Oral Med Pathol 13 (2009) 91

Introduction
Epulis, which is a relatively common tumor-like lesion
of the gingiva, is pathologically classifed into the following
six subtypes: epulis osteoplastica (EO), epulis fbrosa (EF),
epulis granulomatosa, epulis hemangiomatosa, giant cell
epulis and congential epulis (1-4). Although epulis is
considered to be a reactive massive lesion rather than true
neoplasia, details of its pathogenesis and the reasons for the
morphological differences among the subtypes are not fully
understood.
One of the histopathological characteristics of EO is
pathological ossifcation and/or calcifcation with fbroblastic
proliferation (2, 4). Generally, the pathological calcifcation
is divided into dystrophic calcifcation, metastatic calcif-
cation, and pathological ossifcation. Dystrophic and
metastatic calcifcations are due to disturbance of calcium
metabolism, whereas pathological ossifcation is categorized
as progressive change (1, 5). Although ossifcation plays an
important role in various lesions such as infammation and
tumors (6), the mechanism of pathological ossifcation in
epulis remains unclear.
Bone morphogenetic proteins (BMPs), which are
members of the transforming growth factor-beta (TGF-)
superfamily, play important roles in the osteogenesis and
pathogenesis of many lesions (6, 7). BMP-2 and -4 are
closely associated with osteogenesis and bone development,
whereas BMP-6 is related to osteogenesis in metastatic
tumors (8, 9). Osteocalcin (OCN) appears in the later stage
of ossifcation and is implicated in calcifcation. Osteopontin
(OPN) appears in the early stage of osteogenetic maturation
and is also identifed in cementum (10). Osteonectin (ONN)
is an extracellular matrix protein involved in a variety of
physiologic processes. Its primary role is in the mediation of
cell-to-matrix interactions, which become most apparent in
response to injury (11).
Because the relationship between EO and immunohis-
tochemical expression of these antibodies is still unclear, the
present study was undertaken to determine the morphological
nature of pathological ossifcation and its distribution in EO
using histopathological and immunohistochemical analyses
of BMPs, OCN, OPN and ONN as markers of bone
formation in order to understand and clarify their relationship
with pathological ossifcation in comparison with EF.

A histopathological and immunohistochemical study of patholo-
gical ossifcation in epulis osteoplastica

Hideto Tajima
Department of Oral Surgery, Nihon University Graduate School of Dentistry at Matsudo, Matsudo, Japan
Abstract: Epulis osteoplastica shows pathological ossifcation, but details of the mecha-
nism involved are still unclear. In the present study we carried out histopathological
and immunohistochemical analyses to clarify the potential for ossifcation in epulis
osteoplastica (n = 8), in comparison with epulis fbrosa, using several markers. Epulis
osteoplastica showed particulate, massive, trabecular and osteoid ossifcation
surrounded by proliferating fbroblasts and fbrous tissues under the mucosal
squamous epithelium. Epulis fbrosa showed proliferation of fbroblasts and collagen
fbers without ossifcation. Although positive immunoreactivity for bone morpho-
genetic proteins 2, 4 and 6, osteocalcin, osteopontin, and ostenectin was observed in
the fbroblasts of epulis osteoplastica, the cell positivity rates were much higher in
epulis osteoplastica than in epulis fbrosa, suggesting that these substances were
associated with pathological ossifcation in epulis osteoplastica. Furthermore, the
fndings indicated that the fbroblasts in epulis osteoplastica might have a higher
ability to produce bone matrix production than those in epulis fbrosa.
[Oral Med Pathol 2009; 13: 91-98 doi: 10.3353/omp.13.91]
Key words: bone morphogenetic protein, epulis osteoplastica, osteocalcin, osteopontin,
pathological ossifcation
Correspondence: Hideto Tajima, Department of Oral Surgery, Nihon University School of
Dentistry at Matsudo, 2-870-1 Sakaechou-Nishi, Matsudo 271-8587, Japan
Phone: +81-47-360-9627, Fax: +81-47-360-9405,
E-mail: tajima.hideto@nihon-u.ac.jp
92 Tajima Immunohistochemistry of epulis osteoplastica
Materials and methods
Tissue preparation
Eight cases of EO and eight cases of EF, removed
surgically at the Department of Oral Surgery, Nihon
University Hospital, School of Dentistry at Matsudo, were
used for this study. Specimens of jaw cyst were also used for
comparative examination. Consideration was given to
patient privacy, diagnosis, management and prognosis of the
lesions. (Nihon University Ethics Committee recognition
number: EC 07-006). The excised materials were fxed in
10% neutral formalin solution and embedded in paraffn by
the standard method. The paraffn embedded blocks were cut
into sections 4 m thick for histopathological and immuno-
histochemical examinations. The sections were then stained
with hematoxylin and eosin (HE) for histopathological
diagnosis according to Ishikawas classifcation (5).
Immunohistochemical staining
The labeled streptavidin-biotin method (a kit from Dako,
Glostrup, Denmark) was used for immunohistochemistry.
Primary antibodies used in the present study were goat
polyclonal antibodies against BMP-2 and BMP-4 (Santa
Cruz, CA, USA, diluted 1:50), mouse monoclonal antibodies
against BMP-6 (Medac Diagnostica, Hamburg, Germany,
1:10), OCN (Takara Bio Inc, Shiga, Japan, 1:50), rabbit
polyclonal antibodies against OPN (Cosmo Bio, Tokyo,
Japan, 1:50) and ONN (Cosmo Bio, 1:50). After deparafni-
zation and hydration, the specimens were incubated with the
antibodies against BMP-2 and BMP-4 for 60 min at room
temperature. With regard to the antibodies against BMP-6,
OCN, OPN, and ONN, incubation with the sections was
done overnight at 4.
Additional treatments for OCN immunostaining were as
follows: the specimens were subjected to thermal treatment
in 0.1 M citric acid buffer solution (pH 5.9) for 3 min in a
microwave oven, and then allowed to cool to room
temperature. Specimens for OPN and ONN immunostaining
were treated with trypsin solution for 60 min at room
temperature. Peroxidase activity was visualized using
diaminobenzidine. Positive control staining for all antibodies
was done on areas of new bone formation in a jaw cyst, and
the specimen was used for comparison with EO. For control
experiments, the primary antibodies were replaced with
mouse (IgG2a), rabbit and goat (immunoglobulin fraction)
preimmune sera (Dako), and lack of any staining reactions
was confrmed. All of the immunohistochemically stained
sections were counterstained with Mayers hematoxylin.
Cell counting and statistical analysis
To count cells that were immunohistochemicallly
positive for BMP-2, BMP-4, BMP-6, OCN, OPN and ONN,
light micrographs were taken using an optical microscope
(original magnifcation 600) at each of three arbitrarily
chosen areas of ossifcation. The percentage of positive cells
was calculated using the formula as follows:
percentage of positive cells (%) = (numbers of positive
cells / numbers of whole cells) 100
Welchs t-test was applied to determine the signifcance
of differences (P<0.05).
Results
Histopathological fndings
Macroscopically, all cases of epulis appeared as a
polypoid or nodular mass (Fig. 1). EF was composed of
proliferating fbroblasts and collagen fbers with a minimal
degree of infammatory cell infltration and vascular
dilatation, and no evidence of ossifcation (Fig. 2a).
In comparison, pathological ossifcation was observed in
EO (Fig. 2b), which appeared as a polypoid mass showing
four types of ossifcation: particulate (2/8 cases), massive
(6/8), trabecular (2/8) and osteoid (8/8). Particulate
ossifcation and osteoid were surrounded by proliferating
fbroblasts and collagen fbers (Fig. 2c). Two types of
mesenchymal cells -fattened cells and spindle cells- were
evident around the four types of ossifcation (Fig. 2d). The
spindle cells were observed predominantly around the
particulate, massive, and trabecular ossifcations and osteoid,
whereas the fattened cells were located adjacent to massive
ossifcation (Figs. 2e-2f). Osteoclasts did not exist in EO.
In the comparative specimen, an osteoblast lining and
osteoclasts were observed in the bone at the periphery of the
cyst wall (data not shown). The bone showed trabecular
ossifcation, which was classifed as membranous
ossifcation.
Immunohistochemical fndings
The results of immunohistochemistry for BMP-2,
BMP-4, and BMP-6 in EF and EO are summarized and
indicated in Figs. 3-5 and 9.
In EF, the proportions of cells that were positive for
BMP-2 (Figs. 3a-3b), BMP-4 (Figs. 4a-4b), and BMP-6
(Figs. 5a-5b) were 35.6 7.0 %, 35.5 9.7 % and 41.9 7.7
%, respectively (Fig. 9). Positive cells were observed mainly
in the cytoplasm of the proliferating fbroblasts around the
Fig. 1. Gross appearance of epulis, cut surface, on a
premolar tooth.The polypoid mass lesion was
composed of white fbrous tissue and covered
by the gingival mucosa. E, epulis; Th, tooth.
E
Th
Oral Med Pathol 13 (2009) 93
vessels and in the areas of hyperplasia.
In EO, the proportions of cells that were positive for
BMP-2 (Figs. 3c-3f), BMP -4 (Figs. 4c-4f), and BMP -6
(Figs. 5c-5f) were 57.1 11.4 %, 63.0 9.8 % and 58.6 8.0
%, respectively (Fig. 9). Positive reactions were found in the
spindle cells surrounding the particulate, massive and
trabecular ossifcations, but the fattened cells around the
ossifcations were not positive. BMP-4 showed a positive
immunoreaction in areas of bone matrix and osteocyte. The
differences in the proportions of cells positive for BMP-2,
BMP-4, and BMP-6 between EO and EF were signifcant.
Immunoreactivity for OCN (Figs. 6a-6b), OPN (Figs.
7a-7b) and ONN (Figs. 8a-8b) was not observed in EF but
was found in EO, the proportions of cells showing positivity
being 61.2 17.5 %, 53.2 18.2 % and 57.4 1 5.2 %,
respectively (Fig. 9). Cells positive for OCN were found in
the bone matrix, and in the fbroblasts and spindle cells
surrounding the bone in areas of particulate, massive and
trabecular ossifcation (Figs. 6c-6f). Positivity for OPN was
evident in fbroblasts, spindle cells, and fattened cells
around areas of particulate, massive and trabecular
ossifcation, but the matrix of ossifcation and osteocytes
were not positive for OPN (Figs. 7c-7f). Positivity for ONN
was evident in fbroblasts and spindle cells surrounding
areas of particulate, massive and trabecular ossifcation and
partly in osteocytes in areas of trabecular ossifcation (Figs.
8c-8f). There was no difference in immunoreactivity for
BMPs, OCN, OPN and ONN between trabecular ossifcation
and osteoid, but the proportion of cells immunoreactive for
BMPs was higher than that of cells immunoreactive for
OCN, OPN and ONN in areas of particulate and massive
ossifcation (Fig. 10). OCN showed the lowest level of
immunoreactivity, and the difference in the proportion of
cells immunoreactive for BMP-4 and OCN was signifcant.
Specimens of new bone formation around cysts were
examined in the present study for comparison with
pathological ossifcation. Positive immunoreactivity for
BMP-2, BMP-4, OCN, OPN and ONN was identifed in
osteoblasts lining areas of new bone formation, but BMP-6
was not positive (data not shown).
Discussion
EO appeared as a polypoid mass with various levels of
maturation of ossifcation, including particulate, massive,
trabecular and osteoid (3-6), but mainly the former three.
Bone formation involves several processes, including histo-
differentiation, morpho-differentiation, matrix formation
and calcifcation. Generally and histologically, immature
mesenchymal cells differentiate into myofbroblasts, and
then to fbroblasts, which then differentiate further into
osteoblasts, which have ossifcation ability. The
morphological changes in osteoblasts during mesenchymal
differentiation have not been fully elucidated. Generally,
a b
c d
e f
Fig. 2. Histopathological features of epulis
fbrosa (EF) (a) and epulis osteoplastica
(EO) (b-f). Hematoxy lin and eosin (HE)
stain, (a) 200, inset: 600; (b) 40;
(c-f) 600. EF was composed of a
proliferation of fbroblasts and collagen
fbers with minimum infammatory cell
infltration (a). EO showed prolif eration
of fbroblasts and collagen fbers around
pathological ossif cation (b). EO con-
sisted of particu late ossifcation (c). EO
consisted of massive ossifcation (d). EO
consisted of immature trabecular ossif-
cation (e). EO consisted of mature trabe-
cular ossifcation sur rounded by fbro-
blastic proliferation with slightly of
infammatory cell infltration and capil-
lary dilatation. The spindle cell lining
was slightly observed around the
ossifcation (f).
94 Tajima Immunohistochemistry of epulis osteoplastica
osteoblasts have a fattened to round shape and are arranged
at the periphery of bone and matrices (12-15).
In the present study of EO, differentiation of
myofbroblasts to fbroblasts could not be confrmed.
However, after the collagen fbers became hyperplastic,
fbroblasts around the matrix could be identifed. Spindle
cells were evident at the periphery of areas of particulate,
massive and trabecular ossifcation, as well as of osteoid,
and fattened cells were observed in the periphery of massive
ossifcation. The spindle cells were thought to be progenitors
of the fattened cells, and fbroblasts were considered to be
progenitors of the spindle cells. During the development of
ossifcation, osteoid appeared to form initially, followed by
particulate ossifcation and massive ossifcation. Trabecular
Fig. 4. BMP-4 immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin counterstain. (a) 200; (b-f) 600. BMP-4 was positive in most of the
proliferating fbroblasts in EF (a). In higher magnifcation, the fbroblasts showed positive reaction for BMP-4 in
their cytoplasm (b, arrow). In EO, BMP-4 was localized in fbroblasts around particulate ossifed materials (c,
arrow). Positive reactions for BMP-4 were enhanced in fbroblasts, spindle or fattened cells around massive
ossifed matrices in EO (d, arrow). Positive reactions for BMP-4 were not only seen in fbroblasts and spindle-
shaped cells around ossifed matrices but also in osteoid in EO (e, arrow). Positive reactions for BMP-4 were in
ossifed matrices and fbroblasts around the mature trabecular ossifed ones in EO (f, arrow).
a b c
d e f
Fig. 3. BMP-2 immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin counterstain. (a) 200; (b-f) 600. Positive in proliferating fbroblasts in
EF (a). In higher magnifcation, EF was immunolocalized in their spindle-shaped cytoplasm (b, arrow). In EO,
positive reactions for BMP-2 were emphasized in fbroblasts around particulate ossifed matrices (c, arrow).
BMP-2 was defnitely positive in fbroblasts around massive ossifed tissues in EO (d, arrow). Positive reactions
for BMP-2 were more prominent in fbroblasts or spindle-shaped cells around immature ossifed trabecular in EO
(e, arrow). BMP-2 was also positive in fbroblasts around mature ossifed trabecular in EO (f, arrow).
a b c
d e f
Oral Med Pathol 13 (2009) 95
ossifcation was thought to be the fnal morphological form
of ossifcation in EO because osteoid did not form at the
periphery of areas of trabecular ossifcation, and spindle
cells were not observed around these areas.
Embryologically speaking, ossifcation is generally
divided into two types: endochondral and membranous (16).
Endochondral ossifcation gives rise to long bones that
comprise the appendicular skeleton, facial bones, vertebrae,
and the lateral medial clavicles. Membranous ossifcation
occurs in fat bones such as those comprising the cranium
and medial clavicles (13). Both types of ossifcation indicate
osteoblastic differentiation and lining around the bone,
although the biological and morphological processes
involved are quite different. Another type or primary stage
of membranous ossifcation is known as woven bone
formation appearing in fbrous dysplasia of bone and other
fbro-osseous lesions (16). This ossifcation results from
abrupt transformation of a fbroblastic proliferation site into
ossifcation without a typical osteoblastic cell lining. Thus,
fbro-osseous lesions are a type of membranous ossifcation,
Fig. 5. BMP-6 immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin counterstain. (a) 200; (b-f) 600. BMP-6 was positive in most of the
proliferating fbroblasts in EF (a). In higher magnifcation, the fbroblasts showed positive reaction for BMP-6 in
their cytoplasm (b, arrow). In EO, BMP-6 was emphasized in fbroblasts around the particulate ossifed materials
(c, arrow) as well as in those around the massive ossifed materials in EO (d, arrows). Positive reactions for BMP-6
were obtained not only in fbroblasts but also in spindle-shaped cells around massive (e, arrow) or mature
trabecular ossifed materials (f, arrow).
a b c
d e f
Fig. 6. OCN immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin couterstain. (a, b) 200; (c-f) 600. OCN was not positive in EF (a), while
it was clearly positive in ossifed matrices of EO (b). OCN was defnitely positive in particulate (c) or massive (d)
ossifed matrices. It was also emphasized in osteoid (e) as well as in trabecular ossifed matrices (f).
a b c
d e f
96 Tajima Immunohistochemistry of epulis osteoplastica
in which mesenchymal cells differentiate into fbroblasts and
osteoblasts (15-16).
Specimens of new bone forming around jaw cysts were
used in the present study for comparison with the pathological
ossifcation seen in EO. These specimens showed a lining of
osteoclasts and osteoblasts. In contrast, EO appeared to be
different from bone formation around a cyst but had a similar
histopathological appearance to fbro-osseous lesions,
indicating an unremarkable lining of osteoblasts and
osteoclasts at the bone periphery. Furthermore, cartilage was
not evident in EO. These fndings appeared to be characteristic
of pathological ossifcation, i.e. a type of membranous
ossifcation, in EO. There was no evidence of osteoclasts in
areas of particulate, massive, trabecular ossifcation and
osteoid. These results suggested that the pathological
ossifcation in EO differs from that of normal bone
regeneration, since the osteoplastic change might give
priority to bone resorption. Furthermore, this phenomenon
suggested that some inhibitors of osteoblast differentiation
might be present.
Fig. 8. ONN immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin counterstain. (a, b) 200; (c-f) 600. ONN was not positive in EF (a),
while it was defnitely positive in fbroblasts around ossifed matrices (b). ONN positive fbroblasts were found
around particulate ossifed tissue (c, arrow). It was localized in fbroblasts and the spindle-shaped cells around
massive (d, arrow) or trabecular (e, arrow) ossifed tissues as well as around osteoid (f, arrows).
a b c
d e f
Fig. 7. OPN immunohistochemical profles compared between epulis fbrosa (a) and epulis osteoplastica (b-f).
Immunoperoxidase stain, hematoxylin counterstain. (a, b) 200; (c-f) 600. OPN was not positive in EF (a),
while it was localized in fbroblasts around ossifed matrices of EO (b). OPN was faintly positive in fbroblasts
around the particulate ossifed (c, arrow) but was more defnitely positive in fbroblasts (d, arrow) and in spindle-
shaped cells (e, arrows) around massive ossifed tissues. OPN positive fbroblasts were also found around the
mature trabecular ossifed tissue (f, arrows).
a b c
d e f
Oral Med Pathol 13 (2009) 97
As mentioned above, BMPs are associated with
proliferation and differentiation, whereas OPN and ONN are
related to matrix formation during ossifcation. OCN is
related to calcifcation (17-19).
BMPs, which are members of the TGF- superfamily,
are well known as important mediators of bone formation,
and can be divided into several subtypes (6-9). They were
frst isolated from bone extracts, and displayed an ability to
induce ectopic ossifcation. They play roles as multifunctional
regulators of vertebrate development, controlling cell
proliferation, differentiation and apoptosis in different
tissues including skin. The BMP subtypes with the greatest
osteogenic capacity are BMP-2, BMP-4-, BMP-6, BMP-7,
and BMP-9 (20), which play an important role in bone
growth and development. BMPs have the ability to induce
and promote new cartilage and ossifcation at ectopic sites.
In vivo, these proteins are expressed in the cells of
developing bones, in osteoclasts, and in pathological
ossifcation induced by implanted recombinant BMPs (20,
21). BMP-2 and BMP-7 are important regulators of skeletal
tissue formation and repair (20). Gene expression of BMP-2,
BMP-4, and BMP-7 is remarkably increased in the early
period of response after bone fracture (6-8, 20-24). BMP-6
isolated from prostatic carcinoma cells, is closely associated
with osteogenic metastasis to bone (21). Since BMP-2,
BMP-4, and BMP-6 are the main ossifcation markers, they
were used in the present study to reveal histological
differentiation of ossifcation in EO.
Positive immunoreactivity for BMP-2 and BMP-4 was
observed in osteoblasts around bone and osteocyte. Lack of
reactivity for BMP-6 was observed in comparative
specimens. In the epulis specimens, positive immunoreactivity
for BMP-2, BMP-4, and BMP-6 was identifed in fbroblasts,
spindle cells, fattened cells and endothelial cells surrounding
areas of ossifcation in EO. In addition, the proportions of
these cells showing positivity for these proteins were higher
in EO than in EF. These results indicate that fbroblasts,
spindle cells and fattened cells around bone are capable of
matrix production and histologic differentiation for
ossifcation in EO. BMP-6 was not produced in large
amounts, and so did not appear to be associated with
ossifcation.
OCN, OPN and ONN are abundant non-collagenous
proteins of the normal bone matrix (10-11, 22-23). Collagen
protein is well known as a major organic substance of bone
matrix, and the remaining 10% are non-collagenous proteins,
such as OCN, OPN, and ONN. The maturation of osteoblasts
can be determined to some extent by their expression of
OCN, OPN and ONN (10-11, 25-26). OCN, the most
abundant non-collagenous protein, is distributed into normal
bone and has been shown by gene knockout technology to be
a negative regulator of ossifcation (10). OCN, OPN, and
ONN were used to clarify matrix formation.
Positive immunoreactivity for OCN, OPN and ONN
was observed in osteoblasts around bone and osteocytes in
positive comparative specimens. Concerning EO, this
immunoreactivity for OCN, OPN, and ONN was identifed
in fbroblasts, spindle cells and fattened cells around bone
and bone matrix. Contrarily, EF showed non-positive
immunoreactivity for OCN, OPN, and ONN. Positive
immunoreactivity for OCN and ONN were observed in
fbroblasts and spindle cells around bone and mature bone
matrix, whereas that for OPN was identifed in fbroblasts,
spindle cells, and fattened cells around areas of ossifcation
and bone matrix, and in some osteocytes. The fattened cells
were not positive for OCN and ONN and appeared a little
different from the fbroblasts and spindle cells. These results
suggested that the fattened cells have different biological
characteristics from fbroblasts and spindle cells. Comparison
of BMPs, OCN, OPN, and ONN immunoreactivities among
different types of ossifcation, showed no difference between
trabecular ossifcation and osteoid, and reactivity for BMPs
was higher than for OCN, OPN, and ONN in areas of
particulate and massive ossifcation. OCN showed the
lowest among the antibodies, and there was a signifcant
difference in the proportions of cells that were positive for
BMP-4 and OCN. These results suggest that BMPs operate
Fig. 9. Comparative immunopositive cell rates between EO and
EF. :EF, :EO, vertical bar: SD. * P< 0.05. Mean positive
percentages for BMPs, OCN, OPN and ONN were compared
between EF and EO. A statistically signifcant difference
between EO and EF was found in BMP-2, BMP-4, and
BMP-6. There was no positive reaction for OCN, OPN and
ONN in EF.
i
m
m
u
n
o
p
o
s
i
t
i
v
e

c
e
l
l

r
a
t
e
s

(
%
)
0
20
40
60
80
BMP-2 BMP-4 BMP-6 OCN OPN ONN
i
m
m
u
n
o
p
o
s
i
t
i
v
e

c
e
l
l

r
a
t
e
s

(
%
)
0
20
40
60
80
BMP-2 BMP-4 BMP-6 OCN OPN ONN
Fig. 10. Comparative immunopositive cell rates among different
ossifcation types in EO. : trabecular ossifcation, :
massive ossifcation, : particulate ossifcation, : osteoid,
vertical bar: SD. Mean positive percentages for BMPs, OCN,
OPN, and ONN were compared among four types of
ossifcation in EO. BMP-4 was most frequently expressed
among cells around any types of ossifcation, while OCN,
OPN, and ONN were more expressed in cells around
trabecular and osteoids.
98 Tajima Immunohistochemistry of epulis osteoplastica
in the early stage of ossifcation, that OPN and ONN are
involved growth of the bone matrix, and that OCN is related
to mature ossifcation.
The results suggest that, fbroblasts acquire ossifcation
ability with BMPs through histo- and morpho-differentiation,
and that they play a role in formation of the non-collagenous
component of the matrix, including OPN and ONN. Epulis
generally arises from the periosteum, periodontium and
gingiva. EF appears to have the potential to undergo
ossifcation, in view of the positive immunoreaction for
BMPs, but no immunoreaction for OCN, OPN, and ONN.
These fndings suggest that EO and EF arise from mixed
cells of the periodontium, periosteum and gingiva, and that
the major difference between them is that EO is composed of
a high percentage of periosteum cells.
Acknowledgments
The author would like to thank Professor Yoshiaki
Akimoto (Department of Oral Surgery) for direction and
helpful advice, as well as Professor Hirotsugu Yamamoto
(Department of Oral Pathology) for reviewing and editing
the manuscript. The author would like to thank Associate
Professor Tadahiko Utsunomiya (Department of Oral
Pathology) for his direct instruction, support and helpful
advice. Finally, the author is grateful to all the patients who
agreed to participate in the present study.
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Received December 14, 2008 Accepted January 7, 2009