Materials Letters 183 (2016) 405–407

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Materials Letters
journal homepage: www.elsevier.com/locate/matlet

Potential formation of hydroxyapatite in total blood and dicalcium

silicate elaborated from shell and glass powders
A. Bouregba n, A. Diouri
Laboratory of Applied Solid State Chemistry, Faculty of Science, Mohammed V University, Rabat, Morocco

art ic l e i nf o a b s t r a c t

Article history: This paper demonstrates the feasibility of using mussel shells and soda-lime glass powders for preparing
Received 30 March 2016 dicalcium silicate (Ca2SiO4, C2S), by solid state reaction, then to synthesis hydroxyapatite, (Ca10
Received in revised form (PO4)6(OH)2, HAp), by soaking the powder of dicalcium silicate in a biological medium for a variety of
30 June 2016
time periods from 2 h to 5 days. The specimens are characterized by X-ray diffraction (XRD) and Fourier
Accepted 31 July 2016
Available online 1 August 2016
transformed infrared spectroscopy (FT-IR). The results shows that solid state reaction method allows the
synthesis of dicalcium silicate powder by heating mixture of mussel shells and the soda-lime glass
Keywords: powders at 1050 °C. The results also shows that the hydroxyapatite is formed after soaking dicalcium
Dicalcium silicate silicate powder in total human blood for 20 h and growth up to 5 days.
Hydroxyapatite
& 2016 Elsevier B.V. All rights reserved.
Bioactivity
Mussel shells powder
Soda-lime glass powder

1. Introduction The seashells were composed of Ca as the main metal, with a

Bone is a natural composite material primarily consisting of an
organic phase (mostly Type I collagen) as matrix and a mineral
phase (hydroxyapatite crystal) as reinforcement [1]. According to
Kokubo, a bioactive material is a bone-like carbonated apatite
which will form selectively after it is immersed in a serum-like
solution [2]. Hydroxyapatite (HAp)[Ca10(PO4)6(OH)2] is a bio-
compatible polycrystalline ceramic material that has a chemical
composition and a crystal structure similar to the human bone and
teeth [3]. HAp can be derived from natural sources, such as bovine
bones, or corals via hydrothermal transformation, or it can be
synthetically prepared [4].
The dicalcium silicate Ca2SiO4 (Belite) (C2S) varieties have at-
tracted intense attention due to their extensive applications in
several fields like cement, ceramics, drugs and biological materials
[5]. It presents five allotropic forms: α, α’H, α’L, β and ɣ exist be-
tween room temperature and 1500 °C, but their stability ranges on
heating and cooling are different [6]. Variety of studies have de-
monstrated that some CaO-SiO2 based biomaterials such as Bio-
glass (BG) [7] Calcium silicate [8] and hydraulic calcium silicate
cements (HCSCs)) [3] are bioactive, and can enhance bone tissue
formation by inducing the formation of a bone-like apatite in vivo
and in vitro which owe to the CaO-SiO2 components.
n
Corresponding author.
E-mail address: bouregba.fssm@gmail.com (A. Bouregba).
small amount of Mg, Al and Si. Generally, Ca should exist in the
seashells as CaCO3 [9]. Mussel shells are a calcium-rich resource
that can be used to produce calcium oxide (lime) [10]. This lime
can be used in several different technologies it can also be con-
verted into hydroxyapatite (HAp) [10].
Glass is the result of the merger of several inorganic mineral
raw materials, which after undergoing a process of controlled
cooling becomes a hard, homogeneous, amorphous and isotropic
material [11]. For soda lime glass the typical glass composition is
approximately 70% of silica, 13–17% Na2O and 10% CaO [12].
However, glass has a composition that favours the development of
pozzolanic reactions [12].
In this work, mussel shells and soda-lime glass which pre-
dominantly composed of calcium carbonate and silica (respec-
tively) was used as starting materials to elaborate dicalcium sili-
cate (Ca2SiO4, C2S), by solid state reaction, then to synthesis hy-
droxyapatite, (Ca10 (PO4)6(OH)2, HAp), by soaking the powder of
dicalcium silicate in a biological medium for a variety of time
periods from 2 h to 5 days.
2. Materials and methods
2.1. Preparation of C2S powder
Dicalcium silicate powder was synthesized by solid state

http://dx.doi.org/10.1016/j.matlet.2016.07.153
0167-577X/& 2016 Elsevier B.V. All rights reserved.
406 A. Bouregba, A. Diouri / Materials Letters 183 (2016) 405–407

Table 1
Chemical composition of powder mussel shells and glass powder (wt%).

Materials SiO2 Al2O3 Fe2O3 CaO MgO SO3 K2O TiO2 MnO P2O5 LOIa

Mussel shells 0,35 0,12 0,08 55,24 0,08 0,20 0,02 0,00 0,00 0,04 43,51
Glass 75,49 1,60 0,24 10,13 0,53 0,32 0,19 0,02 0,01 0,01 8,53

a
LOI: Loss on ignition.

reaction of mixture of mussel shells and glass powders. The

*
30
mussel shells are collected from the coast of the Atlantic Ocean of
Rabat-Morocco, then shells are dried and finely milled (o 50 mm). * * . CaO
**
25
Soda-lime glass powder is obtained by grinding the glass beverage
bottles and cups glass (o125 mm). The chemical composition of * C 2S
20
the basic materials (Table 1) obtained by the Fluorescent X-ray

Lin(Counts)
analysis shows that the fraction of mussel shell contained a sig-
15
*
nificant quantity of calcium oxide (55.24%). It also contained
smaller quantities of impurities, The loss on ignition can be ob- *
* * .
tained by the test portion before and after calcination of the 10

sample at 1000 °C, the loss on ignition for shells (43.51%) was at-
tributed to the loss of carbon dioxide due to the partial dissocia- 5 *
* *** .***. * * **
tion of calcium carbonate. Soda-lime glass consists essentially of
calcium, aluminum and magnesium. Shell and soda-lime glass 0
10 20 30 40 50 60
powders are mixed with specific proportions (70% and 30% re-
spectively) then the mixture is heated up to 500 °C for 24 h fol- 2-Theta-Scale
lowed by milling with ethanol after rapid removal from the fur- Fig. 1. XRD pattern of C2S powder synthesized at 1050 °C.
nace. This procedure was repeated by increasing the temperature
until attending a heat treatment of 1050 °C for 4 h in which we
have obtained a C2S phase. Samples were characterized by X-ray 0.6
diffraction (XRD) using a Siemens D5000 diffractometer and also
by Fourier Transform Infrared (FT-IR) spectroscopy using a JASCO 0.5
FTIR-4600 on the region 400–4000 cm 1.

0.4 1406.01
2.2. In vitro soaking
3637.59
996.24
To entirely reproduce the living environment requires not only 0.3
plasma of synthesis but also the normal human body temperature.
For this a thermostatic marine bath set at a temperature of 37 °C is 0.2 873.68
used. It corresponds to an immersion thermostat in which is 839.09
placed, for various times from 2 h to 5 days, glass tubes of 5 ml
0.1
(Type K3EDTA), containing human blood and C2S powder. After
499.24
soaking, the samples are filtered and air dried.
0.0
4000 3600 3200 2800 2400 2000 1600 1200 800 400
-1
3. Results and discussion Cm

Fig. 2. FTIR spectra of C2S powder synthesized at 1050 °C.

3.1. Characterization of C2S
The C2S characteristic peaks are identified in Fig. 1. We noted
the presence of small peaks of free lime CaO at 35.40°, 40.38° and
57.06° these XRD peaks have been picked from (JCPDS (CaO) file
No. 01-077-7243) . Also the many sharp peaks and low back-
ground noise in the XRD pattern shows that the powder C2S is
purely crystallized [13].
Fig. 2 shows the FTIR spectra of the powder synthesized at
1050 °C, the main bands corresponding to the silicate groups are
due to the SiO44 − tetrahedral. According to Puertas the absorption
bands of the different polymorphs of dicalcium silicate corre-
sponding to ( ν1) (symmetric stretching) located around 800–
900 cm 1 and ( ν3) (antisymmetric stretching) are typically located
around 800–1000 cm 1 and those of ( ν4) (the triply degenerated
out of plane bending) located around 400–500 cm 1 [14]. The
bands absorptions observed around 3637 and 1406 cm 1 are the
characteristic absorption bands of CaO [15].
3.2. Formation of HAp
After immersion and air drying, each sample was analyzed by
(XRD) and (FT-IR). Fig. 3 shows the appearance of hydroxyapatite
peaks after 20 h of immersion in human blood. The presence of
less intense and broad peaks can be explained by the fact that
much of the hydroxyapatite (HAp) remains in its amorphous form
and only a small amount crystallized. We noted that the C2S
characteristic peaks decrease during soaking in human blood,
whereas the characteristic peaks of HAp appeared after 20 h and
increased up to 5 days [13], we noted also the appearance of a new
peak of Ca3Si2O7
2H2O after 6 h and its disappearance after 1 d of
soaking. The appearance of this peak is explained by hydration of
Ca2SiO4 [13].
FTIR spectra (Fig. 4) shows that the absorption band of CaO at
3637 cm 1 is disappeared in the first two hours of soaking, also
when the soaking time increased the intensity of the absorption
 A. Bouregba, A. Diouri / Materials Letters 183 (2016) 405–407 407

a: HAp immersion of the C2S powder in human blood.

b: CaO .SiO
c: CaO
3.4
d: Ca (Si O ).2H O
3.2
3.0
4. Conclusion
2.8
2.6 b
b This paper demonstrates the feasibility of using mussel shells
2.4 b b
2.2 a a a a and soda-lime glass powders for preparing dicalcium silicate
a a b b b b a
Lin(Counts)

a
2.0 a a (Ca2SiO4, C2S), by solid state reaction without chemical stabilizers.
1.8 5d
1.6
Soaking the C2S powder in tubes containing human blood showed
1d
1.4 a strong hydration which is favorable to the formation of hydro-
1.2 d
1.0
20 h xyapatite. Our tests showed that hydroxyapatite may appear at
0.8 b 6h 20 h and grow with time soaking accompanied with decrease of
0.6 2h C2S peaks which dissolved to promote the growth of
0.4 b b b bb b C hydroxyapatite.
0.2 b b bb C C
0h
0.0 The use of shells and soda-lime glass will bring a very sig-
10 20 30 40 50 60
nificant enhancing to mechanical and physical properties of HAp
2-Tetha-Scale
therefore this study opens the possibility of other studies in-
Fig. 3. XRD pattern of C2S powder before and after immersion. vestigating the mechanical and physical properties of HAp syn-
thesized by immersing the C2S powder elaborated from a mixture
2.0
1442.512 of shells and soda-lime glass in a biological medium.
1635.88 1030.47
1.9
1402.01 872.62
1.8 603.56
1.7
1.6 References
1.5 5d
1.4
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