Journal of Biosciences and Medicines, 2014, 2, 30-35
Published Online April 2014 in SciRes. http://www.scirp.org/journal/jbm
http://dx.doi.org/10.4236/jbm.2014.22005
How to cite this paper: Copatti, C., Camargo, N.H.A. and Gemelli, E. (2014) Incorporation of Carboplatin in Microporous
Granular Calcium Phosphate Biphasic Matrix. Journal of Biosciences and Medicines, 2, 30-35.
http://dx.doi.org/10.4236/***.2014.*****
Incorporation of Carboplatin in Microporous
Granular Calcium Phosphate Biphasic
Matrix
Cleomar Copatti1, N. H. A. Camargo2, E. Gemelli2
1Chemistry Department, Santa Catarina University, Joinville, Brazil
2Mechanical Engineering Department, Santa Catarina University, Joinville, Brazil
Received December 2013
Abstract
The HA/β-TCP biphasic bioceramics stand out on researches in different areas of biomedical ap-
plications. These bioceramics with microporous microstructures also stand out in biomedical ap-
plications on controlled drug release. This study aimed at the synthesis of the biphasic HA/β-TCP
powder, and at the elaboration and characterization of the microporous biphasic HA/β-TCP gra-
nular biomaterial. The microporous granular material was elaborated through the process of ce-
ramic powder sieving (200 µm < d < 500 µm mesh sizes). The granular material was sintered at
1100˚C/2 h, providing the microporous biphasic granular biomaterial. The drug loading in the
biomaterial was performed through the high vacuum method. The results here presented are re-
lated to the synthesis method and elaboration of the biphasic biomaterial. The results obtained
from the drug loading through the high vacuum method conducted the incorporation of the drug
onto the surface and into the microporous granular biomaterial.
Keywords
Elaboration; Biphasic; Biomate ria l; Microporosity; Drug
1. Introduction
The calcium phosphate bioceramics are widely studied and stand out in researches as biomaterials for bone re-
construction and defect repairing. They have also shown to be promising as matrices for controlled localized
drug delivery [1]. This is due to their characteristics of non-toxicity and crystallographic compatibility and simi-
larity with the human skeleton apatite [2]-[5]. The calcium phosphate bioceramics are targeted by tissue and
material engineering, which is associated with the possibility of producing biomaterials with microporous archi-
tectures and physical, chemical and mechanical properties similar to the structure of bone tissue [6]. The cal-
cium phosphate biphasic bioceramics stand out in researches as medication storage matrices to be posteriorly
used as products for controlled drug release [7]-[9]. The first study is on loading of microporous bioceramics in
the 1980s with the introduction of ceramic capsules of aluminum-calcium-phosphorous oxide [10].
It was found in the bibliography that the calcium phosphate bioceramics have been used in the treatment of

C. Copatti et al.
31
bone metastasis, osteoporosis and osteomyelitis [11]-[14]. This is due to the fact that these bioceramics provide
an architecture capable of storing and protecting the medications inside their interconnected microporosities
[15]-[17]. A drug delivery system is a technology which allows the drug to be released in the target site, not only
to reduce the medications’ side effects, but also to maximize the drug action [18]-[20]. There are many drug de-
livery models: polymer-based nanocapsules [21], carbon nanotubes [22] [23]. These dosage vehicles or drug
transporters are subjected to different environmental conditions in the living organism. The major concern is
about the dosage and the permanence time before the drug reaches its destination and localized release [24]. In
general, oncological treatments with drugs are not selective; they reach the target cells, but can also destroy bone
tissues in perfect state during its administration, whether it is oral or associated with the bone graft.
This study aimed at the synthesis of a calcium phosphate powder and at the elaboration of a microporous bi-
phasic granular biomaterial for further incorporation and characterization of carboplatin in the microporosity and
surface of the 60%HA/40% β-TCP calcium phosphate biphasic biomaterial at a 70 mg concentration. The high
vacuum method was used for the deposition of 70 mg of drugs on the surface and in the interior of the micr o-
porous granular biomaterial. The characterization studies were carried out with the use of many different tech-
niques. The scanning electron microscopy technique was used for the characterization of the granular biomateri-
al’s morphology and microstructure. It also served as a support for the drug identification after its incorporation
in the biphasic granular biomaterial. The X-ray diffractometry served as a support for the identification of the
biphasic biomaterial’s phases before and after the loading with the drug. Finally, the infrared spectroscopy tech-
nique was used to identify the vibrational bands of the OH-,
3
4
PO
−
and Pt, NH3 groups.
2. Materials and Methods
The synthesis of the calcium phosphate powder was carried out with the wet chemical method, through the reac-
tion of dissolution/precipitation, which involves a CaO solid/liquid phase and a phosphoric acid solution. The
elaboration of the biphasic biomaterial was carried out with the support of a high energy NETZSCH attrition
mill with 2mm-diameter zirconium beads and distilled water. The process in the attrition mill was carried out
with a solid/liquid concentration of 50%/50% volume. This process was kept under stirring for one hour, as de-
scribed by [25] [26]. The colloidal suspension resulting from the attrition was dried in a rotary evaporator, pro-
viding the clustered HA/β-TCP biphasic nanostructured powders. These powders went through a process of
classification with sieves, going through a #500 mesh and having the accumulated granular material recuperated
by a #200 mesh sieve. The granular material obtained was subjected to sintering at 1100˚C/2 h, providing the
microporous biphasic granular biomaterial.
The 450 mg carboplatin was acquired from Tevacarbo. This drug was incorporated to the microporous bi-
phasic granular biomaterial at a concentration of 70 mg/1g of biphasic biomaterial. The high vacuum method
was used to incorporate the drug to the biomaterial. This is a physical process which inhibits the modification of
the chemical and physical characteristics of the medication and the biomaterial [11].
The studies of morphological and microstructural characterization were carried out with a Jeol JSM6701F
scanning electron microscope, through the secondary electron (SE) system. These studies were carried out on
the carboplatin powder and the microporous biphasic granular biomaterial. The X-ray diffractometry helped in
the identification of the current phases of the biphasic biomaterial and the biomaterial loaded with the drug. The
Fourier tr ans form infrared spectroscopy (FTIR) technique allowed the identification of the drug’s and the bi-
phasic biomaterial’s vibrational bands.
3. Results and Discussions
The results obtained from the morphological and microstructural characterization of the microporous biphasic
granular biomaterials revealed, in their micrographs, the morphology of granules with irregular shapes and sizes
between 200 µm < d < 500 µm, as it can be observed in Figure 1(a). Another observation was the intercon-
nected microporous microstructure of the granular biomaterial, as featured in Figure 1(b).
Figure 2(a) features the morphology of the carboplatin medication, where the morphology of thin rods with
rectangular shapes were found indicating the drug (mannitol) loading, as well as the morphology of small crys-
tals, scattered and agglomerated indicating the carboplatin’s morphology. Figure 1(b) illustrates the micro-
structure of the biphasic biomaterial without drug loading. Figures 2(b) and 2(c) feature the microstructure of
the biphasic biomaterial with 70 mg drug loading. It is also noted, in the micrographs, the presence of a new

C. Copatti et al.
32
(a) (b)
Figure 1. (a) Morphology of the HA/β-TCP biphasic granular biomaterial HA/TCP-β; (b) Micro-
structure of the HA/β-TCP biphasic granule.
(a) (b)
(c)
Figure 2. (a) Morphology of the drug; (b) Drug in the interface; (c) Drug in the interior of the bi-
phasic biomaterial’s microstructure.
morphology in the biphasic biomaterial’s grain interface and in the interior of the biomaterial’s microporosity
(Figures 2(c)), indicating that the incorporation of the drug was successful.
The results obtained from the X-ray diffractometry on the microporous biphasic granular biomaterial clearly
revealed, in their X-ray diffractograms, the presence of the hydroxyapatite and β-calcium phases (Figure 3). In
Figure 3, the presence of well-defined peaks of the stoichiometric hydroxyapatite phase in the composition
Ca10(PO4)6(OH)2 can be noticed, with main diffraction plane of [211]. Peaks of lower intensity for the β-calcium

C. Copatti et al.
33
phosphate (Whitlockite) phase, in the β-Ca3(PO4)2 composition, with Rhombohedral crystal system and main
diffraction plane of [021] were also found.
Figure 4 reveals the spectrogram obtained from the microporous biphasic granular biomaterial. In the spec-
trogram, vibrational bands of the OH-groups in 3,575 cm−1, 1,739 cm−1, 632 cm−1 and 1,080 cm−1, 1,018 cm−1,
594 cm−1, 567 cm−1 representing the PO4 vibrational groups.
Figure 5 shows the spectrogram obtained from the biphasic biomaterial after the drug loading. What can be
observed is the presence of typical hydroxyl and phosphate groups, representative of the biphasic biomaterial,
and the groups representative of the drug, NH3 with multiple peaks between 3,200 - 2,400 cm−1 and Pt between
585 and 380 cm−1. It was also found that there is an overlap in the vibrational bands of the PO4 and OH− groups
with the NH3, C = O, C, CH, CH2, Pt groups. This can be explained by the fact that the drug has functional
groups very close to those of the biphasic biomaterial.
4. Conclusion
The wet chemical synthesis method provided the calcium phosphate biphasic powder. The X-ray diffractometry
evidenced the HA and β-TCP phases in the microporous biphasic granular biomaterial. The high vacuum me-
thod proved to be efficient for the incorporation of the drug onto the grain surface and to the interior of the cal-
cium phosphate biphasic biomaterial’s microporosity. The images obtained with the scanning electron micro-
scopy revealed the presence of the drug on the biphasic biomaterial’s surface and in the interior of the bioma-
terial’s microporosity. The results obtained through the infrared spectroscopy on the microporous biphasic gra-
nular biomaterial with the drug presented an overlap in the vibrational bands, especially the functional groups
OH−, NH3, C = O, C, CH, CH2, Pt, which present themselves as similar between the biphasic biomaterial and the
drug.
15 20 25 30 35 40 45 50 55 60 65
0
200
400
600
800
1000
1200
1400
[021] [211]
º
** **
*
*
* Ca
10
(PO
4
)
6
(OH)
2
?β-Ca
3
(PO
4
)
2
* º**
*
*
*
* *
**º
º
*
*
º
* º
º
*
º
º
º
*
*
*
*
*
Intensity (CPS)
2θ
Figure 3. X-ray diffractogram obtained from the microporous
biphasic granular biomaterial.
4000 3500 3000 2500 2000 1500 1000500
0
20
40
60
80
100
º
º
º
º
*
*
* OH
-
?PO
4
567
594
632
1080
3575
1018
Transmittance (%)
cm
-1
Figure 4. Spectrogram obtained from HA/β-TCP granular bio-
materi al.
4000 3500 3000 2500 2000 1500 1000500
0
20
40
60
80
100
*
*
*
*
*
*
#
*
º
º º
?OH-
* PO4
+ NH3
# Pt
Transmitance (%)
cm
-1
Figure 5. Spectrogram obtained from HA/β-TCP granular bio-
material with drug.

C. Copatti et al.
34
The vacuum incorporation technique has shown to be promising for not presenting modification in the struc-
tures of the biphasic biomaterial as well as of the drug. Another relevant point of this method is associated to its
simplicity and low cost, being easily adapted to a production line at an industrial level.
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