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Interleukin-4 inhibits bone resorption through an effect on osteoclasts and proinflammatory cytokines in an ex vivo model of bone resorption in rheumatoid arthritis.

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ARTHRITIS & RHEUMATISM Volume 37
Number 12, December 1994, pp 1715-1722
8 1994, American College of Rheumatology
Arthritis & Rheumatism
Official Journal of the American College of Rheumatology
INTERLEUKIN-4 INHIBITS BONE RESORPTION THROUGH AN
EFFECT ON OSTEOCLASTS AND PROINFLAMMATORY CYTOKINES
IN AN EX VIVO MODEL OF BONE RESORPTION IN
RHEUMATOID ARTHRITIS
PIERRE MIOSSEC, PASCALE CHOMARAT, JULIE DECHANET, JEAN-FRANCOIS MOREAU,
JEAN-PAUL ROUX, PIERRE DELMAS, and JACQUES BANCHEREAU
Objective. To assess local bone resorption in the
context of rheumatoid synovitis and its modulation by
interleukin-4 (IL-4).
Methods. We developed an ex vivo model of bone
resorption using juxtaarticular samples of bone obtained during joint surgery. We studied the histomorphometric parameters of bone resorption and the regulation of the production of IL-6, leukemia inhibitory
factor (LIF), and the collagen cross-link pyridinoline,
which is released during bone resorption in vivo.
Results. This was a sensitive and dynamic model
of bone resorption. The bone samples produced high
levels of pyridinoline and as much cytokine as synovium
pieces obtained from the same joint. 1L-4induced a 70%
reduction of IL-6 and LIF production by bone pieces
and reduced pyridinoline levels. Histomorphometric
studies performed on bone samples indicated a 35%
Dr. Chomarat’s and Dr. Dechanet’s work was supported by
the Fondation MCrieux.
Pierre Miossec, MD, PhD: HBpital Edouard Herriot and
INSERM U80 and U234, Lyon, France; Pascale Chomarat, BSc:
Schering-PloughLaboratory for Immunological Research, Dardilly,
France; Julie Dechanet, BSc: Schering-Plough Laboratory for Immunological Research; Jean-FranGois Moreau, PhD: CNRS-URA
14563, Bordeaux, France; Jean-Paul Roux: INSERM U234, Hbpital
Edouard Herriot; Pierre Delmas, MD, PhD: INSERM U234, HBpital Edouard Heniot; Jacques Banchereau, PhD: Schering-Plough
Laboratory for Immunological Research.
Address reprint requests to Pierre Miossec, MD, PhD,
Departments of Immunology and Rheumatology, HBpital Edouard
Herriot, Lyon, France.
Submitted for publication December 6, 1993; accepted in
revised form June 21, 1994.
increase in the mean total bone area after 7 days of
treatment with IL-4. More importantly, with IL-4,
osteoclasts were not detectable in the bone sections.
Conclusion. The inhibitory effect of IL-4 on bone
resorption extends our knowledge of its antiinflammatory properties and suggests that the inflammatory
cytokine imbalance in rheumatoid synovium also contributes to defects in bone resorption in RA.
Rheumatoid arthritis (RA) is an inflammatory
joint disease in which the perpetuation of chronic
synovitis leads to bone and cartilage degradation (1).
High levels of proinflammatory cytokines such as
interleukin-1 (IL-1), tumor necrosis factor a (TNFa),
IL-6, and leukemia inhibitory factor (LIF) are produced by the synovitis and are thought to play a major
role in joint degradation (refs. 2-9 and Dechanet J et
al: unpublished observations). These factors have
been characterized as major osteoclast-activating factors, responsible for bone resorption in vitro and in
vivo (10-15).
We have previously described an ex vivo model
of RA in which synovium explants were used (3). This
model conserves the architecture of the synovium and
the dynamic activation of the cells, as reflected in the
sustained production of proinflammatory cytokines. In
the present report, samples of juxtaarticular bone
obtained during joint surgery were used to construct
an ex vivo model of local bone resorption, which
allowed the study of proinflammatory cytokine pro-
1715
1716
MIOSSEC ET AL
Table 1. Clinical parameters of the study patients*
Patient/
sexlage
RA
duration
RF
llMl67
9
2lFl44
+
22
21
15
19
15
12
16
+
+
+
+
+
3lFl64
4lFl57
5lFl.53
61F143
7lMl46
8lFl66
-
Surgical
procedure
Treatment
TKR
wrist
TKR
Wrist
TKR
wrist
MCP
MCP
DP
TP
Pred. 12 mg
MTX
Pred. 8 mg
MTX
MTX + pred. 10 mg
MTX + med. 8 mg
*
* The mean SD age of the rheumatoid arthritis (RA) patients was
55 t 10 years; the mean 2 SD duration of RA was 16 ? 4 years.
TKR = total knee replacement; DP = D-penicillamine; T P =
tiopronin; pred. = prednisone: MTX = methotrexate: MCP =
rnetacarpophalangeal joint.
duction a n d bone resorption. IL-4, a cytokine that is
not produced b y RA synovium (16), was found to
inhibit bone resorption by reducing osteoclast activation and survival, and by producing bone-resorbing
cytokines. These results extend the documented antiinflammatory properties of IL-4 observed in the synovium (3) and suggest a potential clinical application of
IL-4 in t h e control of effects o f chronic inflammation
o n bone resorption (17).
PATIENTS AND METHODS
Patients. Eight patients with RA according to the
revised criteria of the American College of Rheumatology
(formerly, the American Rheumatism Association) (18) were
recruited into this study. The characteristics of the patients
are given in Table 1. Most of the patients were receiving
nonsteroidal antiinflammatory drugs; 4 patients were taking
prednisone ( 4 1 2 mg/day). Two of the 8 patients were also
taking D-penicillamine or tiopronin, and 4 were also taking
methotrexate. Treatment with these latter drugs was stopped
1 week before surgery.
Samples of synovium and bone were obtained at the
time of joint surgery: wrist synovectomy in 3 patients, metacarpophalangeal joint replacement in 2 patients, and total knee
replacement in 3 patients. Juxtaarticular bone samples were
obtained from the site of bone resection and joint replacement.
Normal synovium was obtained from patients who
were undergoing knee arthroscopy because of ligament
symptoms. Their arthroscopy findings were normal.
Preparation and culture of bone and synovium. Bone
and synoviurn samples were obtained under sterile conditions and immediately placed in phosphate buffered saline
(PBS) containing antibiotics. Samples were cut into small
pieces of -2 mm’, and cultured in complete RPMI 1640
medium (Gibco, Grand Island, NY) supplemented with 10%
heat-inactivated fetal calf serum (Gibco), 2 mM glutamine,
100 units/ml penicillin, 50 mg/ml gentamicin, and 20 mM
HEPES buffer at 37°C in a humidified environment of 5%
CO, and 95% air. Cultures were performed in 6-well flat-
bottom plates (Falcon, Oxnard, CA) with 1 section of bone
or synovium in a final volume of 6 ml.
Cultures were incubated in the presence or absence
of 50 unitdm1 of IL-4. This concentration had previously
been determined to yield maximal inhibition of proinflammatory cytokine production by synovium (3). Recombinant
human IL-4 (rHuIL-4) purified from Escherichiu coli was
obtained from Schering-Plough Research (Bloomfield, NJ),
and had a specific biologic activity of 1 x lo7 unitslmg (19).
Supernatants were harvested after 10 days of culture (unless
indicated otherwise). In some experiments, larger sections
of bone were collected after 7 days of culture and were used
for histomorphometry.
Measurement of cytokine levels. IL-6 irnmunoreactivity was measured by double-sandwich enzyme-linked immunosorbent assay (ELISA) as described elsewhere (20). Microtiter plates (96-well) were coated with 2.5 mg/ml of mouse
39C3 anti-IL-6 monoclonal antibody (MAb) in carbonate
buffer, pH 9.8, overnight at 4°C. After washing, wells were
saturated with PBS plus 5% bovine serum albumin for 1.5
hours at room temperature. Wells were washed and serial
dilutions of rHuIL-6 standard (Genzyme, Boston, MA) or
supernatants were added for 30 minutes. IL-6 fixation was
detected with a biotinylated mouse 13A5 anti-IL-6 MAb.
After subsequent incubation with streptavidin-alkaline
phosphatase conjugate and washing, paranitrophenylphosphate (Sigma, St. Louis, MO) in diethanolamine buffer was
added for 30 minutes at 37°C. Optical density was measured
at 405 nm using an ELISA plate reader. Data were processed
by computer and are expressed as nanograms per milliliter.
LIF levels were also measured by ELISA. Mouse
anti-LIF MAb were obtained as described previously (21).
Mouse lFlO anti-human LIF MAb was used for coating, and
biotinylated mouse 7D2 anti-human LIF MAb for detection.
Data are expressed as nanograms per milliliter.
Measurement of collagen cross-link levels. Pyridinoline levels were measured using a specific ELISA (Collagen
Crosslinks kit; Metra Biosystems, Palo Alto, CA), as described (22). Samples of supernatants and standards (20 pl)
were incubated in pyridinoline-coated plates at 4°C overnight. A rabbit antipyridinoline polyclonal antibody was
added to the wells, and free pyridinoline competed with the
bound pyridinoline for the antibody. After washing, alkaline
phosphatase-conjugated goat anti-rabbit IgG was added for
detection. Data are expressed as nanomolar concentrations.
Bone histomorphometry. These studies were performed as previously described (23). Bone pieces were fixed
in 60% alcohol and embedded in methylmethacrylate. Sections (7 pm) were stained with Solochrome cyanin R and by
Goldner’s process. Osteoclasts were stained for acid phosphatase. Histomorphometric parameters were measured
with a semiautomatic image analyzer at a low power field
( x 12.5). The total bone area is expressed as the percentage
of a given bone area occupied by trabeculae and excluding
the medullary space. The number of osteoclasts is expressed
per square millimeter of bone surface and is based on
morphologic identification and on acid phosphatase staining
in approximately 40 high power fields (x200).
Statisticalanalysis. Results are expressed as the mean
2 SEM or as the percentage of control production. Differences between various treatments of samples from the same
1717
IL-4 INHIBITS EX VIVO BONE RESORPTION IN RA
joint were compared with the nonparametric Wilcoxon
paired t-test.
Table 2. Interleukin-4 (IL-4) inhibits IL-6 production by rheumatoid bone and synovium*
1L-6 production (ng/ml)
RESULTS
Spontaneous production of cytokines by RA
juxtaarticular bone pieces: an ex vivo model of bone
resorption. RA is a chronic inflammatory disease with
early juxtaarticular bone loss, in part the consequence
of an increased production of cytokines by the synovium. Bone resorption may also result from the release
of cytokines by the bone microenvironment itself. To
investigate this possibility, an ex vivo model of bone
resorption was established using bone pieces obtained
from the site of joint surgery in patients with RA.
Samples of juxtaarticular bone were cultured in
medium alone for 10 days, and cytokine levels in the
cuIture supernatants were measured. These sections
produced nanogram amounts of IL-6 (mean k SEM
250 r 59 ng/ml, range 62-573; n = 8) and of LIF (mean
5 SEM 74.8 5 37.6 ng/ml, range 4.5-258.8; n = 8).
It was important to elucidate whether the established ex vivo model would also demonstrate persistent and active cytokine production. Kinetic studies
showed that the spontaneous production of these
RAbone
+ RAsynovium
-0Normal synovium
-+3n
E"
30-
I
E
\
UI
C
v
c
0
.c
20-
0
3
'El
2
P
10-
0
2
4
Syn oviu m
Bone
6
8
10
Duration of culture (days)
Figure 1. Kinetics of the production of interleukin-6 (IL-6) by
samples of bone and synovium obtained from a patient with rheumatoid arthritis (RA), compared with that by normal synovium.
Bone and synovium samples were cultured for 10 days. Supernatants were collected at the indicated times and tested for IL-6
production by enzyme-linked immunosorbent assay. Results are
expressed in ng/ml of supernatanthg of sample.
Sample
NO IL-4
IL-4
NO IL-4
IL-4
1
251
260
116
62
96
260
513
319
53
55
62
30
70
21 1
270
164
93
140
294
121
164
96
51
31
13
2
3
4
5
6
1
8
Mean
* SEM
250
* 59
75
42
232
65
f
18t
185
4
20
119
41
154
25
71 4 22t
* Bone and synovium samples from 8 patients were cultured for 10
days in the presence or absence of IL-4 (50 unitshl). IL-6 production in supernatants was measured by enzyme-linked immunosorbent assay.
t P < 0.01 versus no IL-4.
cytokines increases over time in vitro, indicating the
presence of an active process. When compared on a
weight-to-weight basis with synovium samples from
the same joint, the kinetics and magnitude of production of IL-6 by bone samples were similar (mean 2
SEM 185 k 25 ng/ml, range 93-294; n = 8) (Figure 1).
Although the cellular composition of RA and normal
synovium is different, IL-6 production by normal
synovium was found to be 10-fold lower than that of
rheumatoid synovium, which produced medium
amounts of cytokines, when compared on a tissueweight basis. Thus, this culture system appears to be a
dynamic system which may represent an ex vivo
model of cytokine production by juxtaarticular bone in
RA synovitis.
IL-4 inhibits IL-6 and LIF production by bone
from RA patients. IL-4, a cytokine not produced by the
RA synovium, has been shown to inhibit the production of proinflammatory cytokines by samples of RA
synovium (3). Since bone degradation is the major
consequence of the increased production of these
cytokines in chronic rheumatoid synovitis, we used
the model we constructed to test how IL-4 would
regulate the production of these factors. Samples of
bone were incubated in parallel with synovial samples,
in the presence and absence of 50 unitdm1 of IL-4.
Levels of IL-6 were measured after 10 days of culture.
Table 2 shows the results obtained for each pair
of bone and synovium samples. IL-4 induced a very
profound reduction of IL-6 production by bone (mean
2 SEM 250 t 59 ng/ml without IL-4 versus 65 k 18
1718
MIOSSEC ET AL
Table 3. Interleukin-4 (IL-4) inhibits leukemia inhibitory factor
(LIF) production by rheumatoid bone*
Table 4. Interleukin-4 (IL-4) inhibits pyridinoline production by
rheumatoid bone*
LIF production (ng/ml)
Sample
NO IL-4
~
74.8
f
IL-4
Sample
NO IL-4
IL-4
6.0
11.5
12.1
2.5
1.5
37.9
6.5
110.0
1
2
3
4
5
6
7
8
52
67
115
70
165
155
99
65
60
33
60
51
70
86
66
42
~~~~
15.5
26.6
29.8
4.5
7.9
233.2
22.1
258.8
Mean 2 SEM
Pyridinoline production (nM)
37.6
23.5
f
13.0"
Mean
f
SEM
99
re_
15
59
f 6t
* Bone samples from 8 patients were cultured for 10 days in the
presence or absence of IL-4 (50 units/ml). LIF levels in supernatants
were measured by enzyme-linked immunosorbent assay.
t P < 0.01 versus no IL-4.
* Bone samples from 8 patients were cultured for 10 days in the
presence or absence of IL-4 (50 unitdml). Pyridinoline levels in
supernatants were measured by enzyme-linked immunosorbent assay.
t P < 0.02 versus no IL-4.
ng/ml with IL-4; P < 0.01). Similar results were
obtained with synovium (mean SEM 185 +- 25 ng/ml
without IL-4 versus 77 f 22 ng/ml with IL-4; P <
0.01).
Levels of LIF, a proinflammatory cytokine also
involved in bone resorption and produced by the RA
synovium (9), responded similarly. IL-4 induced a
very profound decrease in LIF production by bone
(mean _t SEM 74.8 k 37.6 ng/ml without and 23.5 -+
13.0 ng/ml with IL-4; P < 0.01) (Table 3). As shown in
the kinetic studies (Figure 2), the effect of IL-4 on LIF
production was very potent, rapid, and long-lasting.
IL-4 inhibits collagen cross-link production by
bone. Collagen cross-links such as pyridinoline are
released during bone resorption. Measurement of urinary levels of these markers has proven useful for the
clinical followup of various disorders associated with
bone degradation (22,24). In our cultures of bone
pieces, bone resorption was expected to occur, at least
in part, as a consequence of the high production of
bone-resorbing cytokines. Thus, pyridinoline levels
were measured in the supernatants using a specific
ELISA.
Pyridinoline was detectable in the supernatants
of bone, but not in those of synovium. Over 10 days of
culture, pyridinoline levels were 99 15 nM (n = 8)
(Table 4). The specificity of the measurements was
confirmed by high-performance liquid chromatography purification, which showed peaks at the expected
positions (24). These results indicate that this model
was a sensitive and dynamic model of bone resorption.
In the presence of IL-4 (50 unitsiml), a reduction in
6 nM; P <
pyridinoline levels was observed (59
0.02), as seen in Table 4.
IL-4reduces bone resorption. Bone resorption is
related to the activation of osteoclasts, which leads to
bone loss. To investigate the effect of IL-4 on these
targets, bone pieces were cultured for 7 days and
processed for routine histomorphometry (total bone
area and number of osteoclasts). Bone samples from 4
different patients (Table 5) showed a mean total bone
area of 10.7 -+ 2.0%. In the presence of IL-4, the mean
total bone area was 14.5 ? 2.0%, which is a 35%
increase. Figure 3 is an example of this effect. More
importantly, in the presence of IL-4, osteoclasts were
_+
*
L
Y
C
0
.-@
I
6wi1
I
0
4O0
&
i
&0
0
2
P
I
I
+Ik4
I
*
P
4
6
8
10
12
Duration of culture (days)
Figure 2. Kinetics of the effect of interleukin-4 (IL-4) on the production of leukemia inhibitory factor (LIF) by samples of bone from
a patient with rheumatoid arthritis (RA). Bone samples were cultured for 10 days in the presence or absence of IL-4 (50 unitshl).
Supernatants were collected at the indicated times and tested for
LIF production by enzyme-linked immunosorbent assay.
IL-4 INHIBITS EX VIVO BONE RESORPTION IN RA
Table 5. Interleukin-4 (IL-4) inhibits ex vivo bone resorption*
Bone
area
Sample
4
5
I
8
IL-4
(%I
Osteoclast
number
( x 1O-z/mm2)
-
6.0
8.8
8.7
14.4
11.3
16.6
16.9
18.1
4
0
3
0
4
0
5
0
+
+
+
+
1719
no longer detectable with either acid phosphatase
staining or cell staining (Table 5 ) .
In conclusion, this model using juxtaarticular
bone pieces allowed the study of various parameters of
bone resorption in the context of RA synovitis, which
were sensitive to the inhibitory effect of IL-4.
DISCUSSION
* Bone samples from 4 patients were cultured for 7 days in the
presence or absence of IL-4 (50 unitdml), then processed for
histomorphometric studies. Total bone area represents the % of the
area examined.
The hallmark of rheumatoid arthritis is chronic
synovitis characterized by an uncontrolled proliferation of synoviocytes together with a massive infiltration of mononuclear cells (25-27). The persistence of
the synovitis leads to cartilage and bone destruction;
A
B
Figure 3. Effect of interleukin-4 (IL-4) on histomorphometric features of bone from the metacarpophalangeal joint of a patient with rheumatoid
arthritis. Bone was cultured for 7 days in the presence (A) and absence (B)of IL-4 (50 unitshl). Samples were processed for histomorphometry
and stained with Goldner’s process. Photographs are oriented with the cartilage at the top. (Original magnification x 12.5.)
1720
removal of the synovitis with local and/or systemic
treatment protects joints from destruction. Radiography and bone densitometry studies have indicated that
the first signs of bone loss in RA occur at juxtaarticular
sites. Thus, controlling the production of the factors
involved in the perpetuation of synovitis and joint destruction is one of the major goals of treatment in RA.
Cytokines produced at the site of synovitis are
thought to play a pivotal role in the perpetuation of the
synovitis and the resulting bone destruction. In particular, large amounts of IL-1, TNFa, IL-6, and LIF,
which are potent osteoclast-activating factors, are
seen in RA synovial fluid and are actively produced by
the inflamed synovium (refs. 2, 5-9, 28, and 29, and
Dechanet J: unpublished observations). It has been
thought that the cytokines produced by the synovium
would diffuse locally from the joint space and affect
bone. Results presented in this report demonstrate
that the juxtaarticular bone itself can produce as large
a quantity of cytokines as the inflamed synovium of
the same joint. These levels were 10-fold higher than
those produced by normal synovium (on a similar
weight basis). Unfortunately, it was obviously not
possible to obtain paired bone samples. Nevertheless,
these findings suggest the presence of the same imbalance in bone and in synovium, the combination of
which leads to the increased local production of boneresorbing factors.
As for the synovium, we thought that it would
be valuable to look for potential therapeutic agents
that would block the production of proinflammatory
cytokines and, moreover, their action on the targets
related to bone destruction. We therefore developed an
ex vivo model of bone resorption in the context of RA
synovitis, in which small pieces of bone surgically removed from the joint site are cultured in medium and the
release of cytokines and other factors is measured in the
presence and absence of interfering agents. This model
has already proven useful to our understanding of cytokine production by RA synovitis (3). Immunoreactive
cytokines could be detected in culture supernatants, as
determined by ELISA. In particular, we examined the
production of IL-6 and LIF, and found these levels to
be high. These cytokines were chosen since IL-6 has
been shown to induce osteoclast development and to
be directly involved in bone resorption in an in vivo
model of osteoporosis related to estrogen loss (12).
Similarly, the very high levels of LIF produced in
these cultures extend previous results on the contribution of this cytokine to bone resorption in vivo (15).
Because RA is a chronic inflammatory disease,
MIOSSEC ET AL
it was important to elucidate whether the established
ex vivo model would also demonstrate persistent and
active production of factors involved in bone metabolism. Accordingly, kinetic studies showed that cytokines progressively accumulated in the culture supernatant in an active process. Thus, these bone pieces
represent a dynamic ex vivo model of bone metabolism in RA where the culture system supports the
integrity of the original bone architecture. This is
particularly important, since bone resorption results
from the interaction of various cell types, including the
cross-talk between osteoblasts and osteoclasts, in response to factors produced at a local and a more
diffuse level. Furthermore, this model allowed the
measurement of biologic and histologic markers that
are used routinely in the clinical followup of bone
resorption. This could represent an advantage over
isolated osteoblasts and osteoclasts, which lose some
of their differentiation markers and function in culture.
Indeed, increased levels of collagen cross-links could
be detected in the supernatants, reflecting increased
bone turnover, as has been described in the urine of
patients with various bone loss disorders (22,24). In
addition, histomorphometric measurements could be
performed to study short-term bone resorption. This
was studied by measuring the total bone area and the
number and activity of osteoclasts.
Having established an ex vivo system of bone
resorption related to RA synovitis, we wondered
whether biologic response modifiers would modulate
ongoing cytokine production and bone resorption. We
focused our attention on IL-4 since our previous
results have shown that IL-4 strongly inhibited the
secretion of proinflammatory cytokines by pieces of
RA synovium (3). The present study demonstrates that
IL-4 also strongly inhibited the production of the
proinflammatory cytokines IL-6 and LIF by bone
obtained from the same joint as the synovium. After 10
days, the production of cytokines was blocked by
approximately 70% for both IL-6 and LIF. The major
finding of these studies was the demonstration that
IL-4 was able to inhibit ex vivo bone resorption. The
decrease in proinflammatory cytokine production was
associated with a 40% decrease in the release of
collagen cross-links, which were used as an indirect
marker of ongoing bone resorption. Moreover, in the
presence of IL-4, osteoclasts were not detectable
either by morphology or by staining for acid phosphatase. Thus, the picture was compatible with an
effect of IL-4 on osteoclast activity and survival.
Taken together, these results suggest that IL-4
1721
IL-4 INHIBITS EX VIVO BONE RESORPTION IN RA
might inhibit cytokine-induced bone resorption by
interfering at the level of osteoclasts. The findings are
consistent with the inhibitory effect of mouse IL-4 on
IL-I-induced bone resorption but not parathyroid
hormone-induced bone resorption, as measured by
labeled calcium release (30). Mouse IL-4 has also been
shown to inhibit the formation of osteoclasts (3 I), and
IL-4 was shown to control joint destruction in the
streptococcal cell wall-induced model of arthritis in
the rat (32).
Juxtaarticular bone metabolism in RA can be
described in terms of the association of increased bone
degradation and decreased bone formation. The combination of the two components explains the rapid
joint destruction and bone loss that are prominent
features of RA. Osteoblasts, which are responsible for
bone formation, are mesenchymal cells with the same
origin as fibroblasts and synoviocytes. Results obtained with IL-4 treatment of synoviocytes and fibroblasts in vitro have shown the inhibitory effect of IL-4
on cytokine-induced proliferation, which contrasts
with its enhancing effect on extracellular matrix formation (33,34). Consistent with these results, IL-4 in
cultures of mouse osteoblasts was found to induce the
production of a calcified extracellular matrix, leading
to new bone formation (35). This finding suggests that
IL-4 might also increase bone formation in vivo.
Additional studies with in vivo bone labeling remain to
be performed to demonstrate this critical issue.
Since the control of bone destruction is the
most difficult target in the treatment of RA, these
results provide a new understanding of bone resorption in inflammation which might lead to new
therapeutic approaches. Similarly, postmenopausal
osteoporosis has been associated with an increased
production of proinflammatory cytokines by circulating monocytes, which was preventable with estrogen
treatment (36). Taken together, these results suggest
that the bone loss after menopause and in chronic
inflammation might result from a common mechanism,
a consequence of a pro-/antiinflammatory cytokine
imbalance (37,38).
ACKNOWLEDGMENTS
We thank J. Chiller and J. P. Revillard for their
support and advice, E. Gineyts and A. Bertholin for performing the pyridinoline ELISA, and N. Courbikre for
preparing the manuscript.
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