Prof. dr.

Simona Cavalu
Faculty of Medicine and Pharmacy
University of Oradea
ROMANIA
Motivation
As the average age of population grows, the need for
medical devices to replace damaged or worn tissues
increases.
As patients have become more and more demanding
regarding esthetic and biocompatibility aspects of
their dental restorations .
The ideal ceramic is a high performance biocomposite that combines the
excellent material properties of alumina in terms of chemical stability and
low wear, and of zirconia with its superior mechanical strength and fracture
toughness.
Alumina/zirconia ceramics were successfully used in total hip/knee
arthroplasty in the last decades.
For dental application: root canal posts, orthodontic brackets,
implant abutments and all- ceramic restorations.
Bioceramic
interaction with
living tissue
Bioinert Bioactive
Surface modifications and post synthesis
treatments for better performances
Tough and strong ceramics like zirconia, alumina or alumina-zirconia
composites are not capable of creating a biologically adherent interface
layer with bone due to the chemically inert nature of these two stable
oxides .
Surface
covering
layers/coatings
Biological
response
Cells viability
Cells
attachment
Cells
proliferation
Surface modification: organic coating/
inorganic treatment
Organic: proteins, DNA, sugars.
Inorganic: surface blasting , acid etching ,
fluoride
Goal
In the present study we are focused on the possible
beneficial effect of organic coating (fibrinogen) and
inorganic treatment (fluorination with SnF2 and
NaBF4) with respect to new alumina/zirconia
bioceramics.
The main objective is to analyze the biocompatibility
of alumina/zirconia ceramics upon treatment via in
vitro and in vivo tests.
Materials
Composition: 80%Al
2
O
3
20%YSZ with 5%TiO
2
addition
Spark plasma sintering method at 1350-1400

C.
Structural characterization by FTIR and XRD spectroscopy
Morphological details of the surface investigated by SEM
Mechanical properties:
Fracture toughness 5.3 MPa m

(under a load of 19.6 N)
Vickers hardness 16.7 GPa (under a load of 9.8N).
O. Ormanci, S. Cavalu- Mater Sci Eng C 40 (2014)
FTIR spectroscopy
1200 1000 800 600 400
0
3
5
x
Al
2
O
3
80Al
2
O
3
20ZrO
2
xTiO
2
I
n
t
e
n
s
i
t
y
/

a
.
u
.
Wavenumbers (cm
-1
)
6
4
8
6
1
7
4
6
5
Modifications of stretching
vibration modes AlO
6
octaedra
XRD patterns
Al2O3
80Al2O3-20YSZ
80Al2O3-20YSZ +5TiO2
No monoclinic
phase ZrO
2
SEM
80Al
2
O
3
-20YSZ
80Al
2
O
3
-20YSZ with 5% TiO
2
Al
2
O
3
Texture of protein
(fibrinogen) coating on
alumina/zirconia ceramics-
electrodeposition
Native Fibr
Fibr/specimen 1
Fibr/specimen 2
Native Fibr Fibr /specimen 1
Fibr/specimen 2
F
T
I
R

s
p
e
c
t
r
o
s
c
o
p
y

a
n
d

d
e
c
o
n
v
o
l
u
t
i
o
n
helix % sheet% turns % Random % Side chain%
19.9 9.2
Surface treatment with SnF
2
and NaBF
4
- ATR FTIR evidence
Fig. 1 ATR FTIR spectra of SnF2 and
NaBF4 powders as received from the
supplier .
Fig. 2 ATR FTIR spectra recorded on
specimen surface before and after
treatment using SnF2 and NaBF4.
Al-O
Zr-O
Surface treatment- XPS evidence
1200 1000 800 600 400 200 0
F

1
s
A
l

2
s
Z
r

3
d
A
l

2
p
C

1
s
N

1
s
O

1
s
S
n

4
d


Z
r

4
p

F

2
s

S
n

3
p
1
S
n

3
d
Z
r

3
d
N

1
s
F

1
s
A
l

2
p
N
a

1
s
O

1
s
C

1
s


I
n
t
e
n
s
i
t
y

(
a
.
u
)
Binding Energy (eV)
S
n

3
p
3

A
l

2
s
O

A
u
g
e
r
Z
r

4
p
Specimen 2
SnF
2
NaBF
4
In vitro test: cells culture
Human fibroblast (HLF) seeded in a concentration of 2x10
4
/cm
2
cells on the
surface of each sample (SnF
2
respectively NaBF
4
treated ) and cultured for 3h,
7h and 24h.
Cell nuclei were stained with 5 mMDraq5 diluted 1:1000 in distilled water for 5
min at room temperature.
A B
C D
Visual inspection
demonstrating initial
adherence and proliferation of
fibroblasts.
3h 24 h
SnF2
NaBF4
Fibroblasts
adherence/proliferation
evidence by confocal
microscopy
SnF2
NaBF4
24 h 7 h
SnF2
NaBF4
7 h 24 h 3 h
SEM initial stage of adherence 3h
SnF2
NaBF4
7h
NaBF4
SnF2
24 h
SnF2
NaBF4
MTT assay results showing viable fibroblasts cells
with respect to control and surface treated
alumina/zirconia specimens after 3, 7 and 24 hours of
culture.
The label * indicates p<0.001 versus control, **indicates p<0.01
and *** indicates a p<0.001 with respect to specimen 1.
SnF2
NaBF4
In vivo tests: animal model (rabbit)
Implant 1- SnF2 treatment
Implant 2- NaBF4 treatment
Implant 3- Fibrinogen
50m
Implant
site
Haversian
canal
New bone
proliferation
Interface bone-implant
Haversian
canal
New bone
proliferation
Interface bone-implant
50m
Implant
site
Histology; implant 1 = SnF2 treatment
implant 2 = NaBF4 treatment
1
2
Ca/P= 1.62- 1.80
Haversian canal
Bone morphology after 4 and
8 weeks post -surgery
4 weeks
8 weeks
EDAX
XRD spectrum of the femoral bone
0 20 40 60 80 100
0
100
200
300
400
500
600
700
800
900
*
*
A
Z A
Z
A
Z
A
B
A
Z
A
A
A
Z
A
Z
A
Z
A
A
A
I

(
a
.
u
.
)
2 (deg)
AlZr Biocomposite
Bone/AlZr
Bone
A
T
Z
B
Histology: implant 3- bone
marrow cells interaction
Implant 3- fibrinogen coating
Goldners Trichrome stain
Histology: implant 3-
host bone interaction
Goldners Trichrome stain
Implant 3- fibrinogen coating
SEM/EDX bone-implant interface
Ca/P= 1.62
Ca/P= 1.77
4 weeks
8 weeks
Summary
Ceramic specimens with the composition
80%Al
2
O
3
- 20%3YSZ + 5% TiO
2
processed by SPS were surface treated with SnF2/
NaBF4 respectively fibrinogen by electrodeposition.
The surface modifications/texture were revealed by ATR-FTIR, XPS and SEM; it
was demonstrated that the SnF2 treatment is more effective than NaBF4.
Protein characteristics are preserved upon deposition procedure.
Fibroblasts cells culture in the presence of fluorine-treated specimens allowed
to assay cell adhesion, cell proliferation and colony capability by fluorescence
evaluation. Both inorganic treatments shows similar results, but cell
colonization capability seems to be promoted by the SnF2 treatment (cells
culture for fibrinogen coated is not shown, work in progress..)
Morphological details of the fibroblasts attached on the surface of fluorine
treated samples were emphasized by SEM showing the formation of a shell-like
coating after 24 hours incubation.
Histological images demonstrated the biocompatibility of the treated implants
as no gaps, fibrous tissue, multinucleated cells or inflamation were found at the
bone implant interface. A better bone to implant contact was noticed in the
case of SnF2 treatment.
Animal model- The presence of young, compact lamellar bone and
osteocytes near the implant surface indicated good biocompatibility, and
certainly the presence of the implant did not disturb the processes of bone
formation at the interface, for both organic/inorganic treatment.
Microstructure details (including Haversian canals) of bone and
bone marrow tissue and elemental composition at the interface
indicated Ca/P =1.62 - 1.77
Summary
Conclusions: Organic (proteic) film or fluoride as surface
conditioning might be an alternative approach to induce the
bioactivity and improve the biocompatibility of dense bioceramics
designed to load bearing bone replacement (hip joint, dental
abutments) and to optimize the biological response for specific
applications of biomedical implants.
Related papers:
O. Ormanci, I. Akin, F. Sahin, O. Yucel, V. Simon, Simona Cavalu, G. Goller, Spark Plasma
sintered A2O3-YSZ-TiO2 composites: Processing, characterization and in vivo evaluation,
Materials Science and Engineering C, 40 (2014) 16-23.
Simona Cavalu, C. Ratiu, O. Ponta, V. Simon, D. Rugina, V. Miclaus, I. Akin, G. Goller,
Improving osseointegration of alumina/zirconia ceramic implants by fluoride surface
treatment, Digest Journal of Nanomaterials and Biostructures Vol. 9, No. 2 (2014) 797 808.
Simona Cavalu, V. Simon, F. Banica, I. Akin, G. Goller, Surface modification of
alumina/zirconia bioceramics upon different fluoride-based treatments, Int. J. Appl. Ceram.
Technol., 11 [2] 402411 (2014).
Simona Cavalu, V. Simon, I. Akin, G. Goller, Adherence properties of acrylic bone cement
to alumina ceramics designed for clinical application, Acta Physica Polonica A, nr.2,vol.125
(2014) 603-605
S. Cavalu, V. Simon, C. Ratiu, I. Oswald, R. Gabor, O. Ponta, I. Akin, G. Goller, Correlation
between structural properties and in vivo biocompatibility of alumina/zirconia bioceramics,
Key Engineering Materials vols. 493-494, 1-6(2012)
Acknowledgments:
UEFISCDI project PNII-ID-PCE 2011-3-0441 contract nr. 237/2011 and
Bilateral Cooperation RO-TR.
Prof. dr. Viorica Simon Babes-Bolyai
University, Faculty of Physics & Institute of
Interdisciplinary Research in Bio-Nano-
Sciences, Cluj-Napoca, Romania.
Dr. Cristian Ratiu, Ioan Oswald and
Silviu Vlad, University of Oradea, Faculty
of Medicine and Pharmaceutics, Oradea,
Romania.
Dr. Dumitrita Rugina, USAMV Cluj-
Napoca.
Prof. dr. Gultekin Goller and assist. prof.
Ipek Akin, Istanbul Technical University,
Materials Science Department.