J. Nutr.
The
Effect
of Spiny
Lobster
in Ovariectomized
Shell
Powder
Osteoporotic
Sci.
Vitaminol.,
on Bone Model
38, 555-563,
1992
Metabolism
Rats
Naomi OMI, Naomi MORIKAWA, and Ikuko EZAWA Department of Food and Nutrition, School of Home-economics, Japan Women's University, Bunkyo-ku, Tokyo 112, Japan (Received June 9, 1992)
Summary Calcium has been found to be indispensable in the preven tion of osteoporosis. Recently, there has been a great deal of research into the best way to consume calcium. In this study, the effect of "powdered lobster shell" on bone metabolism was examined in ovariectomized osteo porotic model rats. This powder has a good flavor and taste, and contains high quantities of calcium. Six-week-old SD-strain female rats were ovariectomized and were fed a low Ca diet (0.01% Ca and 0.3% P) for 32days. Thereafter, the rats were divided into two groups; the control group was fed a control diet (0.3% Ca and 0.3% P) and an experimental group, the lobster group, was fed a lobster shell powder diet (0.3% Ca and 0.3% P) ad libitum for 30days. The results were as follows: in comparison with the control group, the lobster group had significant increases in (1) bone mineral density [BMD (DEXA Hologic's QDR -1000] of lumbar spines and tibial proximal metaphyses, which are mainly trabecular bones, and BMD of tibial diaphyses, which is a mainly cortical bone, (2) the breaking force and energy of femur. These results suggests the lobster shell powder could be a valuable source of dietary calcium in increasing BMD, breaking force and energy in osteoporotic model rats. Key Words bone mineral density, breaking force, breaking energy, spiny lobster shell powder, dietary calcium source, osteoporosis, ovariectomy Calcium has been found to be indispensable in the prevention of osteoporosis (1). In Japan, our food habits have changed significantly. We have a large variety of food in abundance. However, it has been shown that specifically the calcium intake has never been sufficient according to the national nutrition survey in 1990 conducted by the Ministry of Health and Welfare (2). Japan is becoming an aging society. It is aging more rapidly than all other countries (3). As a result, the number of patients with osteoporosis is increasing and the higher frequency of bone fractures due to osteoporosis is also increasing (4). Accordingly, there has been a great deal ofspiny research recently way to consume calcium. In this sudy, "powdered lobster shell" into was the usedbest . This powder has a good flavor and taste, 555
556
N.
and
contains
high
lobster
shell"
spiny in
osteoporotic
quantities was
model
of
an
We
source
which
had
The
experimental rats
calcium and
32days.
group
source
of
group
(n=8),
diet
of
the
control
Ca
the
The
are
30days.
in
During
cm).
The
kept
at
distilled
p.m.
and
the
constant
for
darkness
from
Biochemical deprived
of
induction
of
blood
assays food
The tometry
Table
were
level (using
1.
for
anesthesia
samples
of
time,
in the
23•}1•Ž,
remained
of
all
the These
animal
of
serum.
one
Composition
allowed
kept
in
were
ad
separate as
the
end
AA-640-12
lobster
and
follows: 50•}5% from
source
experimen for
libitum
shell
the
the
lasted
the
feeding
next and
(15•~25•~19.5
the
at
powder
lobster but
cages
lighting
of
this
temperature and
was
the
lighting
7:00a.m.
experiment,
measured
all The
samples
2,500rpm
was
shell
spiny
to
7:00
7:00a.m.).
blood (at
of spiny
were
The
Calcium
experimental
identical
(7:00p.m.-9:00a.m.).
ether,
calcium
rats
(Ca)
P.
a lobster
powder
a low
groups.
0.3%
period
maintained
At
night
centrifuged
Shimadzu's
to
two
was
was
fed
calcium
The
fed
shell
experimental
(fluorescent
7:00p.m.
with
serum
was
groups
and
diet
group
SD-strain
0.01% into
Ca
this
lobster
laboratory
humidity both
metabolism
were
(CaCO3).
The
were
of
was
of
each
spiny
and
0.3%
group," source
2.
"powdered
bone
six-week-old
divided
carbonate
of
the
were of
"lobster
1 and
study,
consisted
rats diet
content
water.
conditions
the
Calcium
calcium
this
ovariectomized diet
calcium
P.
Tables
this
In
calcium
the
composition
shown
ion-exchanged
was
the
for
METHODS
were
a control
called
0.3%
Thus,
different.
fed
diet was
and
low
whether
calcium
ovariectomized.
protocol.
Thereafter,
was
which
only.
diets
(P).
examined dietary
AND
These
The
(n=8)
0.3%
powder
tal
for
and
(n=16).
phosphorus
control
was
used
diet
0.3%
animals
were
of
been
MATERIALS
female
et al.
calcium.
effective
rats
OMI
were for
taken
15min) by
Atomic
atomic absorption
from to
the
rats
were
next
day,
after
the
aorta.
The
extract
absorption
the
serum.
spectropho
spectrophotometer).
shell powder.
J. Nutr.
Sci.
Vitaminol.
THE
Table 2.
aCa-
EFFECT
and
P -free
3.22;
•E 5H2O,
0.06;
calcium
pantothenate,
and
SHELL
0.01;
ON
BONE
U.
of
105ƒÊg
KCl,
57.7;
0.113;
METABOLISM
%):
thiamine,
nicotinamide,
2.0;
monohydrate,
cotton of
NaCl,
seed 2-methyl-1,
oil
20 .9;
CoCl2.6H2O,
(NH4)6Mo7O24•E4H2O,
(in
glucose in
%): NaF,
0.44; of 2.8;
vitamins
of ƒÀ-carotene, 525 I.
(in 0.078;
consisted
biotin,
fat-soluble
70ƒÊg
mixture
ZnSO4•E7H2O,
mixture
0.002:
salt
CuSO4•E5H2O,
vitamin
of
LOBSTER
557
Composition of the experimental diets (%).
7H2O,
B12,
OF
0.005. 0.5;
inositol,
20.0;
times
4-naphthoquinone,
week
0.01;
pyridoxine,
acid,
0.02;
received which 875ƒÊg
FeSO4•E MnSO4
water-soluble
0.5; folic
rats a
17.9;
KI, bThe
riboflavin,
73.7. cThe three
MgSO4,
0.004;
0.5; vitamin
a supplement was
supplied
with
of ƒ¿-tocopherol,
vitamin-D3.
Phosphorus was determined by the Fiske-Subbarow methods (5), and total-protein was measured by the biuret method (6). Measurement of bone mineral density. At the dissection, lumbar spines and all tibial bones were isolated. The muscles and connective tissues were carefully removed. Thereafter the bone mineral density (BMD) of the fourth and fifth lumbar vertebrae (L4, L5) were measured by dual energy X-ray absorptiometry [DEXA (Hologic's QDR-1000 X-ray bone densitometer)] as previously reported (7). Moreover, whole tibias were measured by DEXA. In comparison with the BMD of a human, the BMD of a small animal is remarkably low in density. Therefore all scans were performed using the ultra high resolution scan mode (rat mode, Version 2.0 software). And, a detector collimator with a single slit was put on the X-ray generator. Analysis of tibial BMD was practiced as follows. The tibial bones were separated into three parts. The first part (A) is the tibial proximal metaphysis, which is a mainly trabecular bone. The second part (B) is the tibial diaphysis; it is mainly a cortical bone. The third part is the tibial distal metaphysic. These are shown in Fig. 1. In this study, the BMDs of the tibial proximal metaphyses and diaphyses were examined. As well, the lumbar vertebrae were measured as Vol.
38, No.
6, 1992
558
N.
OMI
et al.
Fig. 1. Analysis of tibial bone mineral density (BMD). The first cut was made at the point at which the fibula joins the tibia. And the next cut was made at the upper 1/3. A shows the BMD of tibial proximal metaphysis. This is a trabecular bone. B shows the BMD of tibial diaphysis. This is a cortical bone. C shows the BMD of tibial distal metaphysis.
trabecular
bones.
Measurement isolated
of
and
breaking
were
Ca
the
balance
determine Urine
the
IV, was
two
centrifuged
(at
determinaton,
all
The ("Low
0.334%)
Ca and
Statistical the
control
considered
of Ca
diet," the
statistically
and
to
was
extract
ash
the
(at
dissolved
into
last
"experimental diet"
two
period;
nitric
urine
the
fecal
approximately
acid. same
of
Phase All
In for
the
days diet"
period.
supernatant.
1N
using
to for
hydrochloric
the
550-600•Ž,
measured
6N
the
diet"
out
collected
out
"experimental
"experimental
to
were
of
carried
were
1ml
was
120.0cm/min.
Urinary method
Ca as
the
serum. absorption
Ca
methods. group
ash
the
the
using
days
diaphyses
speed
was were
as
measurement
plunger
urine
carried
two
of
of
burnt
excretion
0.014%;
fecal
first
15min)
were
resulting
the
by
I was
the
the
test
femoral
speed
and
were
Then
The
studies
feces
bones
properties
the
1.0cm,
conditions
days
end for
feces'
Ca
assay
13th
the
2,500rpm,
fecal
intestinal
and
before
the
and
biochemical
12th
daily
Thereafter,
excretion
diet
the days
II,
of
chart
balance
Phase
Phase
breaking
analyzed.
the
phase,
femoral removed.
center
was
four
acidic
the
were
and
each
precipitate.
period;
III,
last
study,
under
Ca
by
space
50.0kg,
this
carefully
energy
sample
Within
collected
diet"
Phase
14h).
In
dissection,
The
and
was
absorption.
was
Ca
the
the
range
this were
tested
DYN-1255).
follows:
preventing
"Low
period;
Ca
were
force
study.
the
thus
(Iio's
load
At tissues
femurs
breaking
as
100.0mm/min,
properties. connective
the
(8)
The
conditions
24h.
of
reported
broken.
acid,
and
properties
previously were
breaking
muscles
calculated diet,"
by
using
0.344%;"
Ca
concentrations
lobster
shell
of powder
each diet,"
excretions. The
the
was "control
t-tests
lobster
were group,
used within
to
analyze each
the
differences
experiment.
between p
The
Effect
of Spiny
Lobster
in Ovariectomized
Shell
Powder
Osteoporotic
Sci.
Vitaminol.,
on Bone Model
38, 555-563,
1992
Metabolism
Rats
Naomi OMI, Naomi MORIKAWA, and Ikuko EZAWA Department of Food and Nutrition, School of Home-economics, Japan Women's University, Bunkyo-ku, Tokyo 112, Japan (Received June 9, 1992)
Summary Calcium has been found to be indispensable in the preven tion of osteoporosis. Recently, there has been a great deal of research into the best way to consume calcium. In this study, the effect of "powdered lobster shell" on bone metabolism was examined in ovariectomized osteo porotic model rats. This powder has a good flavor and taste, and contains high quantities of calcium. Six-week-old SD-strain female rats were ovariectomized and were fed a low Ca diet (0.01% Ca and 0.3% P) for 32days. Thereafter, the rats were divided into two groups; the control group was fed a control diet (0.3% Ca and 0.3% P) and an experimental group, the lobster group, was fed a lobster shell powder diet (0.3% Ca and 0.3% P) ad libitum for 30days. The results were as follows: in comparison with the control group, the lobster group had significant increases in (1) bone mineral density [BMD (DEXA Hologic's QDR -1000] of lumbar spines and tibial proximal metaphyses, which are mainly trabecular bones, and BMD of tibial diaphyses, which is a mainly cortical bone, (2) the breaking force and energy of femur. These results suggests the lobster shell powder could be a valuable source of dietary calcium in increasing BMD, breaking force and energy in osteoporotic model rats. Key Words bone mineral density, breaking force, breaking energy, spiny lobster shell powder, dietary calcium source, osteoporosis, ovariectomy Calcium has been found to be indispensable in the prevention of osteoporosis (1). In Japan, our food habits have changed significantly. We have a large variety of food in abundance. However, it has been shown that specifically the calcium intake has never been sufficient according to the national nutrition survey in 1990 conducted by the Ministry of Health and Welfare (2). Japan is becoming an aging society. It is aging more rapidly than all other countries (3). As a result, the number of patients with osteoporosis is increasing and the higher frequency of bone fractures due to osteoporosis is also increasing (4). Accordingly, there has been a great deal ofspiny research recently way to consume calcium. In this sudy, "powdered lobster shell" into was the usedbest . This powder has a good flavor and taste, 555
556
N.
and
contains
high
lobster
shell"
spiny in
osteoporotic
quantities was
model
of
an
We
source
which
had
The
experimental rats
calcium and
32days.
group
source
of
group
(n=8),
diet
of
the
control
Ca
the
The
are
30days.
in
During
cm).
The
kept
at
distilled
p.m.
and
the
constant
for
darkness
from
Biochemical deprived
of
induction
of
blood
assays food
The tometry
Table
were
level (using
1.
for
anesthesia
samples
of
time,
in the
23•}1•Ž,
remained
of
all
the These
animal
of
serum.
one
Composition
allowed
kept
in
were
ad
separate as
the
end
AA-640-12
lobster
and
follows: 50•}5% from
source
experimen for
libitum
shell
the
the
lasted
the
feeding
next and
(15•~25•~19.5
the
at
powder
lobster but
cages
lighting
of
this
temperature and
was
the
lighting
7:00a.m.
experiment,
measured
all The
samples
2,500rpm
was
shell
spiny
to
7:00
7:00a.m.).
blood (at
of spiny
were
The
Calcium
experimental
identical
(7:00p.m.-9:00a.m.).
ether,
calcium
rats
(Ca)
P.
a lobster
powder
a low
groups.
0.3%
period
maintained
At
night
centrifuged
Shimadzu's
to
two
was
was
fed
calcium
The
fed
shell
experimental
(fluorescent
7:00p.m.
with
serum
was
groups
and
diet
group
SD-strain
0.01% into
Ca
this
lobster
laboratory
humidity both
metabolism
were
(CaCO3).
The
were
of
was
of
each
spiny
and
0.3%
group," source
2.
"powdered
bone
six-week-old
divided
carbonate
of
the
were of
"lobster
1 and
study,
consisted
rats diet
content
water.
conditions
the
Calcium
calcium
this
ovariectomized diet
calcium
P.
Tables
this
In
calcium
the
composition
shown
ion-exchanged
was
the
for
METHODS
were
a control
called
0.3%
Thus,
different.
fed
diet was
and
low
whether
calcium
ovariectomized.
protocol.
Thereafter,
was
which
only.
diets
(P).
examined dietary
AND
These
The
(n=8)
0.3%
powder
tal
for
and
(n=16).
phosphorus
control
was
used
diet
0.3%
animals
were
of
been
MATERIALS
female
et al.
calcium.
effective
rats
OMI
were for
taken
15min) by
Atomic
atomic absorption
from to
the
rats
were
next
day,
after
the
aorta.
The
extract
absorption
the
serum.
spectropho
spectrophotometer).
shell powder.
J. Nutr.
Sci.
Vitaminol.
THE
Table 2.
aCa-
EFFECT
and
P -free
3.22;
•E 5H2O,
0.06;
calcium
pantothenate,
and
SHELL
0.01;
ON
BONE
U.
of
105ƒÊg
KCl,
57.7;
0.113;
METABOLISM
%):
thiamine,
nicotinamide,
2.0;
monohydrate,
cotton of
NaCl,
seed 2-methyl-1,
oil
20 .9;
CoCl2.6H2O,
(NH4)6Mo7O24•E4H2O,
(in
glucose in
%): NaF,
0.44; of 2.8;
vitamins
of ƒÀ-carotene, 525 I.
(in 0.078;
consisted
biotin,
fat-soluble
70ƒÊg
mixture
ZnSO4•E7H2O,
mixture
0.002:
salt
CuSO4•E5H2O,
vitamin
of
LOBSTER
557
Composition of the experimental diets (%).
7H2O,
B12,
OF
0.005. 0.5;
inositol,
20.0;
times
4-naphthoquinone,
week
0.01;
pyridoxine,
acid,
0.02;
received which 875ƒÊg
FeSO4•E MnSO4
water-soluble
0.5; folic
rats a
17.9;
KI, bThe
riboflavin,
73.7. cThe three
MgSO4,
0.004;
0.5; vitamin
a supplement was
supplied
with
of ƒ¿-tocopherol,
vitamin-D3.
Phosphorus was determined by the Fiske-Subbarow methods (5), and total-protein was measured by the biuret method (6). Measurement of bone mineral density. At the dissection, lumbar spines and all tibial bones were isolated. The muscles and connective tissues were carefully removed. Thereafter the bone mineral density (BMD) of the fourth and fifth lumbar vertebrae (L4, L5) were measured by dual energy X-ray absorptiometry [DEXA (Hologic's QDR-1000 X-ray bone densitometer)] as previously reported (7). Moreover, whole tibias were measured by DEXA. In comparison with the BMD of a human, the BMD of a small animal is remarkably low in density. Therefore all scans were performed using the ultra high resolution scan mode (rat mode, Version 2.0 software). And, a detector collimator with a single slit was put on the X-ray generator. Analysis of tibial BMD was practiced as follows. The tibial bones were separated into three parts. The first part (A) is the tibial proximal metaphysis, which is a mainly trabecular bone. The second part (B) is the tibial diaphysis; it is mainly a cortical bone. The third part is the tibial distal metaphysic. These are shown in Fig. 1. In this study, the BMDs of the tibial proximal metaphyses and diaphyses were examined. As well, the lumbar vertebrae were measured as Vol.
38, No.
6, 1992
558
N.
OMI
et al.
Fig. 1. Analysis of tibial bone mineral density (BMD). The first cut was made at the point at which the fibula joins the tibia. And the next cut was made at the upper 1/3. A shows the BMD of tibial proximal metaphysis. This is a trabecular bone. B shows the BMD of tibial diaphysis. This is a cortical bone. C shows the BMD of tibial distal metaphysis.
trabecular
bones.
Measurement isolated
of
and
breaking
were
Ca
the
balance
determine Urine
the
IV, was
two
centrifuged
(at
determinaton,
all
The ("Low
0.334%)
Ca and
Statistical the
control
considered
of Ca
diet," the
statistically
and
to
was
extract
ash
the
(at
dissolved
into
last
"experimental diet"
two
period;
nitric
urine
the
fecal
approximately
acid. same
of
Phase All
In for
the
days diet"
period.
supernatant.
1N
using
to for
hydrochloric
the
550-600•Ž,
measured
6N
the
diet"
out
collected
out
"experimental
"experimental
to
were
of
carried
were
1ml
was
120.0cm/min.
Urinary method
Ca as
the
serum. absorption
Ca
methods. group
ash
the
the
using
days
diaphyses
speed
was were
as
measurement
plunger
urine
carried
two
of
of
burnt
excretion
0.014%;
fecal
first
15min)
were
resulting
the
by
I was
the
the
test
femoral
speed
and
were
Then
The
studies
feces
bones
properties
the
1.0cm,
conditions
days
end for
feces'
Ca
assay
13th
the
2,500rpm,
fecal
intestinal
and
before
the
and
biochemical
12th
daily
Thereafter,
excretion
diet
the days
II,
of
chart
balance
Phase
Phase
breaking
analyzed.
the
phase,
femoral removed.
center
was
four
acidic
the
were
and
each
precipitate.
period;
III,
last
study,
under
Ca
by
space
50.0kg,
this
carefully
energy
sample
Within
collected
diet"
Phase
14h).
In
dissection,
The
and
was
absorption.
was
Ca
the
the
range
this were
tested
DYN-1255).
follows:
preventing
"Low
period;
Ca
were
force
study.
the
thus
(Iio's
load
At tissues
femurs
breaking
as
100.0mm/min,
properties. connective
the
(8)
The
conditions
24h.
of
reported
broken.
acid,
and
properties
previously were
breaking
muscles
calculated diet,"
by
using
0.344%;"
Ca
concentrations
lobster
shell
of powder
each diet,"
excretions. The
the
was "control
t-tests
lobster
were group,
used within
to
analyze each
the
differences
experiment.
between p
