Biological and biomechanical evaluation of interface reaction at conical screw-type implants.

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ssBioMed CentHead & Face Medicine
Open AcceResearch
Biological and biomechanical evaluation of interface reaction at
conical screw-type implants
Andre Büchter*1, Ulrich Joos†1, Hans-Peter Wiesmann†1, László Seper†1 and
Ulrich Meyer†2
Address: 1Department of Cranio-Maxillofacial Surgery, University of Münster, Waldeyerstraße 30, D-48129 Münster, Germany and 2Department
for Cranio- and Maxillofacial Surgery, Heinrich-Heine-University, Moorenstr, 5, D-40225 Dusseldorf, Germany
Email: Andre Büchter* - buchtea@uni-muenster.de; Ulrich Joos - joos@uni-muenster.de; Hans-Peter Wiesmann - wiesmap@uni-muenster.de;
László Seper - seper@uni-muenster.de; Ulrich Meyer - Meyer@med.uni-duesseldorf.de
* Corresponding author †Equal contributors
Abstract
Background: Initial stability of the implant is, in effect, one of the fundamental criteria for
obtaining long-term osseointegration. Achieving implant stability depends on the implant-bone
relation, the surgical technique and on the microscopic and macroscopic morphology of the implant
used. A newly designed parabolic screw-type dental implant system was tested in vivo for early
stages of interface reaction at the implant surface.
Methods: A total of 40 implants were placed into the cranial and caudal part of the tibia in eight
male Göttinger minipigs. Resonance frequency measurements (RFM) were made on each implant
at the time of fixture placement, 7 days and 28 days thereafter in all animals. Block biopsies were
harvested 7 and 28 days (four animals each) following surgery. Biomechanical testing, removable
torque tests (RTV), resonance frequency analysis; histological and histomorphometric analysis as
well as ultrastructural investigations (scanning electron microscopy (SEM)) were performed.
Results: Implant stability in respect to the measured RTV and RFM-levels were found to be high
after 7 days of implants osseointegration and remained at this level during the experimented
course. Additionally, RFM level demonstrated no alteration towards baseline levels during the
osseointegration. No significant increase or decrease in the mean RFM (6029 Hz; 6256 Hz and 5885
Hz after 0-, 7- and 28 days) were observed. The removal torque values show after 7 and 28 days
no significant difference. SEM analysis demonstrated a direct bone to implant contact over the
whole implant surface. The bone-to-implant contact ratio increased from 35.8 ± 7.2% to 46.3 ±
17.7% over time (p = 0,146).
Conclusion: The results of this study indicate primary stability of implants which osseointegrated
with an intimate bone contact over the whole length of the implant.
Introduction with a sufficient amount and quality of bone has been
Published: 21 February 2006
Head & Face Medicine2006, 2:5 doi:10.1186/1746-160X-2-5
Received: 20 November 2005
Accepted: 21 February 2006
This article is available from: http://www.head-face-med.com/content/2/1/5
© 2006Büchter et al; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Page 1 of 9
(page number not for citation purposes)
The long-term success of osseointegrated implants in the
treatment of completely and partially edentulous patients
well documented in the literature [1-14]. Initial stability
of the implant is, in effect, one of the fundamental criteria

Head & Face Medicine 2006, 2:5 http://www.head-face-med.com/content/2/1/5
for obtaining long-term osseointegration. [4,6]. Achieving
implant stability depends on the implant-bone relation,
the surgical technique and on the microscopic and macro-
scopic morphology of the implant used.
The osseointegration mode of implants is influenced by
the features of the implant system. Important aspects of a
fast implant osseointegration include the need to achieve
a primary congruence between the implant and the bone
directly after insertion, the need to insert the implant with
minimal surgical trauma and the capability of the implant
surface to attach directly to the adjacent bone tissue. It has
generally been thought in implant dentistry that
osseointegration requires a healing period of at least 3
months in the mandible and 5 to 6 months in the maxilla
[1-3,14,15]. The rationale for choosing a delayed loading
tion[4,6]. Nevertheless, several protocols for immediate
and early loading have been presented and were found
successful over the last two decades. According to Szmuk-
ler-Moncler et al. (2000) [16] two effective approaches
can be used to reduce time between surgery and prosthetic
reconstruction. One is to reduce micro-motion beneath
the critical threshold by means of rigid fixation of loaded
implants. The other possibility is to optimize the healing
period before a safe functional loading can be exerted.
The importance of the implant geometries and surface
characteristics, in an effort to achieve better bone anchor-
age, has been clear for a long time and, [4,17] in fact, var-
ious implant systems have been introduced over the past
several years in order to achieve a faster bone integration
[18]. In order to fasten the osseointegration process a new
parabolic screw-type implant system was developed. The
gross morphology of the implants was designed with the
help of finite element analysis (FEA). The geometry of the
implant was designed to allow micromovements of a
magnitude between 500 and 3,000 µstrain in the loaded
bone layer adjacent to the implant and to achieve a close
Finite element model of strain distribution under vertical loadigur 2
Finite element model of strain distribution under vertical
load. The model corresponds to the implant and bone anat-
omy at the implant site.
SEM of the implant used in this study (length 10 mm, shoul-der diameter 4.1 mm)Figure 1
SEM of the implant used in this study (length 10 mm, shoul-
der diameter 4.1 mm). Microgrooves were located at the
shoulder and tip of the implant.Page 2 of 9
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period was that premature loading resulted in fibrous tis-
sue encapsulation rather than direct bone apposi-
congruency between the surgically created implantation
bed and the implant surface direct after insertion [19-23].

Head & Face Medicine 2006, 2:5 http://www.head-face-med.com/content/2/1/5
We analysed in a combined approach the histological and
biomechanical outcome of a new implant system. Biolog-
ical investigations (histology, histomorphometry and
scanning electron microscopy (SEM)) as well as biome-
chanical tests (resonance frequency measurements (RFM)
and removal torque tests) were performed at early phases
of implant/bone interaction in order to evaluate the time
course of implant osseointegration.
Materials and methods
Implant System
The implants used in this study were newly developed
parabolic screw-type implants (ILI) with a length of 10
mm and a diameter of 4.1 mm at the shoulder of the
implant (Fig 1). The implants were made of pure titanium
with a characteristic progressive thread design. The
threads as well as the curvature of the implant provided a
homogeneous strain distribution over the whole implant
surface under vertical loading conditions (Fig 2), as
revealed by finite element analysis [20]. The implants pos-
sess a microstructured texture of 20 – 30 µm deep grooves,
where as the titanium surface it is smooth on a nanoscale
level. The implant system consists of two parabolic burrs
of different diameters and morphologies. The burrs are
used subsequently to prepare the bony implantation bed.
The diameter of the second burr is slightly smaller then
the core diameter of the implant. Implants have a trans-
versal core/thread relation of 1:1.2. Implant insertion is
performed by manual tapping of the self cutting implants
into the surgically created bony implantation bed.
Experimental animals
Eight male Göttinger minipigs, 14 to 16 months of age
nial and caudal part of the tibia condoyle (Fig 3). This
study was approved by the Animal Ethics Committee of
the University of Münster under the reference number G
38/2003.
Surgical procedure
All surgery was performed under sterile conditions in a
veterinary operating theatre. The animals were sedated
with an intramuscular injection of ketamine (10 mg/kg),
atropine (0.06 ml/kg) and stresnil (0.03 ml/kg). In the
areas exposed to surgery 4 ml of local anaesthesia (2%
lidocaine with 12.5 µg/ml epinephrine, Xylocain/ Adren-
alin®, Astra, Wedel, Germany) was injected. The tibias
were exposed by skin incisions and via fascial-periosteal
flaps. Thereafter, the implants were placed in the cranial
and caudal part of the tibia. The implant sites were
sequentially enlarged with both drills according to the
standard protocol of the manufacturer. Implants with 10
mm in length and 4,1 mm in diameter were inserted by
using continuous external sterile saline irrigation to mini-
mize bone damage caused by overheating. At the surgical
site, the skin and the fascia-periosteum were closed in sep-
arate layers with single resorbable sutures (Vicryl®4-0,
Ethicon, Norderstedt, Germany). Perioperatively, an anti-
biotic was administered subcutaneously (benzylpenicil-
lin/dihydrostreptomycin, Tardomycel®, BayerVital,
Leverkusen, Germany), 2.5 ml every 48 h for 7days. After
placement, the shoulder of each implant was 1 mm below
the ridge crest to allow circumferential bone growth. Res-
onace frequency measurements (RFM) (Osstell, Integra-
tion Diagnostics, Gothenburg, Sweden) were made for
each implant at the time of fixture placement and after
euthanasia [24-27]. The animals were inspected after the
first few postoperative days for signs of wound dehiscence
or infection and weekly thereafter to assess general health.
Healing periods of 7 days and 28 days were allowed for
half of the implants respectively. After 7 days and 28 days
animals were sacrificed (4 minipigs each) with an over-
dose of T61 given intravenously. Following euthanasia,
tibia block specimens containing the implants and sur-
rounding tissues were dissected from all of the animals.
The block samples were sectioned by a saw to remove
unnecessary portions of bone and soft tissue and were
prepared for the various investigations:
Removal torque testing
The removal torque test was performed by applying a
counter-clockwise rotation to the implant, around its axis
at a rate of 0,1°/s according to the experimental set up of
Li et al. 2002 [28]. For each implant the torque rotation
curve was recorded. The removal torque was defined as
the maximum torque (Nmm) on the curve. The interfacial
stiffness was defined as the slope (Nm/degree of the
Scheme of implant placement (surgical procedure)Figure 3
Scheme of implant placement (surgical procedure).Page 3 of 9
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and with an average body weight of 35 kg were used in
this study. A total of 40 implants were placed into the cra-
torque-rotation curve) calculated from a linear regression
analysis of the data between 0,5° and 3°.

Head & Face Medicine 2006, 2:5 http://www.head-face-med.com/content/2/1/5
Resonance frequency measurements (RFM)
This method, as a non-destructive technique, evaluates
the implant stability in term of interfacial stiffness. Reso-
nance frequency measurements were made on each
implant at the time of fixture placement and after the time
of sacrifice (7 and 28 days) in all animals by attaching a 4-
mm long standard transducer (Osstell, Integration Diag-
nostics, Gothenburg, Sweden) to the implant. The excita-
tion sign was given over a range of frequencies (typically
5 kHz to 15 kHz with a peak amplitude of 1 V) and the
first flexural resonance was measured [24-27]. The fre-
quency responses of the system were measured for each
implant.
Scanning electron microscopy (SEM)
Block samples containing the implants were first divided
into 2 halves, and then each sample was further dissected
with a blade to obtain a sample containing the implant
embedded in the alveolar bone and the corresponding
bone sample detached from the implant (28 days after
implant placement). Samples containing the implant
were used for scanning electron microscopy (SEM). For
SEM, glutaraldehyde-fixed specimens were critical point-
dried. Samples were sputtercoated with gold for histolog-
ical analysis. Specimens were examined under a fieldemis-
sion scanning electron microscopy (LEO 1530 VP,
Oberkochen, Germany).
Histomorphometry
The implants were removed together with the surround-
ing bone and fixed in Schaffer's solution (ethanol (96%),
formaldehyde (37%), ratio: 2:1). The specimens were
dehydrated in a graded series of ethanol. Thereafter, sam-
ples were embedded in methylmetacrylate (Techno-
vit®7200, Heraeus Kulzer, Dormagen, Germany).
Utilizing the 'sawing and grinding' technique, longitudi-
nal sections were grounded to about 43–50µm for con-
ventional microscopy (Exakt Apparatebau, Norderstedt,
Germany). Five samples stained by Alizarin S 1% and Bril-
liant-Kresyl-blue 0,1% were prepared for each implant
site. Histology was analysed by light microscopy (Zeiss,
Axioplan 2, Göttingen, Germany).
Filters of wavelengths of 510–560 nm (green filter), 450–
490 nm (blue filter), 355–425 nm (violet filter) and 340–
380 nm (UV filter) (Zeiss, Göttingen, Germany) were uti-
lized. The bone-to-implant contact ratio was defined as
the length of bone surface border in direct contact with
the implant (× 100 (%)). NIH-Imge software was used for
image processing and analysis (National Institutes of
Health, Bethesda, MD, USA)
Statistical analysis
Mean values and standard deviations (SD) were calcu-
lated for RFM, removal torque testing, interfacial stiffness
and bone-to-implant contact ratio. Multiple comparisons
between all groups were performed using two-way analy-
sis of variance and the t-test. Difference was considered
significant when p < 0,05. All calculations were performed
through the use of SPSS for Windows (SPSS Inc., Chicago,
IL, USA).
Results
Clinical observation
All implants were anchored monocortically. At placement
and during healing, the implants remained clinically
immobile. The animals recovered well after surgery and
no signs of infection were noted at any time during the
observation period.
Removal torque testing
Over the healing periods tested, the mean maximum
torque of implants was 390.00 ± 148.32 Nmm after 7 days
and 300.00 ± 69.22 Nmm after 28 days (Table 1). Statisti-
cally significant differences in the removal torque values
were not observed between day 7 and day 28 (p = 0.351).
The implant stiffness (Table 2), as assessed by the linear
regression analysis, was higher after 7 days (0.3992 ±
0.063) than after 28 days (0.2648 ± 0.0257), but the dif-
ference was statistically not significant (p = 0.086).
Table 2: Implant-bone interfacial stiffness values (Nmm/degree) 7 and 28 days
Treatment group 7 days 28 days Change within group Change 7 days to 28
days
Table 1: Removal torque values (Nmm) at two different healing periods
Treatment group 7 days 28 days Change within group Change 7 days to 28
days
ILI 390.00 ± 148.32 300.00 ± 69.22 90.00 ± 91.97 b p = 0.351Page 4 of 9
(page number not for citation purposes)
ILI 0.3992 ± 0.063 0.2648 ± 0.02257 0.1477 ± 0.039 p = 0.086

Head & Face Medicine 2006, 2:5 http://www.head-face-med.com/content/2/1/5
Resonance frequency measurements
The RFM demonstrated no significant change in the reso-
nance frequency responses during the 28 days the experi-
mental period. Implants had a high primary stability as
revealed by RFM (6029 ± 458 Hz and 6057 ± 423) directly
after insertion. The implant stability remained at this
baseline level through the experimental course (6257 ±
229 Hz at day 7 and 5885 ± 367 Hz at day 28) (Table 3
and 4).
SEM
Dissection of the implant-containing bone by a blade
confirmed the clinical finding that the implants were well
osseointegrated after 28 days. There was intimate bone
contact over the whole length of the implant (Fig 4). Typ-
ically, endosteal bone covered the implant surface. Colla-
gen fibres and osteoblasts made up the bulk of the
adjacent tissue layer. The collagen fibres appeared to be
predominantly oriented perpendicular to the implant sur-
face in the bulk bony tissue (Fig 5). Cells, extracellular
matrix proteins, and mineralized bone tissue were in
direct contact with the implant. In contrast to the collagen
fibres in the original bone, which were oriented perpen-
dicular to the implant, newly synthesized collagen in the
vicinity of the surface appeared to form a felt-like matrix
parallel to the surface. Intimate bone contact was pre-
sented at the neck of implants. Typically, cells (osteob-
lasts) were firmly attached to the implant surface. Probe
processing by sample fracturing for the electron micro-
scopic investigations suggested that the bond between the
implant and the adjacent bone layer seemed to mimic the
bond in the bone tissue itself. On the implant surface,
cells and extracellular matrix remained attached following
separation from the enveloping bone.
Histological and histomorphometric measurements
Direct bone-to-implant contact could be achieved during
the healing period. There were no signs of inflammation.
Histological analysis of the bone/implant interface
revealed an intimate contact between the titanium surface
and the bony implantation bed. At the bone-titanium
interface, a thin tissue layer stained with Alizarin S and
Brilliant-Kresyl-blue was seen in some areas coming into
direct contact with the titanium. The bony apposition
revealed a laminar structure containing individual osteo-
cytes and Haversian canals at the neck of implants (Fig 6).
The laminar bone demonstrated also an intimate contact
between spongiosal trabecula and the implant surface at
the body of implants (Fig 7 and 8). Quantitative histo-
morphometric analysis revealed an enhanced bone-to-
implant contact for every healing period. After 7 days the
bone-to-implant contact ratio was 35.8 ± 7.2% and after
28 days the bone-to-implant contact ratio was 46.3 ±
17.7%, but the difference in the bone to implant contact
did not reach a level of significance (table 5). A direct
bone – implant contact was documented especially at the
cortical bone area (neck of implants) after 7 and 28 days
(Fig 6).
Discussion
Insights into cellular processes occurring at the implant/
bone interface have contributed much to an understand-
ing of osseointegration. The understanding of the com-
plex bone/implant interactions at different levels will
provide an opportunity to evaluate and produce implants
with specific and desired biologic responses [29-31]. The
implant system used in this study was designed to allow
implants to have a high primary stability and a direct
bone/implant contact over the whole implant surface
directly after insertion. Various studies emphasise that the
mode of implant osseointegration and stability is depend-
ent to a large extent on the gross and ultrastructural
implant design [4-7]. However, the role of implant geom-
etry and surface structuring in affecting early tissue heal-
ing and implant stability cannot be determined only from
histological or biomechanical observations. The dynam-
ics of bone physiology can also not be evaluated several
weeks post-implantation after long term bone remodel-
Table 4: Resonance frequencies measurement (kHz) for 28 days
Treatment group 0 day 28 days Change within group Change 0 days to 28
days
Table 3: Resonance frequencies measurement (Hz) for 7 days
Treatment group 0 day 7 days Change within group Change 0 day to 7 days
to 7
ILI 6029 ± 485 6257 ± 229 229 ± 203 p = 0.291Page 5 of 9
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ILI 6057 ± 423 5885 ± 367 171 ± 211 p = 0.435