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EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION 
AND ROTATIONAL STRESS ON ADVEOLAR 
BONE LOSS IN RICE RATS 



OTIC 

ELECTE 
NOV 101993 



M. E. COHEN 

D. M. MEYER 


its ! 


93-27632 

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Naval Medical Research and Development Command 

Bethesda, Maryland 

















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NAVAL DENTAL RESEARCH INSTITUTE 
BUILDING 1-H 

GREAT LAKES, ILLINOIS 60088-5259 


EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION 
AND ROTATIONAL STRESS ON ALVEOLAR 
BONE LOSS IN RICE RATS 


M. E. COHEN 


D. M. MEYER 

Research Progress Report NDRI-PR 93-03 
Work Unit M0095.003 - 0003 
Naval Medical Research and DeveJ cpmcnt Command 
National Naval Medical Center 
8901 Wisconsin Avenue 
Bethesda, Maryland 20889-5606 


The opinions expressed herein are those of the authors and cannot 
be construed as reflecting the views of the Navy Department or 
the Naval Service at large. The use of commercially available 
products does not imply endorsement of these products or 
preference to other similar products on the market. 

This document has been approved for public release; its 
distribution is unlimited. 


Approved and released by: 


1 




S. A. RALLS 
Captain, Dental Corps 
United States Navy 
Commanding Officer 


DTIC QUALITY INSPECTED 5 


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Printed in Great Britain Pergamon Press l td 


EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION 
AND ROTATIONAL STRESS ON ALVEOLAR BONE LOSS 

IN RICE RATS 

M. E. Cohkn and D. M. Mi yir 

Naval Denial Research Institute. Building l-H. Great Lakes. IL 60088, U S A 
(Accepted 0 February /W.?j 


Summary The effect of this supplementation on hone loss (distance from the cementum enamel junction 
to the alveolar crest measured at the midhne of the lingual aspect of each of the mandibular molar roots) 
was studied in rats that were either nm stressed or stressed on a rotational device for 00 days. In the first 
study, neither vitamin F. nor stress condition had statistically significant effects but there was substantial 
bone loss and bone-loss variability in all groups Before the start of the second study, to reduce differences 
in bone loss that might otherwise exist before introduction of the treatments, rats received an antibiotic 
in their drinking water In addition, rotational stress was introduced more abruptly than in the first study 
to reduce the likelihood of adaptation Bone loss and bone-loss variability were substantially reduced in 
the second study. Analysis of these data indicated that vitamin E supplementation had a statistically 
significant protective effect, which was most pronounced at sites most susceptible to loss. Stressed subjects 
tended to lose more bone, but this effect was not significant. These findings suggest some role for vitamin 
E supplementation in the maintenance of periodontal health but also a sensitivity in this effect to initial 
periodontal status. 

Key words: alveolar bone loss, vitamin F.. stress, psychological. 


INTRODUCTION 

Vitamin E functions as a free-radical scavenger that 
inhibits lipid peroxidation and inflammation, pro¬ 
tects ischaemic and hypoxic tissues, and is immuno- 
stimulating (Crary, 1984). Because of this spectrum of 
activities, the relation between vitamin E and peri¬ 
odontal health and disease has been studied; findings 
tend to support a therapeutic role for the vitamin but 
the evidence is inconsistent. 

Periodontal tissues of albino rats were not affected 
by long-term vitamin E deficiency despite early Euro¬ 
pean reports of successful treatment of periodontitis 
with this substance in man (Nelson and Chaudhry, 
1966). However, vitamin E deficiency had negative 
impact on periodontal health in rats in another study 
(Schneider and Pose, 1969). Ligature-induced peri¬ 
odontitis in rats was not affected by vitamin E 
supplementation (Parrish, DeMarco and Bissada. 
1977). but careful inspection of the data suggests that 
supplementation resulted in greater numbers of 
inflammatory cells with, simultaneously, less alveolar 
bone loss. This would be consistent with vitamin E’s 
immunostimulant and antioxidant properties, even 
though sample sizes and methods of analysis did not 
have sufficient power to detect these shifts at statisti¬ 
cally significant levels. Dietary vitamin E supplemen¬ 
tation was shown to accelerate gingival wound 
healing in albino rats (Kim and Shklar. 1983). 

In man, greater dietary intake of vitamin E has 
been associated with fewer reported oral symptoms 


Abbreviations: AN(AC)OVA, analysis of (co-(variance; 
CEJ, cementum-enamel junction. 


(Cheraskin and Ringsdorf. 1970), but circulating 
concentrations of the vitamin were the same in 
patients with and without periodontitis (Slade et ul.. 
1976). Patients with periodontal disease who were 
given vitamin E daily for 21 days to swish in their 
mouths and swallow exhibited a significant decrease 
in fluid flow from the gingival sulcus than in controls 
with disease but no vitamin, E supplementation 
(Goodson and Bowles. 1973). Also, subjects given 
vitamin E supplementation for 12 weeks exhibited a 
reduction in Russell's Periodontal Index (Cerna el al., 
1984). However, topical vitamin E did not reduce 
gingivitis over a 4-week period relative to a placebo 
(Cohen et al., 1991). but the method appeared insen¬ 
sitive in that chlorhexidine similarly had non-signifi 
cant effects on gingivitis. 

Lack of consistent benefits associated with vitamin 
E may be due, in part, to the absence of physiological 
stress manipulation. During stress, vitamin E stores 
are depleted and. once depleted, tissues are at greater 
risk. Without stress, vitamin E may remain at effec¬ 
tive concentrations so that there may be no relation 
between it and periodontal destruction. One may 
argue that in man. insufficient vitamin E and a 
subsequent increase in the likelihood of periodontal 
destruction occur during brief intervals associated 
with stressful life-events. Therefore, the cross-sec¬ 
tional analysis of Slade el al. (1976) might not identify 
a relation. 

A further problem with many rat studies is that this 
species is not usually susceptible to periodontal de¬ 
struction, except when this is ligature induced or is 
the result of other extraordinary interventions. It 
would be difficult, therefore, to observe a beneficial 


601 




602 


M E Cohen and D M Meyer 


effect of vitamin E unless it is superimposed on an 
adverse initial state such as healing of experimental 
gingival wounds, vitamin E deprivation (as con¬ 
trasted to supplementation), or stress. 

Effects of stress on periodontal destruction have 
been documented and are consistent with generally 
held concepts of the relation between stress and 
disease resistance (Troxler, 1086; Dantzer and Kelley, 
1989; Ballieux, 1991). In a series of experiments on 
rodents, increased rates of periodontal destruction 
(Ratcliff, 1956; Fedi Jr, 1958; Shklar and Glickman, 
1959; Gupta. Blechman and Stahl, 1960; Gupta, 
1966) and decreased rates of gingival wound healing 
(Stahl, 1961) were shown to accompany the presence 
of stressful stimuli. While similar experimental de¬ 
signs are not possible in man. there is evidence that 
the same relations hold. Early anecdotal reports 
(Moulton, Ewn and Thiemman, 1952) have been 
supported by correlational studies relating life-stress 
events to rates of periodontal destruction (Manhold, 
Doyle and Weisinger, 1971; Haskell, 1975; Green 
et at., 1986). 

Although stress will affect a variety of physio¬ 
logical processes, activation of the sympathetic- 
adrenal-medullary and hypothalamic pituitary- 
adrenocortical systems (Axelrod and Reisine, 1984) 
appears particularly important in terms of defining 
explanatory mechanisms for enhancement of disease 
progression. Exposure of animals to stressful stimuli 
is accompanied by increases in the synthesis, storage 
and release into the circulation of catecholamines and 
glucocorticoids. The effects of these events and sub¬ 
sequent enzymatic shifts, as indexed by a variety of 
measures, can include: salivary and small-vessel vaso¬ 
constriction, release of cytotoxic free radicals, in¬ 
creased membrane permeability, impaired wound 
healing, decreased resistance to infection, and sup¬ 
pressed cellular immune responses (Matheny, 1988; 
Weiss et at.. 1989). 

In addition to studies relating stress to periodontal 
health, there are others in which stress-related 
hormones have been measured or manipulated di¬ 
rectly. Administration of cortisone caused osteoporo¬ 
sis of alveolar bone in mice (Glickman, Stone and 
Chawla, 1953) and patients with acute necrotizing 
ulcerative gingivitis were found to have higher levels 
of overnight urinary cortisol (Cohen-Cole et at., 
1981). Catecholamines have been shown to reduce 
gingival circulation (Ito et at.. 1973; Clarke, Shephard 
and Hirsch. 1981) and to enhance the virulence of 
gingival crevicular bacteria (Courant and Gibbons, 
1966). 

The presence of oxygen radicals has been sug¬ 
gested as playing a central part in tissue damage 
associated with chronic inflammation (in general) 
and periodontal disease (Hoffeld, 1982). Based on 
vitamin E's ability to protect tissue from oxidative 
damage, Goodson (1975) predicts therapeutic value 
but recognizes that there has been an insufficient 
number of studies in this area. Hence we have 
now sought to evaluate the ability of vitamin E 
supplementation to prevent periodontal destruc¬ 
tion in a species inherently susceptible to such de¬ 
struction. and while exposed to environmental stress 
that could affect the supplement’s physiological 
value. 


MATERIALS AND METHODS 
Study I 

Rice rats (Oryzomys palustris ) were chosen because 
of susceptibility to periodontal destruction (Leonard. 
1979), The study (protocol reviewed by a Laboratory 
Animal Use Committee) had a two-by-two factorial 
design with two levels of dietary vitamin E sup¬ 
plementation and either a control or high-stress en- 
wronment. 

Thirty-two male rice rats (57-84 days of age) were 
switched from standard rodent diet to a synthetic test 
diet (modified Purina Basal Test Diet 5755). This diet 
contains 44% dextrin, 21% casein, 15% sucrose, 5% 
lard, 5% corn oil and the remaining percentage is 
comprised of other necessary additives. One half of 
the animals received a synthetic control diet, which 
included a standard 50 IU of vitamin E per kg feed 
(35 IU attributable to d-oc -tocopherol acetate oil 
supplement and approximately 15 IU due to other 
components, principally corn oil). The remaining 
animals received a synthetic test diet that contained 
5000 IU of the vitamin per kg feed (4985 IU due to 
the tocopheiol additive). This lU/kg dose (60 lU/day 
based on a 12 g daily food ingestion) had been used in 
a similar long-term study of periodontal destruction 
in rats, without reported side-effects (Parrish et at., 
1977). In general, vitamin E is not considered toxic, 
mutagenic, carcinogenic, or teratogenic, even at high 
doses (Bendich and Machlin, 1988). 

After 35 days of feeding on the synthetic diets, rats 
were assigned to normal or high-stress environments. 
Assignment to both diet and stress conditions in¬ 
volved grouping animals by weight and then ran¬ 
domly assigning them within groups to the various 
treatment conditions. 

All rats were housed individually in plastic cages 
(standard multi-mouse cages, approx. 7 in. 
wide x 12 in. long x 5 in. high) with compressed, 
shreddable wood shavings as bedding material. All 
groups were housed in the same quiet room with daily 
12-h light/12-h dark cycles. Non-stressed animals 
were housed in standard racks, with diet groups in 
counterbalanced order relative to location. 

Profoundly painful or exhausting stimuli were n't 
used as stressors. Rather, rats were subjected to cage 
rotation. This method of stress induction is con¬ 
sidered relatively benign but nevertheless is associated 
with shifts in physiological status (Shipov et at.. 1985) 
including circulating concentrations of stress-related 
hormones (Riley, 1981). Stressed animals were 
housed in cages mounted in a star pattern on a large 
platter that was rotated by a quiet electric motor 
controlled by a computerized timing device. The 
platter was approx. 120 cm in diameter and the radius 
of a circle formed by the outer edges of the cages was 
55 cm. Two of these devices (henceforth called 
wheels), each holding eight subjects (four from each 
of the two diet groups), were used. 

Immediately after their assignment to high-stress 
conditions, these animals were subjected to a I-min 
period of 30 rev/min rotation once every 4 h. When 
wheels were spinning, rats preferred to place them¬ 
selves at the far end of their cages, approx. 50 cm 
(radius, r) from the cenUc of the wheel. Therefore, 30 
rev/min corresponds to a gravitational (jj) force of 0.5 




Effects of vitamin E amt stress on hone loss 


Mil 


[where a is acceleration and v is velocity. g = a 980; 
ti = r : r; r = ( 2 nr) (rev min 60 *)]. 

Each wheel was rotated on an independent, con¬ 
tinuous (24 h day) variable-interval schedule (i.e. the 
location of the I-min rotational period within the 4-h 
time interval was random). As rotational stress may 
reduce feeding and cause weight loss, subjects were 
weighed at least once per week. A protocol for 
temporary termination of rotation because of 
catastrophic weight loss was in place but never 
needed to be implemented. 

Over a period of approx. 2 months, rotations were 
increased from the initial 1 min at 30 rev min every 
4 h to 15 min (in a single segment) of 42 rev, min 
(1.0 g) every 30 min. Ninety days after initiation of 
stress manipulations, animals were anaesthetized, or¬ 
bital blood samples drawn to assay for vitamin E, and 
the animals killed by nitrogen asphyxiation. 

Jaws were dissected out and defleshed by boiling. 
Mandibles were dried and mounted on modelling clay 
on small dishes, and lingual alveolar bone loss was 
evaluated using a standard approach (Keyes and 
Gold, 1955; Gupta and Shaw, 1956). Seven measure¬ 
ments taken on each jaw side reflected the distance 
from the CEJ to the alveolar bone crest at the midline 
of each root (three roots for the first molar and tw'o 
roots each for the second and third molars). Measure¬ 
ments were made through a dissecting microscope 
fitted with an optical caliper eyepiece. This method 
yielded a measurement precision of approx. 0.01 mm 
and allowed for minor adjustments in light intensity 
and angulation to improve visualization. Neverthe¬ 
less, the CEJ was sometimes difficult to visualize, even 
after application of eosin stain. In such cases, an 
approximation was made based on other anatomical 
landmarks. To decrease variability and to preclude a 
potential for bias, these measurements were made by 
one person who was blind as to experimental con¬ 
dition. The primary dependent variable of interest 
was the CEJ-bone crest distance averaged across 14 
sites (seven sites on each of two sides). 


RESULTS 
Study I 

One animal in the low vitamin E, high-stress group 
died during the course of the study and was not 
included in the analysis. ANACOVA was made on 
body weight. The covariate was weight on stress day 
0 , the dependent variable was weight at study termin¬ 
ation, and the between-subjects variables were stress 
and vitamin condition. Over the 90 days, rats gained 
6.0 g (from a mean weight of 87.6 g to one of 93.6 g). 
but there was no effect of the independent variables, 
or their interaction, on the covariate adjusted 
weights. 

Of 31 blood samples drawn for vitamin E assay, 
one sample was lost and six were combined into three 
sets of pairs (within treatment groups) to achieve a 
minimum assay quantity. ANOVA with the between- 
subject factors of dietary vitamin E and stress re¬ 
vealed only a significant effect for diet condition 
(F = 222.99, d.f. = 1, 23, p <0.001). Animals given 
high div.U..j vitamin E had more than four times the 


Table I Mean distance, in mm, and SD in parentheses from 
CEJ lo alveolar crest as a function of dietary vitamin E and 
rotational stress in Study I 



Dietary 

Standard 

vitamin E 

Supplemented 

Total 

Stress 

No 

0 711 (0 160) 

0.757 (0.239) 

0.724(0 197) 

Yes 

0.806(0.087) 

0 833(0.260) 

0.820(0.240) 

Total 

0.756(0.197) 

0.785(0.246) 

0.771 (0.221) 


circulating vitamin E than those given low amounts 
(10.55 versus 2.44 mg l). 

Table 1 shows mean alveolar bone loss for the four 
groups. Effects of diet and stress level did not ap¬ 
proach statistical significance (ANOVA Ts = 0.10, 
1.37 and 0.00. d.f. = I. 27. for vitamin E stress, and 
interaction, respectively). 


MATERIALS AND METHODS 

Study 2 

As described above, there was high variability in 
bone loss within ail groups in Study I. This may be 
attributed to animals being 57-84 days old before the 
diet variable was introduced and 92-119 days old 
before the stress variable. Rice rats can have substan¬ 
tial bone loss before these ages (Leonard, 1979) and 
this ‘noise’ may have overwhelmed the diet and stress 
effects. 

To evaluate this possibility, the study was repeated 
with two changes. First, starting from 18 to 31 days 
post-weaning, rats received 1.35 g/1 tetracycline 
hydrochloride in their drinking water (Vetquamycin- 
3241 m phizer. 1.902 g/1) on an approx. 2 weeks on/2 
weeks off schedule. Antibiotics substantially reduce 
bone loss in rice rats (Gupta, Auskaps and Shaw, 
1957) and so should reduce pre-study, between-sub- 
ject variability. Based on estimated daily water intake 
of 6.4 ml for an 80-g rat, tetracycline intake was 
(1.35 mg/ml)(6.4 ml) = 8.64 mg. and dosage was 
8.64 mg/0.08 kg = 108 mg/kg per day. However, giv¬ 
ing the antibiotic orally can reduce absorption by 
more than 50% (Plumb, 1991). Recommended oral 
dosages of tetracycline are about 33- llOmg/kg per 
day (Huber, 1982; Kirk, 1992; Plumb. 1991) and 
5 mg/ml (Kohn and Barthold, 1984). In general, 
tetracycline is considered relatively non-toxic, with 
oral doses of 75-465 mg/kg per day for 8 weeks being 
well tolerated by dogs without evidence of toxicity 
(Huber, 1982). Possible interaction between tetra¬ 
cycline (toxicity) and Vitamin E supplementation was 
considered unreasonable given the clinical acceptance 
of these concentrations, the lack of observed toxic 
effects, and the ending of antibiotic supplementation 
1 week before the introduction of test diets. 

Second, in Study 1 the stress had been instituted 
gradually over a period of 2 months This may have 
attenuated the stress variable by allowing animals to 
adapt. Therefore, in Study 2 the rotational schedule 
reached the end-stage of 15 min rotation at 42 
rev/min per 30 min after 3 days. 

The rats in Study 2 were between 76 and 97 days 
old when they were switched to the synthetic diets 


AOB 38 7— 



604 


M. E Cohen and D M MtvtR 


Table 2. Mean distance, in mm. and SD in parentheses from 
CEJ to alveolar crest as a function of dietary vitamin E and 
rotational stress in Study 2 



Dietary vitamin E 



Standard 

Supplemented 

Total 

Stress 

No 

Yes 

Total 

0.420 (0.082) 
0451 (0.102) 
0.436 (0.091) 

0.359 (0.044) 
0.379(0.023) 
0.369 (0.036) 

0.391 (0 072) 
0417(0083) 
0.404(0.077) 

(antibiotics had been 

discontinued 

1 week pre- 


viously), the stress manipulation was instituted 35 
days later, and again they were killed after 90 days. 


vitamin-stress interaction (/'=0.04, d.f. = 1. 26. 
p = 0.850). A second ANOVA was done with the 
tooth root at which measurements were taken (seven 
levels corresponding to mesial of the first molar 
through distal of the third molar, averaged across the 
left and right sides) added as a withm-subject vari¬ 
able The effect of ‘root’ was statistically significant 
(F = 224.?°. d.f. = 6, 156, p < 0.001) with a pattern 
consistent with published descriptions (Gupta and 
Shaw, 1956), and the root-vitamin E interaction 
might be considered significant (F = 2.66, d.f. = 6. 
156, p = 0.018; but p = 0.066 and p = 0.052 with 
Greenhouse-Geisser and Huynh-Feldt adjustments, 
respectively). The data are shown in Fig. I and 
suggest that vitamin E exerts its protective effects at 
those sites that are most susceptible to bone loss. 


RESULTS 
Study 2 

One stressed subject and one non-stressed subject, 
both vitamin E supplemented, died in the course of 
the study and their data were not analysed. Over the 
90 days, subjects gained 5.1 g (from a mean weight of 
89.5-94.6 g). ANACOVA on body weight, similar to 
that in Study 1, revealed a significant main effect for 
stress condition (F= 10.97, d.f. = 1, 25, p =0.003). 
Covariate adjusted terminal weights were 97.6 g for 
non-stressed and 91.5 g for stressed animals. 

Table 2 shows the results of Study 2. Total bone 
loss and bone-loss variability were substantially re¬ 
duced as compared to Study 1. ANOVA revealed a 
statistically significant main effect for vitamin E 
(F = 6.51, d.f. = I, 26, p = 0.017) but no effect due to 
stress (F = 0.97, d.f. = 1, 26, p = 0.337) or to a 


DISCLSSION 

The findings of Study 2 support the hypothesis that 
dietary vitamin E supplementation can decrease alve¬ 
olar bone loss. This study does not define the protec¬ 
tive mechanism of vitamin E, which may be related 
to its antioxidant, immunostimulant or other activi¬ 
ties. The study does not provide sufficient infor¬ 
mation to assess the relative contributions of 
infective, inflammatory and hormonal mechanisms 
on observed bone loss, and does not define the 
therapeutic route, which may be either topical or 
systemic. As an exploratory study it does suggest, 
however, that despite inconsistent findings in the 
literature, this may be a productive area for research. 

The absence of an observed vitamin E effect in 
Study 1 had been attributed to variability in bone loss 



Fig. 1. Distance from the CEJ to the alveolar crest measured at the midline of the seven mandibular roots 
found in rats and averaged across the left- and right-hand sides. Roots I -7 correspond to the mesial, 
central, and distal roots of the first molar (1-3) and the mesial and distal roots of the second (4 and 5) 
and third (6 and 7) molars, respectively. The protective effects of dietary vitamin E supplementation are 
most apparent at those locations at most risk for bone loss. 



I fleets i>l vitamin E and stress on hone loss 


605 


before the start of the study Decreased bone loss and 
bone-loss variability in Study 2. which incorporated 
antibiotic prophy laxis, supports this contention and 
suggests that a vitamin E effect may be relatively 
sensitive, at leant in the rice rat, to initial periodontal 
status 

It had been anticipated that vi'amin E effects 
would be increased among stressed animals. This was 
not observed. Although they tended to lose greater 
amounts of bone, this effect was not statistically 
significant and stress level did not interact with 
vitamin condition It is possible that for rice rats the 
'non-stressed' environment may have been stressful 
d le to solitary caging and small cage size. Because of 
this, the stress manipulation may not have been as 
dramatic as had been anticipated. An appropriate 
non-stress control group might require large, commu¬ 
nal cages. 

It is also possible that, while cage rotation might be 
an effective acute stressor (Riley. 1981), the relevant 
hormones may adjust in animals subject to chronic 
stress (McCarty, Horwatt and Konarska. 1988). 
Thus, there may be a reduction in some stress-related 
effects. Weight loss was not affected by stress level in 
Study I but was in Study 2, suggesting a more 
effective c !ress manipulation. Nevertheless, more 
definitive evaluation of stress effects over time would 
require repeated assays of blood hormone concen¬ 
trations. 

Acknowledgements We thank Leisa Morgan. Dale King 
and Jacquelene Thompson for technical support. The 
opinions expressed herein are those of the authors and 
cannot be construed as reflecting the views of the Navy 
Department or the Naval Service at large. The use of 
commercially available products does not imply endorse¬ 
ment of these products or preference to other similar 
products on the market. Supported by Naval Medical 
Research and Development Command Project Number 
M0095.003-0003. 


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4 . TtTLE (and Subtitle) 

EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION 
AND ROTATIONAL STRESS ON ALVEOLAR BONE LOSS IN 
RICE RATS 


7. AUTHORW 

M. E. COHEN, AND D. M. MEYER 


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NDRI-PR 93-U3 


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18. SUPPLEMENTARY NOTES 


Archives of Oral Biology 1993; Vol. 38 No. 7; 601-606 


T9. KEY WORDS (Continue on reverse aide If necessary and identify by block number) 

Alveolar Bone Loss, Vitamin E, Stress, Psychological 


20. ABSTRACT (Continue on reverse aide if neceeeary and Identity by block number) 

The effect of this supplementation on bone loss (distance from 
the cementum-enamel junction to the alveolar crest measured at 
the midline of the lingual aspect of each of th3 mandibular molai 
roots) was studied in rats that were either not stressed or 
stressed on a rotational device for 90 days. In the first stud>, 
neither vitamin E nor stress condition had 

statistically significant effects but there was substantial 


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bone loss and bone-loss variability in all groups. Before the 
start of the second study, to reduce differences in bone loss 
that might otherwise exist before introduction of the 
treatments, rats received an antibiotic in their drinking 
water. In addition, rotational stress was introduced more 
abruptly than in the first study to reduce the likelihood of 
adaptation. Bone loss and bone-loss variability were 
substantially reduced in the second study. Analysis of these 
data indicated that vitamin E supplementation had a 
statistically significant protective effect, which was most 
pronounced at sites most susceptible to loss. Stressed 
subjects tended to lose more bone, but this effect was not 
significant. These findings suggest seme role of vitamin E 
supplementation in the maintenance of periodontal health but 
also a sensitivity in this effect to initial periodontal 
status. 


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