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JOURNAL OF OPTOELECTRONICS AND ADVANCED MATERIALS Vol. 9, No. 3, March 2007, p. 690 - 693
Bioactivity evaluation of new silver doped bone cement
for prosthetic surgery
S. CAVALU*
, V. SIMONa
, C. ALBONa
, C. HOZAN
University of Oradea, Faculty of Medicine and Pharmacy, P-ta 1 Decembrie10, Oradea 410068 Romania
a
Babes-Bolyai University, Faculty of Physics, Cluj-Napoca 400084, Romania
Due to the broad antimicrobial spectrum of silver, the composites used as bone substitutes or for coating metallic implants
can be doped with silver. Polymer-ceramic composites are widely applied in orthopaedics due to their biocompatibility and
ability to support bone growth (osteoconductivity) and also due to bioactivity implied by self-assembling of a hydroxyapatite
type layer on their surface. This in vitro study is focussed on surface microstructure changes in simulated body fluid and on
antibacterial silver release from two different types of biocomposites doped with silver. SEM microscopy and ATR-FTIR
spectroscopy were employed in order to evaluate the properties of the surface layer after immersion in simulated body fluid.
(Received November 15, 2006; accepted December 21, 2006)
Keywords: Silver, Antimicrobial, Simulated body fluid, SEM, ATR FTIR
1. Introduction
The prevention of infections disease represents
nowadays a central need, especially for prosthetic surgery.
Many nosocomial bacteria, in fact, show an increasing
resistance towards antibiotics, causing serious infections
that lead to prolonged times of hospitalization. The
development of surfaces with low bacterial adhesion
together with biocompatibility can represent a solution to
prevent infections. Biocompatible composites can be used
to realize bone substitutes as well as coated metallic
devices; moreover such materials can be opportunely
treated to enrich their surfaces with silver ions [1,6] thus
providing a controlled ion release and consequently
antibacterial efficacy. Silver is, in fact, a well-known
broad-spectrum antimicrobial agent, so silver-doped
composites can be produced to realize different devices,
such as bone substitutes or coated metallic implants, with
antibacterial action.
Porous materials have been used in surgical implant
design to fabricate devices or augment soft or hard tissues,
as coatings on prostheses to accommodate tissue ingrowth
for biological fixation and as scaffolds to facilitate the
regeneration of tissue. Polymer-ceramic composite are
widely used in orthopaedics as suture materials and
fixation devices due to their biocompatibility-ability to
support bony growth (osteoconductive) and also bone
bioactive (to form a calcium fosfat layer on its surface)
[2]. The aim of the work is to compare the microstructure,
biocompatibility and antimicrobial activity of two different
types of acrylic bone cements after silver doping, from in
vitro study in simulated body fluid using electrochemical
measurements, SEM microscopy and ATR-FTIR
spectroscopy in order to evaluate the properties of the
surface layer.
2. Experimental
Silver doping procedure was performed with respect
to different acrylic orthopedic cements, commercially
available, the final biocomposites having the following
composition: BIOLOS3®
-Liquid total 16.4 g:
methylmethacrylate (monomer) 84.4%, butylmethacrylate
13.2%, N:N dimethyl p- toluidine 2.4%, hidroquinone 20
ppm. Powder total 40 g: methylmethacrylate (copolymer)
87.3%, polymethyl metacrylate 2.7 %, barium sulphate
10%, AgNO3 20%. ANTIBIOTIC SIMPLEX®
- Liquide
total 20 ml: methyl methacrylate (monomer) 19.5 ml, N:N
dimethyl p- toluidine 0.5 ml, hidroquinone 1.5 mg. Powder
total 41 g: methyl methacrylate (copolymer) 30 g,
polymethyl methacrylate 6 g, barium sulphate 4 g,
erythromycin 0.5 g, colistin sulphomethate sodium 3
million I.U, AgNO3 8g. Samples from both materials were
incubated for 14 days in SBF following the procedure
describe by Kokubo et al [7]. Electrochemical
measurement were performed in static conditions, without
refreshing the fluid, using CONSORT C835 Multimeter
equipped with Na+
, Ca++
and Ag+
selective electrodes,
during different time intervals. The FT-IR spectra of
biocomposites were recorded in the region 4000-500 cm-1
by a Bruker EQUINOX 55 spectrometer OPUS software,
using an Attenuated Total Reflectance accessory with a
scanning speed of 32 cm-1
min-1
and spectral width
2.0 cm-1
. The internal reflection element was a ZnSe ATR
plate (50 × 20 × 2 mm) with an aperture angle of 45°.
Scanning Electron Microscope (SEM) images were taken
with a JEOL- JSM 5510 LV microscope.
3. Results and discussion
Electrochemical measurements carried out on pure
SBF (initial) and after 14 days incubation in SBF at
37 °C, of silver doped materials, reveal a considerable
diminution of Ca2+
concentration (two order of magnitude)
Bioactivity evaluation of new silver doped bone cement for prosthetic surgery 691
after 14 days incubation, for both types of biocomposites
(Fig. 1a,b). By comparing the diagrams a) and b) one can
observe that the silver release is facilitated by the BIOLOS
3®
, wich is an antibiotic free material, as the concentration
of Ag+
measured after five hours incubation is
significantly greater (one order of magnitude) then
released from ANTIBIOTIC SIMPLEX®
. Concomitant,
Na+
content decrease one order of magnitude, as shown in
diagram of Fig.1b. The SBF conductivity measured after
ANTIBIOTIC SIMPLEX®
incubation during this interval
increase continuously from 16.5 mSv to 28.3 mSv,
whereas the conductivity of SBF related to BIOLOS 3®
exhibit a minimum value of 12 mSv after 5 days and a
maximum of 15 mSv after 14 days incubation. The initial
value of the SBF conductivity is 17.5 mSv.
2 40
100
200
300
400
500
Ag
+
Ag
+
Ag
+
Ca
2+
Ca
2+
Na
+
Na
+
Na
+
Ca
2+
Ca
2+
Na
+
14 days5 days5 hoursinitial
concentration(M)
1
10
-1
10
- 2
10
- 3
10
- 4
a
2 40
100
200
300
400
500
Ag
+
Ag
+
Ag
+
Ca
2+
Ca
2+
Na
+
Na
+
Na
+
Ca
2+
Ca
2+
Na
+
14 days5 days5 hoursinitial
concentration(M)
1
10
-1
10
- 2
10
- 3
10
- 4
b
Fig. 1. Effect of SBF incubation on Ca2+
, Na+
and Ag+
concentration, after different incubation times of silver
doped ANTIBIOTIC SIMPLEX®
(a) and silver doped
BIOLOS®
(b).
The silver release mechanism can be described as
follow: In the presence of the sodium or calcium ions
characteristic of body fluids, the antimicrobial agents ion
exchange with these physiological cations. Long chain
hydrophilic polymers incorporated in the coating adsorb
water molecules and facilitate the ion exchange. Rather
than exhibiting a huge “spike” of activity that diminishes
rapidly over time - and that may be toxic to the
surrounding tissues - the new biomaterials prevents
bacterial adherence by its slow ion-exchange release of
anti-microbial agents.
Prior to the doping procedure, the study was focused
on scanning electron microscopic (SEM) analysis before
and after immersion in SBF, as the development of an
active layer is expected [2,5], followed by infrared
spectroscopic measurements on the graded layer structure
of hydroxyapatite type developed after different periods of
immersion in SBF and comparison with those of the native
materials. ATR-FTIR spectroscopy is a non invasive
method for monitoring the biomineralization at different
times intervals, and is an advantageous method as it is
very rapid, does not require sample preparation, is non-
distructive and in the same time it provides information
about the molecular structure of the polymers.
a)
b)
c)
d)
Fig. 2. Morphology of hydroxyapatite-like crystals at
BIOLOS 3®
surface before (a) and after 14 days
incubation (b) compared to ANTIBIOTIC SIMPLEX®
surface in the same conditions (c and d, respectively).
S. Cavalu, V. Simon, C. Albon, C. Hozan692
The SEM images show a nucleation process of
calcium containing crystals, the formation of this layer in
the early stages being considered indicative of the
bioactivity of the materials [8,9]. In Fig. 2 (a, b, c, d) is
displaied comparatively the surface morphology of both
biocomposites, before and after 14 days incubation in
SBF, in the same conditions. The SEM images show the
formation of hydroxyapatite-like crystals, cuasi-spherical
as well as irregularly shaped aggregates, with an average
size of 2.5 μm, grouped homogeneously, the distribution
of the aggregates being densely on the ANTIBIOTIC
SIMPLEX®
surface.
The surface layer structure has been investigated by
ATR- FTIR technique after 14 days incubation of both
materials and the corresponding spectra are displayed in
Figures 3 (a,b) along with the reference spectrum recorded
before the immersion in SBF. The reference spectrum
indicate the details of functional groups in methyl
methacrylate. The peaks from 2994 cm-1
to 2840 cm-1
are
due to CH2 asymmetric and symmetric stretching, a sharp
and intense peak at 1726 cm-1
is due to the presence of
ester carbonyl group stretching vibration, a broad band at
1438 cm-1
due to C-H bending and the peaks in the range
1260-900 cm-1
are assigned to O-C-O, C-CH3 stretching
and C-COO vibrations [10,11]. The features of this
spectrum are very well preserved after 14 days incubation
in SBF. The major modifications are observed in the range
3200-2800 cm-1
, 1600-1540 cm-1
and 1150-1035 cm-1
.
Typical calcium phosphates band is observed at 1035 cm-1
assigned to ν3 PO4
3-
streching, suggesting the presence of
newly formed bone, and according to the literature, the
sharpness of the phosphate band demonstrate increasing
mineralisation [12-15]. Other bands of interest in Fig. 3
are the intense band at 3180 cm-1
assigned to OH stretching
and those at 1630 and 1544 cm-1
corresponding to OH
deformation, demonstrating that after 14 days incubation,
the antibiotic-loaded bone cement absorbs a considerable
amount of water. We assume that the ions belonging to the
surface layer show a relatively high mobility. Thus, as
already reported, the cations as well as the anions can be
readily and reversibly exchanged. The hydrated layer is
not stable and it is progressively replaced by apatite during
ageing in an aqueous media (maturation). The mechanism
of this transformation is not yet well known but it involves
a decrease of the amount of HPO4
2-
ions and an increase of
the calcium content of the mineral phase. Simultaneously
OH-
ions are included in the structure and water is
excluded [14,15].
The calcium phosphate band arises at the same
wavenumber, 1035 cm-1
, in both cases, but the intensity of
this band indicates that mineralization at BIOLOS3®
surface after 14 days is diminished in comparison with
ANTIBIOTIC SIMPLEX®
. On the other hands, the
dinamics of water uptake in time is very different with
respect to the different types of methyl methacrylate
biocomposites.
4000 3500 3000 2500 2000 1500 1000 500
14 days
initial
2948
2360
1726
1438
1246
1144
997
997
1035
1144
1294
1438
1544
1630
1726
2954
3180
Absorbance(a.u.)
W avenum ber (cm
-1
)
a)
4000 3500 3000 2500 2000 1500 1000 500
977
1035
1064
14 days
initial
1144
1544
1630
1726
2954
3180
2948
2360
1726
1438
1246
1144
1064
977
Absorbance(a.u.)
W avenumber (cm
-1
)
b)
Fig. 3. ATR- FTIR spectra of ANTIBIOTIC SIMPLEX®
(a) and BIOLOS3®
(b) before and after 14 days
incubation in SBF.
4. Conclusions
Two different types of silver doped methyl
methacrylate based biocomposites are compared in vitro,
in simulated body fluid, with respect to their
microstructure and bioactivity. Electrochemical
measurements reveal a considerabe diminution of Ca2+
content in SBF during the first two weeks of incubation.
The antibiotic-free bone cement is capable to release more
Ag+
after five hours incubation, as compared to the
antibiotic loaded one. SEM images demonstrate that the
mineralization process after 14 days incubation is more
intense on the surface of antibiotic containing biomaterial.
This result is confirmed by ATR-FTIR data which
evidence calcium phosphate band characteristic for
hydroxyapatite-type compounds and at the same time by
the water uptake during the incubation time.
Bioactivity evaluation of new silver doped bone cement for prosthetic surgery 693
References
[1] E. Vernè, S. Di Nunzio, M. Bosetti, P.Appendino,
C. Vitale Brovarone, G. Maina, M. Cannas,
Biomaterials 26(25), 5111 (2005).
[2] L. L. Hench, J.Am.Ceram.Soc. 81(7),1705 (1998).
[3] A. E. Porter, N. Patel, J. N. Skeppre, S. M. Best,
W. Bonfield, Biomaterials 24, 4609(2003).
[4] S. Verrier, J. J. Blaker, V. Maquet, L. L. Hench,
A. R. Boccaccini, Biomaterials 25, 3013 (2004)
[5] S. Cavalu, V. Simon, J. Optoelectron. Adv. Mater.
8(4), 1520 (2006).
[6] M. Bellentone, H. D. Williams, L. L. Hench,
Antimicrobial Agents and Chemotherapy
46(6), 1940 (2002).
[7] T. Kokubo, S. Ito, Z. T. Huang, T.Hayashi, S. Sakka,
T. Kitsugi, T. Yamamuro, J. Biomed. Mater. Res.
24, 331(1990).
[8] J. Vandiver, N. Patel, W. Bonfield, C. Ortiz, Key
Engineering Materials 284, 497(2005).
[9] L. Pramatarova, E. Pecheva, R. Presker, M.T. Pham,
M. F. Maitz, M. Stutzmann, Eur. Cells Mater.
9, 9(2005).
[10] A. Balamurugan, S. Kannan, V. Selvaraj,
S. Rajeswari, Trends. Biomater. Artif. Organs
18(1) 41(2004).
[11] I. Notinger, J. J. Blaker, V. Maquet, L. L. Hench,
A. Boccaccini, Asian J. Physics 15(2) 221(2006).
[12] S. Kale, S. Biermann, C. Edwards, C. Tarnowski,
M. Morris, M. W. Long, Nature Biotech,
18, 954 (2000).
[13] A. Tinti, P. Tadei, R.Simoni, C. Fagnano,
Asian J. Physics 15(2), 267 (2006).
[14] D. Eichert, H. Sfihi, M. Banu, S. Cazalbou,
C. Combes, C. Rey, CIMTEC 2002 Proceedings of
10th Inter.Ceramics Congress, Florence, Italy( 2002).
[15] P. Sutandar, D. J. Ahn, E. I. Franses,
Macromolecules 27, 7316 (1994).
____________________________
*
Corresponding author: scavalu@rdslink.ro

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  • 1. JOURNAL OF OPTOELECTRONICS AND ADVANCED MATERIALS Vol. 9, No. 3, March 2007, p. 690 - 693 Bioactivity evaluation of new silver doped bone cement for prosthetic surgery S. CAVALU* , V. SIMONa , C. ALBONa , C. HOZAN University of Oradea, Faculty of Medicine and Pharmacy, P-ta 1 Decembrie10, Oradea 410068 Romania a Babes-Bolyai University, Faculty of Physics, Cluj-Napoca 400084, Romania Due to the broad antimicrobial spectrum of silver, the composites used as bone substitutes or for coating metallic implants can be doped with silver. Polymer-ceramic composites are widely applied in orthopaedics due to their biocompatibility and ability to support bone growth (osteoconductivity) and also due to bioactivity implied by self-assembling of a hydroxyapatite type layer on their surface. This in vitro study is focussed on surface microstructure changes in simulated body fluid and on antibacterial silver release from two different types of biocomposites doped with silver. SEM microscopy and ATR-FTIR spectroscopy were employed in order to evaluate the properties of the surface layer after immersion in simulated body fluid. (Received November 15, 2006; accepted December 21, 2006) Keywords: Silver, Antimicrobial, Simulated body fluid, SEM, ATR FTIR 1. Introduction The prevention of infections disease represents nowadays a central need, especially for prosthetic surgery. Many nosocomial bacteria, in fact, show an increasing resistance towards antibiotics, causing serious infections that lead to prolonged times of hospitalization. The development of surfaces with low bacterial adhesion together with biocompatibility can represent a solution to prevent infections. Biocompatible composites can be used to realize bone substitutes as well as coated metallic devices; moreover such materials can be opportunely treated to enrich their surfaces with silver ions [1,6] thus providing a controlled ion release and consequently antibacterial efficacy. Silver is, in fact, a well-known broad-spectrum antimicrobial agent, so silver-doped composites can be produced to realize different devices, such as bone substitutes or coated metallic implants, with antibacterial action. Porous materials have been used in surgical implant design to fabricate devices or augment soft or hard tissues, as coatings on prostheses to accommodate tissue ingrowth for biological fixation and as scaffolds to facilitate the regeneration of tissue. Polymer-ceramic composite are widely used in orthopaedics as suture materials and fixation devices due to their biocompatibility-ability to support bony growth (osteoconductive) and also bone bioactive (to form a calcium fosfat layer on its surface) [2]. The aim of the work is to compare the microstructure, biocompatibility and antimicrobial activity of two different types of acrylic bone cements after silver doping, from in vitro study in simulated body fluid using electrochemical measurements, SEM microscopy and ATR-FTIR spectroscopy in order to evaluate the properties of the surface layer. 2. Experimental Silver doping procedure was performed with respect to different acrylic orthopedic cements, commercially available, the final biocomposites having the following composition: BIOLOS3® -Liquid total 16.4 g: methylmethacrylate (monomer) 84.4%, butylmethacrylate 13.2%, N:N dimethyl p- toluidine 2.4%, hidroquinone 20 ppm. Powder total 40 g: methylmethacrylate (copolymer) 87.3%, polymethyl metacrylate 2.7 %, barium sulphate 10%, AgNO3 20%. ANTIBIOTIC SIMPLEX® - Liquide total 20 ml: methyl methacrylate (monomer) 19.5 ml, N:N dimethyl p- toluidine 0.5 ml, hidroquinone 1.5 mg. Powder total 41 g: methyl methacrylate (copolymer) 30 g, polymethyl methacrylate 6 g, barium sulphate 4 g, erythromycin 0.5 g, colistin sulphomethate sodium 3 million I.U, AgNO3 8g. Samples from both materials were incubated for 14 days in SBF following the procedure describe by Kokubo et al [7]. Electrochemical measurement were performed in static conditions, without refreshing the fluid, using CONSORT C835 Multimeter equipped with Na+ , Ca++ and Ag+ selective electrodes, during different time intervals. The FT-IR spectra of biocomposites were recorded in the region 4000-500 cm-1 by a Bruker EQUINOX 55 spectrometer OPUS software, using an Attenuated Total Reflectance accessory with a scanning speed of 32 cm-1 min-1 and spectral width 2.0 cm-1 . The internal reflection element was a ZnSe ATR plate (50 × 20 × 2 mm) with an aperture angle of 45°. Scanning Electron Microscope (SEM) images were taken with a JEOL- JSM 5510 LV microscope. 3. Results and discussion Electrochemical measurements carried out on pure SBF (initial) and after 14 days incubation in SBF at 37 °C, of silver doped materials, reveal a considerable diminution of Ca2+ concentration (two order of magnitude)
  • 2. Bioactivity evaluation of new silver doped bone cement for prosthetic surgery 691 after 14 days incubation, for both types of biocomposites (Fig. 1a,b). By comparing the diagrams a) and b) one can observe that the silver release is facilitated by the BIOLOS 3® , wich is an antibiotic free material, as the concentration of Ag+ measured after five hours incubation is significantly greater (one order of magnitude) then released from ANTIBIOTIC SIMPLEX® . Concomitant, Na+ content decrease one order of magnitude, as shown in diagram of Fig.1b. The SBF conductivity measured after ANTIBIOTIC SIMPLEX® incubation during this interval increase continuously from 16.5 mSv to 28.3 mSv, whereas the conductivity of SBF related to BIOLOS 3® exhibit a minimum value of 12 mSv after 5 days and a maximum of 15 mSv after 14 days incubation. The initial value of the SBF conductivity is 17.5 mSv. 2 40 100 200 300 400 500 Ag + Ag + Ag + Ca 2+ Ca 2+ Na + Na + Na + Ca 2+ Ca 2+ Na + 14 days5 days5 hoursinitial concentration(M) 1 10 -1 10 - 2 10 - 3 10 - 4 a 2 40 100 200 300 400 500 Ag + Ag + Ag + Ca 2+ Ca 2+ Na + Na + Na + Ca 2+ Ca 2+ Na + 14 days5 days5 hoursinitial concentration(M) 1 10 -1 10 - 2 10 - 3 10 - 4 b Fig. 1. Effect of SBF incubation on Ca2+ , Na+ and Ag+ concentration, after different incubation times of silver doped ANTIBIOTIC SIMPLEX® (a) and silver doped BIOLOS® (b). The silver release mechanism can be described as follow: In the presence of the sodium or calcium ions characteristic of body fluids, the antimicrobial agents ion exchange with these physiological cations. Long chain hydrophilic polymers incorporated in the coating adsorb water molecules and facilitate the ion exchange. Rather than exhibiting a huge “spike” of activity that diminishes rapidly over time - and that may be toxic to the surrounding tissues - the new biomaterials prevents bacterial adherence by its slow ion-exchange release of anti-microbial agents. Prior to the doping procedure, the study was focused on scanning electron microscopic (SEM) analysis before and after immersion in SBF, as the development of an active layer is expected [2,5], followed by infrared spectroscopic measurements on the graded layer structure of hydroxyapatite type developed after different periods of immersion in SBF and comparison with those of the native materials. ATR-FTIR spectroscopy is a non invasive method for monitoring the biomineralization at different times intervals, and is an advantageous method as it is very rapid, does not require sample preparation, is non- distructive and in the same time it provides information about the molecular structure of the polymers. a) b) c) d) Fig. 2. Morphology of hydroxyapatite-like crystals at BIOLOS 3® surface before (a) and after 14 days incubation (b) compared to ANTIBIOTIC SIMPLEX® surface in the same conditions (c and d, respectively).
  • 3. S. Cavalu, V. Simon, C. Albon, C. Hozan692 The SEM images show a nucleation process of calcium containing crystals, the formation of this layer in the early stages being considered indicative of the bioactivity of the materials [8,9]. In Fig. 2 (a, b, c, d) is displaied comparatively the surface morphology of both biocomposites, before and after 14 days incubation in SBF, in the same conditions. The SEM images show the formation of hydroxyapatite-like crystals, cuasi-spherical as well as irregularly shaped aggregates, with an average size of 2.5 μm, grouped homogeneously, the distribution of the aggregates being densely on the ANTIBIOTIC SIMPLEX® surface. The surface layer structure has been investigated by ATR- FTIR technique after 14 days incubation of both materials and the corresponding spectra are displayed in Figures 3 (a,b) along with the reference spectrum recorded before the immersion in SBF. The reference spectrum indicate the details of functional groups in methyl methacrylate. The peaks from 2994 cm-1 to 2840 cm-1 are due to CH2 asymmetric and symmetric stretching, a sharp and intense peak at 1726 cm-1 is due to the presence of ester carbonyl group stretching vibration, a broad band at 1438 cm-1 due to C-H bending and the peaks in the range 1260-900 cm-1 are assigned to O-C-O, C-CH3 stretching and C-COO vibrations [10,11]. The features of this spectrum are very well preserved after 14 days incubation in SBF. The major modifications are observed in the range 3200-2800 cm-1 , 1600-1540 cm-1 and 1150-1035 cm-1 . Typical calcium phosphates band is observed at 1035 cm-1 assigned to ν3 PO4 3- streching, suggesting the presence of newly formed bone, and according to the literature, the sharpness of the phosphate band demonstrate increasing mineralisation [12-15]. Other bands of interest in Fig. 3 are the intense band at 3180 cm-1 assigned to OH stretching and those at 1630 and 1544 cm-1 corresponding to OH deformation, demonstrating that after 14 days incubation, the antibiotic-loaded bone cement absorbs a considerable amount of water. We assume that the ions belonging to the surface layer show a relatively high mobility. Thus, as already reported, the cations as well as the anions can be readily and reversibly exchanged. The hydrated layer is not stable and it is progressively replaced by apatite during ageing in an aqueous media (maturation). The mechanism of this transformation is not yet well known but it involves a decrease of the amount of HPO4 2- ions and an increase of the calcium content of the mineral phase. Simultaneously OH- ions are included in the structure and water is excluded [14,15]. The calcium phosphate band arises at the same wavenumber, 1035 cm-1 , in both cases, but the intensity of this band indicates that mineralization at BIOLOS3® surface after 14 days is diminished in comparison with ANTIBIOTIC SIMPLEX® . On the other hands, the dinamics of water uptake in time is very different with respect to the different types of methyl methacrylate biocomposites. 4000 3500 3000 2500 2000 1500 1000 500 14 days initial 2948 2360 1726 1438 1246 1144 997 997 1035 1144 1294 1438 1544 1630 1726 2954 3180 Absorbance(a.u.) W avenum ber (cm -1 ) a) 4000 3500 3000 2500 2000 1500 1000 500 977 1035 1064 14 days initial 1144 1544 1630 1726 2954 3180 2948 2360 1726 1438 1246 1144 1064 977 Absorbance(a.u.) W avenumber (cm -1 ) b) Fig. 3. ATR- FTIR spectra of ANTIBIOTIC SIMPLEX® (a) and BIOLOS3® (b) before and after 14 days incubation in SBF. 4. Conclusions Two different types of silver doped methyl methacrylate based biocomposites are compared in vitro, in simulated body fluid, with respect to their microstructure and bioactivity. Electrochemical measurements reveal a considerabe diminution of Ca2+ content in SBF during the first two weeks of incubation. The antibiotic-free bone cement is capable to release more Ag+ after five hours incubation, as compared to the antibiotic loaded one. SEM images demonstrate that the mineralization process after 14 days incubation is more intense on the surface of antibiotic containing biomaterial. This result is confirmed by ATR-FTIR data which evidence calcium phosphate band characteristic for hydroxyapatite-type compounds and at the same time by the water uptake during the incubation time.
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