Calcified Tissue International

, Volume 76, Issue 4, pp 272–279 | Cite as

Vitamin E Provides Protection for Bone in Mature Hindlimb Unloaded Male Rats

  • B. J. Smith
  • E. A. Lucas
  • R. T. Turner
  • G. L. Evans
  • M. R. Lerner
  • D. J. Brackett
  • B. J. Stoecker
  • B. H. Arjmandi
Article

Abstract

The deleterious effects of skeletal unloading on bone mass and strength may, in part, result from increased production of oxygen-derived free radicals and proinflammatory cytokines. This study was designed to evaluate the ability of vitamin E (α-tocopherol), a free-radical scavenger with antiinflammatory properties, to protect against bone loss caused by skeletal unloading in mature male Sprague-Dawley rats. A 2 × 3 factorial design was used with either hindlimb unloading (HU) or normal loading (ambulatory; AMB), and low-dose (LD; 15 IU/kg diet), adequate-dose (AD; 75 IU/kg diet), or high-dose (HD; 500 IU/kg diet) vitamin E (DL-α-tocopherol acetate). To optimize the effects of vitamin E on bone, dietary treatments were initiated 9 weeks prior to unloading and continued during the 4-week unloading period, at which time animals were euthanized and blood and tissue samples were collected. Serum vitamin E was dose-dependently increased, confirming the vitamin E status of animals. The HD treatment improved oxidation parameters, as indicated by elevated serum ferric–reducing ability and a trend toward reducing tissue lipid peroxidation. Histomorphometric analysis of the distal femur revealed significant reductions in trabecular thickness (TbTh), double-labeled surface (dLS/BS), and rate of bone formation to bone volume (BFR/BV) due by HU. AMB animals on the HD diet and HU animals on the LD diet had reduced bone surface normalized to tissue volume (BS/TV) and trabecular number (TbN); however, the HD vitamin E protected against these changes in the HU animals. Our findings suggest that vitamin E supplementation provides modest bone protective effects during skeletal unloading.

Keywords

α-Tocopherol Hindlimb unloading Antioxidants COX-2 Osteoporosis 

Notes

Acknowledgments

The authors wish to express their appreciation to Ms. Tracy Riggs and Mrs. Sloan Martin for their assistance with animal care and laboratory analyses. This research was supported by grants from the National Aeronautic and Space Administration (98-HEDS-02) and the Department of Veterans Affairs.

References

  1. 1.
    Aloia, J, Cohn, S, Ostuni, J, Cane, R, Ellis, K 1978Prevention of involutional bone loss by exerciseAnn Intern Med89356358PubMedGoogle Scholar
  2. 2.
    Dalsky, GP 1989The role of exercise in the prevention of osteoporosisComp The153037Google Scholar
  3. 3.
    Rikli, RE, McManis, BG 1990Effects of exercise on bone mineral content in postmenopausal womenRes Quart Exer Sport61243249Google Scholar
  4. 4.
    Barengolts, EI, Curry, DJ, Bapna, MS, Kukreja, SC 1993Effects of endurance exercise on bone mass and mechanical properties in intact and ovariectomized ratsJ Bone Miner Res8937942PubMedGoogle Scholar
  5. 5.
    Chow, JWM, Wilson, AJ, Chambers, TJ, Fox, SW 1998Mechanical loading stimulates bone formation by reactivation of bone lining cells in 13-week-old ratsJ Bone Miner Res1317601767PubMedGoogle Scholar
  6. 6.
    Westerland, KC, Fluckey, JD, Gordon, SE, Kraemer, WJ, Farrell, PA, Turner, R 1998Effect of resistance exercise training on cortical and cancellous bone in mature male ratsJ Appl Physiol84459464PubMedGoogle Scholar
  7. 7.
    Donaldson, CL, Hulley, SB, Vogel, JM, Hattner, RS, Bayers, JH, McMillan, DE 1970Effect of prolonged bed rest on bone mineralMetabolism1910711077PubMedGoogle Scholar
  8. 8.
    Lang, T, Leblanc, A, Evans, H, Lu, Y, Genant, H, Yu, A 2004Cortical and trabecular bone mineral loss from the spine and hip in long-duration spaceflightJ Bone Miner Res1910061012PubMedGoogle Scholar
  9. 9.
    LeBlanc, AD, Schneider, VS, Evans, HJ, Engelbretson, DA, Krebs, JM 1990Bone mineral loss and recovery after 17 weeks of bed restJ Bone Miner Res5843850PubMedGoogle Scholar
  10. 10.
    Baldwin, KM, White, TP, Arnaud, SB, Edgerton, VR, Kraemer, WJ, Kram, R, Raab-Cullen, D, Snow, CM 1996Musculoskeletal adaptions to weightlessness and development of effective countermeasuresMed Sci Sports Exerc1012471253Google Scholar
  11. 11.
    Cavanagh, PR, Davis, BL, Miller, TA 1992A biomechanical perspective on exercise countermeasures for long term spaceflightAviat Space Environ Med63482485PubMedGoogle Scholar
  12. 12.
    Nicogossian, AE, Rummel, JD, Leveton, L, Teeter, R 1992Development of countermeasures for medical problems encountered in space flightAdv Space Res12329337PubMedGoogle Scholar
  13. 13.
    Norman, TL, Bradley-Popovich, G, Clovis, N, Cutlip, RG, Bryner, RW 2000Aerobic exercise as a countermeasure for microgravity-induced bone loss and muscle atrophy in a rat hindlimb suspension modelAviat Space Environ Med71593598PubMedGoogle Scholar
  14. 14.
    Bikle, DD, Harris, J, Halloran, BP, Currier, PA, Tanner, S, Morey-Holton, E 1995The molecular response of bone to growth hormone during skeletal unloading: regional differencesEndocrinology13620992109PubMedGoogle Scholar
  15. 15.
    Bikle, DD, Morey-Holton, ER, Doty, SB, Currier, PA, Tanner, S, Halloran, BP 1994Alendronate increases skeletal mass of growing rats during unloading by inhibiting resorption of calcified cartilageJ Bone Miner Res917771787PubMedGoogle Scholar
  16. 16.
    Turner, RT, Evans, GL, Cavolina, JM, Halloran, BP, Morey-Holton, E 1998Programmed administration of parathyroid hormone (PTH) increases bone formation and reduces bone loss in hindlimb unloaded ovariectomized ratsEndocrinol13940864091Google Scholar
  17. 17.
    Globus, RK, Bikle, DD, Halloran, BP, Morey-Holton, E 1986Skeletal response to dietary calcium in a rat model simulating weightlessnessJ Bone Miner Res1191197PubMedGoogle Scholar
  18. 18.
    Navidi, M, Wolinsky, I, Fung, P, Arnaud, SB 1995Effect of excess dietary salt on calcium metabolism and bone mineral in a spaceflight rat modelJ Appl Physiol787075PubMedGoogle Scholar
  19. 19.
    Azzi, A, Breyer, I, Feher, M, Pastori, M, Ricciarelli, R, Spycher, S, Staffieri, M, Stocker, A, Zimmer, S, Zingg, J 2000Specific cellular responses to alpha-tocopherolJ Nutr13016491652PubMedGoogle Scholar
  20. 20.
    Ricciarelli, R, Zingg, J, Azzi, A 2001Vitamin E: protective role of a Janus moleculeFASEB J1523142325PubMedGoogle Scholar
  21. 21.
    Meydani, SN, Barklund, MP, Liu, S, Meydani, M, Miller, RA, Cannon, JG, Morrow, FD, Rocklin, R, Blumberg, JB 1990Vitamin E supplementation enhances cell-mediated immunity in healthy elderly subjectsAm J Clin Nutr52557563PubMedGoogle Scholar
  22. 22.
    Pathania, V, Syal, N, Pathak, CM, Khanduja, KL 1999Vitamin E suppreses the induction of reactive oxygen species release by lipopolysaccharide, interleukin-1β and tumor necrosis factor-α in rat alveolar macrophagesJ Nutr Sci Vitaminol45675686PubMedGoogle Scholar
  23. 23.
    Wu, D, Hayek, MG, Meydani, SN 2001Vitamin E and macrophage cyclooxygenase regulation in the agedJ Nutr131382S388SPubMedGoogle Scholar
  24. 24.
    Hadjigogos, K 2003The role of free radicals in the pathogenesis of rheumatoid arthritisPanminerva Med45713PubMedGoogle Scholar
  25. 25.
    Golden, TR, Hinerfeld, DA, Melov, S 2002Oxidative stress and aging: beyond correlationAging Cell1117123PubMedGoogle Scholar
  26. 26.
    van’t Hof, RJ, Ralston, SH 2001Nitric oxide and boneImmunology103255261PubMedGoogle Scholar
  27. 27.
    Garrett, IR, Boyce, RF, Oreffo, ROC, Bonewald, L, Poser, J, Mundy, GR 1990Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivoJ Clin Invest85632639PubMedGoogle Scholar
  28. 28.
    Göktürk, E, Turgut, A, Baycu, C, Günal, I, Seber, S, Gülbas, Z 1995Oxygen-free radicals impair fracture healing in ratsActa Orthop Scand66473475PubMedGoogle Scholar
  29. 29.
    Manolagas, SC 2000Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosisEnd Rev21115137Google Scholar
  30. 30.
    Tam, VK, Schotland, S, Green, J 1998Inflammatory cytokines (IL-1α, TNF-α) and LPS modulate the Ca2+ signaling pathway in osteoblastsAm J Physiol274C1686C1698PubMedGoogle Scholar
  31. 31.
    Traianedes, K, Dallas, M, Garrett, IR, Mundy, G, Bonewald, LF 19985-Lipoxygenase metabolites inhibit bone formation in vitroEndocrinology13931782184PubMedGoogle Scholar
  32. 32.
    Xu, H, Watkins, BA, Seifert, MF 1995Vitamin E stimulates trabecular bone formation and alters epiphyseal cartilage morphometryCalcif Tissue Int57293300PubMedGoogle Scholar
  33. 33.
    Arjmandi, BH, Juma, S, Beharka, AA, Bapna, MS, Akhter, M, Meydani, SN 2002Vitamin E improves bone quality in the aged but not in young adult male miceJ Nutr Biochem13543549PubMedGoogle Scholar
  34. 34.
    Arjmandi, BH, Akhter, MP, Chakkalakal, D, Khalil, DA, Lucas, EA, Juma, S, El-Osta, M, Devareddy, L, Stoecker, BJ 2001Effects of isoflavones, vitamin E, and their combination on bone in an aged rat model of osteopeniaJ Bone Min Res16S533Google Scholar
  35. 35.
    Reeves, PG, Nielsen, FH, Fahey, GC 1993AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing on the reformation of the AIN-76 rodent dietJ Nutr12319391951PubMedGoogle Scholar
  36. 36.
    Morey-Holton, ER, Globus, RK 2002The hindlimb unloading rodent model: technical aspectsJ Appl Physiol9213671377PubMedGoogle Scholar
  37. 37.
    Bieri, JG, Tolliver, TJ, Catignani, GL 1979Simultaneous determination of α-tocopherol and retinol in plasma or red cells by high pressure liquid chromatographyAm J Clin Nutr3221432149PubMedGoogle Scholar
  38. 38.
    Benzie, IF, Strain, JJ 1996The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assayAnal Biochem2397076PubMedGoogle Scholar
  39. 39.
    Dobrian, AD, Davies, MJ, Schriver, SD, Lauterio, TJ, Prewitt, RL 2001Oxidative stress in a rat model of obesity-induced hypertensionHypertension37554560PubMedGoogle Scholar
  40. 40.
    Smith, PK, Krohn, RJ, Hermanson, GT, Mallia, AK, Gartner, FH, Provenzano, MD, Fujimoto, EK, Goeke, NM, Olson, BJ, Klenk, DC 1985Measurement of protein using bicinchoninic acidAnal Biochem1507685PubMedGoogle Scholar
  41. 41.
    Buhl, KM, Jacobs, CR, Turner, RT, Evans, GL, Farrell, PA, Donahue, H 2001Aged bone displays an increased responsiveness to low-intensity resistance exerciseJ Appl Physiol9013591364PubMedGoogle Scholar
  42. 42.
    Smith, BJ, King, JB, Lucas, EA, Akhter, M, Arjmandi, BH, Stoecker, BJ 2002Detrimental effects of skeletal unloading and copper depletion on bone in mature ratsJ Nutr132190196PubMedGoogle Scholar
  43. 43.
    Hargens, A, Steskal, J, Johansson, C, Tipton, CM 1984Tissue fluid shift, forelimb loading and tail suspension in tail-suspended ratsThe Physiologist27S37S38Google Scholar
  44. 44.
    Jenkins, JK, Huang, H, Ndebele, K, Salahudeen, AK 2001Vitamin E inhibits renal mRNA expression of COX II, HO I, T and osteopontin in the rat model of syslosporine nephrotoxicityTransplantation71331334PubMedGoogle Scholar
  45. 45.
    Wu, D, Mura, C, Beharka, AA, Han, SN, Paulson, KE, Hwang, D, Meydani, SN 1998Age-associated increase in PGE2 synthesis and COX activity in murine macrophages is reversed by vitamin EAm J Physiol275C661C668PubMedGoogle Scholar
  46. 46.
    Dominguez, JH, Mundy, GR 1980Monocytes mediate osteoclastic bone resorption by prostaglandin productionCaicif Tissue Int312934Google Scholar
  47. 47.
    Kumei, Y, Shimokawa, H, Katano, H, Hara, E, Akiyama, H, Hirano, M, Mukai, C, Nagaoka, S, Whitson, PA, Sams, CF 1996Microgravity induces prostaglandin E2 and interleukin-6 production in normal rat osteoblasts: role in bone demineralizationJ Biotechnology47313324Google Scholar
  48. 48.
    Azzi, A, Stocker, A 2000Vitamin E: non-antioxidant rolesProg Lipid Res39231255PubMedGoogle Scholar
  49. 49.
    Fukuzawa, K, Gebicki, JM 1983Oxidation of alpha-tocopherol in micelles and liposomes by the hydroxyl, perhydroxyl, and superoxide free radicalsArch Biochem Biophys226242251PubMedGoogle Scholar
  50. 50.
    Nishikimi, M, Yamada, H, Yagi, K 1980Oxidation by superoxide of tocopherols dispersed in aqueous media with deoxycholateBiochim Biophys Acta627101108PubMedGoogle Scholar
  51. 51.
    Soung, DY, Droke, EA, Khalil, DA, Arquitt, AB, Hammond, LJ, Smith, BJ, Arjmandi, BH 2002Soy isoflavones and vitamin E reduce precursors of osteoclastsFASEB J16A231Google Scholar
  52. 52.
    Lin, SK, Kok, SH, Kuo, MY, Lee, MS, Wang, CC, Lan, WH, Hsiao, M, Goldring, SR, Hong, CY 2003Nitric oxide promotes infectious bone resorption by enhancing cytokine-stimulated interstitial collagenase synthesis in osteoblastsJ Bone Miner Res183946PubMedGoogle Scholar
  53. 53.
    Ricciarelli, R, Maroni, P, Ozer, N, Zingg, JM, Azzi, A 1999Age-dependent increase of collagenase expression can be reduced by alpha-tocopherol via protein kinase C inhibitionFree Radic Biol Med27729737PubMedGoogle Scholar
  54. 54.
    Ricciarelli, R, Zingg, JM, Azzi, A 2002The 80th anniversary of vitamin E: beyond its antioxidant propertiesBiol Chem383457465PubMedGoogle Scholar
  55. 55.
    Beharka, AA, Wu, D, Han, SN, Meydani, SN 1997Macrophage prostaglandin production contributes to the age-associated decrease in T cell function which is reversed by the dietary antioxidant vitamin EMech Aging Dev935977PubMedGoogle Scholar
  56. 56.
    Mitchell, ME, Stern, LS, Shah, N, Ostrum, R 2001Effect of flurbiprofen on hind-limb suspension-induced bone lossAviat Space Environ Med72790793PubMedGoogle Scholar
  57. 57.
    Fall, PM, Breault, DT, Raisz, LG 1994Inhibition of collagen synthesis by prostaglandins in the immortalized rat osteoblastic cell line Py1a: structure-activity relations and signal transduction mechanismsJ Bone Miner Res919351943PubMedGoogle Scholar
  58. 58.
    Li, X, Okada, Y, Pilbeam, CC, Lorenzo, JA, Kennedy, CR, Breyer, RM, Raisz, LG 2000Knockout of the murine prostaglandin EP2 receptor impairs osteoclastogenesis in vitroEndocrinology14120542061PubMedGoogle Scholar
  59. 59.
    Weinreb, M, Suponitzky, I, Keila, S 1997Systemic administration of an anabolic dose of PGE2 in young rats increases the osteogenic capacity of bone marrowBone20521526PubMedGoogle Scholar
  60. 60.
    Huang, M, Schocken, M, Block, G, Sowers, M, Gold, E, Sternfeld, B, Seeman, T, Greendale, GA 2002Variation in nutrient intakes by ethnicity: results from the Study of Women’s Health Across the Nation (SWAN)Menopause9309319PubMedGoogle Scholar
  61. 61.
    Bonner, LT, Peskind, ER 2002Pharmacologic treatments of dementiaMed Clin North Am86657674PubMedGoogle Scholar
  62. 62.
    Meydani, SN, Meydani, M, Rall, LC, Morrow, F, Blumberg, JB 1994Assessment of the safety of high-dose, short-term supplementation with vitamin E in healthy older adultsAm J Clin Nutr60704709PubMedGoogle Scholar
  63. 63.
    Machlin, LJ, Bagriel, E, Brin, M 1982Biopotency of alpha-tocopherols as determined by curative myopathy bioassay in the ratJ Nutr11214371440PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2004

Authors and Affiliations

  • B. J. Smith
    • 1
  • E. A. Lucas
    • 2
  • R. T. Turner
    • 3
  • G. L. Evans
    • 3
  • M. R. Lerner
    • 4
  • D. J. Brackett
    • 4
  • B. J. Stoecker
    • 2
  • B. H. Arjmandi
    • 2
  1. 1.Department of Surgery Research University of Oklahoma Health Sciences CenterOklahoma CityUSA
  2. 2.Department of Nutritional Sciences, College of Human and Environmental SciencesOklahoma State UniversityStillwaterUSA
  3. 3.Department of Orthopedics, and Biochemistry and Molecular BiologyMayo ClinicRochesterUSA
  4. 4.Department of Surgery, College of MedicineUniversity of Oklahoma Health Sciences Center & Veterans Affairs Medical CenterOklahoma CityUSA

Personalised recommendations