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Beneficial Effects of Tocotrienol and Tocopherol on Bone Histomorphometric Parameters in Sprague–Dawley Male Rats After Nicotine Cessation

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Abstract

This study was conducted to determine the effectiveness of three forms of vitamin E supplements following nicotine treatment on bone histomorphometric parameters in an adult male rat model. Rats were divided into seven groups: baseline (B, killed without treatment), control (C, normal saline for 4 months), nicotine (N, nicotine for 2 months), nicotine cessation (NC), tocotrienol-enhanced fraction (TEF), gamma-tocotrienol (GTT), and alpha-tocopherol (ATF). Treatments for the NC, TEF, GTT, and ATF groups were performed in two phases. For the first 2 months they were given nicotine (7 mg/kg), and for the following 2 months nicotine administration was stopped and treatments with respective vitamin E preparations (60 mg/kg) were commenced except for the NC group, which was allowed to recover without treatment. Rats in the N and NC groups had lower trabecular bone volume, mineral appositional rate (MAR), and bone formation rate (BFR/BS) and higher single labeled surface and osteoclast surface compared to the C group. Vitamin E treatment reversed these nicotine effects. Both the TEF and GTT groups, but not the ATF group, had a significantly higher trabecular thickness but lower eroded surface (ES/BS) than the C group. The tocotrienol-treated groups had lower ES/BS than the ATF group. The GTT group showed a significantly higher MAR and BFR/BS than the TEF and ATF groups. In conclusion, nicotine induced significant bone loss, while vitamin E supplements not only reversed the effects but also stimulated bone formation significantly above baseline values. Tocotrienol was shown to be slightly superior compared to tocopherol. Thus, vitamin E, especially GTT, may have therapeutic potential to repair bone damage caused by chronic smoking.

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References

  1. Church DF, Pryor WA (1985) Free-radical chemistry of cigarette smoke and its toxicological implications. Environ Health Perspect 64:111–126

    Article  PubMed  CAS  Google Scholar 

  2. Pryor WA, Stone K (1993) Oxidants in cigarette smoke. Radicals, hydrogen peroxide, peroxynitrate, and peroxynitrite. Ann NY Acad Sci 28:12–27

    Article  Google Scholar 

  3. Morrow JD, Frei B, Longmire AW, Gaziano JM, Lynch SM, Shyr Y, Strauss WE, Oates JA, Robert LJ (1995) Increase in circulating products of lipid peroxidation (F2-isoprostanes) in smokers. Smoking as a cause of oxidative damage. N Engl J Med 332:1198–1203

    Article  PubMed  CAS  Google Scholar 

  4. Duthie GG, Arthur JR, James WP (1991) Effects of smoking and vitamin E on blood antioxidant status. Am J Clin Nutr 53:1061S–1063S

    PubMed  CAS  Google Scholar 

  5. Garrett IR, Boyce BF, Oreffo ROC, Bonewald L, Poser J, Mundy GR (1990) Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. J Clin Invest 85:632–639

    Article  PubMed  CAS  Google Scholar 

  6. Suda N, Morita I, Kurodo T, Murot S (1993) Participitation of oxidative stress in the process of osteoclast differentiation. Biochim Biophys Acta 1157:318–323

    PubMed  CAS  Google Scholar 

  7. Mody N, Parhami F, Sarafian TA, Demer DL (2001) Oxidative stress modulates osteoblastic differentiation of vascular and bone cells. Free Radic Biol Med 31:509–519

    Article  PubMed  CAS  Google Scholar 

  8. Wynder EL, Hoffmann D (1979) Tobacco and health. A societal challenge. N Engl J Med 300:894–903

    CAS  Google Scholar 

  9. Ramp WK, Lenz LG, Galvin RJ (1991) Nicotine inhibits collagen synthesis and alkaline phosphatase activity but stimulates DNA synthesis in osteoblast-like cells. Proc Soc Exp Biol Med 197:36–43

    PubMed  CAS  Google Scholar 

  10. Fang MA, Frost PJ, Iida-Klein A, Hahn TJ (1991) Effects of nicotine on cellular function in UMR 106-01 osteoblast-like cells. Bone 12:283–286

    Article  PubMed  CAS  Google Scholar 

  11. Henemyre CL, Scales DK, Hokett SD, Cuenin MF, Peacock ME, Parker MH, Brewer PD, Chuang AH (2003) Nicotines stimulates osteoclast resorption in a porcine marrow cell model. J Periondotol 74:1440–1446

    Article  CAS  Google Scholar 

  12. Norazlina M, Nik-Farideh YMK, Arizi A, Faisal A, Ima-Nirwana S (2004) Effects of nicotine on bone resorbing cytokines in male rats. Int Med J 3:1–8. www.e.imj.com/Vol3-No2/Vo13-No2-B10.htm

    Google Scholar 

  13. Ima-Nirwana S, Cheng CT, Norazlina M (2005) Effects of nicotine on bone mineral density and calcium homeostasis in male Sprague–Dawley rats. Curr Top Pharmacol 9:125–129

    CAS  Google Scholar 

  14. Wetscher GJ, Bagchi M, Bagchi D, Perdikis G, Hinder PR, Glaser K, Hinder RA (1995) Free radical production in nicotine treated pancreatic tissue. Free Radic Biol Med 18:877–882

    Article  PubMed  CAS  Google Scholar 

  15. Helen A, Krishnakumar K, Vijayammal PL, Augusti KT (2000) Antioxidant effect of onion oil (Allium cepa. Linn) on the damages induced by nicotine in rats as compared to alpha-tocopherol. Toxicol Lett 116:61–68

    Article  PubMed  CAS  Google Scholar 

  16. Kozlovskis-Wade PL, Smets MJ, Myerburg RJ (1998) The effect of nicotine on DNA repair in adult myocytes. J Mol Cell Cardiol 30:1483–1491

    Article  PubMed  CAS  Google Scholar 

  17. Kalpana C, Menon VP (2004) Protective effect of circumin on circulatory lipid peroxidation and antioxidant status during nicotine-induced toxicity. Toxicol Mech Methods 14:339–343

    Article  CAS  PubMed  Google Scholar 

  18. Ahmad NS, Khalid BAK, Ima-Nirwana S (2004) Effects of vitamin E on interleukin-1 in ferric nitrilotriacetate treated rats. Malays J Biochem Biol 9:43–47

    Google Scholar 

  19. Ahmad NS, Khalid BAK, Luke DA, Ima-Nirwana S (2005) Tocotrienol offers better protection than tocopherol from free radical–induced damage of rat bone. Clin Exp Pharmacol Physiol 32:761–770

    Article  PubMed  CAS  Google Scholar 

  20. Ima-Nirwana S, Kiftiah A, Sariza T, Gapor MT, Khalid BAK (1999) Palm vitamin E improves bone metabolism and survival rate in thyrotoxic rats. Gen Pharmacol 32:621–626

    Article  PubMed  CAS  Google Scholar 

  21. Ima-Nirwana S, Norazlina M, Khalid BAK (2000) Palm vitamin E prevents osteoporosis in orchidectomized growing male rats. Natural Product Sci 694:155–160

    Google Scholar 

  22. Norazlina M, Ima-Nirwana S, Gapor MT, Khalid BAK (2000) Palm vitamin E is comparable to alpha-tocopherol in maintaining bone mineral density in ovariectomised female rats. Exp Clin Endocrinol Diab 108:305–310

    Article  CAS  Google Scholar 

  23. Ima-Nirwana S, Fakhrurazi H (2002) Palm vitamin E protects bone against dexmethasone-induced osteoporosis in male rats. Med J Malaysia 57:136–144

    Google Scholar 

  24. Norazlina M, Ima-Nirwana S, Gapor MT, Khalid BAK (2002) Tocotrienols are needed for normal bone calcification in growing female rats. Asia Pacific J Clin Nutr 11:194–199

    Article  CAS  Google Scholar 

  25. Norazlina M, Ng FW, Ima-Nirwana S (2005) Gamma-tocotrienol is required for normal vitamin D metabolism in female rats. Indian J Pharmacol 37:309–314

    Article  CAS  Google Scholar 

  26. Norazlina M, Chua CW, Ima-Nirwana S (2004) Vitamin E deficiency reduced lumbar bone calcium content in female rats. Med J Malaysia 59:297–304

    Google Scholar 

  27. Fort FL (1991) Drug safety evaluation. In: Swarbrick J, Boylan JC (eds) Encyclopedia of pharmaceutical technology, vol 4. Marcel Dekker, New York, pp 416–421

  28. Meydani SN, Meydani M, Rall LC, Morrow F, Blumberg JB (1994) Assessment of the safety of high-dose, short-term supplementation with vitamin E in healthy older adults. Am J Clin Nutr 60:704–709

    PubMed  CAS  Google Scholar 

  29. Difford J (1974) A simplified method for the preparation of methyl methacrylate embedding medium. Med Lab Tech 31:79–81

    CAS  Google Scholar 

  30. Von Kossa J (1974) Nachweis von Kalk. Beitrage zur pathologischen Anatomie und zur allgemeinen. Pathologie 29:163

    Google Scholar 

  31. Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30:23–38

    Article  PubMed  CAS  Google Scholar 

  32. Freere RH, Weibel ER (1967) Stereologic techniques in microscopy. J R Microsc Soc 87:25–34

    Google Scholar 

  33. Baldock PA, Morris HA, Need AG, Moore RJ, Durbridge TC (1998) Variation in the short-term changes in bone cell activity in three regions of the distal femur immediately following ovariectomy. J Bone Miner Res 13:1451–1457

    Article  PubMed  CAS  Google Scholar 

  34. Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR (1987) Bone histomorphometry: standardization of nomenclature, symbols and units. Report of the ASMBR Histomorphometry Nomenclature Committee. J Bone Miner Res 2:595–610

    Article  PubMed  CAS  Google Scholar 

  35. Frost HM, Jee WS (1992) On the rat model human osteopenias and osteoporosis. Bone Miner 18:227–236

    Article  PubMed  CAS  Google Scholar 

  36. Boran R, Tross R, Vignery A (1984) Evidence of sequential remodeling in rat trabecular bone: morphology, dynamic histomorphometry, and changes during skeletal maturation. Anat Rec 208:137–145

    Article  Google Scholar 

  37. Erben R (1996) Trabecular and endocortical bone surfaces in the rat: modeling or remodeling? Ant Rec 246:39–46

    Article  CAS  Google Scholar 

  38. Wronski TJ, Dann LM, Scott KS, Cintron M (1989) Long-term effects of ovariectomy and aging on the rat skeleton. Calcif Tissue Int 45:360–366

    Article  PubMed  CAS  Google Scholar 

  39. Wing KJ, Fisher GC, O’Connell JX, Wing PC (2000) Stopping nicotine exposure before surgery: the effect on spinal fusion in rabbit model. Spine 25:30–34

    Article  PubMed  CAS  Google Scholar 

  40. Hoidrup S, Prescott E, Sorensen TI, Gottschau A, Lauritzen JB (2000) Tobacco smoking and risk of hip fracture in men and women. Int Epidemiol Assoc 29:253–259

    Article  CAS  Google Scholar 

  41. Szulc P, Garnero B, Claustraf F, Marchand F, Duboeuf F, Delmas PD (2002) Increased bone resorption in moderate smokers with low body weight: the minos story. J Clin Endocrinol Metab 87:666–674

    Article  PubMed  CAS  Google Scholar 

  42. Krall EA, Dietrich T, Nunn ME, Garcia RI (2006) Risk of tooth loss after cigarette smoking cessation. Prev Chronic Dis 3:1–8

    Google Scholar 

  43. Hapidin H, Othman F, Shuid AN, Soelaiman IN, Mohamed N (2007) Negative effects of nicotine on bone-resorbing cytokines and bone histomorphometric parameters in male rats. J Bone Miner Metab 25:93–98

    Article  PubMed  CAS  Google Scholar 

  44. Broulik PD, Jarab J (1993) The effect of chronic nicotine administration on bone mineral content in mice. Horm Metab Res 25:219–221

    Article  PubMed  CAS  Google Scholar 

  45. Akmal M, Kesani A, Anand B, Singh A, Wiseman M, Goodship A (2004) Effect of nicotine on spinal disc cells: a cellular mechanism for disc degeneration. Spine 29:568–575

    Article  PubMed  Google Scholar 

  46. Broulik PD, Rosenkrancova J, Ruzicka P, Sedlacek R, Kurcova I (2007) The effect of chronic nicotine administration on bone mineral content and bone strength in normal and castrated male rats. Horm Metab Res 39:20–24

    Article  PubMed  CAS  Google Scholar 

  47. Syversen U, Nordsletten L, Falch JA, Madsen JE, Nilsen OG, Waldum HL (1999) Effects of lifelong nicotine inhalation on bone mass and mechanical properties in female rats femurs. Calcif Tissue Int 65:246–249

    Article  PubMed  CAS  Google Scholar 

  48. Fung YK, Iwaniec UT, Cullen DM, Akhter MP, Haven MC, Timmins P (1999) Long-term effects of nicotine on bone and calciotropic hormones in adult female rats. Pharmacol Toxicol 85:181–187

    Article  PubMed  CAS  Google Scholar 

  49. Iwaniec UT, Fung YK, Akhter MP, Haven MC, Nespor S, Haynatzki GR, Cullen DM (2001) Effects of nicotine on bone mass, turnover and strength in adult female rat. Calcif Tissue Int 68:358–364

    Article  PubMed  CAS  Google Scholar 

  50. Riesenfeld A (1985) Growth-depressing effects of alcohol and nicotine in two strains of rats. Acta Anat 122:18–24

    Article  PubMed  CAS  Google Scholar 

  51. Cesar-Neto JB, Benatti BB, Manzi FR, Sallum EA, Sallum AW, Nociti FH Jr (2005) The influence of cigarette smoke inhalation on bone density. A radiographic study in rats. Braz Oral Res 19:47–51

    Article  PubMed  Google Scholar 

  52. Cesar-Neto JB, Benatti BB, Sallum EA, Casati ZM, Nociti FH Jr (2006) The influence of cigarette smoke inhalation and its cessation on the tooth-supporting alveolar bone: a histometric study in rats. J Periodontal Res 41:118–123

    Article  PubMed  Google Scholar 

  53. Oncken C, Prestwood K, Kleppinger A, Wang Y, Cooney J, Raizs L (2006) Impact of smoking cessation on bone mineral density in postmenopausal women. J Womens Health 15:1141–1150

    Article  Google Scholar 

  54. Pasco JA, Hendry MJ, Wilkinson LK, Nicholson GC, Schneider HG, Kotowicz MA (2006) Antioxidant vitamin supplements and markers of bone turnover in a community sample of nonsmoking women. J Womens Health 15:295–300

    Article  Google Scholar 

  55. Nafeeza MI, Asma H, Ahmad Asmadi Y, Zaitun Z, Alini M (2002) Role of vitamin E on oxidative stress in smokers. Malays J Med Sci 9:34–42

    Google Scholar 

  56. Xu H, Watkins BA, Seifert MF (1995) Vitamin E stimulates trabecular bone formation and alters epiphyseal cartilage morphology. Calcif Tissue Int 57:293–300

    Article  PubMed  CAS  Google Scholar 

  57. Arjmandi BH, Akhter MP, Chakkalakal D, Khalil DA, Lucas EA, Juma S, El-Osta M, Devareddy L, Stoecker BJ (2001) Effects of isoflavones, vitamin E, and their combination on bone in an aged rat model of osteopenia. J Bone Miner Res 16:S533

    Google Scholar 

  58. Arjmandi BH, Juma S, Beharka A, Bapna S, Akhter M, Meydani SN (2002) Vitamin E improves bone quality in the aged but not in young adult male mice. J Nutr Biochem 13:543–549

    Article  PubMed  CAS  Google Scholar 

  59. Nishida A, Yamaguchi A, Tanizawa T, Endo N, Mashiba T, Uchiyama Y, Suda T, Yoshiki S, Takahashi HE (1994) Increased bone formation by intermittent parathyroid hormone administration is due to the stimulation of proliferation and differentiation of osteoprogenitor cells in bone marrow. Bone 6:717–723

    Article  Google Scholar 

  60. Schmidt IU, Dobnig H, Turner RT (1995) Intermittent parathyroid hormone treatment increases osteoblast number, steady state messenger ribonucleic acid levels for osteocalcin, and bone formation in tibial metaphysis of hypophysectomized female rats. Endocrinology 136:5127–5134

    Article  PubMed  CAS  Google Scholar 

  61. Jiang Y, Zhao JJ, Mitlak BH, Wang O, Genant HK, Eriksen EF (2003) Recombinant human parathyroid hormone (1–34) [teriparatide] improves both cortical and cancellous bone structure. J Bone Miner Res 18:1932–1941

    Article  PubMed  CAS  Google Scholar 

  62. Neer RM, Arnaud CD, Zanchetta JR, Prince R, Gaich GA, Reginster JY, Hodsman AB, Eriksen EF, Ish-Shalom S, Genant HK, Wang O, Mitlak BH (2001) Effect of parathyroid hormone (1–34) on fractures and bone mineral density in postmenopausal women with osteoporosis. N Engl J Med 10:1434–1441

    Article  Google Scholar 

  63. Greenspan SL, Bone HG, Ettinger DA, Lindsay R, Zanchetta JR, Blosch CM, Mathisen AL, Morris SA, Marriott TB (2007) Effects of recombinant human parathyroid hormone (1–84) on vertebral fracture and bone density in postmenopausal women with osteoporosis. Ann Intern Med 146:326–339

    PubMed  Google Scholar 

  64. Soelaiman IN, Ahmad NS, Khalid BAK (2004) Palm oil tocotrienol mixture is better than alpha-tocopherol acetate in protecting bones against free-radical induced elevation of bone-resorbing cytokines. Asia Pacific J Clin Nutr 13 Suppl:S111

  65. Norazlina M, Lee PL, Lukman HI, Nazrun AS, Ima-Nirwana S (2007) Effects of vitamin E supplementation on bone metabolism in nicotine-treated rats. Singapore Med J 48:195–199

    PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Carotech (Ipoh, Malaysia) for supplying the gamma-tocotrienol and the Malaysian Palm Oil Board for providing the tocotrienol-enhanced fraction. We are also grateful to Mr. Rafizul Mohd Yusoff, Mr. Arizi Aziz, Ms. Hairi Ghazalli, Ms. Azizah Osman, and Mr. Faisal Ariffin for their technical assistance. We also thank the Ministry of Science, Technology, and Innovation for funding this project via research grant IRPA/UKM 06-02-02-051-EA243.

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Hermizi, H., Faizah, O., Ima-Nirwana, S. et al. Beneficial Effects of Tocotrienol and Tocopherol on Bone Histomorphometric Parameters in Sprague–Dawley Male Rats After Nicotine Cessation. Calcif Tissue Int 84, 65–74 (2009). https://doi.org/10.1007/s00223-008-9190-x

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