Vitamin E improved bone strength and bone minerals in male rats given alcohol

Document Type: Original Article


Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia


Objective(s): Alcohol consumption induces oxidative stress on bone, which in turn increases the risk of osteoporosis. This study determined the effects of vitamin E on bone strength and bone mineral content in alcohol-induced osteoporotic rats.
Materials and Methods: Three months old Sprague Dawley male rats were randomly divided into 5 groups: (I) control group; (II) alcohol (3 g/kg) + normal saline; (III) alcohol (3 g/kg) + olive oil; (IV) alcohol (3 g/kg) + alpha-tocopherol (60 mg/kg) and (V) alcohol (3 g/kg) + palm vitamin E (60 mg/kg). The treatment lasted for three months. Following sacrifice, the right tibia was subjected to bone biomechanical test while the lumbar (fourth and fifth lumbar) and left tibia bones were harvested for bone mineral measurement.
Results: Alcohol caused reduction in bone biomechanical parameters (maximum force, ultimate stress, yield stress and Young’s modulus) and bone minerals (bone calcium and magnesium) compared to control group (P<0.05). Palm vitamin E was able to improve bone biomechanical parameters by increasing the maximum force, ultimate stress and Young’s modulus (P<0.05) while alpha-tocopherol was not able to. Both alpha-tocopherol and palm vitamin E were able to significantly increase tibia calcium and magnesium content while only alpha-tocopherol caused significant increase in lumbar calcium content (P<0.05).
Conclusion: Both palm vitamin E and alpha-tocopherol improved bone mineral content which was reduced by alcohol. However, only palm vitamin E was able to improve bone strength in alcohol treated rats.


1. World Health Organization. Global status report on alcohol and health 2014. Global status report on alcohol 2014; 1-392.

2. Mutalip MHBA, Kamarudin RB, Manickam M, Abd Hamid HAB, Saari RB. Alcohol consumption and risky drinking patterns in Malaysia: Findings from NHMS 2011. Alcohol Alcohol 2014; 49:593–599.

3. Yun J, Son M, Abdelmegeed M, Banerjee A, Morgan T, Yoo S, et al. Binge alcohol promotes hypoxic liver injury through a CYP2E1-HIF-1alpha-dependent apoptosis pathway in mice and humans. Free Radic Biol Med 2014; 77:183–194.

4. Thayanukulvat C, Harding T. Binge drinking and cognitive impairment in young people. Br J Nurs 2015; 24:401–407.

5. VandeVoort C, Grimsrud K, Midic U, Mtango N, Latham K. Transgenerational effects of binge drinking in a primate model: implications for human health. Fertil Steril 2015; 103:560–569.

6. Turner RT. Skeletal response to alcohol. Alcohol Clin Exp Res 2000; 24:1693–1701.

7. Chakkalakal DA. Alcohol-induced bone loss and deficient bone repair. Alcohol Clin Exp Res 2005; 29:2077–2090.

8. Tønnesen H, Pedersen A, Jensen MR, Møller A, Madsen JC. Ankle fractures and alcoholism. The influence of alcoholism on morbidity after malleolar fractures. J Bone Joint Surg Br 1991; 73:511–513.

9. Wu D, Cederbaum A. Alcohol, oxidative stress, and free radical damage. Alcohol Res Heal 2003; 27(4):277–284.

10. González-Reimers E, Santolaria-Fernández F, Martín-González M, Fernández-Rodríguez C, Quintero-Platt G. Alcoholism: a systemic proinflammatory condition. World J Gastroenterol 2014; 20:14660–14671.

11. Norazlina M, Hermizi H, Faizah O, Nazrun AS, Norliza M, Ima-Nirwana S. Vitamin E reversed nicotine-induced toxic effects on bone biochemical markers in male rats. Arch Med Sci 2010; 6:505–512.

12. Muhammad N, Luke DA, Shuid AN, Mohamed N, Soelaiman IN. Tocotrienol supplementation in postmenopausal osteoporosis: evidence from a laboratory study. Clinics 2013; 68:1338–1343.

13. Feresin R, Johnson S, Elam M, Kim J, Khalil D, Lucas E, et al. Effects of vitamin E on bone biomechanical and histomorphometric parameters in ovariectomized rats. J Osteoporos 2013; 2013:825985.

14. Chin K, Gengatharan D, Mohd Nasru F, Khairussam R, Ern S, Aminuddin S, et al. The effects of annatto tocotrienol on bone biomechanical strength and bone calcium content in an animal model of osteoporosis due to testosterone deficiency. Nutrients 2016; 8:E808.

15. Shuid AN, Mehat Z, Mohamed N, Muhammad N, Soelaiman IN. Vitamin E exhibits bone anabolic actions in normal male rats. J Bone Miner Metab 2010; 28:149–156.

16. Kasai S, Ito A, Shindo K, Toyoshi T, Bando M. High-dose alpha-tocopherol supplementation does not induce bone loss in normal rats. PLoS One 2015; 10: e0132059.

17. Iwaniec UT, Turner RT, Smith BJ, Stoecker BJ, Rust A, Zhang B, et al. Evaluation of long-term vitamin E insufficiency or excess on bone mass, density, and microarchitecture in rodents. Free Radic Biol Med 2013; 65:1209–1214.

18. Chin K-Y, Mo H, Soelaiman I-N. A review of the possible mechanisms of action of tocotrienol - a potential antiosteoporotic agent. Curr Drug Targets 2013; 14:1533–1541.

19. Callaci J, Juknelis D, Patwardhan A, Sartori M, Frost N, Wezeman F. The effects of binge alcohol exposure on bone resorption and biomechanical and structural properties are offset by concurrent bisphosphonate treatment. Alcohol Clin Exp Res 2004; 28:182–191.

20. Sengupta P. The laboratory rat: Relating its age with human’s. Int J Prev Med 2013; 4:624–630.

21. Callaci JJ, Juknelis D, Patwardhan A, Wezeman FH. Binge alcohol treatment increases vertebral bone loss following ovariectomy: Compensation by intermittent parathyroid hormone. Alcohol Clin Exp Res 2006; 30:665–672.

22. Olson KN, Smith SW, Kloss JS, Ho JD, Apple FS. Relationship between blood alcohol concentration and observable symptoms of intoxication in patients presenting to an emergency department. Alcohol Alcohol 2013; 48:386–389.

23. Norazlina M, Ima-Nirwana S, Gapor MT, Khalid BAK. Palm vitamin E is comparable to alpha-tocopherol in maintaining bone mineral density in ovariectomised female rats. Exp Clin Endocrinol Diabetes 2000; 108(4):305–310.

24. Nair AB, Jacob S. A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm 2016; 7:27–31.

25. Stürmer EK, Seidlová-Wuttke D, Sehmisch S, Rack T, Wille J, Frosch KH, et al. Standardized bending and breaking test for the normal and osteoporotic metaphyseal tibias of the rat: effect of estradiol, testosterone, and raloxifene. J Bone Miner Res 2006; 21:89–96.

26. Rai D, Kumar G, Tewari P, Saxena D. Acute and chronic dose of alcohol affect the load carrying capacity of long bone in rats. J Biomech 2008; 41:20–24.

27. Ghanbari R, Anwar F, Alkharfy K, Gilani A, Saari N. Valuable nutrients and functional bioactives in different parts of olive (Olea europaea L.)—A review. Int J Mol Sci 2012; 13:3291–3340.

28. Dong H, Hutchins-Wiese H, Kleppinger A, Annis K, Liva E, Lammi-Keefe C, et al. Effects of omega-3 polyunsaturated fatty acid supplementation on bone turnover in older women. Int J Vitam Nutr Res 2014; 84:124–132.

29. Sampson HW. Alcohol’s harmful effects on bone. Alcohol Health Res World 1998; 22:190–194.

30. Hogan HA, Sampson HW, Cashier E, Ledoux N. Alcohol consumption by young actively growing rats: a study of cortical bone histomorphometry and mechanical properties. Alcohol Clin Exp Res 1997; 21:809–816.

31. Mohamad S, Shuid AN, Mokhtar SA, Abdullah S, Soelaiman IN. Tocotrienol supplementation improves late-phase fracture healing compared to alpha-tocopherol in a rat model of postmenopausal osteoporosis: A biomechanical evaluation. Evidence-based Complement Altern Med. 2012; 2012: 372878.

32. Jiang F, Liao Z, Hu L-H, Du Y-Q, Man X-H, Gu J-J, et al. Comparison of antioxidative and antifibrotic effects of alpha-tocopherol with those of tocotrienol-rich fraction in a rat model of chronic pancreatitis. Pancreas 2011; 40:1091–1096.

33. Suzuki Y, Tsuchiya M, Wassall S, Choo Y, Govil G, Kagan V, et al. Structural and dynamic membrane properties of alpha-tocopherol and alpha-tocotrienol: implication to the molecular mechanism of their antioxidant potency. Biochemistry 1993; 32:10692–10699.

34. Ahsan H, Ahad A, Iqbal J, Siddiqui W. Pharmacological potential of tocotrienols: a review. Nutr Metab 2014; 11:52.

35. Seo S, Chun S, Newell M, Yun M. Association between alcohol consumption and Korean young women’s bone health: a cross sectional study from the 2008 to 2011 Korea National Health and Nutrition Examination Survey. BMJ Open 2015; 5:e007914.

36. Paccou J, Edwards MH, Ward K, Jameson K, Moon R, Dennison E, et al. Relationships between bone geometry, volumetric bone mineral density and bone microarchitecture of the distal radius and tibia with alcohol consumption. Bone 2015; 78:122–129.

37. Keiver K, Duggal S, Simpson ME. Ethanol administration results in a prolonged decrease in blood ionized calcium levels in the rat. Alcohol 2006; 37:173–178.

38. Romani AMP. Magnesium homeostasis and alcohol consumption. Magnesium Research 2008; 21:197–204.

39. Gaddini G, Grant K, Woodall A, Stull C, Maddalozzo G, Zhang B, et al. Twelve months of voluntary heavy alcohol consumption in male rhesus macaques suppresses intracortical bone remodeling. Bone 2015; 71:227–236.

40. Dyer S, Sampson H. Magnesium levels in alcohol-treated rodents using different consumption paradigms. Alcohol 1998; 16:195–199.

41. Preedy V, Baldwin D, Keating J, Salisbury J. Bone collagen, mineral and trace element composition, histomorphometry and urinary hydroxyproline excretion in chronically-treated alcohol-fed rats. Alcohol Alcohol 1991; 26:39–46.

42. Defino H, Vendrame J. Morphometric study of lumbar vertebrae’s pedicle. Acta Ortopédica Bras 2007; 15:183–186.

43. Hammett-Stabler C. Osteoporosis: From Pathophysiology to Treatment: Special Topics in Diagnostic Testing. Washington: AACC Press; 2004.

44. Norazlina M, Ima-Nirwana S, Khalid BAK. Effects of palm vitamin E, vitamin D and calcium supplementation on bone metabolism in vitamin E deficient rats. Med J Islam Acad Sci 1999; 12:89–96.

45. Norazlina M, Rita L, Ima-Nirwana S. The effects of vitamin E or calcium supplementation on bone mineral composition in vitamin E deficient rats. Malaysian J Biochem Mol Biol 2002; 7:1–5.

46. Ferretti M, Cavani F, Smargiassi A, Roli L, Palumbo C. Mineral and skeletal homeostasis influence the manner of bone loss in metabolic osteoporosis due to calcium-deprived diet in different sites of rat vertebra and femur. Biomed Res Int 2015; 2015:304178.

47. Chen J, Lazarenko O, Shankar K, Blackburn M, Badger T, Ronis M. A role for ethanol-induced oxidative stress in controlling lineage commitment of mesenchymal stromal cells through inhibition of Wnt/beta-catenin signaling. J Bone Min Res 2010; 25:1117–1127.

48. Callaci JJ, Himes R, Lauing K, Wezeman FH, Brownson K. Binge alcohol-induced bone damage is accompanied by differential expression of bone remodeling-related genes in rat vertebral bone. Calcif Tissue Int 2009; 84:474–484.

49. Berg KM, Kunins H V, Jackson JL, Nahvi S, Chaudhry A, Harris KA, et al. Association between alcohol consumption and both osteoporotic fracture and bone density. Am J Med  2008; 121:406–418.

50. Ostman B, Michaëlsson K, Helmersson J, Byberg L, Gedeborg R, Melhus H, et al. Oxidative stress and bone mineral density in elderly men: antioxidant activity of alpha-tocopherol. Free Radic Biol Med 2009; 47:668–673.

51. Mata-Granados JM, Cuenca-Acebedo R, Luque De Castro MD, Quesada Gomez JM. Lower vitamin E serum levels are associated with osteoporosis in early postmenopausal women: A cross-sectional study. J Bone Miner Metab 2013; 31:455–460.

52. Michaëlsson K, Wolk A, Byberg L, Ärnlöv J, Melhus H. Intake and serum concentrations of alpha-tocopherol in relation to fractures in elderly women and men: 2 cohort studies. Am J Clin Nutr 2014; 99:107–114.

53. Abukhadir SSA, Mohamed N, Makpol S, Muhammad N. Effects of palm vitamin E on bone-formation-related gene expression in nicotine-treated rats. Evidence-based Complement Altern Med. 2012; 2012: 656025.

54. Wu S, Liu P, Ng L. Tocotrienol-rich fraction of palm oil exhibits anti-inflammatory property by suppressing the expression of inflammatory mediators in human monocytic cells. Mol Nutr Food Res 2008; 52:921–929.