explain initial variations in solution pH during
sions from the disks [172,173]. With time, these spherical forms the transformation of ACP into more crystalline phases of OCP
become the dominant shape for ACP. Although generally smaller and/or CDHA. Further, based on the results of TEM analysis, ACP2
(20–300 nm in diameter) than the disks they supplant, the spherical was identified as a separate amorphous phase with a floccular mor-
forms, like the disks, frequently aggregate into irregularly shaped phology and no electron diffraction pattern, if compared with a
and branching clusters. The progression from disk-shaped to ball- spherular morphology of the initially precipitated amorphous phase
like particles most probably represents a spontaneous desolvation (ACP1) [192–194]. The same terms ACP1 and ACP2 were used in an-
in situ or the initial gel-like flocculates into smaller, denser, less-hy- other study [137], in which the authors studied aging of calcium
drated structures [174]. That the spherules are formed in suspension orthophosphate precipitates in methanol at room temperature, fi-
and are not a drying artifact is supported by the crystallization nally leading to formation of a nanosized b-TCP. Furthermore, in pre-
behavior of ACP preparations. Although the evolution of a spherical cipitation experiments from aqueous solutions containing
morphology would be favored during consolidation as this shape polyethylene glycol, two types of ACPs were detected [142]: one
minimizes interfacial tension with the surrounding solution, it also with a broadened peak centered at 31° and another with a broad-
requires that the contracting surface be isotropic. This is possible for ened peak centered at 32° (Fig. 3.1, spectra 16:1 and 4:1, respec-
uniformly curved surfaces only when the enclosed structure re- tively). According to the authors, the first one was similar to the
mains non-crystalline while desolvating [20]. basic structure of b-TCP, while the second one was similar to the ba-
sic structure of HA [142]. A similar shift in the position of the amor-
3.4. Chemical composition phous maximum but obtained at a different aging time of ACP
precipitates (Table 2) has been detected in another study [126].
ACPs should be recognized as a special class of biomedically rel- Additional analyzes of ACPs prepared from aqueous solutions at
evant calcium orthophosphate salts having variable chemical but pH 7.4 with wide variations (from 5:1 to 1:5) in starting Ca/PO4
essentially identical glass-like physical properties. Presumably, molar ratios showed that the compositional Ca/P ratio decreased
all compounds mentioned in Table 1 might somehow be fabricated only slightly from 1.5 due to the presence of small amounts
in an amorphous state. Therefore, perhaps, some time in the future (<15%) of HPO2 4 ions [169,196–198]. This is an indirect confirma-
people will deal with an amorphous phase corresponding to the tion of the existence of an amorphous TCP. The presence of
chemical composition of MCPM (‘‘amorphous MCPM”), an amor- HPO24 in ACP is easy to detect by infrared spectroscopy, most nota-
phous phase corresponding to the chemical composition of DCPA bly by an absorption peak at about 890 cm1, due to a P–O(H)
(‘‘amorphous DCPA”), an amorphous phase corresponding to the stretching mode of protonated orthophosphate species (Fig. 7)
chemical composition of TTCP (‘‘amorphous TTCP”), etc. (in most [127,132]. Furthermore, in infrared spectra of HPO2 4 -containing
cases, stabilization procedures will become necessary), as well as ACPs, there is a large band of a weak intensity near 2300 cm1,
with various mixtures thereof. Currently most such compounds which corresponds to H–O(P) stretching in HPO2 4 ions [132]. More
4464 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

Table 2 presence of 20% of HPO2 4 in two ACP samples [199]. Furthermore,

Position of the XRD amorphous maximum at different aging times of ACP precipitates orthophosphate units close to water and a third (as yet unknown)
type of orthophosphate group were found in the NMR spectra of
Aging time 2 min 1h 2h 3h 4h 5h 6h the ACPs. Furthermore, substantial differences were noticed be-
2h (°) 29.5 30.1 30.4 30.3 30.5 30.8 29.0 tween the NMR spectra of two ACP samples, donated by two differ-
ent research groups [199]. In other studies, two ACP samples of
Note: The 2h values were derived from the profile analysis of the scattering curves.
Negligible changes were found for longer aged ACPs. The experimental
different chemical composition (since they had been prepared at
error ± 0.5°2h [126]. solution pH 6.5 and 10.0, respectively) were found to give very sim-
ilar extended X-ray absorption fine structure (EXAFS) spectra
[127,200]. The latter indicated that the SRO around calcium ions of
both ACPs is very similar; however, the reasons why the pH differ-
ences were not reflected in the calcium environment remained un-
clear. These experiments confirm the fact that ACPs are not a
single chemical compound but represent a special class of amor-
phous calcium orthophosphate salts. Thus, ACPs cannot be de-
scribed by a single chemical formula; a sketch CaxHy(PO4)znH2O
(Table 1) seems to be the most reasonable element demonstration
of this class of calcium orthophosphate salts. Presumably, ACPs with
the Ca/P ratio exceeding 1.6 should also contain some hydroxide
anions; however, no information on this point has been found in
the literature. However, the presence of CaO is frequently detected
in ACP-containing calcium orthophosphate coatings prepared by
plasma spraying.
Nevertheless, in slightly alkaline aqueous solutions, ACP might
have a well-defined chemical composition. For example, ACP slur-
ries in the pH range 7.4–9.25 appeared to have a nearly constant
solution ion activity product of 1.6  1025 when the solid-phase
composition is postulated to be Ca3(PO4)l.87(HPO4)0.2, i.e. when the
local chemical unit in ACP is postulated to have a Ca/P molar ratio
of 1.45, with 10% HPO2 -
4 , but without OH ions [197]. Thus, in
spite of the absence of LRO, this relative constancy in the composi-
tion over such a wide pH range indirectly suggests that ACP should
have some well-defined local chemical units. However, at solution
pH exceeding 9.25, ACP does not appear to have a nearly constant
solution ion activity product. This breakdown in the solution con-
Fig. 7. Infrared spectra after baseline subtraction for ACPs prepared at: (A) pH 10.0, stancy suggests that the solubility-controlling ions of ACP are sub-
(B) pH 7.0, (C) pH 6.5 and (D) pH 6.0. Reprinted from Ref. [127] with permission. The tly dependent compositionally on the preparation conditions. It is
complete (from 400 to 4000 cm1) infrared spectrum of ACP is available in Ref. [30].
possible that at solution pH > 9.25 the content of HPO2 4 ions no
longer remains constant and gradually decreases with increasing
to the point, the amount of HPO2 pH [122]. Furthermore, at more acidic pH 6.9, ACP precipitates
4 in ACP strongly depends on the
solution pH: the lower the Ca/P ratio for any given ACP precipitate, with Ca/P molar ratios as low as 1.15 have been reported [201].
the greater is its HPO2 These latter precipitates are extremely unstable and rapidly
4 content (Fig. 8).
The presence of HPO2 change into crystalline DCPD. Again, the term ‘‘amorphous DCPD”
4 ions in ACP has been confirmed by solid-
state nuclear magnetic resonance (NMR): the results revealed the was not been used in that study.
Even after lyophilization, solution-matured, spheroidal ACP sol-
ids still retain 15% water by weight [187,202]. A temperature-
programmed description analysis by Sedlak and Beebe indicated
that most (75%) of this retained water was tightly bound inside
the solid, the rest was a more loosely held surface water, with acti-
vation energies of 20.0 and 10.5 kcal mol1, respectively [203].
These results suggest that ACPs do not completely desolvate in
solutions but remain partially hydrated with about three water
molecules per formula unit. Other researchers found that water oc-
curred in regions that were only loosely associated with calcium
cations in ACP [5]. Furthermore, when prepared from carbonate-
containing solutions, ACPs can readily incorporate carbonate an-
ions [34–37,39,204,205]. The amount of carbonate incorporated
at any given pH increases with solution carbonate concentration.
At a given concentration, carbonate uptake also increases with
pH. Incorporating carbonate into ACP does not affect the HPO2 4
content but increases the Ca/P molar ratio. At physiological pH,
the carbonate content of ACP precipitated from solutions contain-
ing 30 mmol l1 carbonates is 3 wt.% [173]. These data suggest
Fig. 8. Acid phosphate content (per cent of total P as (HPO2
that ACP, if present in skeletal tissues, would contain appreciable
4 ) as a function of pH for
washed (dashed line) and unwashed (solid line) ACP precipitates. Reprinted from amounts of carbonates, although less than those present in the apatitic
Ref. [169] with permission. phases of bones [205]. Two other ions that readily incorporate into
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4465

the ACP structure are Mg2+ [36,43,127,170,206–208] and P2 O4 7 characteristic diffuseness resulted from a breakdown in atomic
[205,209,210]. Ions such as P2 O4 7 , carbonate and Mg
increase LRO within the interior of the synthetic ACP [213–215]. The major
the solution stability of ACP and, in the case of the latter two ions, peak positions were found to occur at distances of 2.55, 3.75 and
could possibly play an important role in maintaining the presence 6.40 Å with evidence of a peak at 2.9 Å [213]. Furthermore, any or-
of ACP in skeletal tissue. In addition, other ionic substitutions are derly atomic arrangements did not extend beyond 0.95 nm in
possible; however, such inorganic additives alter the ACP composi- diameter, which corresponds to the smallest values of a far-MRO.
tion, which would enhance the negative effects in the biomedical A contiguous periodic regularity in the distribution of these do-
application of ACP. Besides, with a few important exceptions, mains typical of crystalline materials was absent. EXAFS spectros-
ion-substituted forms of ACP [33–45] are not discussed in this copy [110,216] indicated that possible regularities in the local
review. environment around individual Ca2+ ions were even more circum-
scribed, not extending beyond distances of 0.3 nm (Fig. 9). This is
3.4.2. Other types of ACPs a SRO scale. In addition, an infrared analysis showed a similar lack
Little is known on the chemical composition of ACPs prepared of crystalline order for orthophosphate anions in the ACP structure
by other amorphization techniques. For example, various ACP sam- [30,123,132,218]. The method is based on the observation that a
ples prepared by compressing of several calcium orthophosphates splitting of the P–O antisymmetric bending mode at 550–
at very high pressures revealed collapse of their initial crystal 600 cm1 (Fig. 2.2) increases as crystallinity increases. This appar-
structures but possible changes in their chemical compositions ent lack of crystalline regularity is one of the striking features of
were not investigated [166,167]. Interestingly, the authors found ACP that distinguishes it from other calcium orthophosphates
that in the region below 550 cm1 the infrared spectra of DCPD and provides the structural basis for its name [20].
in amorphous phase resembled that of HA in the crystalline phase, The apparent absence of observable crystalline features in ACPs
and conversely the spectra of DCPD in the crystalline phase resem- does not exclude the possibility that ACPs might have a well-de-
bled that of HA in the amorphous phase [167]. In the case of fined local structural unit. The compositional constancy of ACPs
milling, calcium orthophosphates were found to become amor- over a wide range of preparative solution conditions suggests an
phous; however, no additional phases were detected [149,150]. existence of such a core structure [98]. Several lines of evidence,
Presumably, this means that during amorphization their chemical however, indicate that the local unit of ACPs is not a cryptostruc-
composition remained unchanged. tural variant of one of the crystalline calcium orthophosphates.
Concerning the ACPs formed in plasma-sprayed HA coatings, For example, although in the majority of cases precipitated ACPs
the authors of one study have reported that ‘‘the amorphous phase appear to be closest in composition to TCP, in aqueous solutions
mostly consists of a dehydroxylated calcium phosphate” [163], ACPs transform into either OCP or CDHA but not into TCP. If ACP
which, presumably, meant dehydroxylated HA. If so, the chemical were a cryptocrystalline TCP, direct growth into observably crys-
composition of that particular ACP should be close to amorphous talline TCP by crystal ripening would be expected to occur. The
OA. The authors of another study considered ‘‘that the amorphous aforementioned possibility that ACP may be a cryptocrystalline
phase substance consists of HA molecules” [152]. However, in a CDHA or HA cannot be as easily excluded. Only below pH 9 can
subsequent study, the same authors mentioned that ‘‘the plasma- CDHA be ruled out for the same reason as TCP. However, even
sprayed amorphous phase is an oxyapatite” [153]. No further clar- above this pH, the finding that CDHA forms primarily as an out-
ification has been provided; however, all these authors reached the growth from the surface of ACP suggests that an in situ ripening
same conclusion regarding the apatitic chemical composition of process does not occur. Dissimilarity in composition rules out
the plasma-sprayed amorphous phases. Furthermore, these ACPs ACP as being made up of highly disordered arrays of OCP unit cells.
are definitively anhydrous, contrary to the precipitated ACPs. It is equally unlikely that ACP is an orthophosphate-deficient OCP
To conclude the chemical part, the solubility of ACPs should be as the structural integrity of the latter depends on a full comple-
briefly mentioned. Due to chemical variations, this value cannot be ment of orthophosphate groups in the unit cell. The finding that
measured precisely (Table 1). Several different solubility products the Ca/P molar ratio remains relatively constant at 1.5 over
have been proposed for ACPs and interested readers are referred a wide range of pH (7.4–9.25) appears to exclude ACP being a
to Table 1 of Ref. [30] for details.

3.5. Structure

The atomic arrangement within ACPs depends on the chemis-

try. The first quantitative studies on a synthetic ACP were done
in the mid-1960s on a material precipitated at pH 10 [128,186].
As Watson and Robinson found at neutral pH, the initial phase that
spontaneously formed immediately upon mixing concentrated
alkaline Ca- and PO4-containing solutions was structurally non-
crystalline [92]. The XRD pattern of this rapidly precipitated phase
showed only two very broad and diffuse peaks, typical for sub-
stances that lack periodic LRO [186].
Initially, there were suggestions that a synthetic ACP was, in
fact, HA of such small crystal dimensions that its XRD pattern
was widely broadened to appear amorphous in character. How-
ever, calculated XRD patterns assuming that ACP consisted of small
groups of HA unit cells, or even a single HA unit cell, did not match
the observed ACP diffraction data [211]. The NMR spectra of ACP
are also sufficiently different from HA to suggest that they do not
Fig. 9. Diagrammatic representation of the SRO structure of four phosphate
have the same structural motif [212]. The probability that ACP tetrahedra and two water molecules about calcium ions in an acidic ACP, calculated
was structurally distinct from HA then led to a study of this mate- using the DCPD shell model. The positions of hydrogen atoms are not determined.
rial by the X-ray radial distribution method, which showed that its Reprinted from Ref. [217] with permission.
4466 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

cryptocrystalline mixture of OCP and CDHA. Instead of remaining

constant, an increase in pH would be expected to increase the overall
Ca/P molar ratio of such a mixture as the relative proportion of the
OCP and CDHA components would shift in favor of the latter [20].
Based on the relatively constant Ca/P ratio of ACP formed under
varying conditions, such as different concentrations of calcium and
orthophosphate ions, solution pH and different temperatures, as
well as spectroscopic and structural analyzes, Posner and Betts
hypothesized that the initial solid phase of ACP (more precisely,
of amorphous TCP) precipitated in vitro consisted of spherical par-
ticles ranging from 300 to 1000 Å in diameter with water mole-
cules in the interstices [98,214,215]. In other words, the defining
structural unit of ACP is a spatial subset of the HA unit cell consist-
ing of a central Ca2+ ion coordinated by the oxygens of six sur-
rounding orthophosphate groups, which, in turn, are stabilized
by another eight Ca2+ ions spherically distributed around the outer
boundary of the subset (Fig. 10). This is a neutral ion cluster 9.5 Å Fig. 11. A model of Posner’s cluster (in a circle) showing its relationship with the
in diameter, whose composition is expressed by the formula HA crystal structure. Black lines forming a rhomb delimiting the borders of one unit
cell of HA. Reprinted from Ref. [30] with permission. A similar relationship is also
Ca9(PO4)6. Since then this structure has been defined in the litera- available in Ref. [242]. This correlation between Posner’s clusters and HA structure
ture as a Posner’s cluster. It was later suggested that these clusters was first published in Ref. [98]. One should note that in Ref. [220] another set of
in fact possess a S6 symmetry [219,220]. This subset, which has a atoms of the HA crystal structure has been chosen to represent a Posner’s cluster.
composition close to that of TCP, is linearly expanded by 3% and
its radial distribution function is similar to that calculated from
the diffuse XRD profile of ACP. The authors postulated that at the faces associated with [Ca3(PO4)2]n clusters were analyzed in detail
far-MRO level these slightly enlarged spherical subsets are ran- using ab initio calculations for n = 1–4 [223]. The energy criteria
domly clustered in ACP particles (Fig. 10) with water filling the were found to favor the Posner’s cluster, which is the core of the
intervening spaces [98,214,215]. No data have been found in the actual structural model of ACP (more precisely, of amorphous
literature to suggest that plasma-spayed ACPs cannot contain Pos- TCP). Moreover, the calculations showed that an aggregation of
ner’s clusters as the structural units; however, undoubtedly, they the clusters corresponded to a large energy stabilization irrespec-
cannot contain water molecules in the intervening spaces. Interest- tive of the cluster considered [223], which is in agreement with
ingly, an atomic arrangement of Posner’s clusters appears to be Posner’s hypothesis in which clusters are proposed to be closely
analogous to that existing in several other calcium orthophos- packed in ACP structure. Furthermore, the vibrational spectra of
phates, such as HA, OCP and b-TCP. For example, the relationship ACP have been modeled as well [224]. In spite of this, it is not quite
between the original Posner’s cluster and the atomic structure of clear from what experimental spectroscopic or structural data the
HA is represented in Fig. 11. However, it should be stressed that cluster model (Fig. 10) has been derived, except the TCP composi-
this is only a model because the arrangements of clusters in larger tion. Further, it is not entirely clear whether there are volumetric
structures are not known. regions within the large 300–1000 Å particles that are free of the
In general, an electrostatic interaction, a hydrophobic interac- 9.5 Å clusters. In other words, are there some volumetric regions
tion and a ‘‘cementing” effect of water molecules are among the within the large 300–1000 Å particles where the atoms are com-
main factors holding ions of a cluster together [221]. Again, a pletely random with no SRO? Moreover, the results of EXAFS anal-
‘‘cementing” effect of water molecules is applied for the ACPs pre- ysis indicated that the range of orderly Ca–Ca and Ca–P
cipitated from aqueous solutions only. Theoretical investigations interactions in ACP were much shorter than would be predicted
on the stability of different calcium orthophosphate clusters with from the Betts and Posner model [110,216]. Another weakness in
an increasing number of ions have confirmed that Posner’s clusters the model is that water only serves to fill the interstices between
are the most stable arrangement [222]. The potential energy sur- the HA subsets and is not an essential part of the ACP structure,
contrary to the assumptions by Sedlak and Beebe [203]. Both of
these weaknesses could possibly be remedied if a smaller spatial
domain is carved from a portion of the OCP unit cell that contains
part of the hydration layer. However, this suggestion has not been
examined in detail. It is also possible, however, that the defining
structural unit for ACP is a truly unique entity with no satisfactory
crystallographic model. In fact, there is no a priori need for such a
model, as the defining unit for ACP would not be constrained struc-
turally by the symmetry requirements for crystalline arrange-
ments. Therefore, ACP could have a structural motif not found, or
even permissible, in any of the crystalline calcium orthophos-
phates [20].
More to the point, mechanisms of calcium and orthophosphate
ion association in aqueous solutions have been elucidated by
means of quantum and classical molecular mechanics simulations
[225]. A special focus was dedicated to the role of the protonation
state of orthophosphate ions and depronation of the hydrogen
orthophosphate ions appeared to be necessary during crystal
growth. According to the simulation results, a triple ion [Ca2+(H-
Fig. 10. A model of an ACP particle, showing one Posner’s cluster. Reprinted from PO4)2Ca2+]2+ could form in aqueous solutions and subsequently
Refs. [19,99] with permission. yield another triple ion [Ca2+(PO4)3Ca2+]+ by releasing a proton.
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4467

The latter ion was suggested to be the smallest stable aggregate, of hydroxyl-depleted areas of the amorphous phase to OA and
and to contain an entirely deprotonated orthophosphate ion 230 kJ mol1 for crystallization of hydroxyl-rich areas of the amor-
[225]. Some indirect experimental evidence in support of this phous phase to HA [163]. Therefore, hydroxylated ACP regions were
hypothesis has been obtained recently [226]. Nevertheless, verifi- found to crystallize more readily compared to the dehydroxylated
cation of the aforementioned structural models of ACPs will ones. A heat of recrystallization of ACP to HA was calculated to be
require details of the chemistry and the processing conditions to- 43 kJ mol1 in yet another study [161].
gether with viewing techniques at the atomic scale. Unfortunately, To conclude the thermal part, the results mentioned in this sec-
no additional information on the structure of high-temperature tion appear to be additional indirect confirmation of the fact that
ACP phases has been found in the literature. Thus, for want of any- ACP is not a single chemical compound but represents a special
thing better, one is forced to assume, that except for water mole- class of amorphous calcium orthophosphate salts.
cules and the possible presence of HPO2 4 ions, all the
aforementioned is valid for ACPs produced using plasma-sprayed 3.7. Amorphous-to-crystalline transformations in aqueous solutions
coatings. This assumption should clearly be verified.
To conclude the structural part, determination of the specific As previously stated, in the vast majority of cases, ACPs are the first
surface area of ACPs generally leads to surprisingly low numbers. solid phases to appear upon mixing of calcium- and orthophosphate-
This might be related to the larger spherical associations of Pos- containing aqueous solutions at pH > 7 and concentrations
ner’s clusters and the apparent hindrance to nitrogen adsorption sufficiently high to produce an immediate precipitation. The sponta-
of these inner surfaces [180,202]. neous formation of ACPs is a kinetically driven process. A rapid mixing
of highly concentrated solutions creates sufficiently strong stochastic
3.6. Thermal properties interactions between the ions and they quickly coalesce into irregu-
larly coordinated, highly hydrated clusters large enough to separate
All types of ACP are thermally unstable and sustain neither cal- from solution in a gel-like state before they have a chance to rearrange
cining nor sintering. If precipitated ACPs are heated, they first of into orderly nuclei capable of growth as crystals. This structural
all lose water. Two types of water loss occur, corresponding to arrangement, however, is inherently unstable. In addition to desol-
loosely bound water molecules adsorbed on the surface of ACP vating, ACPs kept in solution eventually disappear, being supplanted
agglomerates and more strongly bound internal water molecules, by more stable crystalline phases such as OCP or CDHA [20].
respectively. The first loss is essentially reversible, whereas the sec- Watson and Robinson in their pioneering study on ACP [92] were
ond is mostly irreversible [203,227]. Furthermore, in experiments the first to observe the transient nature of ACP when kept in contact
where water interactions were minimized by allowing for the with its preparative medium. They found that electron diffraction
escape of volatile components, crystallization of ACP was found to patterns of ACP taken later in the precipitation reaction were no
begin at about 530 °C [187]. Below this temperature, the non- longer diffuse but resembled patterns of a poorly crystalline CDHA.
crystalline features of ACPs seemed thermally stable. The first crys- Further investigations revealed that this amorphous-to-crystalline
talline phase to appear was invariably b-TCP. However, between 600 transition was not gradual but occurred rather precipitously. Ini-
and 800 °C, depending upon the preparation, a-TCP was found to tially, there is a period of a relative stability, where surfaces of the
become generally the favored ignition product, even though b-TCP high-contrast spherules generally remain smooth and regular
is normally the stable phase up to 1200 °C. Neither washing/drying [172]. However, as seen from the data of Table 2 [126], some changes
procedures employed to isolate the amorphous material, nor the occur with solid ACP during this time. Afterwards, the transition fol-
choice of soluble orthophosphate salt used in its preparation, were lows a sigmoid evolution with the solid phase rapidly progressing
found to have any significant effect on the thermocrystallization from being barely crystalline to where the amorphous features dis-
properties of ACP [187]. However, in other studies, both a-TCP appear. Once the first crystals appear on the surface of the spherules,
[139,164,165,180] and carbonated CDHA [133] appeared to be the the transition proceeds rapidly to completion. Simultaneously, dra-
first detectable crystalline phase obtained from heating various matic declines in ionic concentrations of calcium and orthophos-
ACPs to 550–600 °C. Interestingly, the presence of organic solvents phate ions occur in the mother solution. The time it takes to reach
(in that case, polyethylene glycol) at the ACP preparation stage this amorphous-to-crystalline boundary varies considerably with
was found to influence the products formed at elevated tempera- the preparation conditions, being particularly sensitive to tempera-
tures [142]. Namely, when the amount of polyethylene glycol was ture and pH [66,124]. For example, at pH 7.4, ACP converts five
small, a-TCP was formed on heating; when the amount of polyeth- times faster at 37 °C than at 20 °C [229]. The pH dependency is some-
ylene glycol was large, b-TCP was formed on heating; biphasic what more complex than that for temperature. Namely, at 25 °C the
(a-TCP + b-TCP) formulations were formed when the amount of aqueous lifetime of freshly precipitated ACP is less than 0.3 h at pH
polyethylene glycol was average (Fig. 3.2). Similar effects of both 7.4. It increases to a maximum lifetime of over 9 h between pH
aging time and the solution pH were also detected [143]. 10.0 and 10.5, then rapidly decreases until at pH 12.8 the lifetime
ACPs with Ca/P ratios of 1.00 (‘‘amorphous DCPA”) and 1.34 is nearly as short as that at pH 7.4 [197,230]. The solution lifetime
(‘‘amorphous OCP”) were found to remain amorphous after heating of ACP can be greatly extended by inclusion of simple inorganic ions
to 600 °C, while crystalline compounds (b-Ca2P2O7 in the case of Ca/ such as Mg2+, Zr2+, silicates, carbonates and pyrophosphates
P = 1.00 and a-TCP + b-Ca2P2O7 in the case of Ca/P = 1.34) started to [39,40,42,127,170,194,206–209,231,232]. As an extreme example,
appear at 620 °C [132]. In the same study, ACP with Ca/P ratio of 1.51 ACP prepared from Mg2+-containing solutions at pH 10.0 and
(‘‘amorphous TCP”) was found to remain amorphous at heating up to 32.5 °C remained in a gel-like amorphous state for up to 20 weeks
550 °C, while crystalline compounds (b-TCP) started to appear at when the reactant Mg/Ca molar ratio was set at 0.2 [170]. Other sub-
600 °C. Interestingly, heating of a crystalline DCPA (monetite) stances can increase the stability of ACP in aqueous solutions include
leads to c-Ca2P2O7 and this phase is then transformed at 750 °C F [229,230], various polyelectrolytes [65,66], polyalcohols and
to b-Ca2P2O7 [228]. Thus, amorphous DCPA showed a thermal polyglycols [125,136,138], phospholipids [233], dentin phospho-
behavior different from that of crystalline DCPA [132]. protein [234], phosvitin [210,234], glycochenodeoxycholic acid
Furthermore, the crystallization of ACPs is an exothermic pro- [235], biomacromolecules such as casein phosphopeptide [67], as
cess. The heat produced was found to be 21 kJ mol1, while the well as adenosine di- and triphosphates (but not the monophos-
activation energy was 450 kJ mol1 [157]. Other researchers phate) [236,237]. On the other hand, an excess of Ca2+ ions in the
reported activation energy values of 440 kJ mol1 for crystallization solution accelerates the transformation of ACP into a crystalline
4468 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

CDHA [168]. Interestingly, collagen, which is the principal matrix To conclude this part, as ACPs represent a special class of cal-
protein in skeletal tissues, has no effect on ACP stability [229,234], cium orthophosphates, it is reasonably to assume that several
while the presence of gelatin promotes the transformation of ACP amorphous-to-crystalline transformation mechanisms might exist
into crystalline phases [47]. It should be noted that in a wet atmo- due to the initial differences in the investigated ACPs. Furthermore,
sphere solid ACP with a Ca/P ratio of 1.33 (amorphous OCP) was various competitive processes might occur simultaneously and
found to recrystallize into a mixture of CDHA + DCPA [134]. their importance might depend on the conversion conditions. Fur-
Conversion of ACPs to solid phases with XRD patterns similar to ther details on this topic are available in the literature [30,117].
that of CDHA has been studied largely in aqueous suspensions
[65,66,110,113,115,124,126,146,168,176,181,186,208,238–241]. 4. ACP in vivo
Transmission electron micrographs of this transformation are
shown in Fig. 4. The kinetics of this process can be described by As stated in the Introduction, both physical and morphological
the following empirical equation: dC/dt = k1 + k2C, where C is the evidence for the presence of ACPs in skeletal tissue has been diffi-
fraction of ACP converted into the crystalline phase by time t, k1 cult to establish directly, and the validity of inferential evidence for
is a rate constant associated with the nucleation of the first crystals the presence and amounts of ACPs has been the subject of consid-
and k2 is an autocatalytic rate constant indicative of the observa- erable debate. Indirect assessments, such as by XRD methods, have
tion that the transition rate is proportional to the mass fraction al- produced widely varying estimates of the amorphous content of
ready crystallized and not to the fraction of remaining ACP bone mineral, placing it at less than 1% [19] to more than 30%
[128,186]. Numerical values for k2 appear to be much larger than of the total mineral mass, the rest presumably being poorly crystal-
those for k1, reflecting the exponential rapidity of the transition line ion-substituted CDHA (biological apatite). Even if ACPs only
once this has started [186]. occur at a lower percentage, one would expect TEM to reveal some
The amorphous-to-crystalline transformation mechanisms of evidence [9], but, as stated earlier, most TEM studies of bones and
ACPs have not been well elucidated. Chemically, this process is de- teeth do not even mention the existence of amorphous-like struc-
scribed by Eqs. (1) and (2), while in the reality it might proceed tures that could be ascribed to ACPs. However, the absence of such
along several pathways. For example, it might occur via dissolution structures could have been a negative artifact caused by aqueous
of ACP and reprecipitation of crystalline phases (e.g. CDHA) dissolution of more labile ACPs during the sample preparation for
[20,112,115,122,124,146,181,238], internal structural rearrange- examination. A few early TEM studies [102,103] that avoided aque-
ments [168,181,208,219,242,243], development of a LRO without ous processing of bone specimens by directly embedding and
changing the immediate environment of Ca [110], formation of sectioning freeze-dried material revealed a zone of electron dense,
crystalline phases directly within the ACP phases [141,226], or 6–20 lm diameter spheroidal bodies adjacent to crystal-rich areas
self-aggregation and surface-mediated transformations [30]. For of bone tissue. Electron diffraction of these sites revealed a hazy,
example: ‘‘When the density reached a critical value, the random diffuse pattern similar to that observed by Watson and Robinson
arrangement of growth units became disadvantageous in terms [92] in their synthetic, amorphous-like precipitates. Whether or
of total free energy, resulting in a sudden regularization of the not these spheroidal particles represented ACPs has been much de-
structure, which was deposited as HA” [219]. Furthermore, Yin bated, but the labile nature of ACPs suggests the need to maintain
and Stott suggested that, in the transformation from ACP to CDHA, carefully the anhydrous conditions in order to preserve this phase
ACP needed only to dissociate into clusters rather than undergo in calcified material for examination [20].
complete ionic solvatation [244]. Another possibility that could account for the inability to estab-
Having summarized findings of their own and previously pub- lish with certainty the existence of ACPs in bone is that the ACP
lished results by other researchers, Wang et al. [226] depicted does not exist as separate particles but, instead, as an amorphous
the following picture of the main events that take place during layer on crystals of the poorly crystalline phase of biological apa-
the induction period and finally trigger the rapid precipitation of tite. Although there has been no direct evidence for this possibility,
calcium orthophosphates from supersaturated aqueous solutions: similar coatings appear to form in physiological-like solutions
‘‘Calcium and orthophosphate ions form pairs and clusters succes- seeded with HA [245,246]. Under these in vitro conditions, the
sively in the first few seconds. These ions, pairs and clusters then initial accretions appeared to form an amorphous calcium carbon-
compose the initial solid phase that is heavily hydrated and con- ate-ACP coating on the seed crystals. The initial phase also incorpo-
tains hydrogenorthophosphates. Growing and aggregating, the so- rated a small amount of Mg2+ ions from solution that was
lid increases in size and quantity without affecting the solution pH. subsequently released upon the ancillary formation of CDHA crys-
During the induction period, the solid particles exhibit a steady tals. This Mg2+ behavior, consistent with the amorphous-to-crys-
size distribution, which is around 300 nm at the early stage and talline transition, is the most compelling compositional evidence
shifts toward 1000 nm with time. These particles are agglomerates from these studies for the initial coat being an amorphous layer
of primary particles of 60–100 nm in diameter and are originally [245].
amorphous in structure. At multiple sites inside a particle, crystal- A proper assessment of the possibility that some of the mineral
line domains develop from ion pairs and/or clusters by taking up in skeletal tissues is in a free-standing amorphous state is further
calcium and releasing hydrated proton, possibly through a stage complicated by the fact that the minimum ion activity product
at which the more compact cluster Ca9(PO4)6 presents. Since the needed to form ACP de novo in physiological-like synthetic solu-
expansion of crystalline domains consumes surrounding calcium tions at pH 7.4 is considerably greater than that calculated for ser-
and orthophosphate ions (or their pairs and clusters) and releases um [247,120]. If extracellular skeletal fluids were in electrolyte
hydrated protons, the mechanic strength decreases in the inter-do- equilibrium with serum, it would appear unlikely that ACPs could
main regions. Finally, under the action of the shearing strength of form in vivo except possibly as a coating on crystals of biological
the fluid, these primary particles collapse and the liberated crystal- apatite of bones. Without stronger evidence than that described
lites induce the rapid precipitation of calcium orthophosphates, to- above, even this possibility is highly speculative. However, there
gether with the previously trapped hydrated protons in primary is some evidence that suggests that calcifying bone matrix may
particles, resulting in the abrupt pH drop. Indeed, it is the crystal- be compartmentalized with the establishment of an interior milieu
lization at multiple sites inside amorphous particles that finally different from that of serum [248]. Unfortunately, it is not known
triggered the rapid precipitation of calcium phosphate from the whether such compartmentalization results in an extracellular
supersaturated solution” [226]. fluid space capable of initiating de novo ACP formation [20].
S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475 4469

Although the general compartmentalization of bone-producing with considerable amounts of TTCP and trace amounts of a-TCP
conditions favorable for ACP needs to be further established, a con- [264]. Therefore, these cases can hardly count as biomedical appli-
siderable body of evidence suggests that local micro-compart- cations of ACPs.
ments exist in bones that could allow for ACP development. The There is another type of calcium orthophosphate coating, ob-
most thoroughly studied of these micro-spaces are the mem- tained by a biomimetic route. Some of these coatings contain
brane-enclosed aqueous cores of matrix vesicles [249,250]. Most ACP as the major component [265–268]. For example, an ACP coat-
commonly found near osteoblasts in the extracellular regions of ing on polyethersulfone plates was prepared by precipitation from
rapidly mineralizing embryonic bone, these spherical bodies of cel- a simulated body fluid (SBF) at 35 °C. It took the authors 12 days to
lular origin are the sites of initial mineral formation [251]. Preced- obtain a 20 lm thick ACP coating and 28 days for a 50 lm thick
ing appearance of the first crystals at these sites is an accumulation one [265]. Similar results were obtained in other studies
of calcium and orthophosphate ions within the aqueous cores of [266,267]. One should stress that, in the vast majority of cases,
the vesicles to levels that far exceed the threshold level for de novo the biomimetic approach presumes application of SBF, which con-
ACP formation [252,253]. Studies with synthetic liposomes confirm tains ions of sodium, magnesium, potassium, sulfate and hydrog-
that ACP should readily form under such compartmentalized con- encarbonate; thus, ion-substituted ACPs are always formed as a
ditions [254]. However, Raman spectroscopic data indicate that result.
ACP in matrix vesicles is not in a pure chemical state but instead Furthermore, ACP coatings can be deposited on titanium using
calcium and orthophosphate ions are combined in a single-phase an electrochemical technique at 36 °C and solution pH 6.4 [268].
complex with lipid and protein moieties found within the vesicles The obtained ACP coatings appeared to be unstable and trans-
[255]. Also consistent with formation of an amorphous precursor formed into those of CDHA. By choosing the electrochemical
phase are infrared and Raman spectroscopic findings that the first parameters, a homogeneous coating of ACP, CDHA or some inter-
crystals in matrix vesicles are OCP and not CDHA [255,256]. These mediate phases could be achieved, thus allowing formation of
crystals, in turn, penetrate the enclosing membrane and initiate a the coatings with different morphologies and solubilities [268].
chain of crystallization events that appears to trigger the mineral- Among the self-setting calcium orthophosphate cements and
ization of the collagenous matrix. Thus ACP, even when present in concretes [258,259], there is only one ACP-based formulation cur-
small amounts, may be an important initiating factor in the calci- rently on the market, namely BiobonÒ (a-BSMÒ) [269,270]. This ce-
fication of skeletal tissues [20]. ment comprises a mixture of ACP (50 wt.%) and DCPD (50 wt.%)
In addition to the questionable cases of calcified tissue, ACPs are which is mixed with an appropriate amount of aqueous medium
found in mammalian milk [7,8,70]. Clearly, the presence of the (deionized water or saline) with a liquid to solid ratio of 0.8 ml g1
most easily biodegradable calcium orthophosphates in the form at room temperature. An injectable paste is obtained which sets in
of ACPs in milk is necessary to construct skeletons of young less than 20 min at 37 °C. After hardening, the cement is consti-
organisms. tuted of nanocrystalline CDHA with crystal dimensions close to
those in human bones [30]. A similar cement formulation but addi-
5. Biomedical application of ACPs tionally containing mechanoactivated b-cyclodextrins has been
studied as well [147]. Furthermore, an ACP + DCPA cement formu-
Currently, biomaterials and bioceramics of calcium orthophos- lation is known [271]. ACP can also be added as an admixture
phates are available in various physical forms: powders, particles, phase to standard calcium orthophosphate cement formulations
granules, dense blocks, porous scaffolds, injectable formulations, [43,272]. In such cases, addition of ACP resulted in cements exhib-
self-setting cements and concretes, implant coatings, as well as iting shorter setting times, a compressive strength suitable for
composite components of different origin (natural, biological or non-load-bearing applications and full conversion to nanocrystal-
synthetic) often with the specific shapes, such as implants, pros- line CDHA. Moreover, ACP-containing formulations demonstrated
theses or prosthetic devices [31,32,257]. In principle, all these good cell viability, making them suitable candidates for biomedical
physical forms should apply to ACPs; however, not all of them have applications [272]. More to the point, amorphous TCP nanoparti-
yet been realized. It is easy to prepare ACPs in powder form by a cles of 13, 19 and 40 nm diameter were found to be highly reactive
wet-precipitation technique (see Section 3.1); however, manufac- and set to CDHA within minutes, if compared with microcrystalline
turing of other physical forms of ACPs is not so simple. Further- powders of both a-TCP and b-TCP [178,179]. A similar approach
more, for use in surgery, all implantable 3-D constructions must was reported in other studies [148–150].
possess the necessary mechanical properties, which is difficult to Since ACPs do not sustain heating above 600 °C (see Section
achieve in the case of ACPs. For example, both dense blocks and 3.5), the possibility of preparing dense ACP bioceramics was stud-
3-D porous scaffolds made of calcium orthophosphates achieve ied using spark plasma sintering at temperatures ranging from 150
their desired mechanical properties only after sintering at temper- to 200 °C with processing times of <15 min. Unfortunately, the ob-
atures exceeding 1000 °C, which is impossible in the case of ACPs served mechanical strength of the prepared consolidated disks
(see Section 3.5). Furthermore, an ACP powder might be easily (sintered for 6 min at 150 °C) was poor [273]. However, an increase
added as a component to self-setting cements and concretes; how- in the sintering processing time to 13 min indicated a possibility of
ever, no cement formulation is known which results in ACP forma- improving the mechanical properties. The low-temperature condi-
tion as the major end-product [258,259]. More to the point, ACPs tions appeared to be more adaptive to the processing of ACP when
are present as components of various calcium orthophosphate compared with the experimental conditions in conventional sin-
coatings; however, only a few studies are known in which ACP tering. However, the physicochemical characterization of the pre-
coatings have been fabricated [260–263]. pared consolidated bioceramics indicated a crystallization of the
Not many examples of biomedical application of ACPs are cur- initial ACP to an apatitic phase with no other detectable crystalline
rently available. Most cases of plasma-sprayed calcium orthophos- phase [30].
phate coatings should be excluded because the coatings consist of However, the majority of cases of the biomedical applications of
a complex mixture of various phases, of which ACP is just one ACPs comprise various biocomposites and hybrid biomaterials
[9,151–163]. Furthermore, both the amount and the composition containing ACP as one of the phases [47–72,147,274–285]. Several
of amorphous phases cannot be accurately controlled. Even in ACP-containing formulations (e.g. Recaldent™ and Enamelon™)
the case, when ACP was plasma sprayed, the coating was found are now commercially available [285]. As can be seen from these
to be a mixture of a crystalline carbonate-containing apatite phase references, most of the applications are in the field of dentistry.
4470 S.V. Dorozhkin / Acta Biomaterialia 6 (2010) 4457–4475

In all cases, addition of calcium orthophosphates, including ACPs, a SRO) matter? A good attempt to discuss this topic is available in
imparts both biocompatibility and bioactivity to the biocomposites the literature [312], and interested readers are referred to this.
[278]. For example, to improve cell adhesion, a hydrophilic array of To conclude, the biomedical applications of ACPs are currently
ACP was fabricated on a surface of hydrophobic polystyrene [286]. limited due to the great difficulties in consolidating these materials
A similar effect was found when coatings composed of ACP and into 3-D structures, which have to possess sufficient mechanical
hyaluronic acid were used [287]. Furthermore, in the acidic oral properties. As ACPs do not sustain heating at temperatures exceed-
environment the ACP-containing biocomposites take advantage ing 600 °C, they must be consolidated by low-temperature tech-
of the ability of ACPs to release calcium and orthophosphate ions, niques only. Spark plasma sintering is one such method [30,273];
which potentially can take part in enamel remineralization [49– another comprises mixing of thermally unstable calcium orthophos-
64,276,277,179,288–307]. Such ACP-containing biocomposites phates with water-soluble porogens, followed by cold isostatic
and hybrid biomaterials might be coatings [279] and cements pressing of the prepared mixture and dissolving the porogens
[147,274,275]. In dentistry, the ACP-containing formulations are [313]. Obviously, other consolidation approaches are possible, and
used mainly as anticariogenic and/or remineralizing agents [288– these need to be developed.
307], e.g. in chewing gums [291–293], sugar confections [71], var-
ious tooth mousses [294–296], bleaching gels [297,298], various Acknowledgements
drinks [299,300] or even in milk [304,305]. Furthermore, ACP-con-
taining formulations are used in orthodontics [280–284]. Many thanks to various contributors for their kind permissions
to reproduce figures. Special thanks to Dr. Marc Bohner for sending
6. Conclusions me a copy of ‘‘Amorphous calcium phosphate”, a chapter from
Eanes [20].
ACPs can be both found in living organisms (especially inverte-
brates) and synthesized in the laboratory. Due to their chemical
variability, they should be recognized as a special class of calcium Appendix A. Figures with essential colour discrimination
orthophosphates offering a wide variety of compositions. Presum-
ably, all known calcium orthophosphates (see Table 1) might be Certain figures in this article, particularly Figures 1 and 3, are dif-
prepared in an amorphous state; however, not all of them (espe- ficult to interpret in black and white. The full colour images can be
cially, those with Ca/P ratio <1.1) have been prepared. Further- found in the on-line version, at doi:10.1016/j.actbio.2010.06.031.
more, in the available literature nothing has been found on the
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