materials
Article
Diffusion of Copper Ions in the Lattice of Substituted
Hydroxyapatite during Heat Treatment
Natalia V. Bulina 1, * , Natalya V. Eremina 1 , Olga B. Vinokurova 1 , Arcady V. Ishchenko 2 and Marina V. Chaikina 1
Citation: Bulina, N.V.; Eremina, N.V.;
Vinokurova, O.B.; Ishchenko, A.V.;
Chaikina, M.V. Diffusion of Copper
Ions in the Lattice of Substituted
Hydroxyapatite during Heat
Treatment. Materials 2022, 15, 5759.
https://doi.org/10.3390/ma15165759
Academic Editor: Arne Berner
Received: 19 July 2022
Accepted: 11 August 2022
Published: 20 August 2022
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Materials 2022, 15, 5759. https://doi.org/10.3390/ma15165759
1 Institute of Solid State Chemistry and Mechanochemistry, Siberian Branch of the Russian Academy of
Sciences, Kutateladze Str. 18, 630128 Novosibirsk, Russia
2 G.K. Boreskov Institute of Catalysis, Siberian Branch of Russian Academy of Sciences,
Pr. Akad. Lavrentieva 5, 630090 Novosibirsk, Russia
* Correspondence: bulina@solid.nsc.ru
Abstract: The doping of hydroxyapatite with various substituent ions can give this material new and
useful properties. Nonetheless, local distortions of structure after doping can change the properties
of the material. In this work, the thermal stability of copper-substituted hydroxyapatite synthesized
by the mechanochemical method was investigated. In situ diffraction analyses showed that copper
ion diffusion during the heating of Cu-substituted hydroxyapatite promotes phase transformations
in the substituted hydroxyapatite. The behavior of copper ions was studied in samples with ratios
(Ca + Cu)/P = 1.75 and 1.67. It was found that in both cases, single-phase Cu-substituted hydroxyap-
atite with the general formula Ca10−x Cux (PO4 )6−y (CO3 )y (OH)2−y Oy is formed by the mechanochem-
ical synthesis. When heated at approximately 600–700 ◦ C, the lattice loses copper cations, but at
higher temperatures, CuO diffusion into the hydroxyl channel takes place. Cuprate-substituted
hydroxyapatite with the general formula Ca10 (PO4 )6 (OH)2−2x (CuO2 )x forms in this context. At
1200 ◦ C, the sample is single-phase at (Ca + Cu)/P = 1.75. Nonetheless, slow cooling of the material
leads to the emergence of a CuO phase, as in the case of (Ca + Cu)/P = 1.67, where the material
contains not only CuO but also Cu-substituted tricalcium phosphate. In the manufacture of ceramic
products from Cu-substituted hydroxyapatite, these structural transformations must be taken into
account, as they alter not only thermal but also biological properties of such materials.
Keywords: mechanochemical synthesis; hydroxyapatite; copper; copper oxide; substitution; heat treatment
1. Introduction
Hydroxyapatite (HA), i.e., Ca10 (PO4 )6 (OH)2 , is widely used in various fields of
medicine: in traumatology, orthopedics, craniofacial surgery, dental technologies, medical
treatments, and cosmetology, as a means of targeted drug delivery. HA is a suitable material
for the development of biocompatible ceramic products, composites, bone defect fillers,
medical cements, and implant coatings [1–3].
HA (Figure 1a) crystalizes in hexagonal syngony with space group P63 /m [4]. The HA
unit cell contains 10 calcium cations located at two nonequivalent positions: four cations at
the Ca1 site and six cations at the Ca2 site, which are surrounded by nine and seven oxygen
atoms, respectively. In addition to calcium ions, the HA unit cell contains six phosphate and
two hydroxyl groups. The latter are located on the c axis in a hexagonal channel formed by
calcium ions and by oxygen ions from phosphate tetrahedrons. The crystal lattice of HA
is unique in that it offers ample opportunities for substitutions and for the formation of
solid solutions. Some or even all ions can be substituted, and replacement with isovalent
or heterovalent ions of other chemical elements or their chemical groups is possible [5].
Doping of HA with copper ions gives an antibacterial property to HA-based materials,
which reduces the risk of inflammation after implantation [6]. In addition, copper cations
induce protein absorption, osteogenic differentiation, and bone-like apatite nucleation and
https://www.mdpi.com/journal/materials
 °C makes it possible to stabilize copper ions in the hydroxyl channel in the form of linear
oxocuprate groups (Figure 1c) with the formation of the Ca10(PO4)6(OH)2–2x(CuO2)x struc-
ture, where x ≤ 0.6 [10–16]. Thus, the following mechanisms of substitution in HA are
possible for the copper cation:
Materials 2022, 15, 5759 Са2+ → Cu2+ 2 of(1)
15

2OH– → (CuO2)2– (2)

growth at an implant site [7]. These effects stimulate the growth of new bone tissue and
accelerate the healing process, both of which are important for surgical applications.

(a) (b) (c)

Figure
Figure 1.
1. Crystal
Crystal structure
structure of
of stoichiometric
stoichiometric HA
HA (a)
(a) and
and of
of Cu-HA,
Cu-HA, featuring
featuring the
the localization
localization of
of the
the
copper cation either at the site of the calcium cation (b) or in the hydroxyl channel (c). The arrow
copper cation either at the site of the calcium cation (b) or in the hydroxyl channel (c). The arrow
indicates the position of the copper atom.
indicates the position of the copper atom.

The main
There areas
exists of application
a study indicatingofthat
apatite materials
in samples ofnecessitate a heatHA
Cu-substituted treatment
(Cu-HA) (usu-
ob-
ally high-temperature) procedure during the manufacture of a product.
◦
tained by the precipitation technique, after calcination at 400 C, copper ions can re- The authors of
ref. [8] report that after heat treatment at 700 °C, orthophosphates
place calcium ions (Figure 1b) up to substitution degree x = 1.5 with the formation of Са 3 Сu 3(PO 4 ) 4 and
Ca
Ca19 Cu2(PO4)14 form in Cu-HA with a high degree of substitution. The absence of high-
10−x Cux (PO4 )6 (OH)2 [8]. Solid-phase mechanochemical synthesis in a planetary ball
temperature
mill allows thetreatment
obtaining is necessary
of Cu-HAfor thesubstitution
with practical application
degree xof= such
2 [9]. aHeat
material because
treatment at
>1000 ◦
a change in the phase
C makes composition
it possible of copper
to stabilize the material
ions inaffects its properties.
the hydroxyl channelTherefore,
in the form it of
is
necessary to knowgroups
linear oxocuprate the limitations
(Figure 1c)ofwith
the heat treatmentof
the formation ofthe
Cu-HA.
Ca10 (PO4 )6 (OH)2−2x (CuO2 )x
The aim
structure, whereof this
x ≤work is to investigate
0.6 [10–16]. Thus, the the evolution
following of phase composition
mechanisms of substitution during
in HAheat are
treatment of Cu-HA samples
possible for the copper cation: prepared by the mechanochemical method with a low de-
gree of substitution. In situ high-temperatureCa2+ → experiments
Cu2+ on these samples were carried (1)
out for the first time in the present study. − 2−
2OH → (CuO2 ) (2)
2. Materials
The main and Methods
areas of application of apatite materials necessitate a heat treatment (usu-
ally high-temperature) procedure
Schematic representation during
of the the manufacture
experiments is shown in of Figure
a product.
2. AtThe authors
the first of
stage,
ref. [8] report that after heat treatment at 700 ◦ C, orthophosphates Ca Cu (PO ) and
mechanochemical synthesis of three samples and their analyses were carried 3 out.
3 Then,
4 4 in
Ca19high-temperature
situ Cu2 (PO4 )14 form in Cu-HA with
experiments werea high degree on
conducted of substitution.
the synthesizedThesamples.
absence of high-
temperature treatment
The following is necessary
reagents for the
were used as practical application of
starting components forsuch
the amechanochemical
material because
a change in the phase composition of the material affects
synthesis: anhydrous calcium hydrogen orthophosphate CaHPO 4, annealed its properties. Therefore,
calcium itox-
is
necessary to know the limitations of the heat treatment of Cu-HA.
ide CaO, and copper oxide CuO. All reagents were of chemically pure grade and were
The aim of this work is to investigate the evolution of phase composition during heat
treatment of Cu-HA samples prepared by the mechanochemical method with a low degree
of substitution. In situ high-temperature experiments on these samples were carried out
for the first time in the present study.

2. Materials and Methods

Schematic representation of the experiments is shown in Figure 2. At the first stage,
mechanochemical synthesis of three samples and their analyses were carried out. Then, in
situ high-temperature experiments were conducted on the synthesized samples.
The following reagents were used as starting components for the mechanochemical
synthesis: anhydrous calcium hydrogen orthophosphate CaHPO4 , annealed calcium oxide
CaO, and copper oxide CuO. All reagents were of chemically pure grade and were produced
by Vekton (St. Petersburg, Russia). The ratio of components for the synthesis was chosen in
accordance with reactions 1–3 given in Table 1.
 der the assumption that copper ions replace either calcium ions (sample 0.5Cu-Ca) or hy-
droxyl groups (sample 0.5Cu-OH). The mechanochemical synthesis of the samples was
carried out in an AGO-2 planetary ball mill [17] in two water-cooled steel drums (volume:
150 mL), with steel balls weighing 200 g, at a drum rotation speed of 1800 rpm in ambient
air. The weight ratio of the reaction mixture to the balls was 1:20, and the processing time
Materials 2022, 15, 5759 3 ofthe
of the initial mixture in the mill was 30 min. Before the synthesis, the working zone of 15
mill was lined with a reaction mixture of the same composition.

Figure 2. Schematic
Figure 2. Schematic representation
representation of
of the
the experiments.
experiments.

Table 1. Expected reactions of mechanochemical synthesis.

Table 1. Expected reactions of mechanochemical synthesis.

Reaction Reaction K
K Sample
Sample
Reaction Reaction
ID Ca/P (Ca + Cu)/P Name
ID Ca/P (Ca + Cu)/P Name
6CaHPO4 + 4CaO →
1 6CaHPO4 +1 4CaO → Ca10(PO 4)6(OH)2 + 2H2O 1.67
1.67 1.67
1.67 0.0Cu
0.0Cu
Ca10 (PO4 )6 (OH)2 + 2H2 O
2 6CaHPO4 + 3.5CaO + 0.5CuO → Ca9.5Cu0.5(PO4)6(OH)2 + 2H2O 1.58 1.67 0.5Cu-Ca
6CaHPO4 + 3.5CaO + 0.5CuO →
3 6CaHPO4 + 4CaO + 20.5CuO →CaCa10Cu
(PO(PO
4)6(OH)1Cu0.5O + 2.5H2O
) (OH) + 2H O
1.67
1.58 1.75
1.67 0.5Cu-OH
0.5Cu-Ca
9.5 0.5 4 6 2 2

6CaHPO4 + 4CaO + 0.5CuO →

X-ray powder diffraction
3 (XRPD) patterns of the obtained samples
1.67 1.75 were 0.5Cu-OH
recorded on
Ca10 (PO4 )6 (OH)1 Cu0.5 O + 2.5H2 O
a D8 Advance powder diffractometer (Bruker, Germany) with Bragg–Brentano geometry
using СuKα radiation. In situ high-temperature experiments were conducted in an HTK
1200NTo chamber
obtain substituted-HA
(Anton Paar, Graz,samples, the amount
Austria) of the dopant
with a corundum (CuO)
carrier wasairselected
in the atmos-
under the assumption that copper ions replace either calcium ions (sample
phere. Heating was carried out stepwise at a heating rate of 0.5 °C/s with exposure 0.5Cu-Ca)toora
hydroxyl groups (sample
given temperature 0.5Cu-OH).
for 600 The mechanochemical
s. The diffraction synthesiswith
patterns were detected of thea samples was
step of 0.02°
carried out in
and a scan an AGO-2
speed planetary
of 1 s per ball mill
step. X-ray phase[17]analysis
in two water-cooled
of the compoundssteel drums (volume:
was performed
150
using the ICDD PDF-4 database of XRPD patterns (a 2011 release). Refinement of ambient
mL), with steel balls weighing 200 g, at a drum rotation speed of 1800 rpm in unit cell
air. The weight
parameters andratio of the reaction
of crystallite mixture to the
size, calculations balls was
of phase 1:20, and the
concentrations, andprocessing
refinementtimeof
of
structural parameters (atomic coordinates and occupancies) were carried out by the
the initial mixture in the mill was 30 min. Before the synthesis, the working zone of the
mill was lined
Rietveld method with
inathe
reaction
Topasmixture of the(Bruker,
4.2 software same composition.
Germany). The instrumental contri-
X-ray powder diffraction (XRPD) patterns
bution was modeled by the fundamental parameter of themethod.
obtainedInitial
samples were recorded
structural on
data on the
a D8 Advance powder diffractometer (Bruker, Germany) with Bragg–Brentano geometry
Cu-HA phase were borrowed from ref. [11].
using CuKα radiation. In situ high-temperature experiments were conducted in an HTK
Transmission electron microscopy (TEM) and high-resolution TEM (HRTEM) images
1200N chamber (Anton Paar, Graz, Austria) with a corundum carrier in the air atmosphere.
were obtained using a Themis-Z 3.1 microscope (TFS, Waltham, MA, USA) at an acceler-
Heating was carried out stepwise at a heating rate of 0.5 ◦ C/s with exposure to a given
ating voltage of 200 kV. The microscope is equipped with a field emission cathode having
temperature for 600 s. The diffraction patterns were detected with a step of 0.02◦ and a scan
a monochromator and with two aberration correctors. Energy-dispersive X-ray microa-
speed of 1 s per step. X-ray phase analysis of the compounds was performed using the
nalysis of elemental composition of the samples was performed on a four-segment Super-
ICDD PDF-4 database of XRPD patterns (a 2011 release). Refinement of unit cell parameters
and of crystallite size, calculations of phase concentrations, and refinement of structural
parameters (atomic coordinates and occupancies) were carried out by the Rietveld method
in the Topas 4.2 software (Bruker, Germany). The instrumental contribution was modeled
by the fundamental parameter method. Initial structural data on the Cu-HA phase were
borrowed from ref. [11].
Transmission electron microscopy (TEM) and high-resolution TEM (HRTEM) images
were obtained using a Themis-Z 3.1 microscope (TFS, Waltham, MA, USA) at an accelerating
voltage of 200 kV. The microscope is equipped with a field emission cathode having a
monochromator and with two aberration correctors. Energy-dispersive X-ray microanalysis
of elemental composition of the samples was performed on a four-segment Super-X detector
(with an energy resolution of ~120 eV) in scanning dark-field mode with the construction of
Materials 2022, 15, x FOR PEER REVIEW 4 of 15

Materials 2022, 15, 5759 4 of 15

X detector (with an energy resolution of ~120 eV) in scanning dark-field mode with the
construction of maps of distributions of elements by means of characteristic lines of the
spectrum
maps from each point
of distributions in an analyzed
of elements by meansregion. Samples for
of characteristic the
lines ofanalysis were from
the spectrum dispersed
each
by ultrasonication and deposited from alcohol on perforated aluminum
point in an analyzed region. Samples for the analysis were dispersed by ultrasonication grids covered
withdeposited
and a thin carbon
frommesh.
alcohol on perforated aluminum grids covered with a thin carbon
mesh.Fourier transform infrared (FTIR) spectra were acquired with the help of an Infralum-
801 spectrometer (Simex,
Fourier transform Russia).
infrared The spectra
(FTIR) sampleswere
wereacquired
prepared by the KBr
with help pellet method.
of an Infralum-
801 spectrometer (Simex, Russia). The samples were prepared by the KBr pellet method.
3. Results and Discussion
3. Results and Discussion
3.1. Mechanochemical Synthesis
3.1. Mechanochemical Synthesis
Figures 3 and 4 depict electron micrographs of the synthesized samples and distribu-
tionsFigures 3 andacross
of elements 4 depict electroninmicrographs
particles of the
the synthesized synthesized
samples. samples
All the andproved
samples distribu-
to
tions of elements across particles in the synthesized samples. All the samples
be aggregates of nanoparticles ranging in size from 50 to 1000 nm. The nanoparticles are proved to
be aggregates of nanoparticles ranging in size from 50 to 1000 nm. The nanoparticles are
crystalline, with a size of 20–100 nm. X-ray microanalysis of Cu-doped samples showed
crystalline, with a size of 20–100 nm. X-ray microanalysis of Cu-doped samples showed
that calcium and phosphorus are distributed evenly, but copper behaves differently. Most
that calcium and phosphorus are distributed evenly, but copper behaves differently. Most
of the aggregates in samples 0.5Cu-Ca and 0.5Cu-OH contain 3 at.% and 4 at.% of evenly
of the aggregates in samples 0.5Cu-Ca and 0.5Cu-OH contain 3 at.% and 4 at.% of evenly
distributed copper, respectively. At the same time, nanoparticles with a high copper con-
distributed copper, respectively. At the same time, nanoparticles with a high copper content
tent are present (Figure 4b,c). The average Ca/P ratios in samples 0.5Cu-Ca and 0.5Cu-OH
are present (Figure 4b,c). The average Ca/P ratios in samples 0.5Cu-Ca and 0.5Cu-OH
are 1.4 and 2.1, respectively, whereas the average (Ca + Cu)/P ratio is 1.5 and 2.3. These
are 1.4 and 2.1, respectively, whereas the average (Ca + Cu)/P ratio is 1.5 and 2.3. These
values differ from theoretical ones (see Table 1), possibly because this method of analysis
values differ from theoretical ones (see Table 1), possibly because this method of analysis
is local; micron-sized particles are not analyzed in this case. In sample 0.0Cu, where no
is local; micron-sized particles are not analyzed in this case. In sample 0.0Cu, where no
copper was used in the synthesis, 0.2 at.% of copper was detected (Figure 4a), which is a
copper was used in the synthesis, 0.2 at.% of copper was detected (Figure 4a), which is a
background value
background value and
and is
is explained
explained by by the
the presence
presence ofof copper
copper in
in components
components of of the
the optical
optical
part of the microscope.
part of the microscope.

Figure 3.
Figure 3. TEM
TEM (a–c)
(a–c) and
and HRTEM
HRTEM (d–f)
(d–f) images
images of
of synthesized
synthesized samples 0.0Cu (a,d), 0.5Cu-Ca (b,e),
and 0.5Cu-OH
and 0.5Cu-OH (c,f).
(c,f).
Materials 2022, 15, x FOR PEER REVIEW 5 of 15

x FOR PEER REVIEW

Materials 2022, 15, 5759 5 of 15

Figure 4. Distribution maps of elements across particles in synthesized samples 0.0Cu (a), 0.5Cu-Ca
(b), and 0.5Cu-OH (c).
Figure 4.
Figure 4. Distribution
Distribution maps
maps ofof elements
elements across
across particles
particles in
in synthesized samples 0.0Cu
synthesized samples 0.0Cu (a),
(a), 0.5Cu-Ca
0.5Cu-Ca
(b), and 0.5Cu-OH
Figure
(b), and 5 shows
0.5Cu-OH (c).
(c). diffraction patterns of the samples after the mechanochemical syn-
thesis. In the diffraction patterns, there are reflections characteristic of the HA phase, cor-
Figureto
Figure
responding 55shows
shows
card diffraction
diffraction
PDF patterns
patterns
40-11-9308, of of
thethe
indicating samples
samples
that theafter after the mechanochemical
the mechanochemical
synthesized substances are syn-
synthesis.
single-
thesis.
In the The
phase. In the diffraction
diffraction
absence of anypatterns,
patterns, there are
reflections there are reflections
reflections
of impurity characteristic
characteristic
phases of the HA
confirmed of the
that theHA
phase, phase,sub-
correspond-
expected cor-
responding
ing to card to
PDF card PDF
40-11-9308, 40-11-9308,
indicating indicating
that the that the
synthesized synthesized
stances were obtained in accordance with the above reactions (Table 1). From Table 2,The
substances substances
are are
single-phase. single-it
phase.
absence
can The absence
of any
be concluded of any
reflections
that reflections
the of impurity
lattice of impurity
phases
parameters phases
confirmed
of HA confirmed
that
in the that the
the expected
Cu-HA samples expected
substances
undergowere sub-
the
stances were
obtained
dynamics in obtained
accordance
similar to thosein accordance
with
of the above with the above
reactions
unsubstituted-HA (Table reactions
1). From
parameters. (Table
Table
In both 2, 1). From
it can Tablesam-
be concluded
substituted-HA 2, it
can be concluded
that parameters
ples, that
the lattice parameters the lattice parameters
of HA inless
a and c are slightly thethan
Cu-HA of HA
thosesamplesin the Cu-HA
undergo the
of unsubstituted samples
HA.dynamics undergo the
similar
Such a situation
dynamics
to
can those ofsimilar
be observed to those
unsubstituted-HA
when of unsubstituted-HA
calcium parameters. In both
cations are substituted parameters.
substituted-HA
[8,9] and Inisboth substituted-HA
samples,
explained byparameters sam-a
the smaller
ples,
and c parameters
are slightly a and
less c are
than slightly
those of less than
unsubstituted those of
HA. unsubstituted
Such
ion radius of the dopant: rCа2+ = 0.100 nm versus rCu2+ = 0.073 nm. From Table 2, it alsoa HA.
situation Such
can be a situation
observed
can becalcium
when
follows observed when
that thecations calcium cations
are substituted
introduction of are
[8,9]substituted
the dopant and [8,9]crystallinity
is explained
reduces the and byisthe
explained
smaller
of the by
ionthe smaller
radius
material of
in
ion
the radius
dopant:
question. In the ofr the dopant:
= 0.100 r
nm =
versus0.100
lattice, the substituent Cu2+
Ca2+ Cа2+ r nm =versus
0.073 rnm. = 0.073
From nm.
Table From
2, it
ions represent point defects that complicate crys-
Cu2+ Table
also 2,
follows it also
that
the introduction
follows
tallite that the of
growth. the dopant of
introduction reduces the crystallinity
the dopant reduces the of crystallinity
the material in of question.
the material In thein
lattice,
question. theInsubstituent
the lattice,ionsthe represent
substituent point
ionsdefects
representthatpoint
complicate
defectscrystallite growth.crys-
that complicate
tallite growth.

XRPDpatterns
Figure 5.5.XRPD
Figure patternsofof samples
samples 0.0Cu
0.0Cu (1),(1), 0.5Cu-Ca
0.5Cu-Ca (2),(2),
andand 0.5Cu-OH
0.5Cu-OH (3) after
(3) after synthesis
synthesis ac-
cording to reactions
according (1–3)
to reactions (Table
(1–3) 1),1),
(Table as as
detected
detectedat at
room temperature.
room temperature.
Figure 5. XRPD patterns of samples 0.0Cu (1), 0.5Cu-Ca (2), and 0.5Cu-OH (3) after synthesis ac-
cording to reactions (1–3) (Table 1), as detected at room temperature.
 Materials 2022, 15, x FOR PEER REVIEW 6 of 15
Materials 2022, 15, 5759 6 of 15

Table 2. Structural characteristics of the synthesized samples.

Table 2. Structural characteristics of the synthesized samples.
Sample Name a (Å) c (Å) Crystallite Size (nm)
Sample0.0Cu
Name 9.437
a (Å)(1) 6.892c (1)
(Å) 24.9 (2)
Crystallite Size (nm)
0.5Cu-Ca
0.0Cu 9.435 (1)
9.437 (1) 6.881 (1)
6.892 (1) 20.2 (2)
24.9 (2)
0.5Cu-OH 9.432 (2) 6.887 (1) 20.2 (2)
0.5Cu-Ca 9.435 (1) 6.881 (1) 20.2 (2)
Note: The estimated standard deviations of the refined values are in parentheses.
0.5Cu-OH 9.432 (2) 6.887 (1) 20.2 (2)
Note: The estimated standard deviations of the refined values are in parentheses.
FTIR spectra of the synthesized samples are identical (Figure 6). They contain an ab-
sorption patternofcorresponding
FTIR spectra the synthesized to HA structure:
samples absorption
are identical bands
(Figure 6).ofThey
the phosphate
contain anion
(572, 602, 960, 1048, and 1089 cm −1) and of the hydroxyl group (630 and 3573 cm−1). The
absorption pattern corresponding to HA structure: absorption bands of the phosphate
ionsame
(572, intensity
602, 960, of the and
1048, hydroxyl group’s
1089 cm absorption
−1 ) and bands among
of the hydroxyl group all(630theand
synthesized
3573 cm−sam-
1 ).

Theples
same(Figure 6c) indicates
intensity hydroxyl’s
of the hydroxyl group’sequal concentrations;
absorption therefore,
bands among substitution
all the synthesizedof the
hydroxyl group did not occur in the 0.5Cu-OH sample, where a twofold
samples (Figure 6c) indicates hydroxyl’s equal concentrations; therefore, substitution of the decrease in the
hydroxyl group did not occur in the 0.5Cu-OH sample, where a twofold decrease in the 3).
concentration of OH groups was expected according to the reaction (Table 1, reaction
Hence, for the
concentration 0.5Cu-Ca
of OH groupssample, the reaction
was expected proceeded
according to theas expected
reaction (Table
(Table 1, reaction
1, reaction 3). 2).
By contrast,
Hence, the synthesis
for the 0.5Cu-Ca sample,of the
the0.5Cu-OH sample, where
reaction proceeded hydroxyl
as expected (Tablegroup substitution
1, reaction 2).
By was assumed
contrast, (Table 1),ofdid
the synthesis thenot match the
0.5Cu-OH desired
sample, reaction
where (Table
hydroxyl 1, reaction
group 3). To was
substitution under-
stand what
assumed (Tablehappened
1), did notinmatch
the 0.5Cu-OH
the desired sample, we created
reaction (Table 1,the following
reaction line
3). To of reasoning.
understand
what happened in the 0.5Cu-OH sample, we created the following line of reasoning.

Figure
Figure 6. FTIR
6. FTIR spectra
spectra of of samples
samples 0.0Cu(1),
0.0Cu (1),0.5Cu-Ca
0.5Cu-Ca(2),
(2),and
and0.5Cu-OH
0.5Cu-OH(3).
(3). (a) General
General view;
view; (b)
(b) enlarged view of the 1300–1600 cm− region; (c) enlarged view of the 3300–3750 cm−region.
−11region; (c) enlarged view of the 3300–3750 cm−1 1 region.

In accordance
In accordance with
with the
theliterature
literaturedata
datagiven
given above, the thecopper
coppercation
cationintroduced
introduced dur-
during the synthesis can occupy
ing the occupythe theposition
positionofofeither
eitherthethehydroxyl
hydroxyl group
group or or
thethe
phosphate
phosphate
group
group(Figure 1b,c).
(Figure If the
1b,c). former
If the former scenario
scenariodoes notnot
does materialize
materialize in the 0.5Cu-OH
in the 0.5Cu-OH sample,
sample,
then thethe
then latter oneone
latter does. When
does. When copper
copperions areare
ions located at the
located calcium
at the calciumsite, coefficient
site, k for
coefficient k for
thisthis
sample
samplecancan
be calculated
be calculatedas follows: k=k
as follows: (Ca + Cu)/P
= (Ca + Сu)/P = 1.75, which
= 1.75, whichexceeds
exceeds k necessary
k necessary
for for
thethe
formation
formationof HA
of HAstructure
structure(1.67); therefore,
(1.67); such
therefore, suchapatite cannot
apatite cannotform. Nonetheless,
form. Nonetheless,
k ofk 1.67 can be obtained by increasing the concentration of anions
of 1.67 can be obtained by increasing the concentration of anions by means by means of carbonate
of carbonate
groups,
groups,so that k = (Ca
so that k = +(CaCu)/(P
+ Сu)/(P+ C)+=С)1.67. This This
= 1.67. idea is
ideasupported by knowledge
is supported by knowledgeabout the
about
process of HA of
the process mechanochemical
HA mechanochemical synthesis [18]. If[18].
synthesis the synthesis is carried
If the synthesis out inout
is carried ambient
in ambi-
air ent
withair calcium oxide asoxide
with calcium the calcium source, then
as the calcium the then
source, following reactions reactions
the following take placetake
on the
place
particle surface:
on the particle surface:
CaO + H2 O = Ca(OH)2 (3)
СаО + Н2О = Са(ОН)2 (3)
Ca(OH)2 + CO2 = CaCO3 + H2 O (4)
Са(ОН)2 + СО2 = СаСО3 + Н2О (4)
Materials 2022, 15, 5759 7 of 15

The second reagent used—the phosphate source CaHPO4 —reacts with calcium car-
bonate, and as a result, carbonate ions have an opportunity to become integrated into the
HA crystal lattice:

(6 − x)CaHPO4 + (4 − x)CaO + xCaCO3 = Ca10− x (PO4 )6− x (CO3 )x (OH)2− x + 2H2 O (5)

where x ≤ 1.
Since air access during the synthesis is limited (the synthesis is performed in closed
drums), only a small amount of carbonate gets incorporated [19,20]. These anions occupy
the position of phosphate groups, as indicated by FTIR spectroscopy data (Figure 6b). In the
spectra of samples 0.0Cu and 0.5Cu-Ca, low-intensity absorption bands at 872, 1420, and
1470 cm−1 are seen. According to ref. [21], these bands belong to the carbonate ion at the
position of a phosphate group. In the case of the 0.5Cu-OH sample, absorption bands of the
carbonate ion are more intense (Figure 6b); hence, there is another factor stimulating the car-
bonate incorporation. The reason is the necessity to increase the amount of anions to reach
the same value as HA has: k = (Ca + Cu)/(P + C) = 1.67. In this regard, the general chemical
formula for the 0.5Cu-OH sample should be Ca10−x Cux (PO4 )6−y (CO3 )y (OH)2−y Oy , where
y = x/1.67.
From the above findings, we can conclude that after 30 min of the mechanochemical
processing of the mixtures according to reactions 2 and 3 (see Table 1), in both cases,
the type of Cu-HA forms, where copper ions substitute for calcium ions. Under the
tested conditions of mechanochemical synthesis, the replacement of hydroxyl groups with
copper cations was unsuccessful. A possible reason is that a vacancy must be present
to accommodate the oxocuprate group in the hydroxyl channel; this means that an OH
group must be removed, which is possible only at high temperatures [22,23]. Under our
conditions of mechanochemical synthesis (power of the mill, speed of drum rotation,
and cooling conditions), an insufficient amount of energy is released via the collision and
friction of the balls. A local increase in temperature can reach 600 ◦ C [24]; however, effective
dehydroxylation in ambient air requires a temperature of at least 1000 ◦ C [22], consistent
with data from refs. [10–16], where a temperature of 1100 ◦ C was ensured for the synthesis
of Cu-HA containing oxocuprate groups in the hexagonal channel (Figure 5 and Table 2).

3.2. In Situ Diffraction

3.2.1. The 0.0Cu Sample
Examination of the synthesized samples by high-temperature in situ XRPD analysis
revealed that they behave differently when heated to 1200 ◦ C. According to the in situ XRPD
data, the unsubstituted-HA sample (0.0Cu) is stable over the entire analyzed temperature
range (Figure 7a). With an increase in temperature, rising intensity of reflections was
observed, as was a decrease in their half-width, indicating the growth of crystallites during
the heating of the sample. Reflections of any additional phases were not detectable. Due to
thermal expansion, the lattice parameters of the unsubstituted-HA sample enlarged with
the increasing temperature (Figure 7a); parameter c deviated from a linear dependence
in the temperature range 1000–1200 ◦ C. The process stopped being linear because in this
temperature range, HA begins to rapidly lose hydroxyl groups located on the c axis [22];
these groups leave vacancies and O2− behind in accordance with the equation:

Ca10 (PO4 )6 (OH)2 → Ca10 (PO4 )6 (OH)2−2x Ox x + xH2 O (6)

where  is a vacancy. In this context, two hydroxyl groups give rise to one water molecule:

2OH− → H2 O + O2− (7)

 Materials 2022, 15, x FOR PEER REVIEW 8 of 15
Materials 2022, 15, 5759 8 of 15

Figure 7. XRPD patterns recorded during in situ heating of samples 0.0Cu (a,b), 0.5Cu-Ca (c,d), and
Figure 7. XRPD patterns recorded during in situ heating of samples 0.0Cu (a,b), 0.5Cu-Ca (c,d), and
0.5Cu-OH (e,f) in ambient air. —TCP reflections; *—CuO reflections.
0.5Cu-OH (e,f) in ambient air. ↓—TCP reflections; *—CuO reflections.
3.2.2.
3.2.2. TheThe 0.5Cu-Ca
0.5Cu-Ca Sample
Sample
As As
forfor
thethe 0.5Cu-Ca
0.5Cu-Ca sample,
sample, where
where thethe dopant
dopant waswas introduced
introduced under
under thethe assumption
assumption
of substitution
of substitution of calcium
of calcium ionsions
[k =[k(Ca
= (Ca + Сu)/P
+ Cu)/P = 1.67],
= 1.67], this
this material
material is less
is less thermally
thermally stable.
stable.
Materials 2022, 15, 5759 9 of 15

At 700 ◦ C, a CuO release was observed, whose concentration increased at 800 ◦ C and then
began to diminish with the increasing temperature up to 1000 ◦ C (Figure 7, Table 3). At
higher temperatures, no CuO reflections were detectable in the diffraction pattern.

Table 3. Concentrations (C) of impurity phases and changes in crystallite size (CS) of the substituted-
HA samples during the in situ experiment.

Impurity Phases in the Samples

0.5Cu-Ca 0.5Cu-OH
(Ca + Cu)/P = 1.67 (Ca + Cu)/P = 1.75
Temp. (◦ C)
β-Ca3 (PO4 )2 CuO β-Ca3 (PO4 )2 CuO
C (wt%) CS (nm) C (wt%) CS (nm) C (wt%) CS (nm) C (wt%) CS (nm)
500 – – – – – – – –
600 – – – – – – 2.8 (2) 15.8 (5)
700 – – 1.1 (3) 52 (17) – – 3.5 (3) 19.2 (4)
800 17.0 (8) 134 (9) 2.4 (4) 45 (8) – – 3.5 (2) 32.1 (4)
900 25.6 (8) 220 (26) 2.0 (3) 105 (22) – – 3.5 (2) 32.0 (4)
1000 27.0 (8) 288 (22) 0.7 (4) 77 (49) – – 2.9 (2) 56.2 (8)
1100 30.9 (5) 444 (43) – – – – – –
1200 33.7 (5) 593 (66) – – – – – –
500 – – – – – – – –
Note: The estimated standard deviations of the refined values are in parentheses.

In addition, starting from 800 ◦ C, a large amount of another impurity phase emerged–
β-Ca3 (PO4 )2 —the concentration of which rose with the increasing temperature and reached
a maximum at 1200 ◦ C, namely 33.7 wt% (Table 3). Evidently, the drop of the CuO
concentration at high temperatures down to complete disappearance points to the diffusion
of copper ions into the lattice of the observed calcium phosphates [HA and β-Ca3 (PO4 )2 ].
It is known that substitution of some cations with copper is possible, not only in HA but
also in Ca3 (PO4 )2 [8,25].
Judging by the obtained data, the copper ions introduced into the sample under the
assumption of substitution of calcium ions (see Table 1) indeed occupied the positions of
calcium ions after the synthesis, thereby forming the structure of substituted HA. This
crystal lattice is stable up to 600 ◦ C. At 700 ◦ C, copper ions begin to depart from the HA
lattice, thus leaving vacancies behind (former positions of calcium ions) and forming the
CuO phase, possibly by combining with anion O2− from the hydroxyl channel. In this
scenario, the process should be accompanied by dehydration and additional formation
of vacancies in the hydroxyl channel. When copper cations leave the Cu-HA lattice, the
k value, (Ca + Cu)/P, becomes less than 1.67. A slight increase in lattice parameter a of
the 0.5Cu-Ca sample, as compared with the behavior of this parameter in unsubstituted
HA, is evidently attributable to the presence of vacancies in the HA lattice (Figure 8a).
The highest concentration of CuO in the 0.5Cu-Ca sample is observed at 800 ◦ C. There-
fore, the maximum number of vacancies should also be seen at this temperature. The
high concentration of vacancies stimulates structural transformation; thus, at 800 ◦ C, the
β-Ca3 (PO4 )2 phase appears, which has k = 1.5. This observation is consistent with the
results of Destainville et al. [26], who have demonstrated that HA with k = 1.5 transforms
into β-Ca3 (PO4 )2 at 750 ◦ C. In our experiment, a complete transition of HA to β-Ca3 (PO4 )2
was not observed. The formation of 17 wt% β-Ca3 (PO4 )2 was detected at 800 ◦ C because k
is 1.58 for 0.5Cu-Ca.
Materials 2022, 15, x FOR PEER REVIEW 10 of 15
Materials 2022, 15, 5759 10 of 15

Figure 8. 8.
Figure Shifts inin
Shifts lattice parameters
lattice a and
parameters c (a)
a and and
c (a) inin
and the coherent
the coherentscattering region
scattering (b)(b)
region of of
thethe
HAHA
phase in the synthesized samples during heat treatment in the air atmosphere.
phase in the synthesized samples during heat treatment in the air atmosphere.

Starting ◦ C, the concentration of released CuO begins to diminish until it

at 900Sample
3.2.3. The 0.5Cu-OH
disappears ◦
In the inatsitu
1100XRPD C. This observation
experiment on theindicates
0.5Cu-OH that sample,
copper ions
wherereturn
(Ca to the HA
+ Сu)/P lattice
= 1.75,
and probably to β-Ca (PO ) . It is known that prolonged 1100 ◦ C annealing of a mixture of
CuO separated out as 3well,4 which 2 began at a lower temperature: starting from 600 °C
reagents containing CuO causes
(Figure 7, Table 3). Perhaps the reason the formation of HA having
is the presence linearconcentration
of a higher oxocuprate groups
of thein
the hydroxyl channel [12,13]; this arrangement enlarges the lattice
carbonate ion in the apatite structure of the 0.5Cu-OH sample, as reported above; the elim- parameters of HA [12].
In our case, the lattice parameters of the 0.5Cu-Ca sample also increase in comparison
ination of this ion is accompanied by a release of CO2 starting from 600 °C [22]. Reflections
with unsubstituted HA (Figure 8a). Accordingly, it can be assumed that copper ions of
of the β-Ca3(PO4)2 phase in the 0.5Cu-OH sample were not detectable in the entire tem-
the 0.5Cu-Ca sample at 900–1000 ◦ C are localized to the hydroxyl channel owing to the
perature range. This is because in this sample, copper ions are introduced at a greater-
presence of vacancies of hydroxyl groups, thereby forming the Ca (PO4 )6 (OH)2−2x Cux O2x
than-stoichiometric ratio (as an additive to the reagents used for10reaction 1 in Table 1).
structure. The substitution mechanism will be as follows:
Hence, there is no shortage of calcium cations in this sample (k = Ca/P = 1.67) after the exit
of copper cations. At 1000 °C,the 0.5Cu-OH 2− sample manifests
→ (O-Cu-O) 2−an enlargement of the lat-
OH + O + CuO (8)
tice parameters of apatite relative to those of unsubstituted HA (Figure 8a) as well as a
decline in the
3.2.3. The CuO concentration
0.5Cu-OH Sample (Table 3). These observations match the behavior of the
0.5Cu-Ca sample.
In the in situConsequently,
XRPD experiment in 0.5Cu-OH, copper ions
on the 0.5Cu-OH at high
sample, temperatures
where (Ca + Cu)/Palso= re-
1.75,
turn to the HA lattice and end up in the hydroxyl channel according
CuO separated out as well, which began at a lower temperature: starting from 600 ◦ C to the mechanism
described
(Figure 7, inTable
Formula3). (6).
Perhaps the reason is the presence of a higher concentration of the
carbonate ion in of
The behavior thethe coherent
apatite scattering
structure of the region in the sample,
0.5Cu-OH HA phase as is almost above;
reported identical the
among
elimination of this ion is accompanied by a release of CO2 starting from 600the
the studied samples (Figure 8b). When the samples are heated to 600 °C, ◦ Csize
[22].
ofReflections
crystallites ofstays
the virtually
β-Ca3 (PO unchanged. Next, there is intensive growth of crystallites up
4 )2 phase in the 0.5Cu-OH sample were not detectable in the
toentire
a size temperature
of ~400 nm. The range.slightThis difference
is because in crystallite size atcopper
in this sample, 1200 °Cionscan be
areexplained
introduced byat
the
a greater-than-stoichiometric ratio (as an additive to the reagents used for reaction 1inin
presence of the β-Ca 3 (PO 4 ) 2 phase, which prevents the growth of Cu-HA crystallites
sample 0.5Cu-Ca.
Table 1). Hence, there is no shortage of calcium cations in this sample (k = Ca/P = 1.67)
Thus,
after from
the exit ofthe datacations.
copper in TableAt3,1000 we can◦ C, deduce that thesample
the 0.5Cu-OH synthesized
manifestsCu-HA samples,
an enlargement
which
of thewere
latticesingle-phase
parameters of before
apatite heating,
relativeunderwent phase transformations
to those of unsubstituted HA (Figure during
8a) asthewell
heating. In contrast
as a decline in the toCuO0.5Cu-Ca,
concentrationthe 0.5Cu-OH
(Table 3).sample
These returned to the
observations single-phase
match stateof
the behavior
atthe
1100 °C.
0.5Cu-Ca sample. Consequently, in 0.5Cu-OH, copper ions at high temperatures also
return to the HA lattice and end up in the hydroxyl channel according to the mechanism
described in Formula (6).
The behavior of the coherent scattering region in the HA phase is almost identical
among the studied samples (Figure 8b). When the samples are heated to 600 ◦ C, the size of
crystallites stays virtually unchanged. Next, there is intensive growth of crystallites up to
a size of ~400 nm. The slight difference in crystallite size at 1200 ◦ C can be explained by
the presence of the β-Ca3 (PO4 )2 phase, which prevents the growth of Cu-HA crystallites in
sample 0.5Cu-Ca.
Materials 2022, 15, x FOR PEER REVIEW 11 of 15

Materials 2022, 15, 5759 11 of 15

Table 3. Concentrations (C) of impurity phases and changes in crystallite size (CS) of the substi-
tuted-HA samples during the in situ experiment.

Thus, from theImpurity

data in Table
Phases3, we can Samples
in the deduce that the synthesized Cu-HA samples,
which were0.5Cu-Casingle-phase before heating, underwent phase transformations during the
0.5Cu-OH
Temp. (°C) heating. In contrast
(Ca + Сu)/P = 1.67 to 0.5Cu-Ca, the 0.5Cu-OH sample returned to the
(Ca + Сu)/P single-phase state at
= 1.75
1100 ◦ C.
β-Ca3(PO4)2 CuO β-Ca3(PO4)2 CuO
С (wt%) 3.3.CS (nm)Structure
Crystal С (wt%)
of the Cu-HACS (nm) С (wt%)
Samples after CS (nm) Treatment
High-Temperature С (wt%) CS (nm)
500 – – – – – –
In situ XRPD patterns indicated that in contrast to 0.5Cu-Ca, samples 0.0Cu – – and
600 – 0.5Cu-OH – are single-phase– at 1200 – ◦ C (Figure–9a). After cooling – of 2.8
the (2)
samples, 15.8
one(5)may
700 – – 1.1 (3) 52 (17) – –
observe that in 0.5Cu-Ca, there is a notable broadening of reflections of both the apatite3.5 (3) 19.2 (4)
800 17.0 (8) phase 134and(9) of tricalcium
2.4 (4) phosphate45 (8)[β-Ca3 (PO – 4 )2 ; Figure–9b]. In the 3.5 0.5Cu-OH
(2) 32.1 (4)
sample,
900 25.6 (8) which220is(26)
single-phase at 1200 ◦105
2.0 (3) (22)
C, splitting –
of some – of the 3.5
reflections HA(2) phase is32.0 (4)
observed
1000 27.0 (8) (Figure 9b). Rietveld
288 (22) 0.7refinement
(4) (Figure
77 (49) 10) shows – that during – the cooling
2.9 (2)of the 56.2
0.5Cu-OH
(8)
1100 30.9 (5) sample, the
444 (43) Cu-HA phase
– decomposes– into two – phases having
– different– lattice parameters
–
1200 33.7 (5) (Table
5934). The concentration
(66) – of copper
– ions was – determined – based on the – assumption – that
500 – linear oxocuprate
– groups
– are formed– in the hydroxyl
– channel
– [11]. The– refinement–results
uncovered
Note: differences
The estimated among
standard the apatite
deviations of thephases
refinedinvalues
occupancy
are in rates of copper ions (Table 4),
parentheses.
in agreement with ref. [10]. The higher the copper content in the hydroxyl channel, the
greater
3.3. theStructure
Crystal lattice parameters
of the Cu-HA of Cu-HA.
Samples after High-Temperature Treatment
It is known that during the cooling of dehydroxylated HA in ambient air, the lost
In situ XRPD patterns indicated that in contrast to 0.5Cu-Ca, samples 0.0Cu and
hydroxyl groups are restored, i.e., rehydroxylation takes place [23]. This is because re-
0.5Cu-OH are single-phase at 1200 °C (Figure 9a). After cooling of the samples, one may
action 7 is reversible; hence, higher water vapor pressure and a decrease in the temper-
observe that in 0.5Cu-Ca, there is a notable broadening of reflections of both the apatite
ature promote the incorporation of water into the crystal lattice. It is probable that the
phase and of tricalcium phosphate [β-Ca3(PO4)2; Figure 9b]. In the 0.5Cu-OH sample,
rehydroxylation, which proceeds most actively on the surface of particles, leads to the
which is single-phase at 1200 °C, splitting of some reflections of the HA phase is observed
displacement of oxocuprate groups from surface layers of the particles, concurrently with
(Figure 9b). Rietveld refinement (Figure 10) shows that during the cooling of the 0.5Cu-
the formation of nanosized CuO particles. Accordingly, reaction 8 is also reversible and
OH sample,
depends on the Cu-HAvapor
saturated phase decomposes
pressure. into two phases
As a consequence having
of this different
process, a phaselattice pa-
emerges
rameters (Table 4). The concentration of copper ions was determined
having a lower copper content (probably the surface layer of the particle) and a higher based on the as-
sumption
copper content (the core of the particle); as a result, the coherent scattering regionre-
that linear oxocuprate groups are formed in the hydroxyl channel [11]. The of
finement results particles
substituted-HA uncovered differences
diminishes among thewhile
considerably, apatite phases size
crystallite in occupancy
decreases. rates
Judgingof
copper
by the ions (Table of
occupancy 4),copper
in agreement
ions, inwith ref. [10]. The
the 0.5Cu-OH higher
sample theheating
after copper to content
1200 ◦in C,the
the
hydroxyl channel, the greater the lattice parameters of Cu-HA.
cooled sample contains the following phases of Cu-HA: Ca10 (PO4 )6 (OH)1.72 (CuO2 )0.14 and
Ca10 (PO4 )6 (OH)0.76 (CuO2 )0.62 .

XRPDpatterns
Figure9.9.XRPD patternsofofsamples
samples0.0Cu
0.0Cu(1),
(1),0.5Cu-Ca
0.5Cu-Ca(2),
(2),and
and0.5Cu-OH
0.5Cu-OH(3)
(3)atat1200 ◦ C (a) and
1200°C
Figure (a) and
after cooling to 30 ◦
°C (b).
after cooling to 30 C (b).
Materials 2022, 15, x FOR PEER REVIEW 12 of 15
Materials 2022, 15, 5759 12 of 15

Figure
Figure10.
10.The
TheRietveld
Rietveldrefinement
refinementplot
plotfor
forthe
the 0.5Cu-OH
0.5Cu-OH sample.
sample.

TableIt4.isConcentrations
known that during of impurity the phases
cooling and ofchanges
dehydroxylated HA scattering
in the coherent in ambient air,within
region the lost
the
hydroxyl
cooled samples groups in theareinrestored, i.e., rehydroxylation takes place [23]. This is because reac-
situ experiment.
tion 7 is reversible; hence, higher water vapor pressure and a decrease in the temperature
promote the incorporation of water into the crystal Sample lattice.
Name It is probable that the rehydrox-
ylation, which proceeds most actively on the surface of particles, leads 0.5Cu-OH
0.0Cu 0.5Cu-Ca to the displacement
HA-1/HA-2 * of oxocuprate groups
C (wt%) 100from surface layers 70.2 of the(6) particles, concurrently 58.2with the(4)formation
(4)/40.8
a of
(Å)nanosized CuO 9.4224 (1)
particles. Accordingly,9.4274 (3) 8 is also reversible
reaction 9.4281
and (6)/9.4318
depends (4) on sat-
c (Å) 6.8843 (1) 6.9006 (3) 6.8922 (5)/6.9058 (4)
urated vapor pressure.
Crystallite size (nm) 384 (25)
As a consequence 139 of this
(3)
process, a phase emerges having a lower
199 (8)/362 (28)
copper
Cu occupancy content (probably
– the surface layer of
0.34 (2) the particle) and a higher
0.14 (2)/0.62 (4) content
copper
Calculated chemical
(the core of the Ca 10 of4 )substituted-HA
(PO 6 (OH)1.72 Cu0.14 O0.28par-
/
Caparticle);
10 (PO4 )6 (OH)2
as a result, the4 )coherent
Ca10 (PO scattering region
6 (OH)1.32 Cu0.34 O0.68
formula Ca10 (PO4 )6 (OH)0.76 Cu0.62 O1.24
ticles diminishes considerably, while crystallite size decreases. Judging by the occupancy
β-Ca3 (PO4 )2 C (wt%)
of copper ions, in the – 0.5Cu-OH sample after 29.2 (6)heating to 1200 °C, the cooled – sample con-
a (Å) – 10.4050 (7) –
tains
c (Å) the following – phases of Cu-HA:
37.426 (3) Ca 10(PO4)6(OH)1.72(CuO2)0.14
– and
Ca 10 (PO
Crystallite size (nm) 4 ) 6 (OH) 0.76 (CuO– 2 ) 0.62 . 127 (7) –
CuO C (wt%) A similar phenomenon – of separation of 0.6Cu-substituted
(3) apatite into phases1.0 (4) containing
Crystallite size (nm) copper concentrations,
different – accompanied 170 (65) by a release of the CuO 58phase,
(16) has been
Reliability factor Rwp documented by the5.2authors of ref. [11], where 5.8 the material was also slowly 4.8 cooled in a
high-temperature
* If two HA phases are present chamber. Ex situ
in a sample, heating
then parameters with simultaneous
of the second HA phase airarequenching enablesslash.
given after a forward re-
searchers to obtain a single-phase material [12]. On the other hand, Karpov et al. have
statedAthat similar the heating
phenomenon of a sample of certain
of separation ofcomposition
Cu-substituted to 1300 °C with
apatite subsequent
into phases slow
containing
cooling
differentdown copper to concentrations,
room temperature makes it possible
accompanied by a releaseto grow
of the stand-alone
CuO phase, crystals
has been of
Ca (PO ) Cu O H [13].
documented by the authors of ref. [11], where the material was also slowly cooled in
10 4 6 0.27 0.86 y

In the 0.5Cu-Ca sample,

a high-temperature chamber. which contained
Ex situ heatingthe β-Ca
with 3(PO4)2 phaseair
simultaneous at 1200 °C, the enables
quenching apatite
phase did not
researchers tosplit
obtain in two during cooling
a single-phase material after[12].
the in Onsitu
theexperiment,
other hand,likely Karpov owing
et al.tohave
the
low
stated concentration
that the heating of copper cations present
of a sample of certain in composition
it (Table 4) because
to 1300some ◦ C with of them are lo-
subsequent
slow cooling down to room temperature makes it possible to grow stand-alone crystals of
Ca10 (PO4 )6 Cu0.27 O0.86 Hy [13].
Materials 2022, 15, 5759 13 of 15

In the 0.5Cu-Ca sample, which contained the β-Ca3 (PO4 )2 phase at 1200 ◦ C, the apatite
phase did not split in two during cooling after the in situ experiment, likely owing to the
low concentration of copper cations present in it (Table 4) because some of them are located
in the β-Ca3 (PO4 )2 phase, as mentioned above. Upon cooling, copper oxide is released
within this sample as well. The crystal lattice parameters of the β-Ca3 (PO4 )2 phase are
less than those usually seen in unsubstituted β-Ca3 (PO4 )2 [27,28], thereby confirming the
presence of copper cations in β-Ca3 (PO4 )2 at the positions of calcium cations, which possess
a larger ion radius. During the cooling, less CuO separated out in the 0.5Cu-Ca sample
than in the 0.5Cu-OH sample (Table 3). The calculated chemical formula of the Cu-HA
phase in sample 0.5Cu-Ca in its cooled state is Ca10 (PO4 )6 (OH)1.32 (CuO2 )0.34 .
Colors of the samples changed substantially after the in situ experiments. Samples
containing copper ions before the heating had a gray tint, whereas after heating to 1200 ◦ C
and subsequent cooling, they turned dark brown with a lilac tint. The brown color of the
samples can probably be ascribed to the presence of copper (II) oxide, and the lilac hue is
due to the oxocuprate groups in the Cu-HA lattice, as also demonstrated in ref. [29].

4. Conclusions
In the presented work, for the crystal lattice of HA, two types of substitution with
copper ions were investigated. It was shown that with the help of soft mechanochemical
synthesis, one can obtain Cu-HA with doping ions taking the place of calcium ions. After
30 min of mechanochemical processing of an appropriate mixture of initial reagents in a
planetary ball mill, Cu-HA was successfully prepared with the degree of substitution (x) of
calcium cations by copper cations at 0.5. Substitution of the hydroxyl group by this method
of synthesis was unsuccessful.
For the first time, in situ diffraction analyses were performed on Cu-HA samples
with (Ca + Cu)/P ratios of 1.75 and 1.67. It was established that at (Ca + Cu)/P = 1.75,
the sample after the synthesis is single-phase Cu-HA, in which copper cations replace
calcium cations. During heat treatment, starting from 600 ◦ C, copper cations leave the
HA structure with a release of a copper (II) oxide phase, while the crystal lattice of HA
is preserved. The amount of the released CuO starts to diminish at 1000 ◦ C, and CuO is
undetectable at 1100 ◦ C. Under these conditions, dehydroxylation of HA takes place, giving
rise to OH group vacancies in the hydroxyl channel, thereby driving a reverse process: CuO
diffusion into the HA lattice. Oxocuprate groups (O–Cu–O)2− are formed in the hydroxyl
channel. The resultant substance is cuprate-substituted HA with the general chemical
formula Ca10 (PO4 )6 (OH)2−2x (CuO2 )x . After the cooling of this material, the CuO phase
appears, indicative of the reverse process of diffusion of copper oxide in conjunction with
the rehydroxylation of apatite. Nonetheless, some copper cations remain in the HA lattice.
The cooled sample is a mixture containing 58 wt% Ca10 (PO4 )6 (OH)1.72 (CuO2 )0.14 , 41 wt%
Ca10 (PO4 )6 (OH)0.76 (CuO2 )0.62 , and 1 wt% CuO.
At (Ca + Cu)/P = 1.67, the product of the mechanochemical synthesis is single-
phase Cu-HA, where copper cations substitute for calcium cations. When this material is
heated, starting from 700 ◦ C, copper cations leave the HA lattice with a release of the CuO
phase. Starting from 800 ◦ C, a third phase begins: Cu-substituted tricalcium phosphate
β-Ca3 (PO4 )2 . With a further increase in temperature, the concentration of β-Ca3 (PO4 )2
increases, while the CuO concentration declines because reverse diffusion of CuO into the
HA lattice occurs, with the formation of oxocuprate groups (O–Cu–O)2− in the hydroxyl
channel of HA and the emergence of a mixed phosphate. The cooled sample is a three-
phase system containing 70 wt% Ca10 (PO4 )6 (OH)1.32 (CuO2 )0.34 , 29 wt% Cu-substituted
β-Ca3 (PO4 )2 , and 1 wt% CuO.
Thus, by in situ diffraction analyses of cuprate-substituted HA, it is shown for the
first time that the highest concentration of copper cations in the HA lattice is attained at a
high temperature. Slow cooling of the material is accompanied by rehydroxylation, which
promotes the exit of some proportion of the dopant in the oxide form.
Materials 2022, 15, 5759 14 of 15

Author Contributions: Conceptualization, N.V.B.; methodology, M.V.C.; investigation, N.V.B., N.V.E.,

A.V.I. and O.B.V.; writing—original draft preparation, N.V.E.; writing—review and editing, M.V.C.
and N.V.B. All authors have read and agreed to the published version of the manuscript.
Funding: This research was carried out within the State Assignment to the Institute of Solid State
Chemistry and Mechanochemistry SB RAS (project No. 121032500064-8).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The raw/processed data required to reproduce these results are in-
cluded in the Section 2.
Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design
of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or
in the decision to publish the results.

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