r e v b r a s o r t o p . 2 0 1 5;5 0(2):232–238

www.rbo.org .br

Original Article

Effects  of  long-term  administration  of omeprazole
on bone  mineral  density  and  the mechanical
properties of the  bone�

Gabriela Rezende Yanagiharaa,∗, Aline Goulart de Paivaa, Maurílio Pacheco Netoa,b,
Larissa  Helena Torresc, Antônio Carlos Shimanod, Mário Jefferson Quirino Louzadae,
Raquel Annonia, Álvaro César de Oliveira Penonia

a Pouso Alegre School of Medical Sciences, Universidade do Vale do Sapucaí, Pouso Alegre, MG, Brazil
b Department of Clinical Pathology, School of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
c Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil
d Ribeirão Preto School of Medicine, Universidade de São Paulo, Ribeirão Preto, São Paulo, SP, Brazil
e Araçatuba School of Veterinary Medicine, Universidade Estadual Paulista Júlio Mesquita Filho, Araçatuba, São Paulo, SP, Brazil

a  r  t  i  c  l  e  i  n  f  o

Article history:

Received 6 April 2014

Accepted 19 May 2014

Available online 14 March 2015

Keywords:

Bone

Bone density

Omeprazole

Rats

a  b  s  t  r  a  c  t

Objectives: Epidemiological studies have shown a relationship between long-term use of

proton pump inhibitors and bone metabolism. However, this relationship has not yet become

established. The aim of the present study was to analyze the mechanical properties and bone

mineral density (BMD) of rats that were subjected to long-term omeprazole use.

Methods: Fifty Wistar rats weighing between 200 and 240 g were divided equally into

five groups: OMP300 (omeprazole intake at a dose of 300 �moL/kg/day); OMP200

(200 �moL/kg/day); OMP40 (40 �moL/kg/day); OMP10 (10 �moL/kg/day); and Cont (control

group; intake of dilution vehicle). The solutions were administered for 90 consecutive days.

After  the rats had been sacrificed, their BMD, the mechanical properties of the dissected

femurs  and their serum Ca++ levels were analyzed.

Results: The BMD of the OMP300 group was lower than that of the controls (p = 0.006). There

was  no difference on comparing the OMP200, OMP40 and OMP10 groups with the controls.

The  maximum strength and rigidity of the femur did not differ in the experimental groups

in  comparison with the controls. The OMP300 group had a statistically lower serum Ca++

concentration than that of the controls (p = 0.049), but the other groups did not show any

difference in relation to the controls.

Conclusion: Daily intake of 300 �moL/kg/day of omeprazole decreased the BMD of the femur,
but without changes to the rigidity and strength of the femur in adult rats.

©  2015 Sociedade Brasileira de Ortopedia e Traumatologia. Published by Elsevier Editora

Ltda. All rights reserved.

� Work developed at the Pouso Alegre School of Medical Sciences, Universidade do Vale do Sapucaí, Pouso Alegre, MG, Brazil.
∗ Corresponding author.

E-mail: gabrielayanagihara@hotmail.com (G.R. Yanagihara).
http://dx.doi.org/10.1016/j.rboe.2015.03.002
2255-4971/© 2015 Sociedade Brasileira de Ortopedia e Traumatologia. Published by Elsevier Editora Ltda. All rights reserved.

dx.doi.org/10.1016/j.rboe.2015.03.002
http://www.rbo.org.br
http://crossmark.crossref.org/dialog/?doi=10.1016/j.rboe.2015.03.002&domain=pdf
mailto:gabrielayanagihara@hotmail.com
dx.doi.org/10.1016/j.rboe.2015.03.002


r e v b r a s o r t o p . 2 0 1 5;5 0(2):232–238 233

Efeitos  da  administração em  longo  prazo  do  omeprazol  sobre  a  densidade
mineral  óssea  e  as  propriedades  mecânicas  do  osso

Palavras-chave:

Osso

Densidade óssea

Omeprazol

Ratos

r  e  s  u  m  o

Objetivos: Estudos epidemiológicos mostram uma relação entre o uso em longo prazo de

inibidores de bomba de prótons e o metabolismo ósseo, porém essa relação ainda não está

estabelecida. O objetivo deste estudo foi analisar as propriedade mecânicas e a densidade

mineral óssea (DMO) de ratos submetidos ao uso de omeprazol em longo prazo.

Métodos: Cinquenta ratos Wistar, entre 200 e 240 g, foram divididos igualmente em

cinco grupos: OMP300 (ingestão de omeprazol na dose de 300 �moL/Kg/dia), OMP200 (200

�moL/Kg/dia), OMP40 (40 �moL/Kg/dia), OMP10 (10 �moL/Kg/dia) e Cont (grupo controle;

ingestão do veículo de diluição). A administração das soluções ocorreu durante 90 dias segui-

dos. Após a eutanásia, foram analisadas a DMO, as propriedades mecânicas dos fêmures

dissecados e a dosagem de Ca++ sérico.

Resultados: A DMO do grupo OMP300 foi menor do que a do Cont (p = 0,006). Não houve

diferença  na comparação entre os grupos OMP200, OMP40 e OMP10 em relação ao Cont. A

força  máxima e rigidez do fêmur não foram diferentes nos grupos experimentais quando

comparados ao Cont. O grupo OMP300 teve concentrações séricas de Ca++ estatisticamente

menores do que o grupo Cont (p = 0,049) sem diferença entre os demais grupos em relação

ao  Cont.

Conclusão: A ingestão diária de 300 �moL/Kg/dia de omeprazol diminuiu a DMO do fêmur,

porém sem alterações na rigidez e na força do fêmur de ratos adultos.

© 2015 Sociedade Brasileira de Ortopedia e Traumatologia. Publicado por Elsevier

Editora Ltda. Todos os direitos reservados.

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ntroduction

roton pump inhibitors (PPIs) are the main drugs used
or treating diseases such as duodenal ulcers and reflux
sophagitis.1,2 Because these drugs present few adverse
ffects when administered correctly, they have come to be
sed not only for acute symptoms in clinical practice, but
lso for long-term purposes, even though such indications are
ighly debatable.3–5

PPIs act mainly toward suppressing gastric acid secretion
y the parietal cells of the stomach, since they inhibit the
nzyme H + K + ATPase, and this acid suppression may last for
p to 48 h.6

Epidemiological studies have indicated that there is
 relationship between prolonged use of PPIs and bone
etabolism,7–9 although this relationship is still not totally

stablished. Yang et al.4 reported that administration of
meprazole (20 mg/day), which is one of the most impor-
ant PPIs, is capable of significantly diminishing bone mineral
ensity (BMD). It is believed that the mechanism responsi-
le for this consists of elevation of gastric pH, which would
nterfere with calcium absorption.4,10,11 This occurs because
ome salts, such as calcium, are insoluble in basic pH and
ould therefore be absorbed less readily.8 However, a study

y Hyun et al.3 suggested that using omeprazole would
end to diminish bone reabsorption and impede the progres-
ion toward osteoporosis. Therefore, the relationship between

sing PPIs and bone demineralization and the risk of frac-
ures associated with prolonged use of omeprazole remains
nclear.9
Given that there is evidence that prolonged use of PPIs may
change the behavior of bone cells, our objective here was to
analyze the bone mineral density and mechanical properties
of rat femurs that were subjected to long-term use of omepra-
zole.

Materials  and  methods

Type  of  study

This was an experimental study using an animal model.

Animals

The procedures used in this study followed the standards
described by the Brazilian College for Animal Experimentation
(COBEA) in 1991 and the International Guiding Principles for
Biomedical Research Involving Animals12 and were approved
by the Ethics Committee for Animal Research of the University
of Vale do Sapucaí.

Fifty male adult rats of Wistar lineage, weighing 200–240 g,
were used in this study. The animals were kept under normal
environmental and temperature conditions (21 ± 2 ◦C; humid-
ity of 55–60%; and light/dark cycles of 12 h). They received
water ad libitum and feed suitable for rats. Fasting of 6 h was
maintained during the daytime period, before the daily proto-

col was started.

The rats were divided equally into five groups: (1) OMP300
– omeprazole intake at a dose of 300 �mol/kg; (2) OMP200



234  r e v b r a s o r t o p . 2 0 1 5;5 0(2):232–238

Administration of omeprazole or dilution vehicle

Start of protocol

Day 1 Day 66 Day 90

Weighing Sacrifice

n du

points was 30 mm and an accommodation time of 30 s was
used.
Fig. 1 – Experimental desig

– 200 �mol/kg; (3) OMP40 – 40 �mol/kg; (4) OMP10 – 10 �mol/kg;
and (5) control (Cont) – intake only of the dilution vehicle.

Solutions

All the experiments were performed using grade I purified
water from the Milli-Q system and using analytical grade
reagents. The drug solutions were prepared as described by
Larsson et al.13 The omeprazole granules (8.5%; Pharma Nos-
tra, Lavras, Minas Gerais, Brazil) were ground up with the
aid of a mortar and were dispersed in a vehicle containing
0.25% hydroxyethyl cellulose 4400 (Galena Química, Lavras,
Minas Gerais, Brazil) in a 0.1 M solution of sodium bicarbonate
(pH ≈ 7.4). The suspensions were prepared at concentrations
of 10, 40, 200 and 300 �mol/kg.

Experimental  protocol

Over a 90-day period, the animals of the experimental groups
received their respective doses of omeprazole and the animals
of the control group received the dilution vehicle. The doses
were administered orally by means of gavage. Fig. 1 shows
the experimental design over the course of the 90 days of the
experiment.

At the end of the 90 days of receiving the compounds, the
animals were again weighed. They were then anesthetized by
means of intramuscular injection of xylazine hydrochloride
(95 mg/kg) and ketamine hydrochloride (12 mg/kg) and sacri-
ficed by means of exsanguination, using cardiac puncture.14,15

The right and left femurs of each animal were extracted from
their body segments and the muscle and connective tissues
were totally removed. The animals and the specimens were
weighed simultaneously. After weighing and measurement,
the structures were kept in chilled saline solution until the
day of the densitometric analysis and mechanical tests (not
more than 48 h after the dissection).

Densitometric  analysis

The right femurs were subjected to analysis in a dual-emission
X-ray densitometer for small animals (Lunar® DPX Alpha).
The images were all acquired with the femurs in the same
position, immersed in water at a depth of 2 cm. The fol-

lowing imaging options were selected: appendicular; type I
bone; high-resolution mode; 76 kVp; 150 �A; collimation at
the fine adjustment; and areas of standardized size com-
prising 40 mm in width and 20 mm in length. In accordance
ring the exposure period.

with the manufacturer’s recommendations, the apparatus
was calibrated using a phantom that the manufacturer
supplied.

The analyses on the imaging examinations were performed
with the aid of the same computer software as had been used
for image  acquisition. Using a selection tool, the bones were
fully demarcated and information on their mass and area, and
consequently their mineral density, was gathered.

Mechanical  analysis

The animals’ left femurs were analyzed mechanically by
means of a three-point mechanical flexion test (Fig. 2). To per-
form these mechanical tests, a universal test machine was
used (EMIC®, model DL 10000), with a load cell of capac-
ity 500 N (certified by EMIC®), both of which belonged to
the Bioengineering Laboratory of the Ribeirão Preto School of
Medicine, University of São Paulo, along with accessories spe-
cially developed for three-point flexion tests on the bones of
small animals.16–18

The mechanical properties analyzed in the three-point
flexion tests were the maximum strength (Fmax) and rigidity
(Rig). The velocity at which the force was applied in the three-
point flexion tests was 1 mm/min. The span between the test
Fig. 2 – Three-point mechanical flexion test: (A) bone
structure; (B) apparatus for applying force. Ribeirão Preto
School of Medicine, USP.



r e v b r a s o r t o p . 2 0 1 5;5 0(2):232–238 235

Bone mineral density in the experimental groups

OMP 300

B
M

D
 (

g/
cm

2 )

OMP 200

* P=.006
OMP300 group compared with control group (Cont)

OM P 40 CONTOMP 10

0.3

0.2

0.1

0.0

Fig. 3 – Graph showing the bone mineral density results
from each experimental group. Values are expressed as
mean ± standard deviation. OMP300 (omeprazole,
300 �mol/kg), OMP200 (omeprazole 200 �mol/kg), OMP40
(omeprazole 40 �mol/kg), OMP10 (omeprazole 10 �mol/kg),
Cont (control); BMD,  bone mineral density.

S

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a

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g

Serum calcium concentration in the experimental groups

OMP 300

C
a+

+
 (m

m
ol

/L
)

OMP 200

* P=.049.
OMP300 group compared with control group (Cont)

OMP 40 OMP 10 CONT
0

1

2

3

4

5

6

Fig. 4 – Serum calcium concentration (Ca++). Values are
expressed as mean ± standard deviation. OMP300
(omeprazole, 300 �mol/kg), OMP200 (omeprazole
200 �mol/kg), OMP40 (omeprazole 40 �mol/kg), OMP10

the group that received the highest dose (OMP300) presented
erum  Ca++ assay

he blood that had been collected through cardiac puncture
as used to perform a serum calcium assay, by means of a
iagnostic kit supplied by the company Doles (Goiânia, Brazil).

After centrifugation (at 1000 g, for 5 min) of the blood col-
ected by means of cardiac puncture, the serum was collected
nd the serum calcium assays were performed by means of a
iagnostic kit supplied by the company Doles (Goiânia, Brazil).
he method was based on formation of a stained complex
etween calcium and cresolphthalein in an alkaline medium.
or the analyses, a Femto 650® spectrophotometer was used
Femto Ind. e Com. de Instrumentos Ltda., São Paulo, Brazil),
djusted to a wavelength of 570 nm.

tatistical  analysis

he statistical analysis on the data was performed using the
PSS software (SPSS, Chicago, USA), version 15.0. To test the
ormality of the sample, the Kolmogorov–Smirnov test was
sed. Because the data distribution was found to be normal,
omparisons of parameters between the control group and the
ther experimental groups were made using Student’s t-test.
o compare the animals’ weights during the experiment and
he weights of the femurs, the one-way ANOVA test was used.
he significance level of 5% (p ≤ 0.05) was used and the results
ere expressed as means ± standard deviations.

esults
uring the experiment, one animal belonging to the OMP300
roup was lost.
(omeprazole 10 �mol/kg), Cont (control).

The animals’ mean weight did not differ between the
groups on day 1 (p = 0.52), day 66 (p = 0.74) or day 90 (p = 0.45) of
the experiment (Table 1).

The bone density in the OMP300 group (0.20 ± 0.008 g/cm2)
was lower than that of the Cont group (0.22 ± 0.107 g/cm2;
p = 0.006). The other groups did not show any differ-
ences in comparison with the control group: OMP200
(0.21 ± 0.019 g/cm2; p = 0.644), OMP40 (0.21 ± 0.015 g/cm2;
p = 0.305) and OMP10 (0.21 ± 0.016 g/cm2; p = 0.410) (Fig. 3).

There were no differences in the results relating to the max-
imum strength of the femurs, in comparing the Cont group
with the other groups: OMP300 (p = 0.09); OMP200 (p = 0.55);
OMP40 (p = 0.11) and OMP10 (p = 0.62). Likewise, there were no
differences relating to the rigidity values, in comparing the
Cont group with the other groups: OMP300 (p = 0.98); OMP200
(p = 0.84); OMP40 (p = 0.08) and OMP10 (p = 0.72). The strength
and rigidity values are laid out in Table 1.

The serum concentration of Ca++ in the OMP300 group
(5.03 ± 0.39 mmol/L) was lower than that of the Cont
group (5.39 ± 0.35 mmol/L; p = 0.049). The other groups did
not show any differences in comparison with the con-
trol group: OMP200 (5.37 ± 0.26 mmol/L; p = 0.860), OMP40
(5.40 ± 0.25 mmol/L; p = 0.970) and OMP10 (5.41 ± 0.26 mmol/L;
p = 0.910) (Fig. 4).

Discussion

In the present study, the bone characteristics of rats that
received four different daily doses of omeprazole (300, 200,
40 and 10 �mol/kg/day) for 90 consecutive days were evalu-
ated. The results from this study demonstrate the influence
of the dosage of this drug on bone mineral density, given that
lower bone mineralization in comparison with the control
group (Cont). This result may have been related to the decrease



236  r e v b r a s o r t o p . 2 0 1 5;5 0(2):232–238

Table 1 – Initial, mean and final weights of the animals, maximum force (Fmax) and rigidity (Rig).

Group Initial weight (g) Mean weight (g) Final weight (g) Fmax (N) Rig (N/mm)

OMP300 196.88 ± 14.90 357.55 ± 39.61 379.44 ± 43.94 92.35 ± 11.61 154.92 ± 33.52
OMP200 213.80 ± 36.06 365.90 ± 44.71 391.70 ± 49.79 97.19 ± 12.09 152.34 ± 26.73
OMP40 223.40 ± 39.90 355.10 ± 57.83 382.10 ± 59.283 91.22 ± 14.93 130.30 ± 18.50
OMP10 221.90 ± 40.39 378.80 ± 35.04 413.60 ± 33.59 97.01 ± 15.93 149.52 ± 34.91
CONT 215.70 ± 39.20 362.30 ± 22.63 383.50 ± 35.83 99.76 ± 6.10 155.23 ± 37.24

g), OM
OMP300 (omeprazole, 300 �mol/kg), OMP200 (omeprazole 200 �mol/k
(control). Results are presented as mean ± standard deviation.

in serum Ca++ concentration in this group. However, use of
omeprazole was not seen to influence the mechanical proper-
ties of the bone tissue.

Osteoporotic fractures are considered to be a worldwide
public health problem. The mortality rate during the first year
after hip fractures is around 30%.19 It has been estimated that
in the USA, 1.5 million people every year are affected by frac-
tures relating to osteoporosis. This is an event that may reduce
quality of life and increase the risk of death.5 However, the
relationship between the risks of hip fracture and prolonged
use of PPIs remains unclear.

In a study conducted in 1993, Mizunashi et al.20 evaluated
the effect of PPI therapy on the serum levels of parathormone
(PTH) in a small group of patients with gastric ulcers (seven
men  and 12 women, with a mean age of 67 ± 13 years). The
study showed that after eight weeks of therapy with omepra-
zole, the PTH level had increased by 28% in relation to the
baseline among these patients, accompanied by increases in
osteocalcin, alkaline phosphatase and tartrate-resistant acid
phosphatase, which are markers for bone formation. Thus, it
was suggested that in that small group, there was an increase
in bone formation, contrary to the results from the present
study.

However, the urinary excretion of hydroxyproline and cal-
cium among the patients in the study by Mizunashi et al.20

decreased, which may suggest that there was less calcium
absorption. The possibility for comparison with the present
study is limited, given that this was an experimental study,
while the study by Mizunashi et al. was conducted among
humans. Moreover, PTH was only measured at one time point,
which may not reflect the dynamic effect of PTH over a 24-h
period; and also, the omeprazole dose and calcium supple-
mentation among those patients was questionable.

On the other hand, several previous studies have demon-
strated associations between PPI use and a variety of fractures.
Yang et al.4 stated that using PPIs could cause deleterious
effects in bone tissue and could increase the risk of osteo-
porotic fractures, especially after four years of continual use.
Vestergaard et al.21 corroborated the previous study and con-
firmed the relationship between PPI use and osteoporotic
fractures. Furthermore, Gray et al.22 conducted a prospective
analysis that included 161,806 women, with the aim of corre-
lating PPI use with the risk of clinical fractures. The authors
concluded that there were associations between PPI use and
occurrences of clinical fractures of the wrist, vertebrae and

forearm. The present study corroborates the previous find-
ings, given that a relationship between omeprazole use and
decreased bone mineral density was demonstrated. This can
P40 (omeprazole 40 �mol/kg), OMP10 (omeprazole 10 �mol/kg), Cont

be taken to be a predictive sign indicating that fractures may
occur as a result of long-term PPI use.

Kaye and Jick23 conducted a retrospective study in which
PPI use and the risk of hip fractures were evaluated among
patients without significant risk factors. These authors con-
cluded that patients in the age group of 50–79 years who  made
use of PPIs did not have risk factors and so would not have
higher frequency of hip fractures. In that study, the epidemi-
ological data used came from the same data source that Yang
et al.4 used in 2006. However, Yang et al.4 observed that the
risks of hip fractures increased with PPI use, especially when
used for four years or more.  Kaye and Jick took the view that
the results of Yang et al.4 referred only to patients with some
form of risk factor for hip fractures, given that they did not
exclude patients with significant risk factors.

Targowniket al.9 conducted a cross-sectional retrospective
study followed by a longitudinal phase. In the cross-sectional
study, cases of hip or lumbar vertebral osteoporosis were com-
pared with controls presenting normal bone mineral density.
In the longitudinal analysis, bone mineral density alterations
among patients who either were or were not using PPIs were
successively evaluated. The results from their study indicated
that chronic use of PPI was not associated with an increased
likelihood of occurrences of fractures. In addition, the authors
concluded that higher intensity of exposure was not associ-
ated with increased risk of presenting osteoporosis. In other
words, they believed that the risk of fractures was related to
other variables that were independent of those correlated with
osteoporosis.

Our study cannot be fully compared with that of Tar-
gowniket al.,9 since ours was experimental. Nonetheless,
it is known that in addition to the mineral characteristics
that can be assessed using DXA, bones have important vis-
coelastic properties that allow this tissue to fulfill its specific
functions.24,25 These properties are best evaluated through
experimental studies, since these enable ex vivo analyses. To
our knowledge, our study was the first to analyze the effects
of PPIs on the viscoelastic properties of the bone tissue of
animals subjected to different doses of omeprazole for a pro-
longed time.

Osteoporotic fractures are caused by diminished bone min-
eral content. When the bone mass is diminished, i.e. in
situations of osteopenia or osteoporosis, it can be expected
that the mechanical properties of the bone will be altered and
thus, that the bone will break under smaller loads.26 Following

this reasoning, Peng et al.27 analyzed different experimental
models for bone loss and changes to mechanical resistance
such that the bones only withstood smaller breaking loads,



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r e v b r a s o r t o p . 2

specially among ovariectomized female rats. It can therefore
e concluded that the bones of osteopenic female rats present
ltered mechanical properties.

Despite the high dosages of medications and the long
eriod over which the animals remained exposed to omepra-
ole intake, the mechanism of action of the PPIs did not
nfluence any of the mechanical properties of the bones in
ur study. We  believe that the highest dose stipulated or the

ength of use was insufficient to alter the mechanical proper-
ies of the bones, even though these factors diminished the
one mineralization.

We  did not determine the dose of PPIs in these animals
hat could be stipulated for simulating their use in humans.
he dose of this medication that is generally administered to
umans is 20 mg  per day, although it may reach up to 90 mg.
owever, it is not known what the equivalent dose in animals
ould be. It is known that omeprazole binds to around 95% of

he plasma in humans28 and, according to Regårdh et al.,29 the
roportion of binding would be lower in dogs and rats (90% and
7%, respectively). This suggests that a simple calculation of
ose per kilogram would not reflect the dose used in humans.
oreover, it has been seen that while the mean half-life of

his drug is 1 h in humans and dogs, half-lives in the range of
–15 min  have been recorded in rats.29

Some dose–response studies on PPIs in animals have been
onducted, but with objectives that differed from those of our
tudy. Londonget al.30 conducted a dose–response study using
he doses of 30, 60 and 90 mg/kg and a second study used a
ose of 1–10 mg/kg.31 Adhikay et al.32 used a dose of 3 mg/kg

n rats with a view to curing gastric ulcers that had been
nduced. In 2003, Melo et al.33 subjected Wistar rats to intake
.2 mg/kg, with the aim of evaluating the effect of this intake
n partial hepatectomy. Even though many  dosages have been
bserved, there is no consensus in the literature on animal
xperiments regarding the PPI dose that would be equiva-
ent to those used in humans, for the purpose of long-term
esting.

The present study was based on the work by Cui et al.,1

n which the objectives were similar to ours, i.e. to study the
ong-term effects of PPIs. Cui et al.1 treated their animals for
7 days, at a dose of 400 �mol. The highest dose of the present
tudy (300 �mol) was stipulated as slightly lower than the dose
sed by those authors, but was used for 90 days, i.e. for slightly

onger than in the study by Cui et al.1 It could be seen that
he effects were similar, since those authors also identified
ecreased bone mineral density among their animals.

In summary, the present study demonstrated that long-
erm PPI use did not alter the mechanical properties of the
emurs of adult rats. However, it was observed that the femurs
f the rats subjected to daily administration of omeprazole
t a dose of 300 �mol/kg/day developed bone demineraliza-
ion, which may suggest that this gives rise to predisposition
o bone fractures. Further experimental studies using pro-
ocols similar to ours would be useful for confirming these
esults.
onflicts  of  interest

he authors declare no conflicts of interest.
1

;5 0(2):232–238 237

Acknowledgments

Our sincere thanks to Messrs. Lúcio Ivan de Melo, José Lopes
and Marcos Martins for their assistance in caring for the
animals, and to Mrs. Nelsi Nara Vieira Marchette for her con-
tribution relating to the medications. Also to the postgraduate
student Bruna Rezende, for her help in the densitometry anal-
yses.

 e  f  e  r  e  n  c  e  s

1. Cui GL, Syversen CM, Zhao D, Chen HL. Long-term
omeprazole treatment suppresses body weight gain and bone
mineralization in young male rats. Scand J Gastroenterol.
2001;36(10):1011–5.

2. Guimarães EV, Marguet C, Camargos PAM. Tratamento da
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