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DOI 10.1007/s00774-004-0524-0
Springer-Verlag 2004
Department of Clinical Sciences Colorado State University, 300 West Drake Road, Fort Collins, CO 80523, USA
Independent Consultant, Fort Collins, CO, USA
Introduction
Osteoporosis is a common and severe metabolic
disease, such that the lifetime risk of having an
Offprint requests to: J.M. MacLeay
(e-mail: jmacleay@colostate.edu)
Received: January 13, 2004 / Accepted: April 19, 2004
osteoporotic-related fracture is close to 40% in postmenopausal women residing within the United States
[1]. Osteoporosis is a multifactorial disorder, being
influenced by lifestyle, life-stage, genetic, and dietary
factors. While all factors influencing the development
of osteoporosis are important, dietary acid has gained
attention recently. Several authors have implicated a
dietary-induced metabolic acidosis as a contributing
factor in the development of osteoporosis [25].
Metabolic acidosis induces increased calcium excretion
without an adequate compensatory increase in calcium
absorption from the gut, and therefore an overall decrease in total body calcium ensues. This is different
from the rat model, where there appears to be a concomitant decrease in intestinal calcium excretion to
compensate for increased urinary calcium loss in response to metabolic acidosis [6]. Therefore, dietaryinduced metabolic acidosis, in combination with other
factors, appears to have a serious impact on the development of osteoporosis for susceptible individuals.
Osteopenia induced by metabolic acidosis is typically
thought of in the context of pathologic conditions such
as chronic renal insufficiency; however, a relative excess
of strong anions compared to strong cations in the diet
may also induce metabolic acidosis in the face of normal
renal function. This is because strong ions are absorbed
directly from the intestine and therefore immediately
impact acid-base balance. Depending on the composition of the diet, the imbalance may be slight or substantial, leading to a corresponding degree of metabolic
acidosis [7]. Overall blood pH is maintained within the
normal physiologic range due to the combined effect of
renal excretion of excess ions and mobilization of calcium from bone to act as a buffer. The influence of diet
on acid-base balance is accentuated in older human
patients, where decreased or impaired renal function is
common. However, the effect of diet on inducing metabolic acidosis and calciuria in individuals with normal
renal function is well documented [4].
562
563
Diet
Compositions
Estimated amount
consumed/ day per
sheep at 3.3% of
body weight/day
MA
Grain mix
Grass hay
Grass hay
1.26 kg
1.0 kg
2.7 kg
ND
No. of
kcal offered/
day per sheep
(total)
1900
1100 (3000)
2970
765
308
308
Dietary cation-anion
difference of total diet
consumed/ day per sheep
if consumed between 2600
and 3300 kcal/day
150175 mEq
9001100 mEq
MA, low dietary cation-anion difference; ND, normal dietary cation-anion difference; DM, dry matter
Unknown
izing urine and serum values for calcium and phosphorus to urine and serum values for creatinine.
Statistical analysis
0.26%
0.18%
27
6
Unknown
14.5
3.5
5.24
6
0.5a
3.6
Sodium (g)
Sulfur (g)
Chloride (g)
0.18%
0.38%
0.8%
8
4
3.6
5.5
4
2.8
31
45
16
0 Days
180 Days
1.200
1.150
1.100
BMD g/cm2
0.82%
304
208
122
8
11
2.7
MA
ND
Sheep nutrient
requirements
for 80 kg BW
Sheep nutrient
requirements
% of diet dry
matter
Calcium (g)
Phosphorus (g)
Magnesium (g)
Potassium (g)
Results
Adjusted
crude
Protein (g)
Table 2. Major mineral content for each diet, assuming sheep on the MA and ND diets consumed the offered amounts listed in Table 1. Sheep nutrient requirements are
offered for comparison. Sheep nutrient requirements listed are those established by the Subcommittee on Sheep Nutrition, Committee on Animal Nutrition, Board of
Agriculture, National Research Council, and published as Nutrient requirements of sheep. 6th Edn; 1985, by National Academy Press
564
1.050
1.000
0.950
0.900
0.850
0.800
NoOVX ND
OVX ND
No OVX MA
OVX MA
Fig. 1. Mean bone mineral density (BMD [g/cm2]) for the last
four lumbar vertebrae, as measured by dual-energy X-ray
absorptiometry (DEXA) for each group of sheep on day 0 and
day 180 (July to January). Standard deviation bars are shown.
OVX, ovariectomy; ND, control diet; MA, metabolic acidosis
induction diet. Unlike letters are significantly (P 0.05) different from each other
565
Mean Fractional Excretion of Phosphorus (%)
18.000
Day 90 BAP
16.000
16.000
14.000
FE P Day 90
14.000
12.000
12.000
10.000
10.000
8.000
8.000
6.000
6.000
FE P Day 0
18.000
20.000
FE P Day 180
4.000
4.000
2.000
2.000
0.000
0.000
No OVX ND
No OVX ND
OVX ND
No OVX MA
OVX ND
No OVX MA
OVX MA
OVX MA
Fig. 2. Serum bone alkaline phosphatase (BAP [U/l]), as measured on days 0, 90, and 180 for each group of sheep. Standard
deviation bars are shown. OVX, ovariectomy; ND, control
diet; MA, metabolic acidosis induction diet. There was no
significant influence of time, ovariectomy status, or dietary
treatment on BAP
12.000
FE Ca Day 0
Day 90
14
Day 180
12
FE Ca Day 90
10.000
FE Ca Day 180
8.000
10
6.000
8
4.000
6
4
2.000
0.000
0
No OVX ND
NoOVX ND
OVX ND
noOVX MA
OVX ND
No OVX MA
OVX MA
OVX MA
Discussion
The sheep has been shown to be an adequate animal
model to study estrogen depletion and the onset of
566
7.56
7.54
7.52
7.50
7.48
7.46
7.44
7.42
7.40
7.38
noOVX ND
OVX ND
noOVX MA
OVX MA
BAP during winter months [28]. As significant differences were not found, it is important not to overinterpret apparent trends in the BAP and DPD results.
The variance may also reflect the relatively small group
size of this study. Neither serum BAP nor urine DPD
appeared to adequately reflect the changes in BMD that
were observed by DEXA, and as such may not be ideal
markers for use in the ovine model.
Urinary fractional excretions of calcium and phosphorus did increase and were found to be significantly
associated with the MA diet and, therefore, may be
better indicators of calcium and phosphorus balance
within the body. Whether the mechanism by which calcium and phosphorus were liberated from bone was
primarily physicochemical or cell-mediated has yet to
be determined and is an area of future research. Other
markers of bone turnover, such as osteocalcin, parathyroid hormone (PTH), and vitamin D were not examined
in this study and may be determined to be useful markers in future studies.
One other drawback of this study was the relatively
larger proportion of sodium in the MA diet compared
to the control diet. The role of sodium in calcium wasting is controversial [29,30]. Some studies have found
that increased salt intake is associated with calciuria.
However, in these studies it is unclear whether the
overall diet induced metabolic acidosis or not. Other
researchers argue that, as sodium intake is generally
consumed as sodium chloride, that it is the other cations
and anions in the diet that are more important influencers of acid-base balance and, therefore, calciuria is
more important than actual NaCl intake [31]. Further
studies are necessary to settle this controversy.
It is difficult in human studies to examine the effect of
diet on bone metabolism without considering other confounders. Using this ovine animal model, we were able
to examine the influence of diet and ovariectomy status
in a similarly treated, housed, and aged population. We
found that diet was a potent influence on BMD. We did
not see strong evidence for a synergistic effect between
diet and ovariectomy status, but one limit of this study
was its short duration of 180 days. Previous studies in
the sheep typically required at least 12 months to witness a significant decline in BMD of the lumbar spine in
the face of estrogen depletion alone [27]. Nevertheless,
the potent influence of diet on BMD in a large animal
model has heretofore not been described in the literature. It is interesting to note that the calcium content
of both diets was in excess of minimum requirements
for this species and considerably higher than that recommended for humans, and yet significant calcium
wasting still occurred. Therefore the mechanism by
which dietary metabolic acidosis causes calcium wasting
appears to be at least partially independent of calcium
intake.
567
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