Naval Dental Research Institute AD-A272 235 NDRI-PR 93-03 EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION AND ROTATIONAL STRESS ON ADVEOLAR BONE LOSS IN RICE RATS DTIC ELECTE M. E. COHEN NOV 10 1993 D. M. MEYER ( fors u(cid:127) bi' - s and sale its i 1Is3 utui.n fzmIred 93-27632 iHI EII EIll l 1 111I I Naval Medical Research and Development Command Bethesda, Maryland , '~ , III I Best Available Copy 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 n~ex'eopmcnt Ccmrr-mnd 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: ,J.(cid:127)(cid:127)'.. . ,AccecI;c -n,. .(cid:127) or 0 S. A. RALLS CfAB Captain, Dental Corps aunced United States Navy Commanding Officer _ . ly f DTIC QUALITY INSPECTED 8 L " Jft-inm0& .4rh, oral Biol Vol. IX. No, 7, pp N)i W11169.9 , 9 H 9l t(cid:127) q-9';t4,S OW(cid:127) .I JI, V Printed in (ireat Britain Pergamo~n Pre,., I (d EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION AND ROTATIONAL STRESS ON ALVEOLAR BONE LOSS IN RICE RATS M. E. CorH-N and D. M. MEYER Naval Dental Research Institute, Building I-H1, Great Lakes. IL 6008N, U.S.A. (A t tepted 9 February 199.?) Summary --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 ':spect of each of the mandibular molar roots) was studied in rats that were either no, stressed or stressed on a rotational device for 90 days. In the first study, neither vitamin F nor stress condition had statisticallk, 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 E, stress, psychological. INTRODUCTION (Cheraskin and Ringsdorf, 1970), but circulating concentrations of the vitamin were the same in Vitamin E functions as a free-radical scavenger that patients with and without periodontitis (Slade et al., inhibits lipid peroxidation and inflammation, pro- 1976). Patients with periodontal disease who were tects ischaemic and hypoxic tissues, and is immuno- given vitamin E daily for 21 days to swish in their stimulating (Crary, 1984). Because of this spectrum of mouths and swallow exhibited a significant decrease activities, the relation between vitamin E and peri- in fluid flow from the gingival sulcus than in controls odontal health and disease has been studied: findings with disease but no vitamin E supplementation tend to support a therapeutic role for the vitamin but (Goodson and Bowles, 1973). Also, subjects given the evidence is inconsistent, vitamin E supplementation for 12 weeks exhibited a Periodontal tissues of albino rats were not affected reduction in Russell's Periodontal Index (Cerna et al., by long-term vitamin E deficiency despite early Euro- 1984). However, topical vitamin E did not reduce pean reports of successful treatment of periodontitis gingivitis over a 4-week period relative to a placebo with this substance in man (Nelson and Chaudhry, (Cohen et al., 1991), but the method appeared insen- 1966). However. vitamin E deficiency had negative sitive in that chlorhexidine imni!arly h',d non-signifi impact on periodontal health in rats in another study cant effects on gingivitis. (Schneider and Pose, 1969). Ligature-induced peri- Lack of consistent benefits associated with vitamin "odontitis in rats was not affected by vitamin E E may be due, in part, to the absence of physiological * supplementation (Parrish, DeMarco and Bissada, stress manipulation. During stress, vitamin E stores 1977), bit careful inspection of the data suggests that are depleted and, once depgeted, tissues are at greater supplementation resulted in greater numbers of risk. Without stress, vitamin E may remain at effec- inflammatory cells with. simultaneously, less alveolar five concentrations so that there may be no relation bone loss. This would be consistent with vitamin E's between it and periodontal destruction. One may immunostimulant and antioxidant properties, even argue that in man, insufficient vitamin E and a though sample sizes and methods of analysis did not arguett incmas, insufficenitami f anda hav suficte ndtpeoewcr thse hifs a sttisi- subsequent increase in the likelihood of periodontal have sufficient power to detect these shifts at statisti- destruction occur during brief intervals associated cally significant levels. Dietary vitamin E supplemen- with stressful life-events. Therefore, the cross-sec- tation was shown to accelerate gingival wound tional analysis of Slade et al. (1976) might not identify healing in albino rats (Kim and Shklar, 1983). In man, greater dietary intake of vitamin E has a relation, been associated with fewer reported oral symptoms 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 Abbreviations: AN(AC)OVA. analysis of (co-)variance; the result of other extraordinary interventions. It CEJ, cementum-enamel junction. would be difficult, therefore, to observe a beneficial 601 602 M. E. ('_. and D. M MLYER effect of vitamin E unless it is superimposed on an MATERIALS AND METHODS adverse initial state such as healing of experimental Study I gingival wounds, vitamin E deprivation (as con- trasted to supplementation), or stress. Rice rats (Oryzomys palustri(cid:127)) were chosen because Effects of stress on periodontal destruction have of susceptibility to periodontal destruction (Leonard, been documented and are consistent with generally 1979). The study (protocol reviewed by a Laboratory held concepts of the relation between stress and Animal Use Committee) had a two-by-two factorial disease resistance (Troxler, 1986; Dantzer and Kelley, design with two levels of dietary vitamin E sup- 1989; Ballieux, 1991). In a series of experiments on plementation and either a control or high-stress en- rodents, increased rates of periodontal destruction ironment. (Ratcliff, 1956; Fedi Jr, 1958; Shklar and Glickman, Thirty-two male rice rats (57-84 days of age) were 1959; Gupta, Blechman and Stahl, 1960; Gupta, switched from standard rodent diet to a synthetic test 1966) and decreased rates of gingival wound healing diet (modified Purina Basal Yest Diet 5755). This diet (Stahl, 1961) were shown to accompany the presence contains 44% dextrin, 21/% casein, 15% sucrose, 5% of stressful stimuli. While similar experimental de- lard, 5% corn oil and the remaining percentage is signs are not possible in man, there is evidence that comprised of other necessary additives. One half of the same relations hold. Early anecdotal reports the animals received a synthetic control diet, which (Moulton, Ewn and Thiemman, 1952) have been included a standard 50 IU of vitamin E per kg feed supported by correlational studies relating life-stress (35 IU attributable to d-2-tocopherol acetate oil events to rates of periodontal destruction (Manhold, supplement and approximately 15 IU due to other Doyle and Weisinger, 1971; Haskell, 1975; Green components, principally corn oil). The remaining el al., 1986). animals received a synthetic test diet that contained Although stress will affect a variety of physio- 5000 IU of the vitamin per kg feed (4985 IU due to logical processes, activation of the sympathetic- the tocopheiol additive). This IU/kg dose (60 IUiday adrenal-medullary and hypothalamic- pituitary- based on a 12 g daily food ingestion) had been used in adrenocortical systems (Axelrod and Reisine, 1984) a similar long-term study of periodontal destruction appears particularly important in terms of defining in rats, without reported side-effects (Parrish et al., explanatory mechanisms for enhancement of disease 1977). In general, vitamin E is not considered toxic, progression. Exposure of animals to stressful stimuli mutagenic, carcinogenic, or teratogenic, even at high is accompanied by increases in the synthesis, storage doses (Bendich and Machlin, 1988). and release into the circulation ofcatecholamines and After 35 days of feeding on the synthetic diets, rats glucocorticoids. The effects of these events and sub- were assigned to normal or high-stress environments. sequent enzymatic shifts, as indexed by a variety of Assignment to both diet and stress conditions in- measures, can include: salivary and small-vessel vaso- volved grouping animals by weight and then ran- constriction, release of cytotoxic free radicals, in- domly assigning them within groups to the various creased membrane permeability, impaired wound treatment conditions. healing, decreased resistance to infection, and sup- All rats were housed individually in plastic cages pressed cellular immune responses (Matheny, 1988; (standard multi-mouse cages, approx. 7 in. Weiss et al., 1989). wide x 12 in. long x 5 in. high) with compressed, In addition to studies relating stress to periodontal shreddable wood shavings as bedding material. All health, there are others in which stress-related groups were housed in the same quiet room with daily hormones have been measured or manipulated di- 12-h light/12-h dark cycles. Non-stressed animals rectly. Administration of cortisone caused osteoporo- were housed in standard racks, with diet groups in sis of alveolar bone in mice (Glickman. Stone and counterbalanced order relative to location. Chawla, 1953) and patients with acute necrotizing Profoundly painful or exhausting stimuli were n-,t ulcerative gingivitis were found to have higher levels used as stressors. Rather, rats were subjected to cage of overnight urinary cortisol (Cohen-Cole et al., rotation. This method of stress induction is con- 1981). Catecholamines have been shown to reduce sidered relatively benign but nevertheless is associated gingival circulation (Ito et al., 1973; Clarke, Shephard with shifts in physiological status (Shipov et al.. 1985) and Hirsch, 1981) and to enhance the virulence of including circulating concentrations of stress-related gingival crevicular bacteria (Courant and Gibbons, hormones (Riley, 1981). Stressed animals were 1966). housed in cages mounted in a star pattern on a large The presence of oxygen radicals has been sug- platter that was rotated by a quiet electric motor gested as playing a central part in tissue damage controlled by a computerized timing device. The associated with chronic inflammation (in general) platter was approx. 120cm in diameter and the radius and periodontal disease (Hoffeld, 1982). Based on of a circle formed by the outer edges of the cages was vitamin E's ability to protect tissue from oxidative 55 cm. Two of these devices (henceforth called damage, Goodsoi (1975) predicts therapeutic value wheels), each holding eight subjects (four from each but recognizes that there has been an insufficient of the two diet groups), were used. number of studies in this area. Hence we have Immediately after their assignment to high-stress now sought to evaluate the ability of vitamin E conditions, these animals were subjected to a I-min supplementation to prevent periodontal destruc- period of 30 revimin rotation once every 4 h. When tion in a species inherently susceptible to such de- wheels were spinning, rats preferred to place them- struction, and while exposed to environmental stress selves at the far end of their cages, approx. 50 cm that could affect the supplement's physiological (radius, r) from the cen:.; of the wheel. Therefore, 30 value. rev/min corresponds to a gravitational (g) force of 0.5 Etlects of %itamin E and stress on bone loss 603 [where a is acceleration and r is velocity, g = a 980; Table I Mean distance. in nn, and SD in parentheses from a = V2 r; r = (2nr) (rev min 60 ')]. CEJ to alveolar crest as a function of dietary'i tamin F and Each wheel was rotated on an independent. con- rotational stress in Study I tinuous (24 h/day) variable-interval schedule (i.e. the Dietar, vitamin F location of the I -min rotational period within the 4-h time interval was random). As rotational stress may Standard Supplemented Total reduce feeding and cause weight loss, subjects were tress weighed at least once per week. A protocol f r No 0711(0160) 0.737 (0.239S 0.724 0s197s temporary termination of rotation because of Yes 0.806 (0.087) 0.833 (0.260) 0.820(0.240) catastrophic weight loss was in place but never Total 0.756(0.197) 0.785 (0.246) 0.771 (0.221) needed to be implemented. Over a period of approx. 2 months, rotations were increased from the initial I min at 30 rev min every circulating vitamin E than those given low amounts 4 h to 15 min (in a single segment) of 42 rev, min (10.55 versus 2.44 mg! 1). (1.0 g) every 30 min. Ninety days after initiation of Table I shows mean alveolar bone loss for the four stress manipulations, animals were anaesthetized. or- groups. Effects of diet and stress level did not ap- bital blood samples drawn to assay for vitamin E. and proach statistical significance (ANOVA Fs =0.10. "the animals killed by nitrogen asphyxiation. 1.37 and 0.00, d.f. = 1. 27, for vitamin E stress, and Jaws were dissected out and defleshed by boiling, interaction, respectively). Mandibles were dried and mounted on modelling clay on small dishes, and lingual alveolar bone loss was evaluated using a standard approach (Keyes and MATERIALS AND METHODS Gold, 1955; Gupta and Shaw, 1956). Seven measure- ments taken on each jaw side reflected the distance Study 2 from the CEJ to the alveolar bone crest at the midline As described above, there was high variability in of each root (three roots for the first molar and two bone loss within all groups in Study I. This may be roots each for the second and third molars). Measure- atttibuted to animals being 57-84 days old before the ments were made through a dissecting microscope diet variable was introduced and 92-119 days old fitted with an optical caliper eyepiece. This method before the stress variable. Rice rats can have substan- yielded a measurement precision of approx. 0.01 mm tial bone loss before these ages (Leonard, 1979) and and allowed for minor adjustments in light intensity this 'noise' may have overwhelmed the diet and streýss and angulation to improve visualization. Neverthe- effects. less, the CEJ was sometimes difficult to visualize, even To evaluate this possibility, the study was repeated after application of eosin stain. In such cases, an with two changes. First, starting from 18 to 31 days approximation was made based on other anatomical post-weaning, rats received 1.35 gil tetracycline landmarks. To decrease variability and to preclude a hydrochloride in their drinking water (Vetquamycin- potential for bias, these measurements were made by 32 "1, Phizer, 1.902 g/l) on an approx. 2 weeks on,/2 4 one person who was blind as to experimental con- weeks off schedule. Antibiotics substantially reduce dition. The primary dependent variable of interest bone loss in rice rats (Gupta, Auskaps and Shaw, was the CEJ-bone crest distance averaged across 14 1957) and so should reduce pre-study, between-sub- sites (seven sites on each of two sides). ject variability. Based on estimated daily water intake of 6.4 nil for an 80-g rat, tetracycline intake was (!.35 mg/ml)(6.4ml) = 8.64 mg, and dosage was 8.64 mg/0.08 kg = 108 mg/kg per day. However, giv- RESULTS ing the antibiotic orally can reduce absorption by more than 50% (Plumb, 1991). Recommended oral dosages of tetracycline are about 33-110 mg/kg per One animal in the low vitamin E, high-stress group day (Huber, 1982; Kirk, 1992; Plumb, 1991) and died during the course of the study apd was not 5 mg/ml (Kohn and Barthold, 1984). In general, included in the analysis. ANACOVA was made on tetracycline is considered relatively non-toxic, with body weight. The covariate was weight on stress day oral doses of 75-465 mg/kg per day for 8 weeks being 0, the dependent variable was weight at study termin- well tolerated by dogs without evidence of toxicity ation, and the between-subjects variables were stress (Huber, 1982). Possible interaction between tetra- " and vitamin condition. Over the 90 days, rats gained cycline (toxicity) and Vitamin E supplementation was 6.0 g (from a mean weight of 87.6 g to one of 93.6 g), considered unreasonable given the clinical acceptance but there was no effect of the independent variables, of these concentrations, the lack of observed toxic or their interaction, on the covariate adjusted effects, and the ending of antibiotic supplementation weights. I week before the introduction of test diets. Of 31 blood samples drawn for vitamin E assay, Second, in Study I the stress had been instituted one sample was lost and six were combined into three gradually over a period of 2 months. This may have sets of pairs (within treatment groups) to achieve a attenuated the stress variable by allowing animals to minimum assay quantity. ANOVA with the between- adapt. Therefore, in Study 2 the rotational schedule subject factors of dietary vitamin E and stress re- reached the end-stage of 15 min rotation at 42 vealed only a significant effect for diet condition rev/min per 30 min after 3 days. (F = 222.99, d.f. = 1, 23, p < 0.001). Animals given The rats in Study 2 were between 76 and 97 days high di,',-., vitamin E had more than four times the old when they were switched to the synthetic diets AOB8 IN7 604 M. E. CO_'ou ,nd D M MLNLR T(cid:127);hte 2. Mean distance, in mm, and SD in parentheses from vitamin-stress interaction (F" 0.04, d. I 26, CEJ to alveolar crest as a function of dietary vitamin E and p =0.850). A second ANOVA was done with the rotational stress in Study 2 tooth root at which measurements were taken tseven Dietary vitamin E levels corresponding to mesial of the first molar through distal of the third molar, averaged across the _- Standard Supplemented Total left and right sides) added as a within-subject vari- Stress able. The effect of 'root' was statistically significant No 0.420(0.082) 0.359 (0.044) 0.391 (0072) (F = 224.""' d.f. = 6, 156, p < 0.001) with a pattern Yes 0.451 (0.102) 0.379(0.023) 0.417(0.083) consistent with published descriptions (Gupta and Total 0.436(0.091) 0.369(0.036) 0.404(0.077) 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 (antibiotics had been discontinued I week pre- Greenhouse-Geisser and Huynh-Feldt adjustments. viously), the stress manipulation was instituted 35 respectively). The data are shown in Fig. I and days later, and again they were killed after 90 days. suggest that vitamin E exerts its protective effects at those sites that are most susceptible to bone loss. RESULTS Stud)y 2 DISCUSSION One stressed subject and one non-stressed subject, The findings of Study 2 support the hypothesis that both vitamin E supplemented, died in the course of dietary vitamin E supplementation can decrease alve- the study and their data were not analysed. Over the olar bone loss. This study does not define the protec- 90 days, subjects gained 5.1 g (from a mean weight of tive mechanism of vitamin E, which may be related 89.5-94.6 g). ANACOVA on body weight, similar to to its antioxidant, immunostimulant or other activi- that in Study 1, revealed a significant main effect for ties. The study does not provide sufficient infor- stress condition (F = 10.97, d.f. = I, 25, p = 0.003). mation to assess the relative contributions of Covariate adjusted terminal weights were 97.6 g for infective, inflammatory and hormonal mechanisms non-stressed and 91.5 g for stressed animals. on observed bone loss. and does not define the Table 2 shows the results of Study 2. Total bone therapeutic route, which may be either topical or loss and bone-loss variability were substantially re- systemic. As an exploratory study it does suggest, duced as compared to Study 1. ANOVA revealed a however, that despite inconsistent findings in the statistically significant main effect for vitamin E literature, this may be a productive area for research. (F=6.51, d.f. = 1, 26,p = 0.017) but no effect due to The absence of an observed vitamin E effect in stress (F = 0.97, d.f. = 1, 26, p = 0.337) or to a Study I had been attributed to variability in bone loss 1.2- -- 0-- Normal E, Stressed - Normal E, Stressed 1.0. High E, Not Stressed -- S'- High E, Stressed ! 0.8" 0. 6' I 0.4- 0.2 0.0 p * p p 1 2 3 4 5 6 7 Root Number 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 1-7 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. Etf'cts ,lt tlamin F and %tresso n bone los 605 before the start of the study. Decreased bone loss and potentiation of response to gingisal crevice bacteria. bone-loss variability in Study 2. which incorporated Arnis oral BioI II, 737 739. antibiotic prophylaxis, supports this contention and ('rars F J 119K4) Potential clinical apphications for high- suggests that a vitamin F effect may be relatisel dose nutritional antioxidants. lMed Hlipoth 13, 77 98 sensitive, at least in the rice rat. to initial priodontaal Daannt zienrt eRgr. ataendd v.Kie elleo. f Kre.l a\tVio, n(s1h9i8p9s) bSettwreeses na nthde imbmranin.u natn:d ",talus. the immune ssstem. Lile Sci. 44, 1995 20XIX It had been anticipated that vi'amin E effects FediP. F. Jr (195' The effects of stress on the periodontium would be increased among stressed animals. This was of the S\rian hamster .J Pertodoni. 29. 292 3(X). not observed. Although they tended to lose greater Giickman I . Stone 1. C. and Chawla T. N. ( 1953) The effect amounts of bone, this effect was not s!atisttcally of the systemic administration of cortisone upon the significant and stress level did not interact with periodontium of white mice. J. Periodoni 24, 161 166. vitamin condition. It is possible that for rice rats the Goodson J. M. (1975) Vitamin F therapy and periodontal "non-stressed' environment may have been stressful disease. In Diet, Nutrjijon and Periodontal Di.%eas.e (Eds d te to solitary"- caging and small cage size. Because of' Hazen S. P. and Cowean E. B. Jr). pp. 53 66. American Society for Preventive Dentistry, Chicago, IL. tlts, the stress manipulation may not have been as Goodson J. M. and Bowles D. (1973) The etffct of a-toco- dramatic as had been anticipated. An appropriate pherol on sulcus fluid flo, in periodontal disease. J. dent. non-stress control group might require large, commu- Res. 52, 217 (abstract). ""n0alg ecs. Green L. W., "Iryon W. W., Marks B. and Hlurvn J. f1986) "It is also possible that, while cage rotation might be Periodontal disease as a function of life events stress. an effeci i,,e acute stressor (Riley. 1981). the relevant J. !tuman Stress 12, 32 36. hormones may adjust in anitnals subject to chronic Gupta 0. P. (1966) Psychosomatic factors in periodontal stress (McCarty, Horwatt and Konarska, 1988). disease. Dent. ClinN. Anm.1 0, II 19. Thus, there may be a reduction in some stress-related Gupta 0. P. and Shaw J. H. (1956) Periodontal disease in effects. Weight loss was not affected by stress level in tohfe preircieo draotn: ta1lI .d Miseeathseo.d sO froarl tShuer ge.v a9l,u a7t2i7o n7 3o5f. the extent Study I but was in Study 2, suggesting a more Gupta 0. P., Auskaps A. M. and Shaw J. H. (1957) eltective !tress manipulation. Nevertheless, more Periodontal disease in the rice rat: IV. The effects of definitive evaluation of stress cffects over time would antibiotics on the incidence of periodontal lesions. Oral require repeated assays of blood hormone concen- Sur,. 10, 1169 1175. trations. Gupta O. P., Blechman H. and Stahl S. S. (1960) Effects of stress on the periodontal tissues of young adult male rats Acknowledgernenis --We thank Leisa Morgan. Dale King and hamsters. J. Periodot. 31. 413 417. and Jacquelene Thompson for technical support. The Haskell B. &. (1975) Association of aircraft noise stress to opinions expressed herein are those of the authors and periodontal disease in aircrew members..4A tat. Spaie cannot be construed as reflecting the views of the Navy Ent'ir. Mead. 46, 1041 1043. Department or the Naval Service at large. The use of HofTeld J. T. (1982) Oxygen radicals in inflammation and commercially available products does not imply endorse- immunity In Host Parasite l'iteractions inz Periodontal ment of these products or preference to other similar Diseases (Eds Genco R. J. and Mergenhagen S. E.), products on the market. Supported by Naval Medical pp. 343 -353. American Society for Microbiology. Wash- Research and Development Command Project Number ington, DC. M0095.003-0003. Huber W. G. (1982) Tetracyclines. In Veterinary Pharma- cology and Therapeutic.s 5th edn (Eds Booth N. H. and McDonald L. E.), pp. 740 747. Iowa State University REFERENCES Press, Ames, IA. Ito H., Matsukawa H.. Takahashi K. and Cho Y. W. (1973) Axelrod J. and Reisine T. D. (1984) Stress hormones: their The effects of catecholamine3. acetylcholine and interaction and regulation. Science 224, 452 459. bradykinin on gingival circulation in dogs. Arch/s oral Ballieux R. E. (1991) Impact of mental stress on the immune Biol. 18, 321 328. response. J. Clin. Periodont. 18, 427 430. Keyes P. H. and Gold H. S. (1955) Periodontal lesions in the Bendich A. and Machlin L. J. (1988) Safety of oral intake Syrian hamster. 1. A method of evaluating alveolar bone of vitamin E. Ant. J. Clin. NAur. 48, 612 619. resorption. Oral Surg. 8, 492 499. Cerna H., Fiala B., Fingerov H., Pohl.nka J. and Szwarcova Kim J. E. and Shklar G. (1983) The effect of vitamin F on E. (1984) Contribution to indication of total therapy with the healing of gingival wounds in rats. J. Perirdont. 54, vitamin E in chronic periodontal disease. Acta Unir. 305 308. Palacki Olomuc Fac. Med. 107, 167 170. Kirk R. W. (1992) Current Veterinary Therapy Xl, p. 1248. Cheraskin E. and Ringsdorf W. M. (1970) Relationship of W. B. Saunders, Philadelphia. PA. * reported oral symptoms and signs versus daily vitamin E Kohn D. F. and Barthold S. W. (1984) Biology and diseases consumption. Oral Surg. 29, 361-364. of the rat. In Laboratory Animal Medicine (Eds Fox J. G.. Clarke N. G., Shephard B. C. and Hirsch R. S. (1981) The Cohen B. J. and Loew F. M. Ill), p. 105. Academic Press, effects of intra-arterial epinephrine and nicotine on gingi- Orlando, Fl. val circulation. Oral Surg. 52, 577-5821 Leonard E. P. (1979) Animal model of human disease: Cohen R. E., Ciancio S. G., Mather M. L. and Curro, F. periodontitis in the rice rat (Oryzomys palustris). Ant. A. (1991) Effect of vitamin E gel, placebo gel, and J. Path. 96, 643 646. chlorhexidine on periodontal diseases. Clin. Prey. Dent. Manhold J. H., Doyle J. L. and Weisinger E. H. (1971) 13, 20-24. Effects of social stress or, oral and other bodily tissues. I. Cohen-Cole S., Cogen R., Stevens A.. Kirk K., Gaitan E., Results offering substance to a hypothesis for the for- Hain J. and Freeman A. (1981) Psychosocial, endocrine, mation of periodontal pathology. J. Periodont. 42, and immune factors in acute necrotizing ulcerative 109-111. gingivitis ("trenchmouth"). Psychosom. Med. 43, 91 (ab- Matheny J. L. (1988) Adrenocorticosteroids: systemic and stract). topical therapy. In ClinicalP harmacology in Dental Prac- Courant P. R. and Gibbons R. J. (1966) Epinephrine rice, 4th edn (Eds Holroyd S. V., Wynn R. L. and McCarty R., Horwatt K. and Konarska M. (198h) Chronic in sitamin E. Arth.s oral Bu~d. 14. 431 433. stress and sympathetic-adrenal mnedullary responsiseness. Shipo% A. A.- Shsets V N.. labakosa L. A. and Kabitskaid So~cS. o. Mted. 26, 333 341. 0. E. (1985) Changes in the physical condition. sestibular Moulton R. Ewn S. and Thiemnman W. (1952) Emo- function and bone system of rats exposed to long-term tional factors in periodontal disease. Oral Surg. 5, rotation. Kosin Biol, At iako~sr f.,d. 19, 46 53. 833 860. Shklar Gi. and Glickman 1. f 1959) The effect of' cold as a Nelson M. A. and Chaudhry A. P. (1966) Effects of' stressor agent upon the periodontium of albino rats Oral tocopherol (vitamin E)-deficient diet on some oral, para- Surg. 12, 1111 1320. oral, and hematopoietic tissues of the rat. J. dent. Res. 45, Slade Jr E. W., Bartuska D)., Rose L. F and Cohen D. W. 1072 1077. (1976) Vitamin E and periodontal disease. J. Periodowt Parrish Jr J. H.. DeMarco T. J. and Bissada N. F (1977) 47, 352 354. Vitamin E and periodontitis in the rat. Oral Surg. 44, Stahl S. S. (1961) Healing gingisal injury in normal and 210-218. systemically stressed young adult male rats. J. Periodont. Plumb D. C.) (1991) feterinarY Drug Handbook pp. 501 504. 32, 63 73. Pharmavet Publishing, White Bear Lake, MN. Troxler R. Ci. ( 1986) Human stress response and its posstble Rateliff P. A. (1956) The relationship of the general adap- relationship to disease. Dent. ('in.-N.lAn. 30(4), SI I S27. tation syndrome to the periodontal tissues in the rat. Weiss J. M., Sundar S. K.. Becker K. J. and Cierpial M. A. J. Periodont. 27, 40 43. (1989) Behavioral and neural influences on cellular im- Riley V. (1981) Psychoneuroendocrine influences on im- mune responses: effects of stress and interleukin- 1. J. ('tn. munocompetence and neoplasia. Science 212, 1100 1109. Ptch. 50, 43 53. UNCLASSIFIED SECURITY CLASSIFICATION OF THIS PAGE (When Data Entered) READ INSTRUCTIONS REPORT DOCUMENTATION PAGE BEFORE COMPLETING FORM 1 REPORT NUMBER 12. GOVT ACCESSION NO. 3 RECIPIENT'S CATALOG NUMBER 4. TITLE (and Subtitle) S. TYPE OF REPORT & PERIOD COVERED EFFECT OF DIETARY VITAMIN E SUPPLEMENTATION AND ROTATIONAL STRESS ON ALVEOLAR BONE LOSS IN RICE RATS 6. PERFORMING ORG. REPORT NUMBER NDRI-PR 93-o3 7. AUTHOR(s) 8. CONTRACT OR GRANT NUMBER(s) M. E. COHEN, AND D. M. MEYER 9. PERFORMING ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT, PROJECT, TASK Naval Dental Research Institute AREA & WORK UNIT NUMBERS Building 1-H M0095.003-0003 Great Lakes, IL 60088-5259 11. CONTROI LING OFFICE NAME AND ADDRESS 12. REPORT DATE Naval Medical Research and Developmenr Command August 1993 National Naval Medical Center, 8901 Wisconsion Ave 13. NUMBER OF PAGES Bethesda, MD 20889-5606 (6) 14. MONITORING AGENCY NAME & ADDRESS(if different from Controlling Office) 15ý SECURITY CLASS. (of this report) Chief, Bureau of Medicine and Surgery UNCLASSIFIED Navy Department 2300 E Street NW 1s. DECLASSIFICATION DOWNGRADING SCHEDULE Washington, DC 20372-5300 16. DISTRIBUTION STATEMENT (of this Report) '[his document has been approved for public release; distribution unlimited. 17. DISTRIBUTION STATEMENT (of the abatract entered in Block 20, If different from Report) This document has been approved for publ.ic release; distribution unlimited. IB. SUPPLEMENTARY NOTES Archives of Oral Biology 1993; Vol. 38 No. 7; 601-606 19. KEY WORDS (Continue on reverse aide It necessary and identify by block number) Alveolar Bone Loss, Vitamin E, Stress, Psychological 20. ABSTRACT (Continue on reverse aide It neceeeary and Identify 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 thý mandibular molai roots) was studied in rats that were either not stressed or stressed on a rotational device for 90 days. In the first study, neither vitamin E nor stress condition had statistically significant effects but there was substantial DD JAN 73 1473 EDITION OF I NOV 65 IS OBSOLETE UNCLASSIFIED S/N 0102- LF- 014- 6601 SECURITY CLASSIFICATION OF THIS PAGE (When Date Entered)