Effect of regular resistance exercise, vitamin D, and calcium supplements on the bone mineral content and density in postmenopausal model of rats: An experimental study


Background: Postmenopausal osteoporosis progressively occurs due to alteration in the estrogen level during the menopause period, and subsequently elevates the risk of fractures.

Objective: To evaluate the effect of regular resistance exercise, vitamin D, and calcium supplements on bone mineral content and density, postmenopausal rats used.

Materials and Methods: In this experimental study, 72 female Sprague-Dawley rats (8-10 wk: 250 ± 15 gr) were ovariectomized and randomly divided into nine groups (n = 8/each): control, placebo, exercise (EX), exercise with vitamin D supplement (EX + D), exercise with calcium (EX + Ca), exercise with calcium and vitamin D (EX + Ca + D), vitamin D administration (D), calcium administration (Ca), and calcium and vitamin D (Ca + D) groups. Finally, the tail, hip, and lumbar bone mineral content, bone mineral density, bone thickness, and bone cells were evaluated in each group.

Results: The tail, hip, and lumbar bone mineral density was increased significantly in the EX + Vit D group compared to the control group (p = 0.004, p = 0.007, p = 0.003, respectively). However, there were no significant changes in the bone mineral content of the hips and lumbar among the groups. Besides, bone thickness in the Ex + Vit D group was more than the other groups (p = 0.02). The number of osteoclast cells decreased in the Ca + Vit D, Ex + Ca, Ex + Vit D, and Ex + Vit D + Ca groups compared to the control group. Osteocyte numbers were increased only in the Ex + Vit D group.

Conclusion: Resistance exercise in combination with vitamin D and calcium have a positive effect on the bone mineral density and bone mineral content and might be able to prevent or delay the osteoporosis among elderly women. However, additional researches are needed to assess the molecular pathways of this process.

Key words: Menopause, Vitamin D, Exercise, Calcium, Bone mineral density.

[1] Bjelica A, Ćirilović VV, Todorović ST, Filipović K. Postmenopausal osteoporosis. Med Pregl 2018; 71: 201–205.

[2] Dalal PK, Agarwal M. Postmenopausal syndrome. Indian J Psychiatry 2015; 57 (Suppl.): S222–S232.

[3] Muka T, Oliver-Williams C, Kunutsor S, Laven JS, Fauser BC, Chowdhury R, et al. Association of age at onset of menopause and time since onset of menopause with cardiovascular outcomes, intermediate vascular traits, and all-cause mortality: A systematic review and meta-analysis. JAMA Cardiol 2016; 1: 767–776.

[4] Hernández-Angeles C, Castelo-Branco C. Early menopause: A hazard to a woman’s health. Indian J Med Res 2016; 143: 420–427.

[5] Sathyapalan Th, Aye M, Rigby AS, Fraser WD, Thatcher NJ, Kilpatrick ES, et al. Soy reduces bone turnover markers in women during early menopause: A randomized controlled trial. J Bone Miner Res 2017; 32: 157–164.

[6] Levin VA, Jiang X, Kagan R. Estrogen therapy for osteoporosis in the modern era. Osteoporos Int 2018; 29: 1049–1055.

[7] Florencio-Silva R, da Silva Sasso GR, Sasso-Cerri E, Simões MJ, Cerri PS. Biology of bone tissue: Structure, function, and factors that influence bone cells. BioMed Res Int 2015; 2015: 1–17.

[8] Nakamichi Y, Udagawa N, Suda T, Takahashi N. Mechanisms involved in bone resorption regulated by vitamin D. J Steroid Biochem Mol Biol 2018; 177: 70–76.

[9] Khalil A, Youssef GA. Effect of aerobic exercise, vitamin K and vitamin D on bone metabolism in ovariectomized adult rats. Nat Sci 2015; 13: 1–11.

[10] Reid IR, Horne AM, Mihov B, Gamble GD, Al−Abuwsi F, Singh M, et al. Effect of monthly high−dose vitamin D on bone density in community−dwelling older adults substudy of a randomized controlled trial. J Intern Med 2017; 282: 452–460.

[11] Eastell R, O’Neill TW, Hofbauer LC, Langdahl B, Reid IR, Gold DT, et al. Postmenopausal osteoporosis. Nat Rev Dis Primers 2016; 2: 16069.

[12] Fung JL, Hartman TJ, Schleicher RL, Goldman MB. Association of vitamin D intake and serum levels with fertility: Results from the lifestyle and fertility study. Fertil Steril 2017; 108: 302–311.

[13] Gowder SJT. A Critical Evaluation of Vitamin D-Basic Overview [Internet]. UK: Intech Open; 2017. Chapter 13, Elhusseini H, Lizneva D, Gavrilova-Jordan L, Eziba N, Abdelaziz M, Brakta S, et al. Vitamin D and Female Reproduction. Available at: Https://www.intechopen.com/books/ a-critical-evaluation-of-vitamin-d-basic-overview/ vitamin-d-and-female-reproduction.

[14] Foroozanfard F, Jamilian M, Bahmani F, Talaee R, Talaee N, Hashemi T, et al. Calcium plus vitamin D supplementation influences biomarkers of inflammation and oxidative stress in overweight and vitamin D−deficient women with polycystic ovary syndrome: A randomized double−blind placebo−controlled clinical trial. Clin Endocrinol 2015; 83: 888–894.

[15] Chon SJ, Koh YK, Heo JY, Lee J, Kim MK, Yun BH, et al. Effects of vitamin D deficiency and daily calcium intake on bone mineral density and osteoporosis in Korean postmenopausal woman. Obstet Gynecol Sci 2017; 60: 53–62.

[16] Tai V, Leung W, Grey A, Reid IR, Bolland MJ. Calcium intake and bone mineral density: Systematic review and meta-analysis. BMJ 2015; 351: h4183.

[17] Purdue-Smithe AC, Whitcomb BW, Szegda KL, Boutot ME, Manson JE, Hankinson SE, et al. Vitamin D and calcium intake and risk of early menopause. Am J Clin Nutr 2017; 105: 1493–1501.

[18] Kemmler W, Engelke K, von Stengel S. Long−term exercise and bone Mineral density changes in postmenopausal women-are there periods of reduced effectiveness? J Bone Miner Res 2016; 31: 215–222.

[19] Segev D, Hellerstein D, Dunsky A. Physical activity-does it really increase bone density in postmenopausal women? A review of articles published between 2001-2016. Curr Aging Sci 2018; 11: 4–9.

[20] Muir JM, Ye Ch, Bhandari M, Adachi JD, Thabane L. The effect of regular physical activity on bone mineral density in post-menopausal women aged 75 and over: A retrospective analysis from the Canadian multicentre osteoporosis study. BMC Musculoskeletal Disord 2013; 14: 253–261.

[21] Xu J, Lombardi G, Jiao W, Banfi G. Effects of exercise on bone status in female subjects, from young girls to postmenopausal women: An overview of systematic reviews and meta-analyses. Sports Med 2016; 46: 1165–1182.

[22] Bilek LD, Waltman NL, Lappe JM, Kupzyk KA, Mack LR, Cullen DM, et al. Protocol for a randomized controlled trial to compare bone-loading exercises with risedronate for preventing bone loss in osteopenic postmenopausal women. BMC Women’s Health 2016; 16: 59–70.

[23] Chen C, Noland KA, Kalu DN. Modulation of intestinal vitamin D receptor by ovariectomy, estrogen and growth hormone. Mech Ageing Dev 1997; 99: 109– 122.

[24] Prestes J, Leite RD, Pereira GB, Shiguemoto GE, Bernardes CF, Asano RY, et al. Resistance training and glycogen content in ovariectomized rats. Int J Sports Med 2012; 33: 550–554.

[25] Khoo BCC, Beck ThJ, Qiao QH, Parakh P, Semanick L, Prince RL, et al. In vivo short-term precision of hip structure analysis variables in comparison with bone mineral density using paired dual-energy X-ray absorptiometry scans from multi-center clinical trials. Bone 2005; 37: 112–121.

[26] Beck TJ, Ruff CB, Warden KE, Scott JW, Rao GU. Predicting femoral neck strength from bone mineral data. A structural approach. Invest Radiol 1990; 25: 6–18.

[27] DeFina LF, Leonard D, Willis BL, Barlow CE, Finley CE, Jenkins MR, et al. High cardiorespiratory fitness is associated with reduced risk of low bone density in postmenopausal women. Journal Women’s Health 2016; 25: 1073–1080.

[28] Willems HME, van den Heuvel EGHM, Schoemaker RJW, Klein-Nulend J, Bakker AD. Diet and exercise: A match made in bone. Current Osteoporosis Reports 2017; 15: 555–563.

[29] Beavers KM, Beavers DP, Martin SB, Marsh AP, Lyles MF, Lenchik L, et al. Change in bone mineral density during weight loss with resistance versus aerobic exercise training in older adults. J Gerontol A Biol Sci Med Sci 2017; 72: 1582–1585.

[30] Veldurthy V, Wei R, Oz L, Dhawan P, Jeon YH, Christakos S. Vitamin D, calcium homeostasis and aging. Bone Res 2016; 4: 16041–16047.

[31] Fischer V, Haffner-Luntzer M, Prystaz K, vom Scheidt A, Busse B, Schinke Th, et al. Calcium and vitamin- D deficiency marginally impairs fracture healing but aggravates posttraumatic bone loss in osteoporotic mice. Sci Rep 2017; 7: 7223.

[32] Liguori C, Romigi A, Izzi F, Mercuri NB, Cordella A, Tarquini E, et al. Continuous positive airway pressure treatment increases serum vitamin D levels in male patients with obstructive sleep apnea. J Clin Sleep Med 2015; 11: 603–607.

[33] Agostini D, Donati Zeppa S, Lucertini F, Annibalini G, Gervasi M, Ferri Marini C, et al. Muscle and bone health in postmenopausal women: Role of protein and vitamin D supplementation combined with exercise training. Nutrients 2018; 10: 1103–1123.

[34] Tachiki T, Kouda K, Dongmei N, Tamaki J, Iki M, Kitagawa J, et al. Muscle strength is associated with bone health independently of muscle mass in postmenopausal women: the Japanese populationbased osteoporosis study. J Bone Miner Metab 2019; 37: 53–59.

[35] Kemmler W, Haberle L, von Stengel S. Effects of exercise on fracture reduction in older adults: A systematic review and meta-analysis. Osteoporos Int 2013; 24: 1937–1950.

[36] Polidoulis I, Beyene J, Cheung AM. The effect of exercise on pQCT parameters of bone structure and strength in postmenopausal women–a systematic review and meta-analysis of randomized controlled trials. Osteoporos Int 2012; 23: 39–51.

[37] Peterson SE, Peterson MD, Raymond G, Gilligan C, Checovich MM, Smith EL. Muscular strength and bone density with weight training in middle-aged women. Med Sci Sports Exerc 1991; 23: 499–504.

[38] Mason C, Xiao L, Imayama I, Duggan CR, Bain C, Foster-Schubert KE, et al. Effects of weight loss on serum vitamin D in postmenopausal women. Am J Clin Nutr 2011; 94: 95–103.

[39] Toumi H, Best TM, Cesaro A, Lespessailles E. Exercise and anti-osteoporotic medication combined treatment for osteoporosis. J Yoga Phys Ther 2016; 6: 1000232–1000235.

[40] Huang TH, Su IH, Lewis JL, Chang MSh, Hsu AT, Perrone CE, et al. Effects of methionine restriction and endurance exercise on bones of ovariectomized rats: a study of histomorphometry, densitometry and biomechanical properties. J Appl Physiol 2015; 119: 517–526.

[41] Li W, Zhang Y, Xu X, Wang K, Ding W. Relationship between osteogenesis and angiogenesis in ovariectomized osteoporotic rats after exercise training. Int J Clin Exp Pathol 2017; 10: 11438–11449.

[42] Silbermann M, Bar-Shira-Maymon B, Coleman R, Reznick A, Weisman Y, Steinhagen-Thiessen E, et al. Long-term physical exercise retards trabecular bone loss in lumbar vertebrae of aging female mice. Calcif Tissue Int 1990; 46: 80–93.

[43] Wu J, Wang X, Chiba H, Higuchi M, Nakatani T, Ezaki O, et al. Combined intervention of soy isoflavone and moderate exercise prevents body fat elevation and bone loss in ovariectomized mice. Metabolism 2004; 53: 942–948.

[44] Zhang L, Chen X, Wu J, Yuan Y, Guo J, Biswas S, et al. The effects of different intensities of exercise and active vitamin D on mouse bone mass and bone strength. J Bone Miner Metab 2017; 35: 265–277.