Materials and Design 87 (2015) 445–449

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Materials and Design

journal homepage: www.elsevier.com/locate/jmad

Synthesis of hydroxyapatite nanorods from abalone shells via

hydrothermal solid-state conversion
Jingdi Chen a,⁎, Zhenliang Wen a, Shengnan Zhong a, Zihao Wang a, Jiulin Wu a, Qiqing Zhang a,b,⁎⁎
a
Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002, China
b
Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192, China

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

Article history: Hydroxyapatite (HAP) has been widely applied as a biomaterial for repairing or substituting human hard tissues.
Received 8 June 2015 In this paper, HAP nanorods were successfully produced from abalone shell powders via hydrothermal solid-state
Received in revised form 11 August 2015 conversion without surfactants or complex agents. The field emission scanning electron microscopy (FESEM)
Accepted 13 August 2015
showed that the produced HAP exhibited the typical rod-like structure. Based on the X-ray diffraction (XRD)
Available online 18 August 2015
and thermal analysis (thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)), these
Keywords:
samples contain a small amount of aragonite and calcite crystals, and their content gradually decreases by
Hydroxyapatite nanorods prolonging the hydrothermal time. However, this decrease only slightly changes for the longer times. The Fourier
Abalone shell transform infrared spectroscopy (FTIR) revealed that organic matter was detected in the samples without adding
Solid-state conversion surfactants or complex agents. This study provides a solution to the resource waste and environmental pollution
Hydrothermal caused by abandoned abalone shells, and we also synthesized HAP for potential bone repair materials.
© 2015 Elsevier Ltd. All rights reserved.

1. Introduction and osteoconductivity [11–14]. For this reason, synthetic nano-HAP

has been widely applied as a biomaterial for repairing or substituting
As a type of marine mollusk, abalone (Haliotis) is a typical single human hard tissues [15–17]. In recent years, methods to prepare
shell shellfish. Because of its delicious taste and high nutrition values, nano-HAP using different forms of the raw material have been reported,
abalone meat is a rare ingredient of traditional Chinese food and one including solid-phase synthesis [12,18], sol–gel method [19,20], tem-
of the necessary dishes for Chinese banquets. However, the abalone plate synthesis [12,21,22], and the hydrothermal method [12,23–25],
shell is a great burden to the environment after consumption of the ab- among others. However, the solid-phase synthesis method requires
alone meat. Thousands of tons of shells were found abandoned on an is- processing with high temperature. Template synthesis is necessary to
land and around a town (shown in Fig. 1). This not only resulted in add organic templates; however, it is not easy to degrade after
waste of natural resources, but also polluted the environment. Abalone implanting into the organism, which limits the clinical applications. For-
shell is a natural composite material. It is composed of more than tunately, the hydrothermal synthesis method can make use of the orig-
95 wt.% calcium carbonate [1,2] and less than 5 wt.% organic matter inal mineral that is poorly soluble or insoluble. Additionally, ion
[3–5]. Thus, abalone shell is a good source of calcium. Until now, there exchange and recrystallization occur, which make a smooth solid-
have been few reports on the efficient use of abalone shells. state conversion from one mineral phase to another realizable under
Hydroxyapatite (HAP, Ca10(PO4)6(OH)2) is the most stable form of hydrothermal conditions. Moreover, solid-state conversion reduces
calcium phosphate and is slightly soluble in water [6–8]. It is the main the calcination and milling steps. Hence, it is easy to control the reaction
inorganic component of mammalian hard tissues, such as bones and conditions. Thus far, some scholars have reported the synthesis of HAP
teeth [9,10]. The HAP content of human enamel is about 96 wt.%. Gener- using calcium carbonate with different forms [24,26], but it is still a chal-
ally, synthetic biomaterials have excellent bioactivity, biocompatibility lenge to synthesize well-organized nano-HAP with an expected compo-
sition and mechanical properties using abalone shell powders under
hydrothermal conditions.
⁎ Correspondence to: J. Chen, Institute of Biomedical and Pharmaceutical Technology, Herein, we utilized abalone shell powders as a calcium source for
Fuzhou University, No. 523 Gongye Road, Fuzhou 350002, China.
⁎⁎ Correspondence to: Q. Zhang, Institute of Biomedical Engineering, Chinese Academy
HAP. HAP nanorods were successfully synthesized via hydrothermal
of Medical Science & Peking Union Medical College, Tianjin 300192, China. solid-state conversion. The crystal phase and morphology of the sam-
E-mail addresses: [email protected] (J. Chen), [email protected] (Q. Zhang). ples were characterized using X-ray diffraction (XRD), field emission

http://dx.doi.org/10.1016/j.matdes.2015.08.056
0264-1275/© 2015 Elsevier Ltd. All rights reserved.
446 J. Chen et al. / Materials and Design 87 (2015) 445–449

For FTIR, (Nicolet 360 intelligent spectrometer, Thermo Nicolet

Corporation, US) studies, we used the KBr pellet method, which was
performed on dried samples.
The microstructure and surface potential of the sample were ana-
lyzed using FESEM, (Nova NanoSEM, FEI, Netherlands) and a potential
instrument (Malvern Zetasizer Nano-ZS90).
A thermogravimetric analyzer (TGA, DIL402C, NETZSCH, Germany)
further determined the composition of the obtained HAP. The samples
were heated to 1000 °C in flowing air at a heating rate of 10 °C/min.

3. Results and discussion

Fig. 2 shows the XRD patterns of the abalone shell powders (A) and
the samples exposed to different hydrothermal times of (B) 6 h,
(C) 18 h, and (D) 72 h. In Fig. 2A, the abalone shell was determined to
be composed of aragonite and calcite. After the hydrothermal treat-
ment, most of the diffraction peaks of the calcium carbonate (aragonite
and calcite) disappeared, whereas the HAP phase diffraction peaks were
Fig. 1. Abandoned abalone shells. detected at 2θ of 25.77°, 31.81°, 32.12°, 34.01°, 39.97°, 46.74°, and
49.38°. Moreover, the intensity of the diffraction peaks is increased as
a function of the prolonged hydrothermal treatment time at 2θ of
scanning electron microscopy (FESEM), Fourier transform infrared 25.77°, and 31.81° (shown in Fig. 2D). This suggested that the crystallin-
spectroscopy (FTIR), zeta potential measurements (ZP) and a thermo ity of the samples also increased. Moreover, the weak diffraction peaks
gravimetric analyzer. of the calcite and aragonite (at the 2θ of 29.38° and 33.08°, respectively)
were observed even after hydrothermal treatment for 72 h. This may be
2. Materials and methods due to the natural structure and special composition of the abalone shell
[1,27,28].
2.1. Materials To further investigate the composition of the samples exposed to dif-
ferent hydrothermal times of (B) 6 h, (C) 18 h, and (D) 72 h, FTIR anal-
The shells of Wrinkles Abalone were collected from the official dock ysis was conducted (Fig. 3). The bands at 1789 cm−1, 1464 cm−1,
Ocean Development Co., Ltd (Lianjiang Country in Fujian Province, CN). 862 cm−1 and 710 cm−1 represent CO2− 3 (shown in Fig. 3A), which is
Diammonium phosphate ((NH4)2HPO4, ≥ 99%) was purchased from due to the composition of the abalone shell that contains calcium car-
Sinophar Chemical Reagent Co., Ltd, China. Sodium hydroxide (NaOH, bonate (aragonite and calcite). After the hydrothermal treatment, the
98%) and acetic acid (CH3COOH, ≥ 99.5%) were purchased from components of all the samples were similar, which showed the pres-
Shanghai Lianshi Chemical Co., Ltd (Shanghai, CN). ence of the characteristic groups of the HAP. The characteristic vibration
of the PO3−4 groups was detected at 1037 cm−1, 960 cm−1, 608 cm−1
and 567 cm . The band at 3567 cm−1 was assigned to the stretching
−1
2.2. Preparation of the samples vibration of the OH−. Additionally, the vibration peaks at 1462 cm−1,
1418 cm−1, and 874 cm−1 represent CO2− 3 . These bands are due to cal-
First, the abalone shell was immersed in 10 vol.% acetic acid solution cium carbonate entrapped inside the HAP, which is consistent with the
to clean off the surface impurities. After rinsing with distilled water and XRD results. C\\H asymmetric stretching vibration and symmetric
naturally dying them, the abalone shell was crushed and screened using stretching vibration were detected at 2929 cm−1 and 2856 cm−1, re-
a pulverizer (SF-130, Zhong Cheng Pharmaceutical Machinery Factory, spectively. These bands demonstrated that organic matter exists in the
CN). This produced the abalone shell powders. synthesized HAP. This particular organic matter originates from the
For the synthesis, abalone shell powders and diammonium phos-
phate ((NH4)2HPO4) were used as the calcium and phosphorous
sources, respectively. The samples were prepared as follows: 2 g of ab-
alone shell powders and 1.5847 g of (NH4)2HPO4 were mixed (Ca/P in
a molar ratio of 1.67:1), and they were dissolved in 20 ml of deionized
water. The pH of the solution was adjusted to 10 using sodium hydrox-
ide solution (NaOH, 1 M) and the mixture was stirred at room temper-
ature for 30 min using a magnetic stirrer. Then, the mixed solution was
transferred to an autoclave, and heated at 150 °C at hydrothermal times
of 6 h, 18 h, and 72 h. After completing the reaction, the final precipitate
was washed with deionized water via centrifugation several times. Fi-
nally, the white latex-like precipitant was freeze-dried to yield white
powders. This produced the samples.

2.3. Characterization of the samples

The phase of the samples was examined using X-ray diffraction

(XRD, X'Pert PRO Panalytical diffractometer) with Cu Kα (λ =
0.154056 nm) incident radiation at a working voltage of 40 keV in the
range of 10° to 70° at a 0.02° step size. The phases were identified by
comparing the data with data reported in the Joint Committee of Powder Fig. 2. XRD patterns of the abalone shell powders (A) and the samples exposed to different
Diffraction Standards database (JCPDS no. 09-0432). hydrothermal times: (B) 6 h, (C) 18 h, and (D) 72 h.
 J. Chen et al. / Materials and Design 87 (2015) 445–449 447

450 °C. The TGA curve also showed a loss of adsorbed H2O at 100 °C.
After the hydrothermal treatment, the TGA–DSC patterns of all of the
samples were similar (shown in Fig. 4B–D). Thus, compared with
Fig. 4A, in all of the samples, we detected the mass loss of adsorbed
H2O, organic matter and carbon dioxide. This suggests that the sample
contained a small amount of calcium carbonate and organic matter,
which is similar to the results of XRD and FTIR. Particularly, slightly
below 700 °C, the sample exhibits an endothermic peak (shown in
Fig. 4B–D). This may be due to the HAP crystallized from the amorphous
calcium phosphate (ACP) of the sample, which is consistent with the re-
sults reported by Suchanek et al. [36].
Fig. 5 shows the FESEM images of the abalone shell cross-section
(A) and the samples exposed to different hydrothermal times of
(B) 6 h, (C) 18 h, and (D) 72 h. In Fig. 5A, the abalone shell consists of
aragonite sheets and prismatic calcite, this composition was consistent
with the XRD analysis (Fig. 2A). After 6 h of hydrothermal treatment,
the morphology of the sample is rod-like or flake-like (shown in
Fig. 5B), similar to the structure of the precursor, but the morphology
and the size of the samples are not uniform. As the hydrothermal time
Fig. 3. FTIR spectra of the abalone shell powders (A) and the samples exposed to different
increased to 18 h, the size and morphology of the samples became
hydrothermal times: (B) 6 h, (C) 18 h, and (D) 72 h.
more regular (Fig. 5C). A good rod-like structure and uniform sizes
were obtained for the nano-HAP after a hydrothermal time of 72 h
abalone shell [3,29], and it can play an important role in biological min- (shown in Fig. 5D). Fig. 5E is an enlarged image of the red area in
eralization [4,30–35]. Fig. 5D. As clearly shown, the structure of the obtained HAP is a hexag-
The thermal analysis (TGA and DSC) is illustrated in Fig. 4. A weight onal rod-like structure, which is similar to the structure reported by
loss in the TGA curve corresponds to an absorption peak of the DSC Zhan et al. [6]. Furthermore, the crystal growth schematic diagram of
curve, which represents a phase transition process. In Fig. 4A, we ob- the HAP hexagonal crystallites is showed in Fig. 5F. The sheet structure
serve that the major mass loss of the abalone shell powder occurred at of the HAP gradually reduces, whereas the rod-like structure increases
approximately 800 °C, which is due to the thermal decomposition of cal- as a function of the extended hydrothermal time. This result may be at-
cium carbonate to calcium oxide and carbon dioxide. Additionally, the tributed to the composition transformation of both aragonite and calcite
organic matter degraded in one or several steps between 200 °C and to HAP. At the beginning of the hydrothermal process, the amorphous

Fig. 4. TGA–DSC patterns of the abalone shell powders (A) and the samples exposed to different hydrothermal times: (B) 6 h, (C) 18 h, and (D) 72 h.
448 J. Chen et al. / Materials and Design 87 (2015) 445–449

Fig. 5. FESEM images of the abalone shell cross-section (A) and the samples exposed to different hydrothermal times: (B) 6 h, (C) 18 h, and (D) 72 h; The inset (E) is a magnified view of the
selected red area. (F) Schematic diagram of the HAP hexagonal crystallites. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this
article.)

calcium phosphate of the HAP precursor is deposited around the sur-

faces of the aragonite and calcite crystals. With more time, the forma-
tion of the outside layer HAP prevents the phase transformation of the
inside aragonite and calcite. Thus, the content of the internal aragonite
and calcite did not significantly change even after the long hydrother-
mal time. This result agrees with the slight change observed in the ara-
gonite and calcite in Fig. 2B–D and can explain the existence of the small
amount of aragonite and calcite in the produced HAP even though the
hydrothermal treatment time was 72 h (Fig. 2D). According to
previous research, monocrystalline HAP typically grows following the
c-axis of the HAP structure [37]. Hence, the rod-like structure of HAP
was finally formed.
Fig. 6 shows the zeta potential (ZP) value of the abalone shell pow-
ders (A) and the samples exposed to different hydrothermal times of
(B) 6 h, (C) 18 h, and (D) 72 h. We visually observe that the surface po-
tential of the abalone shell powders and the samples exposed to differ-
ent hydrothermal times are very similar, which all have a negative value
of approximately −22 mv. Smith et al. [38] reported that HAP particles
with a negative surface charge favor bone growth and osteoblast forma-
tion. Thus, the samples with a negative surface may be used as potential
bone repair materials to promote bone growth and osteoblast forma-
tion. Moreover, for particles that are small enough, a high zeta potential Fig. 6. Zeta potential patterns of the abalone shell powders (A) and the samples exposed to
will confer stability; thus, the particles will resist aggregation. However, different hydrothermal times: (B) 6 h, (C) 18 h, and (D) 72 h.
 J. Chen et al. / Materials and Design 87 (2015) 445–449 449

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