CHAPTER -1

INTRODUCTION
1.1. Biomaterials
Biomaterials are materials used for making devices that can interact with
biological systems to coexist for longer service with minimal failure. “Biomaterial
is defined as any systemically, pharmacologically inert substance or combination of
substances utilized for implantation within living system to support function of
living tissue” [1].

Biomaterials are widely used in repair, replacement or augmentation of

diseased or damaged parts of the musculoskeletal system such as bones, joints and
teeth. The fundamental requirement of a biomaterial is that the material and the
tissue environment of the body should coexist without having any undesirable or
inappropriate effect on each other. Biocompatibility is an essential requirement for
any biomaterial, implies the ability of the material to perform effectively with an
appropriate host response for the desired application. Common medical devices
made of biomaterials include hip replacements, prosthetic heart valves and the less
common neurological prostheses and implanted drug delivery systems.

Biomaterials can be classified into three categories by their macroscopic

surface characteristics or by their chemical stability in the body environment. They
are bioinert, bioactive and bioresobable.

Bioinert refers to any material that once placed within the human body has
minimal interaction with its surrounding tissue. Examples of bioinert materials are
stainless steel, titanium, alumina, partially stabilized zirconia, and ultra high
molecular weight polyethylene. Generally a fibrous capsule forms around a bioinert
implant.

Bioactive refers to a material, which upon being placed within the human
body interacts with the surrounding bone and in some cases, even with the soft
tissue. An ion exchange reaction between the bioactive implant and surrounding
body fluids results in the formation of a biologically active carbonate apatite layer
on the implant that is chemically and crystallographically equivalent to the mineral
phase of bone. Examples of bioactive materials are hydroxyapatite and bioglass.

1
 Bioresorbable refers to material that upon placement within the human body
starts to dissolve (resorbed) and is slowly replaced by tissue such as bone.
Examples of bioresorbable materials are synthetic tricalcium phosphate and
polylactic-polyglycolic acid copolymers.

Materials including metals and alloys, polymers, ceramics and composites

with the appropriate physical properties and biocompatibility are chosen for the
fabrication of medical device.

1.1.1 Metals and alloys

Metals and alloys have a wide range of applications including as devices for
fracture fixation, partial and total joint replacement, external splints, braces and
traction apparatus as well as dental amalgams. Although metals exhibit high
strength and toughness, they are susceptible to chemical and electrochemical
degradation. The implant materials may corrode or wear, leading to the generation
of particulate debris, which may in turn aggravate the body environment and elicit
both local and systemic biological responses. Metals and alloys are very important
in implant applications due to its good mechanical properties.

Example: Stainless steel, gold and Ni-Cr alloy [1,2].

1.1.2 Polymers

Polymers are large molecules made up by the repetition of small, simple

chemical units termed as monomers. Polymers are considered for implant
applications in various forms such as fibres, textiles, rods and viscous liquids.
Recently, polymers have been introduced for hip socket replacement in orthopedic
implant applications due to its close resemblance to natural polymeric tissue
components. However, polymers undergo degradation in the body environment due
to biochemical and mechanical factors. This results in ionic attack and formation of
hydroxyl ions and dissolved oxygen, leading to tissue irritation and decrease in
mechanical properties.

Example: Polyethylene, Polyesters and Silicones [2].

2
1.1.3 Composite

Composites are mixtures of two or more materials, which in combination,

offer superior properties to the material alone. Composites usually refer to the use
of fibers which are embedded in a plastic. This composites offer high strength with
very little weight [3].

Example: BUS-GMA-quartz/silica filler, PMMA-glass fillers

1.1.4 Ceramics

Ceramics are inorganic compounds between metallic and nonmetallic

elements that can be classified into three categories of biomaterials by their
macroscopic surface characteristics or by their chemical stability in the body
environment. They are nearly inert (carbon, alumina and zirconia), surface reactive
(glass ceramics) and completely resorbable (hydroxyapatite, calcium phosphate).
The limitations of ceramic materials are their low tensile strength and fracture
toughness.

Example: HA, TCP, and Alumina.

Table 1 gives a list of biomaterials used in body implants with their

properties [1,2,3].

Among which, hydroxyapatite (HA), Ca10(PO4)6(OH)2 was recognized as

the most suitable biocompatible materials since it resembles the mineral component
of bone and teeth.

3
 Table.1 List of Biomaterials and Applications
Material Applications
Metals and Alloys:
316L Stainless steel Fracture fixation, stents, surgical
instruments
Co-Cr-Mo, Cr-Ni-Cr-Mo Bone and joint replacement dental
implants, dental restoration, heart valves
Gold alloys Dental restorations
Polymers:
Polyethylene Joint replacement
Polyurethanes Blood-contacting devices
Composites:
BUS-GMA-quartz/silica filler Dental restorations
PMMA-glass fillers Dental restorations
Ceramics:
Alumina Joint replacement, dental implants
Zirconia Joint replacement
Calcium phosphates Bone repair and augmentation, surface
coatings on metals
Bioactive glasses Bone replacement
Carbons Heart valves, precutaneous devices,
dental implants

1.2 Structure of bone and teeth

1.2.1 Bone
Bone is the most typical calcified tissue in mammals. It comes in all sorts of
shapes and sizes in order to achieve the various functions of protection and
mechanical support for the body.

4
 The major inorganic component of bone mineral is a biological apatite,
which can be defined as a non-stoichiometric, ion-substituted calcium deficient
HA [4]. From the material point of view, bone can be considered as an assembly of
distinct levels of seven hierarchical structural units, from the macro- to the micro-
and nanoscales (Fig.1), to meet numerous functions [5]. At the nanostructural level,
tiny plate-like crystals of biological apatite in bone occur within the discrete spaces
within the collagen fibrils and grow with specific crystalline orientation along the c-
axes, which are roughly parallel to the long axes of the collagen fibrils.

The apatite growth is somehow limited by these small intercollagenous

spaces, which are approximately 50 nm in length, 25 nm in width and 2–3 nm thick.
Type I collagen molecules are self-assembled into fibrils with a periodicity of 67
nm and with 40 nm gaps between the ends of their molecules, into which the apatite
crystals are placed. A composite of these two constituents forms mineralized fibrils.
This is why bone is usually termed a fiber-reinforced composite of biological
origin, in which nanometer sized hard inclusions are embedded in a soft protein
matrix. Though the size of biological apatite crystals reported in the literature varies
due to different treatment methods and analysis techniques, it is generally around
the nanometric level, with values in the ranges of 30–50 nm (length), 15–30 nm
(width) and 2–10 nm (thickness) [6].

5
Fig.1 Microstructure of bone

6
1.2.2. Teeth
All teeth consist of two parts, the crown and the root. The root is placed in a
socket called he alvesouls in the maxillary (upper) or mandibular (lower) bones,
being covered by a layer of cementum and attached to the bone by the periodontal
membrane (a layer of fibrous connective tissue). A schematic cross section of a
tooth is showin Fig. 2

Fig.2 Schematic diagram of a teeth

The enamel is the hardest substance in the body and consists of 97 wt % (92
vol.%) of relatively large HA crystals (25 nm thick, 40-120 nm wide, 160-1000 nm
long). The remaining 3 wt% (7 vol.%) consists of organic substance and water [7].
The HA crystals in enamel form well-defined rod or prism-like structures of about 4
µm in diameter [8]. Dentine is a mineralized tissue whose distribution of organic
matrix and minerals is similar to that of regular compact done. Dentinal tubules (3-5
µm in diameter) radiate from the pulpcavity toward the periphery and penetrate
every part of the dentine [9]. Tubules in the longitudinal direction and the interface
is cemented by a protein-polysaccharide complex substance. Pulp is a soft tissue
containing thin collagenous fibers, nerve cells, blood vessels, etc [10].

The layer of cementum surrounding the root varies from 20-50 µm at the
cervix to 150-200 µm at the apex. Approximately half of the cementum is inorganic
and half is composed of organic material and water [11]. The periodontal
membrane is made of mostly collagenous fibers and glycoprotins (protein-
polysaccharide complex) [12].

7
1.3 Hydroxyapatite

Calcium phosphate ceramics are attractive materials for bone replacement,

dental defect filling, bone tissue engineering and drug delivery applications. Table 2
shows several calcium phosphates arranged according to their Ca/P ratio [13].
Among the several calcium phosphates, Hydroxyapatite (HA) with the chemical
formula Ca10(PO4)6(OH)2 is one of the most important bioceramic material.
Hydroxyapatite, also called hydroxylapatite, is a mineral. It is a naturally occurring
form of calcium apatite with the formula Ca5(PO4)3(OH), but is usually written
Ca10(PO4)6(OH)2 to denote that the crystal unit cell comprises two molecules. Pure
hydroxyapatite powder is white. Naturally occurring apatites can however also have
brown, yellow or green colorations, comparable to the discolorations of dental
fluorosis. The theoretical density of hydroxyapatite is 3.16 gcm-2. HA has been used
for various biomedical applications like matrices for controlled drug delivery, bone
cements, tooth paste additive, monolithic implants or coatings on metallic implants,
such as Ti alloys, Co-Cr alloys, stainless steels [14]. But its usage at high load
bearing conditions was restricted because of its brittleness property, poor fatigue
resistance and strength. Hence there is a need for improving the mechanical
properties of these materials suitable for clinical applications. Non-medical
applications of HA include packing media for column chromatography, gas sensors,
catalysis and host materials for lasers [15].

Table 2 Calcium phosphates arranged by Ca: P ratio

Name Abbreviation Formula Ca:P

Tetracalcium phosphate TetCP Ca4O(PO4)2 2.0

Hydroxyapatite HA Ca10(PO4)6(OH)2 1.67

Tricalcium phosphate (α, β, γ) TCP Ca3(PO4)2 1.50
Octacalcium phosphate OCP Ca8H2(PO4)6.5H2O 1.33

8
 Dicalcium phosphate dihydrate DCPD CaHPO4.2H2O 1.0
(brushite)

Dicalcium phosphate (monetite) DCP CaHPO4 1.0

Calcium phosphate (α, β, γ) CPP Ca2P2O7 1.0
Calcium pyrophosphate dihydrate CPPD Ca2P2O7.2H2O 1.0
Heptacalcium phosphate HCP Ca7(P5O16)2 0.7

Tetracalcium phosphate diacid TDHP Ca4H2P6O20 0.67

Calcium phosphate monohydrate MCPM Ca(H2PO4)2.H2O 0.5

1.3.1. Structure of hydroxyapatite

Of the two known crystal forms of HA-monoclinic, space group P21/b, and
hexagonal, space group P63/m-only the hexagonal phase is of practical importance
because the monoclinic form is destabilized by the presence of even small amounts
of foreign ions [16]. The hexagonal structure contains two different cation sites,
Ca(I) and Ca(II), but only one phosphate environment (Ca(I), Ca(II) are used for
stoichiometric apatite; M(I), M(II) are the general symbols for substituted apatites).
The structure can be roughly described as a phosphate assembly crossed by parallel
channels filled by OH- ions and parallel to the crystallographic c-axis (Fig. 3). The
channel walls are formed of Ca(II) atoms arranged in staggered triangular arrays.
Ca(I) atoms have a different environment and are positioned in columns parallel to
the OH- channels. A unit cell accommodates a formula unit Ca10(PO4)6(OH)2.
Among the 10 cations, the 4 Ca(I)s are tightly bonded to 6 oxygens and less
strongly to the other 3 oxygens (mean Ca(I)–O distance 0.255 nm), whereas the 6
Ca(II) atoms are surrounded by 7 oxygens (mean Ca(II)–O distance 0.245 nm).
Ca(I) atoms are strictly aligned in columns and any small change in the metal–
oxygen interactions affects the entire lattice [17].

9
 Fig.3 Structure of HAp

1.3.2. Synthetic Hydroxyapatite

HA is a class of bioceramic material frequently used in bone grafting and in
bone drug delivery. It is one of the most widely used biomaterials for various bone
applications because it possesses most of the qualities required for bone tissue
formation. There are a number of methods for processing HA either from natural
sources or from synthetic sources, but these are studies focused on conventional
microphase HA. Recently, nanophase HA has received much attention owing to its
superior surface functional properties over its microphase counterpart, particularly
surface area and surface roughness, which are the most imperative properties
required to promote cell adhesion and cell-matrix interactions immediately after
implantation.
Nanostructured materials are defined as materials containing structure
elements (e.g. cluster, crystallites or molecules) with dimentsions in the range 1-100
nm. It is worth mentioning that the nanophase HA is one of the basic building
blocks of natural bone that occupies about 60 wt.%. It has also been proved that the
nanophase HA, compared to conventional HA, promotes osteoblast adhesion,
differentiation and proliferation, which leads to enhanced formation of new bone
tissue within a short period. The nanophase HA is therefore perceived to be
beneficial in bone tissue engineering as a new generation scaffolding system. HA at

10
nano level would have amazing functional properties due to its grain size, large
surface area to volume ratio and ultrafine structure similar to biological apatite,
which would have a great impact on implant cell interaction in body environment.
There are several methods for the synthesis of nanophase HA, which include
wet chemical method [18], solid state reaction [19], hydrothermal method [20],
micro emulsion method [21], mechanochemical method [22] and sol-gel method
[23]. Among these methods, sol-gel synthesis of HA ceramics has recently attracted
much attention, due to its many advantages, which include high product purity,
homogeneous composition and low synthesis temperature.

1.3.3 Sol-gel method

Sol–gel is a useful process of self-assembly for the synthesis of
nanoparticles. Colloids are suspensions with molecules of 20–100 lm in diameter in
a solvent. The colloid that is suspended in a liquid is the ‘sol’, and the suspension
that keeps its shape is the ‘gel’. Thus, ‘sol–gels’ are suspensions of colloids in
liquids that keep their shape. The sol–gel process involves the evolution of
networks through the formation of a colloidal suspension and gelation of the sol to
form a network in continuous liquid phase. The precursors for synthesising these
colloids normally consist of ions of a metal, but also
sometimes of other elements surrounded by various reactive species, -i.e., the
‘ligands’.
The sol–gel formation occurs in four stages: (a) hydrolysis, (b) condensation and
polymerisation of monomers to form particles, (c) growth of the particles, (d)
agglomeration of the particles followed by the formation of networks that extend
throughout the liquid medium resulting in thickening, which forms a gel. Upon
drying, trapped volatiles are driven off and the network shrinks as further
condensation may occur. These processes are basically affected by the initial
reaction conditions. By controlling these factors, it is possible to vary the structure
and the properties of the sol–gel derived inorganic network. For instance, with
hydrolysis under controlled conditions, dispersed spherical nanoparticles can be
synthesized.[23]

11
1.3.3.1 Microwave irradiation:
Microwave synthesis of materials offers the advantages of heating
throughout the volume and very efficient transformation energy. Microwave
heating is due to the high penetration depths of microwaves its coupling with lossy
materials results in rapid and uniform heating of the entire bulk of the reacting
substance. This turn, minimizes the thermal gradients reduces the time for particle
diffusion and hence the product can be obtained in relatively short time. Compared
with conventional method, microwave synthesis has the advantages of very short
time, small particle size, narrow particle size distribution, and high purity [24].

Microwave heating is fundamentally different from conventional

heating in that the heat is generate internally within the material heating in that is
the generated internally within the material instead of originating from an external
heating and subsequent radioactive transfer [25].
In chemical point of view microwave heating is of higher spatial
distribution of heat or transfer rate in addition it is a very sensitive function of the
material being processed and depends upon such factors size, geometry, and mass
of the sample
Advantages such as rapid heating rates, reduced processing time,
substantial energy savings, and being environmentally cleaner improve properties,
including fine microstructure of products. Consequently HA should via precipitated
in very small nanosized crystallites [26]

1.4. Scope of the work

The objective of present study is to synthesis HA nanoparticle in the
presence of citric acid via microwave heating and its characterization.

12
 Chapter -2

Experimental

2.1 Chemicals

The chemicals used were analar grade calcium nitrate tetrahydrate

(Ca(NO3)2·4H2O, 98%), di-ammonium hydrogen phosphate (NH4)2HPO4, 99%),
citric acid monohydrate (C6H8O7.H2O, 99.5%) and ammonia solution (NH4OH,
25%) obtained from Merck. All reagents were used without further purification.
Deionized water was used as the solvent.

2.2 Synthesis
The nanocrystalline HA was prepared by using sol-gel method via microwave
irradiation.[23-25] Briefly, solutions of 1M Ca(NO3)2·4H2O (23.615 g of
Ca(NO3)2·4H2O in 100 ml deionized water), 1M C6H8O7.H2O (21.014 g of
C6H8O7.H2O in 100 ml deionized water) and 0.6M (NH4)2HPO4 (7.923 g of
(NH4)2HPO4 in 100 ml deionized water) were separately brought to pH above 10 by
adding NH4OH solution. The calcium nitrate solution was stirred vigorously at
room temperature and the citric acid solution and di-ammonium hydrogen
phosphate solution was added drop wise into this calcium nitrate solution one by
one. The reaction mixture was put into a household type LG microwave oven of
600W power under air atmosphere for 15 min. The final suspension was allowed to
cool to room temperature and then the precipitate was dried in vacuum oven at 90ºC
for 48h. Finally, the product was ground into a fine powder using a mortar and
pestle. Flow chart for synthesis of nanocrystalline HA is shown in Fig. 3.

13
 1M citric acid
0.6 M (NH4)2HPO4 pH > 10
pH > 10

1M Ca(NO3)2.4H2O
pH > 10

Mixing under
vigorous stirring

Sol-gel

Microwave heating
(600 W, 15 min)

White precipitate

Dried (90ºC, 48h)

Crushed

Powder

Fig. 3 Flow chart for synthesis of nanocrystalline HA

2.3 Calcination

14
 The as-synthesized sample was annealed at 1100° C for three hours at the
heating rate of 2°C/min. After 3 hours the sample was cooled at the rate of 3°C/min
until reaching 30°C.

2.4 Characterization:
2.4.1 X-ray diffraction:
X-ray diffraction (XRD) is a highly developed and documented technique
that is widely used to identify crystalline materials. It can provide quantitative
information about the chemical composition of a sample, containing different
crystalline phase. The technique involves the generation of X-rays within a sealed
tube that is under vacuum. A current is applied to heat the filament contained within
the tube and the number of electrons emitted corresponds to the strength of the
current. The higher the voltage applied, the greater the number of electrons emitted
from the filament. Typically a voltage of 15–60 keV is used to accelerate the
electrons towards a target commonly made of copper and upon impact the
production of X-rays that is of a wavelength characteristic of the target is then
collimated and directed onto a material sample.
The sample can be in finely ground powder form or a smoothly surfaced
processed disc. The X-ray signal is recognized by a detector and processed by a
microprocessor or electronically and converted to a count rate. An X-ray scan of the
sample can be produced by changing the angle between the X-ray source, the
sample and the detector at a controlled rate between preset limits.
The distances between planes of atoms that constitute the sample can be
measured and provide a characteristic set of data corresponding to the chemical
composition of the material. Comparison against standard reference patterns and
measurements allows for identification and analysis of the material. Other
advantageous information such as degree of crystallinity and any deviation from
ideal composition or the presence of impurities can be obtained.[27]
XRD pattern of synthesized sample was carried out using a Rigaku D/MAX
ULTIMA diffractometer, with voltage and current setting of 40 kV and 30 mA,
respectively and uses of Cu-Kα radiation (1.5406 Å). Crystallographic
identification of the phases of synthesized sample was accomplished by comparing

15
the experimental XRD patterns to standards compiled by the International Center
for Diffraction Data (ICDD).

2.4.2 FT-IR study

IR spectroscopy is an analysis technique that provides information on the
vibrations of atomic and molecular units and is a standard analytical method
employed to reveal information of the specific chemistry and the molecular
structure and orientation of a material specimen.[28] The region of the
electromagnetic spectrum that gives rise to IR radiation promotes transitions in a
molecule between vibrational (from near- and midinfrared radiation) and rotational
(far infrared radiation) energy levels of the lowest electronic energy state. The
molecule can absorb the radiation by increasing its own vibrational energy. IR
spectroscopy relates to the ability of the molecule to absorb radiation energy at
certain specific frequencies which match the natural vibrational frequencies of the
molecule occurring in the IR region of the electromagnetic spectrum, during
irradiation with a whole range of IR frequencies. Therefore, chemical bonds and
groups of bonds within a specimen will vibrate at specific frequencies and upon
exposure to IR rays will only absorb the energy at frequencies characteristic to the
material. The transmittance and reflectance of the IR rays by the sample are
translated into peaks producing a spectral pattern that can be compared and
analysed to identify the material.
The FT–IR spectra of as-synthesized samples were recorded on Perkin
Elmer spectrophotometer in the wavenumber range of 4000–400 cm−1 with 1 cm−1
resolution by using KBr pellet technique.

2.4.3 Scanning electron microscopy (SEM)

This widely used and much written about technique functions by focusing
and rastering a relatively high energy electron beam (5–100 keV) on a material
surface. Low energy electrons are emitted from each impact spot of the focused
beam. The intensity of the secondary electron emission is a function of the atomic
composition of the sample and the geometry of the features under observation. The
surfaces are imaged by spatially reconstructing the intensity of the secondary
electron emission on a phosphour screen. Only the secondary electrons generated

16
near the surface of the bulk material can detected because the shallow penetration
depth of the low energy secondary electrons produced by the primary electron
beam, leading to surface analysis only. Nonconductive specimens are coated with a
thin electrically grounded layer of metal or sputter coated with gold (Au) to
minimise the accumulation of electric charge by the electron beam. Being that the
metal layer is always 300 A° thick, the electrons being emitted from the sample
underneath cannot penetrate and it is the surface coating of the metal that is actually
being monitored. A truly conformal metal coat is therefore required to convey a
good representation of the surface geometry. Low voltage SEM offers a lower
electron accelerating voltage of 1 keV so that charge accumulation is unlikely and
nonconductive specimens do not require coating.[29] The synthesized sample was
coated with a thin layer of Gold (Au) by sputtering and then the microstructure of
the samples was examined using a Hitachi S-3000H scanning electron microscope
(SEM).

Chapter -3

Results and Discussion

3.1 XRD analysis

XRD technique is found to be a powerful tool to determine the phase

composition of materials. The XRD pattern of as-synthesized sample is shown in

the Fig . 4. The obtained data form XRD pattern was compared with JCPDS data

17
for calcium phosphates. The observed data did not match with the JCPDS data of

any calcium phosphate phases, which indicate the formation of calcium phosphate

phases was inhibited by addition of citric acid during the synthesis. However, as-

synthesized sample contain calcium carbonate and ammonium nitrate as the

crystalline phase along with amorphous calcium phosphate.

The XRD pattern of 1100ºC calcined sample is shown in the Fig. 5.

This XRD pattern indicates the sample calcined at 1100ºC consists of HA, β-TCP

and CaO. The composition of the present phase in the sample is given in table 4.

18
Fig. 4 The XRD pattern of as-synthesized sample

19
Fig. 5 The XRD pattern of the 1100ºC calcined sample

Table 4 The phase composition of calcined sample

Phase present Composition (%)

HA 78.33

20
 β-TCP 15.43

CaO 6.24

The obtained from XRD results indicate that appropriate calcination

is needed for achieving pure calcium phosphate phase.

3.2 FT-IR analysis

The FT-IR spectrum of as-synthesized sample by microwave irradiation is

shown in the Fig. 6.

• The peak at 435 cm-1 is assigned to a triply degenerated bending mode, v2 ,

of the O-P-O.[30)

• Bands at 606 cm-1 and 1072 cm-1 are assigned to vibration of phosphate

group, PO42-.

• The peak at 542 cm-1 is assigned to racking vibration of COO-.

• The Bond at 668 cm-1 is assigned to O-H…..O out of plane vibration.

• The peaks at 832 cm-1 and 1382 cm-1 are due to N-O bending vibration

modes of NO3- ions.

• The peak at 892 cm-1 is assigned to characteristic bending mode (v6 or v2) of

the CO32- group.

• The peak at 1268 cm-1 is due to OH deformation of vibration of COO-.

• The bond at 1613 cm-1 is assigned to asymmetry CO2- stretching vibration.

21
 • The peaks between 2400 cm-1 to 3926 cm-1 are assigned to stretching mode

(vS), librational mode (vL) and translational mode (vT) of the hydroxyl group,

OH.

100

435
3926

2400

668
80

738
Transmittance (%)

832 892

542
606
60

1174
1072
40

1268
3035

20
1613
1553
3158

1382

0
4000 3500 3000 2500 2000 1500 1000 500
-1
Wavenumber (cm )

Fig. 6 FT-IR spectrum of as-synthesized sample

Table 3 The FT-IR assignment for as-synthesized sample

Wavenumber Assignment

22
 (cm-1)
435 O-P-O bending modes (v2) of the PO4 group
542 Racking vibration of COO-
606 The triply degenerated v4 bending vibration PO43- ions
668 O-H…….O out of plane vibration
738 C-O deformation vibration of COO-
832 N-O bending vibration modes of NO3-
892 Characteristic bending mode (v6 or v2) of the CO32- group
1072 Due to presence of HPO42-
1176 C-O Stretching
1268 O-H deformation vibration of COO-
1382 N-O Stretching vibration of NO3- ions
1613 Asymmetry CO2- Stretching
2400-3926 H2O and OH vibration

3.3 SEM-analysis

The SEM pictures of as-synthesized sample with different

magnification are shown in the Fig. 7 and 8. The SEM image indicates that the as-

synthesised sample consist of micrometric particles with irregular morphology.

The irregular or inhomogeneous morphology of as-synthesised sample

is due to the presence of different calcium/citrate/phosphate species. These results

were support the XRD and FT-IR result of as-synthesised sample.

23
Fig. 7 The SEM image of as-synthesized sample

24
Fig. 8 The SEM image of as-synthesized sample

25
 Fig. 9 The SEM image of the 1100ºC calcined sample

The SEM image of calcined sample at 1100ºC is shown in Fig. 9. This SEM image

indicates the morphology of 1100ºC calcined sample. Two different morphology

were observed in SEM image of calcined sample which indicate the presence of HA

and β-TCP. This result supports the XRD of calcined sample.

26
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