Ceramics International 44 (2018) 16844–16850

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Ceramics International
journal homepage: www.elsevier.com/locate/ceramint

Synthesis of hydroxyapatite nanoparticles using surface carboxyl- T

functionalized carbon dots as template
⁎
Huifang Yang, Jiadan Hong, Luxi Wei, Chunlin Deng
National Engineering Research Center for Tissue Restoration and Reconstruction, Guangdong Key Laboratory of Biomedical Engineering, School of Materials Science and
Engineering, South China University of Technology, Guangzhou 510641, PR China

A R T I C LE I N FO A B S T R A C T

Keywords: Carbon dots (CDs), which are discrete, nearly spherical nanoparticles with sizes below 10 nm and large amounts
D. Hydroxyapatite of carboxylic acid moieties on the surface, have been proposed as an ideal template candidate for heterogeneous
D. Carbon dots nucleators to regulate hydroxyapatite (HAp) nucleation and growth. In this paper, small HAp nanoparticles
Template formed on carboxyl-functionalized CDs in situ were fabricated via the hydrothermal method. Investigation for
the corresponding morphologies and detailed formation mechanisms of samples were conducted by Fourier
transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), transmission electron micro-
scopy (TEM), high-resolution transmission electron microscopy (HR-TEM) and Rietveld refinement. The op-
timum size and crystallinity of HAp particle had been obtained in the preparation when the additive content of
CDs was 1.11 g/L. Moreover, results also suggested that CDs were served as nucleators in the HAp particles.
Therefore, a novel synthetic strategy is presented for small HAp nanoparticles using CDs as template.

1. Introduction ethylene glycol), PS-PAA-PEG) micelles as templates to synthesize HAp

Hydroxyapatite (HAp) is a bioactive material with good bio-
compatibility and a similar composition to bone mineral, making it
promising for in vivo applications in human medicine [1]. Small-par-
ticulate forms of HAp, especially nanoscale hydroxyapatite, possess the
properties of both nanomaterials and bulk hydroxyapatite, and have
been used widely in biomedicine as carriers to deliver drugs, proteins,
and even nucleic acids [2–4]. Interestingly, several studies have re-
ported that nanoscale hydroxyapatite may be able to act as a novel anti-
tumor drug to significantly inhibit the proliferation of cancer cells, in-
cluding breast cancer, hepatoma, gastric cancer and melanoma cells
[5–8]. Dong et al. [9] reviewed the therapeutic effects on brain diseases
of nanomaterials with various sizes, shapes and surface charges. Yuan
et al. [10] found that the anti-tumor activity of hydroxyapatite nano-
particles was closely related to their size, such that particles in the
range of 20–80 nm were translocated into the nucleus of HepG2 cells
and induced size-dependent cytotoxicity and apoptosis. Therefore, the
synthesis of small hydroxyapatite nanoparticles is vital for studying the
potential relationship between size and anti-tumor activity in vitro and
in vivo.
A variety of surfactants and copolymer micelles have been used as
templates to prepare small HAp particles. Bastakoti et al. [11] reported
the use of anionic triblock copolymer (poly(styrene-acrylic acid-
particles with sizes of about 30 nm. The PAA shells captured Ca2+ ions
to enable mineralization of HAp on the spherical PS core, which in-
volved coordination bonding between Ca2+ and the COO- groups of
PAA. Cai et al. [12] described the synthesis of HAp in the presence of
cetyltrimethyl ammonium bromide (CTAB) and obtained products with
average sizes of 20 ± 5, 40 ± 10 and 80 ± 12 nm depending on the
CTAB concentration. The positively charged CTAB took part in co-
ordinate bonding with phosphate by electrostatic interaction. The shape
of the templates and the existence of coordination bonding between the
templates and mineralized ions play an important role in the con-
trollable formation of HAp. Based on this concept, we resolved to search
for suitable template candidates for the synthesis of small hydro-
xyapatite particles, as reported in this paper.
Carbon dots (CDs) are nearly spherical, discrete carbon nano-
particles with sizes below 10 nm. Typically, the surfaces of CDs contain
a large amount of carboxylic acid moieties, making them readily dis-
persible in water and suitable for subsequent functionalization with
organic, polymeric or inorganic species [13–15]. Thus, CDs is an ideal
heterogeneous nucleator or template to synthesize HAp as a result of
their shape, size and surface features. However, for this practical use of
CDs it is necessary to increase their surface content of carboxyl groups.
Surface carboxyl-functionalized CDs can be obtained either by physical
methods followed by acid oxidative treatment or through thermal

⁎
Correspondence to: No. 381, Wushan Road, Tianhe District, Guangzhou 510641, PR China.
E-mail address: chldeng@163.com (C. Deng).

https://doi.org/10.1016/j.ceramint.2018.06.120
Received 25 May 2018; Received in revised form 13 June 2018; Accepted 13 June 2018
Available online 15 June 2018
0272-8842/ © 2018 Elsevier Ltd and Techna Group S.r.l. All rights reserved.
H. Yang et al.
oxidation of appropriate molecular precursors. Bourlinos et al. [16]
fabricated ionically charged CDs by an extension of the citrate method,
in which amino carboxylate species served as a modifier and carboxyl
groups were successfully grafted onto the carbon core surfaces.
The aim of this study was to synthesize small HAp hydrothermally
by using surface carboxyl-functionalized CDs as heterogeneous nu-
cleator. A possible mechanism of nucleation was proposed on the basis
of transmission electron microscopy (TEM) images. A series of HAp
samples with different CDs content were prepared, and their size and
crystallinity were investigated in detail.
2. Materials and methods
2.1. Synthesis of CDs
Surface carboxyl-functionalized carbon dots were synthesized by
reference to a literature method [16] with slightly modification. All
chemicals were of analytical grade reagents and obtained from Aladdin
and utilized without any further purification. Briefly, 0.2 g H2N
(CH2)10COOH was dissolved in water, and 0.45 g solid NaOH was then
added for neutralization. The precipitate was discarded, then 25.0 mL
citric acid solution (0.4 M) was added drop by drop, accompanied with
vigorous stirring. The resulting white precipitate was collected. The wet
paste was dried at room temperature for 24 h, subsequently further
dried at 85 °C for 2 h. The dried product was crushed into powder, then
heated in a muffle oven from room temperature to 300 °C at a heating
rate 10 °C min−1, maintained at that temperature for 2 h, and cooled
from 300 °C to room temperature naturally.
A quantity of water was added to the thermal oxidation product and
the product was dissolved by ultrasonication. The insoluble particles
were then removed by centrifugation and the aqueous supernatant was
collected. To adjust the pH of the aqueous supernatant, 37% HCl was
added until a bulk precipitate appeared. The product was then rinsed
with water, and 40.0 mL water and 0.2 g NaOH were added and the
product was then centrifuged to remove minor insoluble particles.
Finally, a brown-black dispersion solution was obtained. The final
concentration of CDs was determined to be about 24.4 g/L by the
weighing method. To purify the CDs, the product was placed into a
dialysis bag of 3500 D for 2 days and the solid powder was collected by
vacuum freeze-drying for subsequent tests.
2.2. Preparation of HAp by using CDs
A series of HAp samples with different CDs content were synthesized
hydrothermally. Ca(NO3)2·4H2O (analytical reagent grade, AR) and
Na3PO4·10H2O (AR) were used as Ca and P sources respectively, and
the above-mentioned CDs were used as heterogeneous nucleators. The
actual amounts of reagents employed for synthesis are listed in Table 1
in detail. The samples were denoted as S1-S5.
To be brief, a quantity of water was added to Ca(NO3)2 (0.5 M) and
the pH of the diluted solution was adjusted to 10 using NaOH (2 M).
Subsequently the CDs (24.4 g/L) were added slowly and a brown pre-
cipitate (denoted as P1) appeared. After stirring for a period of time so
that the precipitate became evenly distributed, Na3PO4 (0.1 M) was
introduced drop by drop such that the Ca/P molar ratio was maintained
Table 1
The actual amounts of reagents employed for synthesis of HAp samples.
Samples Ca(NO3)2 H2O (mL) CDs (mL) Na3PO4 (mL) CDs content
(mL) (mL) (g/L)
S1 10 15 – 30 – with the evidence from FTIR. The XPS spectra in Fig. 3(a) reveal that
S2 10 14.5 0.5 30 0.22
S3 10 13.5 1.5 30 0.67
S4 10 12.5 2.5 30 1.11
S5 10 10 5 30 2.22
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at 1.67. The resulting mixed precipitate solution (denoted as P2) was
transferred into a sealed stainless-steel Teflon-lined autoclave. After
reacting at 180 °C for 8 h it was cooled naturally to room temperature.
The precipitates were collected and washed several times with water
and ethanol, and the obtained wet paste was dried at 50 °C for one day
and crushed into powder. Finally, a series of HAp samples with various
shades of brown were obtained.
2.3. Characterization
Fourier transform infrared (FTIR) spectroscopy (Nicolet 360 spec-
trometer, USA) was performed in the wavenumber range from 400 to
4000 cm−1 to characterize the functional groups of the CDs and HAp
samples. The elemental compositions of the surfaces of the CDs and
HAp samples were recorded by X-ray photoelectron spectroscopy (XPS,
Escalab 250Xi, Thermo Scientific, USA). The morphologies of the CDs,
P1, P2 and HAp samples were observed by transmission electron mi-
croscopy (TEM, JEM-1400Plus, Japan), with more detailed images for
lattice characterization obtained by high-resolution TEM (HR-TEM,
JEM-2100F, Japan). The crystallinity and phase compositions of both
the CDs and HAp samples were recorded by X-ray diffraction (XRD,
X′Pert PRO, PANalytical, The Netherlands) collected in the 2θ range
from 10° to 70° with a step size of 0.013° and a scanning rate of
6° min−1.
2.4. Rietveld refinement
Rietveld refinement was performed to quantitatively analyze the
crystal structures by fitting the slow-scanning XRD diffraction data. The
GSAS and EXPGUI software packages were used to refine the structures
of the HAp samples, for which the diffraction data were collected with a
step size of 0.0065° and a scanning rate of 0.05° s−1. The space group of
the hydroxyapatite structure (p63/m, No.176) was used as the standard
crystallographic reference for the HAp samples.
3. Results
3.1. Analysis of CDs
3.1.1. Morphology and size analysis of CDs
The TEM images of the CDs in Fig. 1(a) distinctly show discrete
spherical-like crystalline nanoparticles with sizes below 10 nm. The HR-
TEM image depicts that the CDs have clear lattice fringes with a spacing
of about 0.21–0.22 nm. Additionally, the bar chart (Fig. 1(b)) displays
the statistical size distribution of the CDs in the picture, showing that
their average sizes are roughly distributed around 3.42 nm. Fig. 1(c) is
the representative structure of carboxyl-functionalized CDs.
3.1.2. Crystallinity, functional groups and elemental analysis of CDs
The XRD patterns in Fig. 2(a) exhibit that the as-prepared CDs have
a rough, broad peak at 2θ = 20°, indicating an amorphous carbon phase
[17]. The FTIR spectra, recorded to analyze the functional groups of the
carboxyl-functionalized CDs, are displayed in Fig. 2(b). The sharp peaks
at 2925 and 2852 cm−1 represent the existence of –CH2– groups, the
strong band at 1701 cm−1 is attributed to the carbonyl (C˭O) stretching
vibration [18], and the C-O stretching vibration peaks at 1306 and
1282 cm−1 indicate the functional group of –COOH. The peaks at 1584
and 1439/1404 cm−1 are associated with the C˭O stretching vibration
of carboxylate (–COO-) [19]. The peaks at 1636 [20] and 1404 cm−1
are assigned to the C˭O and C-N stretching vibrations of amide
(–CONH–) groups. The results of XPS elemental analysis are consistent
the as-prepared CDs are mainly composed of C (77.66%), N (3.81%)
and O (18.53%). As shown in Fig. 3(b), the C1s signals are composed of
five peaks: C-C at 284.62 eV, C-N at 285.21 eV, C-O at 286.02 eV and
286.80 eV, and C˭O at 287.99 eV [21,22]. The signals of C-O and C˭O
H. Yang et al.
Fig. 1. (a) HR-TEM images of CDs and a single particle. (b) Statistic size distribution of CDs from HR-TEM images. (c) Schematics of carboxyl-functionalized CDs.
demonstrate the presence of carbonyl and carboxyl groups on the CDs.
While the binding energy peaks at 285.21 eV imply the existence of
amide groups. In addition, the N1s signals of C-N-C at 399.6 eV and N-H
at 400.2 eV [23,24] confirm the existence of amide groups (Fig. 3(c)).
The analysis also reveals the presence of Na, a consequence of the
partial deprotonation of the –COOH groups by Na+ ions originating
from NaOH, forming an aqueous solution of the COO-Na+ salt. Both the
FTIR and XPS analyses therefore confirm the presence of –COOH,
–CONH– and –COO- groups on the CDs. All the above results demon-
strate that the structure of as-prepared CDs is consistent with Fig. 1(c).
3.2. Analysis of HAp samples
3.2.1. Mechanism of nucleation for HAp
Fig. 4(a) illustrates the formation of discrete CD clusters by the
aggregation of several individual CDs. Fig. 4(b) is an image of the P1
sample, which vividly shows that precipitates emerge on the surface of
Fig. 2. (a) XRD patterns of CDs. (b) FTIR spectra of CDs.
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the CD clusters, while other, monodisperse CDs scatter around these
precipitates, indicating the partial formation of CDs–Ca complexes. The
image of P2 in Fig. 4(c) reveals that the phosphate groups bond with P1
in situ through ionic interactions. However, some isolated CDs still
remain, demonstrating that a small proportion of the CDs do not act as
nucleators but are left as impurities in the aqueous solution.
3.2.2. Morphology and size analysis of HAp samples
The changes in morphology and size of the HAp samples were ob-
served directly from the TEM images. Fig. 5 shows that the HAp sam-
ples are well crystallized, short, rod-like particles with average lengths
below 60 nm. From S1 to S4, the average lengths of the HAp rods
gradually decrease from 58 nm to 40 nm, indicating the influence of the
CDs on the growth of HAp. The sample prepared using the largest
amount of CDs, S5, shows a thin rod-like morphology with slightly
poorer crystallinity than the others, consistent with the XRD patterns
(Fig. 9). Fig. 6(a) displays some small particles attach to HAp, which
H. Yang et al.
Fig. 3. (a) XPS spectra of CDs. (b) C1s signals of CDs from XPS analysis. (c) N1s signals of CDs from XPS analysis.
smaller than CDs. To investigate the growth of HAp, the spacing of the
lattice fringes S4 was measured, as shown in Fig. 6(b). The measured
distances of 0.226 nm, 0.272 nm and 0.281 nm correspond to the 310,
300 and 211 planes respectively. From the HR-TEM images of HAp, it is
difficult to distinguish recognizable individual CDs or their lattice
fringes. Consequently, a certain area was selected from S4 for elemental
detection and the results are shown in Fig. 6(c) and (d). From Fig. 6(d),
we conclude that N is contained in these samples, specifically in the CDs
within the interior of HAp.
3.2.3. Functional groups and surface elements analysis of HAp samples
The functional groups of HAp samples with different CDs content
were analyzed by FTIR, and their spectra are shown in Fig. 7. The small,
but sharp peaks at 3571 and 634 cm−1 represent the hydroxyl-group
stretching and bending bands of HAp. The bands at 1092 and
1031 cm−1 correspond to the P-O antisymmetric stretching vibrations,
and that at 962 cm−1 corresponds to the P-O symmetric stretching vi-
bration of the phosphate groups. The strong peaks at 601 and 564 cm−1
are attributed to antisymmetric O-P-O variable-angle vibrations, and
that at 473 cm−1 is attributed to symmetric O-P-O variable-angle vi-
bration. From S1 to S5, the peaks in the range 1400–1460 cm−1 and at
about 874 cm−1, which correspond to CO32- (ν3b, ν3a and ν2) [25]
groups respectively, revealing that the HAp samples have type B car-
bonate substitution [25–28] and interact closely with the CDs. The
additional peaks of the CDs still remain in the spectra of S2-S5. Speci-
fically, for S5, the bands located at 2925 and 2852 cm−1 are ascribed to
–CH2- groups, while those at 1700 and 1584 cm−1 are assigned to
carbonyl and carboxyl functional groups of the CDs, respectively, in-
dicating that the CDs exist in the interior of the HAp. Comparing S1
with S2-S5, the additional characteristic peaks of the CDs become
gradually more intense in the latter, demonstrating the increasing
presence of CDs (or their derivatives) in HAp.
To verify whether CDs were also present on the surface of the HAp
samples, XPS spectra were recorded. As shown in Fig. 8, the char-
acteristic N peaks of CDs are not detected, indicating the absence of CDs
on the HAp surface. The particles of S1 are white, while those of S2-S5
Fig. 4. TEM images of HAp nucleation process on CDs in situ. (a) CDs alone (scale bar, 20 nm). (b) CDs–Ca complexes P1 (scale bar 50 nm). (c) CDs–Ca–Pi complexes
P2 (scale bar, 100 nm).
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are various shades of brown, suggesting that the CDs occupy the in-
terior of the HAp particles. The Ca/P (2p) ratios in S1-S5 are 1.079,
1.24, 1.58, 1.51 and 1.62, respectively, as measured by XPS.
3.2.4. Crystallinity and phase composition analysis of HAp samples by XRD
The crystallinity and phase compositions of the HAp samples were
analyzed by XRD. The XRD patterns of the five HAp samples are shown
in Fig. 9, wherein each lattice plane is well matched with standard
hydroxyapatite (JCPDS No.09-0432). With increasing CDs content, the
diffraction peaks at 10.82° (100), 25.87° (002), 31.77° (211), 32.19°
(112) and 32.90° (300) become progressively weaker, which indicates
that the crystallinity of the HAp samples was impaired by the CDs.
3.2.5. Quantitative analysis of the crystal structure of HAp by Rietveld
refinement
Rietveld refinement was utilized to analyze the structural difference
among the HAp samples, and the final refined data are displayed in
Table 2. With increasing CDs content, the a lattice parameter and unit
cell volume remained almost invariant, whereas the c lattice parameter
and unit cell density of S2-S5 progressively increased.
4. Discussion
In this work, a series of HAp samples with different CDs content
were synthesized by the hydrothermal method. The effect of the CDs on
the size and morphology of the HAp samples was assessed. Moreover, a
potential mechanism of nucleation and growth was inferred via TEM,
while several other analytical techniques were used to verify the ex-
istence of CDs within the HAp samples.
4.1. The possible role of CDs in the formation of HAp samples
The surface carboxyl-functionalized CDs have small sizes below
10 nm and a large amount of carboxylate groups on their surface, with
sodium ions balancing the negative charge, as confirmed by the FTIR,
HR-TEM and XPS results. The carboxylate groups, enable exchange of
H. Yang et al. Ceramics International 44 (2018) 16844–16850

Fig. 5. TEM images of HAp samples with different CDs content and their statistic size distribution: (a) S1 without CDs, (b) S2 contain 0.22 g/L CDs, (c) S3 contain
0.67 g/L CDs, (d) S4 contain 1.11 g/L CDs, (e) S5 contain 2.22 g/L CDs. ( The scale bar of all above mentioned TEM images is 100 nm).

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Fig. 9. XRD patterns of HAp samples with different CDs content.

Table 2
Structural parameters of HAp samples fitted by Rietveld refinement.
Fig. 6. (a) HR-TEM image of S4 (scale bar, 10 nm). (b) One single particle of S4 Samples Refinement factors Structural parameters
(scale bar, 5 nm). (c) A special area of S4 for elemental detection. (d) The
distribution of N in the region depicted in (c). Rwp χ2 a (Å) c (Å) ρ (g/cm3) V (Å3)

S1 0.0496 2.293 9.3830 6.8466 3.282 522.027

S2 0.0484 2.209 9.3835 6.8469 3.583 522.108
S3 0.0484 2.169 9.3840 6.8479 3.402 522.245
S4 0.0493 2.172 9.3835 6.8490 3.436 522.273
S5 0.0477 2.407 9.3831 6.8474 3.793 522.111

possibly formed by coordination bonding between Ca2+ and the COO-

groups of the CDs. The TEM image in Fig. 4(b) reveal that the pre-
cipitates P1 emerge on the surface of CDs clusters in situ, while other,
monodisperse CDs scatter around the precipitates, indicating the partial
formation of CDs–Ca complexes. In principle the interaction between
ions enables phosphate (PO43-) to bind with P1 in situ, and this can be
observed in Fig. 4(c). However, there still remain several unbound CDs,
demonstrating that a small fraction of the CDs did not act as nucleators
but were left as impurities in the final aqueous solution.

Fig. 7. FTIR spectra of HAp samples with different CDs content. 4.2. Effect of CDs content on the size and morphology of HAp samples

Fig. 8. XPS of HAp samples with different CDs content.
the sodium ions with multivalent inorganic cations [16], leading to the
emergence of a precipitate. For the P1 sample, a brown precipitate
appeared rapidly in calcium nitrate solution in the presence of CDs,
The short rod-like HAp particles with sizes mostly below 100 nm are
vividly shown in Fig. 5 together with a histogram of their statistical size
distribution. With increasing CDs content, the average lengths of HAp
reduced slightly from 58 nm (S1) to 40 nm (S4). However, the particles
of S5 are similar in length to S1, revealing that the CDs content has a
non-linear influence on the growth of HAp. In addition, the FTIR
spectra suggest that carbonate ions substitute for phosphate ions. Wo-
penka et al. [29] found that B-type carbonate substitution reduced the
overall crystallite size and increased the c-axial length, which agrees
with the results of Rietveld refinement herein (Table 2) and may ex-
plain why the size of the HAp samples generally decreased with CDs
content. The CDs may either play a role as CO32- sources or be in-
corporated intact into HAp to provide CO32-.
When the content of CDs was further increased, the resulting sample
S5 showed similar average size and lattice parameters to those of S1,
despite differences in crystallinity and color. From the viscosity of so-
lution and the stability of the CDs–Ca complexes, it can be inferred that
a large number of CDs took part in binding with Ca2+, which hindered
the ability of PO43- ions to interact with free Ca2+ and slowed down the
process of nucleation. The XRD results demonstrate that all the samples
have similar crystal phases and diffraction patterns to pure standard

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H. Yang et al.
hydroxyapatite. Nevertheless, with increasing CDs content, the crys-
tallinity and particle size of HAp decreased slightly. The intensity of the
diffraction peaks at 10.82° (100), 25.87° (002), 31.77° (211), 32.19°
(112) and 32.90° (300) weakened, indicating that the CDs damaged the
crystallinity of HAp. Moreover, Fig. 6(a) displays that some extremely
small particles, smaller than the CDs, are attached to HAp. However, in
Fig. 6(b), it is difficult to distinguish recognizable individual CDs or
their lattice fringes within the interior of HAp. Hence, we have no
evidence to confirm the formation of HAp on the CDs in situ. None-
theless, Fig. 6(c) and (d) reveal the presence of N of CDs in the HAp
samples.
In the context of the interaction between CDs and HAp samples, we
propose that CO32- ions substitute phosphate ions. The CDs have been
assembled from nano-sized carbon cores and amide polymer clusters
(see Fig. 1(c)) shells through covalent bonding [30,31]. In basic
medium accompanied by high temperature and pressure, the polymer
clusters shells of the CDs are likely to separate from the carbon cores
and decompose into small molecules, such as CO32- ions, then partly
incorporate into HAp. This probably explains why there are some ex-
tremely small particles attached to HAp but with no clear CD lattice
fringes.
Furthermore, the XPS results reveal that the C content and the Ca/P
ratio both increase with higher CDs content, which may be caused by
the substitution of CO32- ions. With increasing CDs content, more and
more polymer clusters separate from the carbon cores in the hydro-
thermal environment and attach to HAp, causing the diffraction peaks
to widen, implying that the crystallinity decreases.
5. Conclusion
Small hydroxyapatite (HAp) nanoparticles formed on surface car-
boxyl-functionalized carbon dots in situ were obtained hydrothermally.
In the first step, calcium ions replaced sodium ions on the CDs in situ.
The resulting electrostatic force enabled phosphate ions to bond with
CDs–Ca precipitates in situ. Finally, in the condition of the high tem-
perature and high pressure, the HAp continued to nucleate and the
polymer clusters shells separated from the CDs cores through the dis-
ruption of covalent bonds. With increasing CDs content, the substitu-
tion of phosphate ions by carbonate ions increased. Moreover, the c axis
lengthened and the overall particle size decreased. However, an ex-
cessive amount of CDs slowed the nucleation process, and impaired the
crystallinity of HAp through the presence of CDs as impurities in the
final product. In the future, we believe that small, carbonate-substituted
apatite particles could act as promising anti-tumor drug candidates to
inhibit the proliferation of cancer cells. Further research is needed to
develop such particles for medical applications.
Acknowledgements
This work was supported by the National Natural Science
Foundation of China (Grant nos.51372086 and 51772105).
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