Copyright r Blackwell Munksgaard 2003

J Clin Periodontol 2003; 30: 809818

Printed in Denmark. All rights reserved

Dynamics of bone tissue

formation in tooth extraction sites
An experimental study in dogs

G. Cardaropoli1, M. Araujo1,2
and J. Lindhe1
1
Department of Periodontology, Faculty of
Odontology, The Sahlgrenska Academy at
Goteborg University, Goteborg, Sweden;
2
Department of Dentistry, Maringa State
University, Brazil

Cardaropoli G, Araujo M, Lindhe J: Dynamics of bone tissue formation in tooth

extraction sites. An experimental study in dogs. J Clin Periodontol 2003; 30, 809818.
r Blackwell Munksgaard, 2003.
Abstract
Objectives: The aim of the present experiment was to study events involved in the
healing of marginal, central and apical compartments of an extraction socket, from the
formation of a blood clot, to bone tissue formation and remodeling of the newly
formed hard tissue.
Material and Methods: Nine mongrel dogs were used for the experiment. The fourth
mandibular premolars were selected for study and were divided into one mesial and
one distal portion. The distal root was removed and the socket with surrounding soft
and mineralized tissue was denoted experimental unit. The dogs were killed 1, 3, 7,
14, 30, 60, 90, 120 and 180 days after the root extractions. Biopsies including the
experimental units were demineralized in EDTA, dehydrated in ethanol and embedded
in paraffin. Serial sections 7 mm thick were cut in a mesio-distal plane. From each
biopsy, three sections representing the central part of the socket were selected for
histological examination. Morphometric measurements were performed to determine
the volume occupied by different types of tissues in the marginal, central and apical
compartments of the extraction socket at different intervals.
Results: During the first 3 days of healing, a blood clot was found to occupy most of
the extraction site. After seven days this clot was in part replaced with a provisional
matrix (PCT). On day 14, the tissue of the socket was comprised of PM and woven
bone. On day 30, mineralized bone occupied 88% of the socket volume. This tissue
had decreased to 15% on day 180. The portion occupied by bone marrow(BM) in the
day 60 specimens was about 75%, but had increased to 85% on day 180.
Conclusion: The healing of an extraction socket involved a series of events including
the formation of a coagulum that was replaced by (i) a provisional connective tissue
matrix, (ii) woven bone, and (iii) lamellar bone and BM. During the healing process a
hard tissue bridge cortical bone formed, which closed the socket.

The healing of an extraction socket

following tooth removal was studied in
different animal models (e.g. Schram
1929, Claflin 1936, Simpson 1960,
Kuboki et al. 1988, Lin et al. 1994).
The experiments demonstrated that during the process of healing a series of
events occurred, such as (i) formation
and maturation of a blood clot, (ii)
infiltration of fibroblast to replace the
coagulum, and eventually (iii) establishment of a provisional matrix (PCT) that
allowed for bone tissue formation.
Unfortunately, most of the studies
referred to were of comparatively short
duration and, thus, included limited

information related to later phases of

socket healing including the process of
remodeling of the newly formed bone
tissue in various parts of the alveolus.
The formation of soft and hard tissue
following tooth extraction was also
studied in specimens obtained from
humans (e.g. Mangos 1941, Christopher
1942, Amler 1969). The frequently cited
study by Amler (1969) The time
sequence of tissue regeneration in human extraction wounds reported on
new tissue formation in fresh extraction
sockets in human volunteers. In this
study, following the removal of a tooth,
socket healing was monitored and soft

Key words: bone healing; extraction sockets;

tooth extraction; socket healing; bone
formation
Accepted for publication 26 November 2002

tissue biopsies were harvested from the

extraction sites after varying intervals;
from 48 h to 32 days. Since the tissue
sampled was not demineralized prior to
sectioning, only events that preceded
hard tissue formation could have been
analyzed. From his observations, Amler
(1969) concluded that a blood clot
formed within the socket soon after the
removal of a tooth. This clot was
replaced first by a granulation tissue
(GT) and subsequently by an osteoid.
The illustrations, published by Amler
(1969), however, described only tissues
from the marginal portions of the
socket. It must be anticipated, therefore,

810

Cardaropoli et al.

that the biopsy was restricted to the

superficial region of the wound. Hence,
events that preceded or were involved in
hard tissue formation in the central and
apical compartments of the extraction
site were not included in the investigation.
The aim of the present experiment
was to study the time sequence and
various biological events that are involved in the healing of all compartments (marginal, central, apical) of an
extraction socket, from the formation of
a blood clot, to bone tissue formation
and remodeling of the newly formed
hard tissue.

Material and Methods

The research protocol was approved by

the Regional Ethics Committee for
Animal Research, Maringa State University, Brazil. Nine mongrel dogs,
about 12 months old and weighing
about 10 kg each, were used for the
experiment. The animals were fed a soft
pellet diet throughout the period of
observation. During surgery, the animals were anesthetized with intravenously s administered
Pentothal
Natrium (30 mg/ml; Abbot Laboratories, Chicago, IL, USA). The fourth
mandibular premolars (4P4; experimental teeth) were selected for study. The
pulp of the experimental teeth was
extirpated and the canal of the mesial
root filled with gutta-percha. Subsequently, with the use of a fissure bur,
the fourth premolars were divided in the
furcation fornix into one mesial and one
distal portion. The distal portion was
carefully extracted while the mesial
portion was retained. The buccal and
lingual soft tissues of the extraction site
were stabilized with interrupted sutures
(Fig. 1). The distal socket with surrounding soft and mineralized tissue
was denoted experimental unit.
The dogs were placed in a careful
plaque control program, which called
for tooth cleaning with toothbrush and
dentifrice three times a week. The root
extractions were planned in such a way
that biopsies of two experimental
units could be obtained from each of
the following intervals of healing: after
1, 3, 7, 14, 30, 60, 90, 120 and 180 days.
The dogs were killed with
an overs
dose of Pentothal Natrium and perfused with a fixative through the carotid
arteries. The fixative consisted of a
mixture of glutaraldehyde (5%) and

Fig. 1. Clinical view of the experimental site immediately after extraction of the distal root
of P4 and the placement of a suture.

formaldehyde (4%), buffered to pH 7.2

(Karnovsky 1965). The mandibles were
removed and placed in the fixative. The
experimental units, together with the
mesial roots, were dissected
using a
s
diamond saw (Exact Apparatebeau,
Norderstedt, Hamburg, Germany).
The biopsies were demineralized in
EDTA, dehydrated in increasing concentrations of ethanol and embedded in
paraffin. Serial sections were cut in the
mesio-distal plane and parallel to the
long axis of the mesial root. The
microtome was set at 7 mm. From each
biopsy, three sections representing the
central part of the socket, about 20 mm
apart, were selected for histological
examination. The sections were stained
in hematoxylin and eosin or the Van
Gieson connective tissue stain.

Histological examination

The tissue within and immediately

adjacent to the extraction socket was
examined
(obj  10, 40, 100) in a
s
Leitz DM-RBE microscope (Leica,
Wetzlar, Germany) equippeds with an
image system (Q-500 MC ; Leica,
Wetzlar, Germany).
The overall characteristics of the
healing socket were examined and
described. Morphometric measurements were performed by a blinded
(with respect to time), trained and
calibrated examiner (CG), according
to a modification of the method described by Schroeder & MunzelPedrazzoli (1973). A lattice comprising

100 light points was superimposed over

the experimental unit; the relative
volumes occupied by blood clot (mainly
blood cells, network of fibrin), GT
(highly vascularized tissue with an
inflammatory cell infiltrate), provisional
matrix (PCT) (connective tissue including a multitude of mesenchymal cells
embedded in a fibrous matrix), mineralized bone (MB) (woven bone, parallel
fibered bone and lamellar bone) and
bone marrow (BM) (intraosseous space
mainly occupied by adipocytes) in the
extraction socket were determined in
three different zones, A, B C.
Zone A: the marginal border of this
zone was located 1 mm apical of a line
connecting the mesial and distal borders
of the extraction socket.
Zone B: this zone was located in the
mid-portion of the socket.
Zone C: the apical border of this zone
was located 1 mm coronal of the apical
extension of the socket.
Each of the zones selected was 1 mm
high (apico-coronal direction) and covered the mesio-distal width of the
socket. The area occupied by the
periodontal ligament (PDL) was excluded from the morphometric measurements.

Data analysis

Mean values and standard deviations

were calculated for each experimental
unit, variable and interval of healing
using descriptive statistics. Due to the
aim of the study and the limited number

Bone healing dynamics

811

of animals, statistical analysis between

the units was considered unnecessary.

Results

All extraction sites healed uneventfully.

Gross histological observations

In specimens representing 1 day of

healing, a coagulum was found to reside
in most of the space previously occupied by the distal root of the fourth
premolar (Fig. 2a). The marginal portion of the coagulum was covered with a
layer of inflammatory cells, mainly
neutrophilic granulocytes (Fig. 3). Also
the gingival connective tissue adjacent
to the extraction site harbored inflammatory cells. The clot was comprised
mainly of erythrocytes and platelets that
were trapped in a network of fibrin (Fig.
4). Isolated neutrophils were seen to be
present in the central and apical compartments of the blood clot. Immediately lateral to the hard tissue wall, i.e.
the bundle bone of the socket, the
severed PDL was found to contain large
numbers of mesenchymal cells, fibers
and a multitude of large, apparently
dilated, vascular units. The principal
fibers invested as Sharpeys fibers in the
bundle bone and were, in the central
direction, found to be in direct contact
with the coagulum in the socket (Fig. 5).
In the marginal portion of the experimental unit representing day 3 of
healing (Fig. 2b), small segments of the
coagulum had been replaced by a richly
vascularized GT. In the center of the
coagulum within zones A, B and C,
areas could be identified in which
erythrocytes had undergone lysis (coagulative necrosis; Fig. 6). The membranes of the blood cells in these
compartments had lost their integrity
and the area had a hyaline appearance.
The severed PDL contained a large
number of fibroblasts and vessels. The
principal fibers (i) ran a course perpendicular to the surface of the hard tissue
wall, (ii) were invested in the bundle
bone, and (iii) made contact with the
coagulum (Fig. 7).
After 7 days of healing (Fig. 2c), the
wound in the experimental unit had
undergone a marked change in comparison to the day 3 specimen.
The number of principal fibers (PF)
of PDL that invested in the bundle bone
was comparatively small (Fig. 8), but
the PFs appeared elongated and were

Fig. 2. Mesio-distal sections illustrating the extraction socket after different intervals of
healing: (a) 1 day, (b) 3 day, (c) 7 day, (d) 14 day, (e) 30 day, (f) 60 day, (g) 90 day, (h) 120
day, (i) 180 day. H&E staining; original magnification  16.

included in a PCT that approached the

center of the extraction socket. The PM
(i) was comprised of newly formed
blood vessels, immature mesenchymal
cells, various types of leukocytes and
collagen fibers and (ii) had apparently in
part replaced the fiber bundles of the
PDL as well as residues of the coagulum
and GT (Figs 7 and 8).
In the central and apical zones of the
socket, large areas of the coagulum
exhibited signs of coagulative necrosis.
Several marrow spaces within the bone
walls, bundle bone, lining the socket

harbored osteoclasts. Such multinucleated cells were also seen within the
Volkmann canals and indicated that the
process of remodeling of this particular
bone tissue was ongoing (Fig. 9).
After 14 days of healing, the marginal portion of the extraction socket was
covered by a connective tissue rich in
vessels and inflammatory cells. This
mesenchymal tissue was in part lined
with epithelial cells.
The most conspicuous features characterizing this interval, however, was (i)
the absence of a periodontal ligament

812

Cardaropoli et al.

I-cells

CLOT

Fig. 3. Inflammatory cells located in the upper portion of the blood clot (CLOT) in the
marginal zone of the socket on day 1. I-cells: inflammatory cells. H&E staining; original
magnification  400.

Fig. 4. A well-organized blood clot from a specimen representing day 1 of healing. Note the
large number of erythrocytes that are trapped in a fibrin network. H&E staining; original
magnification  400.

and (ii) the presence of large amounts of

new hard tissue. Thus, the bundle bone
of the extraction socket was in most
areas absent and communications
seemed to exist between the BM spaces
of the neighboring interdental septa and
the newly formed tissue in the experimental unit (Fig. 2d). The woven bone
extended from the old bone of the
socket walls towards the center of the
wound. This woven bone was rich in
cells, and occurred adjacent to newly
formed blood vessels (Fig. 10). A

provisional connective tissue was still

present in the central part of the socket
(Fig. 2d).
In sections representing 30 days of
healing (Fig. 2e), it was observed that
the marginal soft tissue compartment
harbored a well-organized fibrous connective tissue that was lined with a
keratinized epithelium. Further, most
parts of the extraction socket were filled
with newly formed bone. This bone
contained a large number of primary
osteons (Fig. 11), and was continuous

with the old bone of the socket walls. In

some areas, the woven bone was undergoing osteoclastic resorption (Fig. 12).
This indicated that the process of
modeling/remodeling of the newly
formed bone had begun. Osteoclasts
were also observed on the surface of the
old lamellar bone of the crestal region
lateral to the extraction socket.
In all specimens representing days 60
and 90 of healing (Fig. 2f, g), a newly
formed hard tissue bridge, mainly
composed of woven bone, was seen to
separate the marginal mucosa from the
extraction socket (Fig. 13a, b). Further,
most of the woven bone in the socket
apical of the bridge had been replaced
with BM. This BM included large blood
vessels, inflammatory cells and adipocytes (Fig. 14a). In sections from day
90, it could be observed that woven
bone in several areas was being replaced with lamellar bone. Also the old
bone of the socket walls exhibited signs
of remodeling.
After 120 and 180 days of healing
(Fig. 2h, i), the marginal hard tissue
bridge covering the previous socket
had become reinforced by layers of
lamellar bone that had deposited on the
top of the previously formed woven
bone (Fig. 13c, d). Concomitantly,
collagen fibers from the lining mucosa
had became inserted in the new cortical bone and, hence, a periosteum
like structure had been established.
After 180 days of healing, this bridge
of MB was comprised of a mixture of
woven bone and lamellar bone (Fig. 15).
The entire region of the extraction
socket that was located apical of the
marginal bone bridge was characterized by its content of BM. This BM
was well organized and contained
large numbers of adipocytes and only
few inflammatory cells (Fig. 14b).
Within the BM a limited number of
trabeculae of lamellar bone were also
present.
Morphometric measurements

The results of the morphometric measurements are presented in Table 1

(overall) and Table 2a (zone A 5 marginal), Table 2b (zone B 5 middle) and
Table 2c (zone C 5 apical).

Overall composition
The data describing the overall composition of the tissues of the extraction
socket at various time intervals follow-

Bone healing dynamics

813

mandibular premolar. In zone A, a GT

could be identified that occupied about
1% of the tissue volume.
CLOT
PDL

Days 7, 14, 30

BB

Fig. 5. A blood clot from day 1 that is in direct contact with the severed periodontal ligament
fibers in zone B. CLOT 5 blood clot; BB 5 bundle bone; PDL 5 periodontal ligament. H&E
staining; original magnification  400.

In zone A, the GT made up about 13%

(day 7) and 10% (day 14) of the tissue
volume, while in zones B and C no
highly vascularized tissue with a dense
inflammatory cell infiltrate could be
observed at these intervals. A PCT was
observed in sections representing 7 days
of healing (zone A 5 8%, zone
B 5 36%, zone C 5 100%). MB first
occurred as woven bone and could be
observed in the day 14 biopsies (zone
A 5 15%,
zone
B 5 56%,
zone
C 5 72%) and was in all three zones
the dominating tissue in the sections
representing 30 days of healing (zone
A 5 78%,
zone
B 5 90%,
zone
C 5 95%).

Days 60, 90, 120, 180

In the interval between days 90 and 180,
the volume occupied by MB in the
experimental unit was found to
decrease (zone A: from 46% to 27%;
zone B: from 41% to 13%; zone C: from
24% to 5%). BM that included large
amounts of adipocytes was first found in
sections representing 60 days of healing
(zone A 5 61%, zone B 5 80%; zone
C 5 91%). The proportion of the tissue
volume occupied by BM gradually
increased during healing and amounted
to 73% (zone A), 87% (zone B) and
95% (zone C) in the 180 day specimens.

Discussion
Fig. 6. Day 3; zone B, central portion of the coagulum. Note the presence of erythrocytes in
various stages of disintegration. The site has a hyaline appearance (coagulate necrosis). H&E
staining; original magnification  400.

ing removal of the roots are presented in

Table 1. During the first 3 days of
healing most of the socket was filled
with a blood clot (CLOT) that after 7
days had in part been replaced with a
PM. The first signs of MB were noted in
the 14 day specimens, and on day 30,
the extraction socket was more or less
entirely filled with bone (88%). In
specimens obtained from days 60, 90,
120 and 180, a non-mineralized BM
was the dominating tissue and MB

occupied only between 15% and 23%

of the socket volume.
Tissues in zones A, B and C

Days 1 and 3
In specimens representing days 1 and 3
of healing, the experimental unit was
dominated by a coagulum (clot) that
filled most of the space previously
occupied by the distal root of the fourth

The findings of the present investigation

demonstrated that the healing of an
extraction socket involved a series of
events including the formation of a
coagulum that was replaced by (i) a
PM, (ii) woven bone, and (iii) lamellar
bone and BM. Further, in the process of
healing, a hard tissue bridge formed,
which closed the socket. Thus, the
healing of an extraction socket appears
to have many features in common with
events that characterize new tissue
formation in a fracture of a long bone
(Hollinger & Wong 1996).
In the current study, it was also noted
that the PDL fibers and the bundle bone
of the socket wall gradually disappeared, and that the socket site was

814

Cardaropoli et al.

CLOT

PM

Fig. 7. Day 7; the provisional matrix (PM) had invaded the blood clot (CLOT). Note the
presence of the mesenchymal cells and vessels among the erythrocytes of the blood clot.
PM 5 provisional matrix. H&E staining; original magnification  400.

BB

PM

BB

Volkmanns Canal

BB

Fig. 8. Day 7; the lateral wall of the healing

socket. Provisional matrix (PM) had in part
replaced the periodontal ligament. Compare
with Fig. 5 that describes a similar area after
1 day of healing. BB 5 bundle bone. H&E
staining; original magnification  400.

Fig. 9. Day 7; the wall of the alveolus with

a penetrating Volkmanns canal. Note the
presence of several osteoclasts that reside on
the internal surface of the Volkmanns
canal. BB 5 bundle bone; arrows indicate
osteoclasts. H&E staining; original magnification  200.

eventually characterized by its large

content of BM.
The observation that a blood clot
formed and became properly retained in
the socket within the first 24 h after
tooth extraction is in agreement with
findings previously reported (Amler
1969, Lin et al. 1994). This coagulum
was in the marginal region (zone A) of
the socket during the first week found to
be in part replaced with a GT (rich in
vessels and inflammatory cells), while
in the remaining zones (zones B and C)

a PM during this interval replaced the

blood clot. It is suggested that the GT in
zone A formed in response to the
presence of infectious material in the
oral cavity and that the inflamed tissue
served as a barrier that protected the
more apical parts of the alveolus. The
validity of this assumption is supported
by the fact that when the mucosa after 1
month of healing was covered with a
keratinized epithelium, the inflammatory cells and the vascular units in zone
A had markedly decreased in number.

The suggestion that the GT in zone A

formed in response to the presence of
bacterial contaminants in the orifice of
the socket is also in agreement with data
presented by Araujo et al. (1997). They
studied tissue events that led to bone
formation in degree III (through and
through) furcation defects in the dog.
The defects were mechanically produced and plaque was allowed to
accumulate for 2 weeks in the experimental site. Treatment included flap
elevation as well as scaling and root
planning. Barrier membranes were adjusted to the buccal and lingual entrances of the furcation. The wound was
covered with mucosal flaps. Biopsies
were obtained at different intervals
during healing. The authors reported
that the furcation defect in this open
model, after 2 weeks of healing, was
occupied by a GT rich in inflammatory
cells. In the current experiment, a
corresponding (2 weeks) GT was seen
only in zone A. As stated above, a PM
had at this interval replaced the clot in
zones B and C. This finding is in
agreement with the findings by Lin et
al. (1994), who from experiments in the
rat reported that . . . a large portion of
the coagulum . . . in the socket. . .
was replaced by dense connective
tissue. . ..
In the present study, it was observed
that hard tissue formation had started
already after 2 weeks of healing. Thus,
in the 2 weeks specimens more than
50% of the tissue residing in zones B
and C was comprised of woven bone.
The corresponding figure representing
zone A is only 15%. There are reasons
to suggest that the proximity of zone A
to the oral cavity may explain the
delay of hard tissue formation in
this marginal compartment of the alveolus.
Cells (PDL cells) and fibers of the
remaining PDL could be identified in all
specimens sampled between days 1 and
7 in the current experiment. A large
number of PDL cells were seen close to
the coagulum, and PDL-like cells were,
in addition, seen within the coagulum
and to be part of the PM. It is suggested
that PDL cells contributed not only
to the formation of the PM but also
to hard tissue formation in the healing
socket. This observation supports data
by Lin et al. (1994) from their experiment on bone formation in extraction
sockets of rats. The authors used
a cell labeling technique to follow
the fate of PDL fibroblasts. Lin et al.
(1994) concluded that PDL fibroblasts

Bone healing dynamics

WB

WB

WB

Fig. 10. Day 14; newly formed woven bone (mineralized bone) in the central portion of the
healing socket. The deposition of bone had occurred around blood vessels. WB: woven bone,
V: blood vessels. H&E staining; original magnification  400.

SO

PO

Fig. 11. Day 30; newly formed bone located in zone B. Note the presence of primary and
secondary osteons in the bone tissue. PO: primary osteon; SO: secondary osteon. H&E
staining; original magnification  200.

proliferated and migrated into the

center of the extraction socket, where
they differentiated into osteoblasts that
became involved in the formation of
new bone.
In specimens obtained after 60, 90,
120 and 180 days of healing, a hard
tissue bridge was consistently found to
cover the marginal portion of the
extraction site. This so-called corticalization (Ohnishi et al. 2000) of the
socket included a series of proliferative
and resorptive events through which a

cortical bone wall was eventually developed and to which the lining mucosa
became firmly attached. This process
included (i) the formation of woven
bone to fill the alveolus, including
its marginal compartment, (ii) the remodeling, i.e. removal of woven
bone and the formation of lamellar
bone, (iii) the deposition of additional
(incremental) layers of lamellar bone
to reinforce the hard tissue bridge,
and (iv) the establishment of a periosteum that allowed an attachment be-

815

tween the lining mucosa and the

newly formed cortical bone. In many
respects, the above findings are in
agreement with data previously reported
by Schenk et al. 1994). The authors
produced large bone defects in the
edentulous ridge of dogs. The treatment
of these defects included the placement
of a barrier membrane (GBR) that
protected the site during healing. The
authors stated that the corticalization of
the newly formed bone was not only
based on remodeling activities but
also on the direct deposition of lamellar
bone over a primary spongiosa (i.e.
woven bone).
In sections representing 180 days of
healing, it was observed that a large
volume of the previous extraction site
below the cortical bone bridge was
occupied by BM (85%). In other words,
between days 30 and 180, the tissue
within the socket changed from being
dominated by woven bone to a mature
tissue that contained only small
amounts of MB. Similar observations
were reported in an experiment evaluating the remodeling process in the
healing of through and through
furcation defects in dogs (Araujo et al.
1999), treated according to the principles of GTR (Gottlow et al. 1984).
The authors observed that after 2
months of healing the furcation defect
was filled with woven bone, while
after 6 months the healed defect was
characterized by its content of BM
(55%). Araujo et al. (1999) suggested
that the newly formed attachment
apparatus the cementum, the PDL
and the bundle bone was not a load
carrying tissue, a fact that would
explain the large amount of BM observed in the marginal compartment
of the healed furcation. In the
current experiment, the extraction site
was exposed to minimal load, and
hence, once the cortical bridge had
formed, there was apparently no
obvious demand for mineralized
tissue (trabecular bone) in the compartment previously occupied by the
distal root of the premolar. Similar
findings were reported by Buser
et al. (1995), who studied the
tissue formed in augmented (GBR)
areas of the mandibular ridge of dogs.
They observed that at sites that during
healing following the ridge augmentation procedure were without apparent
load, a large marrow space occupied the
central portion of the newly formed
bone.

816

Cardaropoli et al.

V
WB
V

WB

Fig. 12. Day 30; woven bone in zone B of the socket. Note the large number of osteoclasts
(arrows) that are present on the bone surface. WB: woven bone; V: blood vessel. H&E
staining; original magnification  400.

Zusammenfassung
Dynamik der Knochenneubildung in Bereichen
von Zahnextraktionen.
Eine experimentelle Studie an Hunden.
Ziele: Das Ziel des vorliegenden Experiments
war es, Ereignisse die mit der Heilung einer
Extraktionsalveole in marginalen, zentralen und
apikalen Abschnitt verbunden sind zu studieren.

Von der Bildung des Blutkoagulums zur

Knochenneubildung und dem Remodellieren
des neu gebildeten Hartgewebes.
Material und Methoden: 9 Mischlingshunde
wurden fur das Experiment verwendet. 4.
mandibulare Pramolaren wurden fur die Studie
ausgewahlt und wurden in einen mesialen und
in einen distalen Bereich aufgeteilt. Die distale
Wurzel wurde entfernt und die Alveole mit dem

LM
LM

LM
LM

umgebenden Weichgewebe und dem mineralisierten Gewebe wurde als experimentelle

Einheit angegeben. Die Hunde wurden 1, 4,
7, 14, 30, 60, 90, 120 und 180 Tage nach der
Wurzelextraktion geopfert. Die Biopsien, die
die experimentellen Einheiten einschlossen,
thanol
wurden in EDTA entmineralisiert, in A
entwassert und in Parafin eingebettet. Serienschnitte von 7 mm Dicke wurden in der
mesio-distalen Ebene angefertigt. Von jeder
Biopsie wurden 3 Schnitte, welche den zentralen Bereich der Alveole reprasentierten fur
die histologische Untersuchung ausgewahlt.
Zur Bestimmung des Volumens, welches von
den verschiedenen Arten des Gewebes in
den marginalen, zentralen und apikalen
Abschnitten der Extraktionsalveole nach unterschiedlichen Zeitabschnitten eingenommen
wird, wurden morphometrische Messungen
durchgefuhrt.
Ergebnisse: Wahrend der ersten 3 Tage der
Heilung wurde ein Blutkoagulum vorgefunden,
welches den groten Teil der Extraktionswunde
eingenommen hatte. Dieses Koagulum wurde
nach 7 Tage teilweise durch eine provisorische
Matrix ersetzt. An Tag 14 beinhaltete das
Alveolengewebe provisorische Matrix und Geflechtknochen. An Tag 30 bestand das Alveolenvolumen zu 88% aus mineralisiertem
Knochen. Dieses Gewebe hatte an Tag 180
auf 15% abgenommen. Der Anteil, der an Tag
60 von Knochenmark eingenommen wurde
betrug 75% und nahm aber bis Tag 180 auf
85% zu.

WB

WBWB

aa

LB
LB

LM

LM

WB

WB

Fig. 13. A cortical bone bridge formed in the marginal portion of the socket; day 60 (a), day 90 (b), day 120 (c) and day 180 (d). Note the
lamellar bone and incremental lines on days 120 (c) and 180 (d). LB: lamellar bone; LM: lining mucosa; WB: woven bone: WB: arrows
indicate the location of the incremental line. H&E staining; original magnification  400.

Bone healing dynamics

817

Ad
Ad

V
Ad

Fig. 14. Bone marrow present in zones B and C on day 60 (a) and day 180 (b). Note that in (a) the bone marrow includes large vessels (V),
poorly organized adipocytes (Ad) and inflammatory cells, while in (b) the adipocytes are well organized. H&E staining; original
magnification  400.

SO
PO

Fig. 15. Day 180; newly formed bone from the marginal portion of the socket (cortical
bone). Note the presence of primary and secondary osteons. PO: primary osteon; SO:
secondary osteon. H&E staining; original magnification  400.
Schlussfolgerung: Die Heilung einer Extraktionsalveole umfasst eine Reihe von Ereignissen, einschlielich der Bildung eines
Koagulums, welches ersetzt wird durch: 1. Eine
provisorische Bindegewebsmatrix, 2. Geflechtknochen und 3. Lamellaren Knochen und
Knochenmark. Wahrend des Heilungsprozesses
bildete sich eine Hartgewebsbrucke kortikaler
Knochen welcher die Alveole verschloss.

Resume
Dynamiques de la formation tissulaire osseuse
dans les sites davulsion dentaire. Une etude
experimentale chez le chien
Le but de letude presente a ete detudier les
etapes de la guerison des compartiments
apicaux, centraux et marginaux dune alveole
davulsion, depuis la formation du caillot

sanguin jusqua` la formation tissulaire osseuse

et le remodelage du tissu dur neoforme. Neuf
chiens batards ont servi pour cette experience.
Les quatrie`mes premolaires mandibulaires ont
ete selectionnees pour letude et ont ete divisees
en une portion mesiale et une distale. La racine
distale a ete enlevee, et lalveole entouree de
ses tissus mineralises et mous ont ete nommee
unite experimentale. Les chiens ont ete
euthanasies 1, 4, 7, 14, 30, 60, 90, 120 et 180
jours apre`s les avulsions radiculaires. Les
biopsies comprenant les unites experimentales
ont ete demineralisees dans lEDTA, deshydratees dans lethanol et enrobees dans la parafine.
Des coupes en serie de 7 mm depaisseur ont ete
coupees dans un plan mesio-distal. De chaque
biopsie, trois coupes representant la partie
centrale de lalveole ont ete selectionnees pour
lexamen histologique. Les mesures morphometriques ont ete effectuees pour determiner le
volume occupe par les differents types de tissus
dans les compartiments marginaux, centraux et
apicaux de lalveole davulsion aux differents
moments. Durant les trois premiers jours de
guerison, un caillot sanguin occupait la plus
grande partie du site davulsion. Ce caillot,
apre`s sept jours, etait en partie remplace par une
matrice provisoire. Au jour quatorze, le tissu de
lalveole comprenait la matrice provisoire et de
los spongieux. Au jour 30, los mineralise
occupait 88% du volume de lalveole. Ce tissu
diminuait a` 15% au jour 180. La proportion
occupee par de la moelle osseuse dans les
specimens du jour 60 etait denviron 75% mais
avait augmente a` 85% au jour 180. La guerison
dune alveole davulsion comprend une serie
devenements comportant la formation dun

Table 1. Overall proportion (%) of the various tissues in the healing socket during the different time intervals, mean (SD)

CLOT
GT
PCT
MB
BM

1 day

3 days

7 days

14 days

30 days

60 days

90 days

120 days

180 days

99.5 (0.6)
0.5 (0.6)
0
0
0

99.5 (0.6)
0.5 (0.6)
0
0
0

48 (41.9)
4 (7.5)
48 (47.1)
0
0

0
3 (5.7)
49 (23.8)
48 (29.4)
0

0
0
12 (8.6)
88 (8.7)
0

0
0
0
23 (15.2)
77 (15.1)

0
0
0
37 (12)
63 (11.4)

0
0
0
29 (14.7)
71 (14.3)

0
0
0
15 (11.1)
85 (11.2)

CLOT: blood clot; GT: granulation tissue; PCT: provisional matrix; MB: mineralized bone; BM: bone marrow.

818

Cardaropoli et al.

Table 2. Proportion (%) of the various tissues in the healing socket during the different time intervals, mean (SD)
1 day

3 days

7 days

14 days

30 days

60 days

90 days

120 days

180 days

Zone A
CLOT
GT
PCT
MB
BM

99 (2.2)
1 (1.7)
0
0
0

99 (1.2)
1 (1.1)
0
0
0

79 (6.4)
13 (3.8)
8 (11.1)
0
0

0
10 (4.7)
75 (14.4)
15 (11.1)
0

0
0
22 (7.4)
78 (7.1)
0

0
0
0
39 (10.4)
61 (14.1)

0
0
0
46 (12.8)
54 (6.3)

0
0
0
37 (3.7)
63 (2.8)

0
0
0
27 (2.5)
73 (2.5)

Zone B
CLOT
GT
PCT
MB
BM

100 (1.8)
0
0
0
0

100 (0.3)
0
0
0
0

64 (3.4)
0
36 (1.0)
0
0

0
0
44 (5.7)
56 (7.3)
0

0
0
10 (2.2)
90 (2.1)
0

0
0
0
20 (8.8)
80 (1.8)

0
0
0
41 (3.4)
59 (12.0)

0
0
0
38 (1.9)
62 (2.5)

0
0
0
13 (1.1)
87 (1.3)

Zone C
CLOT
GT
PCT
MB
BM

100 (3.6)
0
0
0
0

100 (5.3)
0
0
0
0

0
0
100 (4.9)
0
0

0
0
28 (10.4)
72 (11.2)
0

0
0
5 (2.4)
95 (2.7)
0

0
0
0
9 (4.5)
91 (3.8)

0
0
0
24 (12.1)
76 (12.2)

0
0
0
12 (2.9)
88 (4.1)

0
0
0
5 (0.3)
95 (1.8)

CLOT: blood clot; GT: granulation tissue; PCT: provisional matrix; MB: mineralized bone; BM: bone marrow.

coagulum qui etait remplace par 1) une matrice

tissulaire conjonctive provisoire, 2) de los
spongieux et 3) de los lamellaire et de la
moelle osseuse. Durant le processus de guerison, un pont de tissu dur-os cortical- se formait
pour fermer lalveole.

References
Amler, M. H. (1969) The time sequence of
tissue regeneration in human extraction
wounds. Oral Surgery Oral Medicine and
Oral Pathology 27, 309318.
Araujo, M. G., Berglundh, T., Albrekstsson, T.
& Lindhe, J. (1999) Bone formation in
furcation defects. An experimental study in
the dog.. Journal of Clinical Periodontolology 26, 643652.
Araujo, M. G., Berglundh, T. & Lindhe, J.
(1997) On the dynamics of periodontal tissue
formation in degree III furcation defects. An
experimental study in dogs. Journal of
Clinical Periodontolology 24, 738746.
Buser, D., Ruskin, J., Higginbottom, F., Hardwick, R., Dahlin, C. & Schenk, R. K. (1995)
Osseointegration of titanium implants in
bone regenerated in membrane-protected
defects: a histologic study in the canine
mandible. International Journal of Oral &
Maxillofacial Implants 10, 666681.
Christopher, E. R. (1942) Histological study of
bone healing in relation to the extraction of
teeth. Northwestern University Bulletin 45, 5.

Clafin, R. S. (1936) Healing of disturbed and

undisturbed extraction wounds. Journal of
American Dental Association 23, 945959.
Gottlow, J., Nyman, S., Karring, T. & Lindhe J,
. (1984) New attachment formation as the
result of controlled tissue regeneration.
Journal of Clinical Periodontolology 8,
494503.
Hollinger, J. & Wong M, E. (1996) The
integrated processes of hard tissue regeneration with special emphasis on fracture
healing. Oral Surgery Oral Medicine Oral
Pathololy Oral Radiololy and Endodontic 82,
594606.
Karnovsky, M. J. (1965) A formaldehyde
glutaraldehyde fixative on high osmolarity
for use in electron microscopy. Journal of
Cell Biology 27, 137A138A.
Kuboki, Y., Hashimoto, F. & Ishibashi, K.
(1988) Time-dependent changes of collagen
crosslinks in the socket after tooth extraction
in rabbits. Journal of Dental Research 67,
944948.
Lin, W. L., McCulloch, C. A. & Cho, M. I.
(1994) Differentiation of periodontal ligament fibroblasts into osteoblasts during
socket healing after tooth extraction in the
rat. Anatatomical Records 1240, 492506.
Mangos, J. F. (1941) The healing of extraction
wounds. An experimental study based on
microscopic and radiographic investigations..
New Zealand Dental Journal 37, 424.
Ohnishi, H., Fujii, N., Futami, T., Taguchi, N.,
Kusakari, H. & Maeda, T. (2000) A histochemical investigation of the bone formation

process by guided bone regeneration in rat

jaws. Effect of PTFE membrane application
periods on newly formed bone. Journal of
Periodontology 71, 341352.
Schram, W. R. (1929) A histologic study of
repair in the maxillary bone following
surgery. Journal of American Dental Association 16, 19871997.
Schenk, R. K., Buser, D., Hardwick, W. R. &
Dahlin, C. (1994) Healing pattern of bone
regeneration in membrane-protected defects:
a histologic study in the canine mandible.
International Journal of Oral & Maxillofacial Implants 9, 1329.
Schroeder, H. E. & Munzel-Pedrazzoli, S.
(1973) Correlated morphometric and biochemical analysis of gingival tissue. Morphometric model, tissue sampling and test of
stereologic procedures. Jounal of Microscopy
99, 301329.
Simpson, H. E. (1960) Experimental investigation into the healing of extraction wounds in
macacus rhesus monkeys. Journal of Oral
Surgery 18, 391399.

Address:
Giuseppe Cardaropoli
Department of Periodontology
The Sahlgrenska Academy at Goteborg
University Box 450
40530 Goteborg
Sweden
Fax: 146 31 773 3791
E-mail: Giuseppe.Cardaropoli@odontologi.gu.se