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Effects of muscular strength training and growth hormone (GH) supplementation on femoral bone tissue: analysis by Raman spectroscopy, dual-energy X-ray absorptiometry, and mechanical resistance

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The aim of the present study was to verify the effects of muscular strength training and growth hormone (GH) supplementation on femoral bone tissue by Raman spectroscopy (Raman), dual-energy X-ray absorptiometry (DXA), and mechanical resistance (F-max) analysis. A total of 40 male Wistar animals, 60 days old, were used. The animals were distributed into four groups: control (C), control with GH (GHC), muscular strength training (T), and muscular strength training with GH (GHT). Blood samples were collected for the quantification of creatine kinase (CK-MB) and the femurs were removed for analysis by Raman, DXA, and F-max. A more pronounced increase in the bone mineral components was verified in the T group, for all the variables obtained by the Raman (calcium, phosphate, amide, and collagen). In addition, for animals submitted to GH supplementation, there was a reduction in the variable bone mineral density (BMD) obtained by the DXA (p < 0.05). Finally, the animals that received GH supplementation presented a higher F-max, but without statistical significance (p > 0.05). It was concluded that animals that received GH supplementation demonstrated a decrease in BMD. In addition, T alone was able to promote increased calcium, phosphate, amide, and collagen compounds in bone tissue.

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  1. 1.

    Castoldi RC, Teixeira GR, de Malheiro OCM et al (2015) Effects of 14 weeks resistance training on muscle tissue in wistar rats. Int J Morphol 33:446–451. https://doi.org/10.4067/S0717-95022015000200007

  2. 2.

    Lui JC, Colbert M, Cheung CSF et al (2019) Cartilage-targeted IGF-1 treatment to promote longitudinal bone growth. Mol Ther 38:6–12. https://doi.org/10.1016/j.ymthe.2019.01.017

  3. 3.

    Qi Z, Liu W, Lu J (2016) The mechanisms underlying the beneficial effects of exercise on bone remodeling: roles of bone-derived cytokines and microRNAs. Prog Biophys Mol Biol 122:131–139. https://doi.org/10.1016/j.pbiomolbio.2016.05.010

  4. 4.

    Kjær M, Magnusson P, Langberg H (2006) Extracellular matrix adaptation of tendon and skeletal muscle to exercise tendon and collagen synthesis. J Anat 2:1–5. https://doi.org/10.1111/j.1469-7580.2006.00549.x

  5. 5.

    Patrocínio-Silva TL, de SAMF, Goulart RL et al (2016) Low-level laser therapy associated to a resistance training protocol on bone tissue in diabetic rats. Arch Endocrinol Metab 60:457–464. https://doi.org/10.1590/2359-3997000000190

  6. 6.

    Hong AR, Kim SW (2018) Effects of resistance exercise on bone health. Endocrinol Metab 33:435–444. https://doi.org/10.3803/EnM.2018.33.4.435

  7. 7.

    Watson SL, Weeks BK, Weis LJ et al (2018) High-intensity resistance and impact training improves bone mineral density and physical function in postmenopausal women with osteopenia and osteoporosis: the LIFTMOR randomized controlled trial. J Bone Miner Res 33:211–220. https://doi.org/10.1002/jbmr.3284

  8. 8.

    Dror A, Virk K, Lee K et al (2018) Resistance training threshold for elevating bone mineral density in growing female rats. Int J Sports Med 39:382–389. https://doi.org/10.1055/s-0043-125447

  9. 9.

    Ozaki GAT, Koike TE, Castoldi RC et al (2014) Physical exercise remobilization effects on bone density in adults and elderly rats. Motricidade 10:71–78. https://doi.org/10.6063/motricidade.10(3).2725

  10. 10.

    Peres-Ueno MJ, Stringhetta-Garcia CT, Castoldi RC et al (2017) Model of hindlimb unloading in adult female rats: characterizing bone physicochemical, microstructural, and biomechanical properties. PLoS One 12:1–15. https://doi.org/10.1371/journal.pone.0189121

  11. 11.

    Gupta V (2011) Adult growth hormone deficiency. Indian J Endocrinol Metab 15:197–202. https://doi.org/10.4103/2230-8210.84865

  12. 12.

    Sundström K, Cedervall T, Ohlsson C et al (2014) Combined treatment with GH and IGF-I: additive effect on cortical bone mass but not on linear bone growth in female rats. Endocrinology 155:4798–4807. https://doi.org/10.1210/en.2014-1160

  13. 13.

    Al Herbish AS, Almutair A, Bin Abbas B et al (2016) Diagnosis and management of growth disorders in Gulf Cooperation Council (GCC) countries: current procedures and key recommendations for best practice. Int J Pediatr Adolesc Med 3:91–102. https://doi.org/10.1016/j.ijpam.2016.07.002

  14. 14.

    Díez JJ, Sangiao-Alvarellos S, Cordido F (2018) Treatment with growth hormone for adults with growth hormone deficiency syndrome: benefits and risks. Int J Mol Sci 19:1–19. https://doi.org/10.3390/ijms19030893

  15. 15.

    McGrath RP, Ottenbacher KJ, Vincent BM et al (2017) Muscle weakness and functional limitations in an ethnically diverse sample of older adults. Ethn Health 0:1–12. https://doi.org/10.1080/13557858.2017.1418301

  16. 16.

    Loche S, Carta L, Ibba A, Guzzetti C (2014) Growth hormone treatment in non-growth hormone-deficient children. Ann Pediatr Endocrinol Metab 19(1). https://doi.org/10.6065/apem.2014.19.1.1

  17. 17.

    Crowe BJ, Rekers-Mombarg LTM, Robling K et al (2006) Effect of growth hormone dose on bone maturation and puberty in children with idiopathic short stature. J Clin Endocrinol Metab 91:169–175. https://doi.org/10.1210/jc.2005-0891

  18. 18.

    Siebert DM, Rao AL (2018) The use and abuse of human growth hormone in sports. Sport Heal A Multidiscip Approach 10:419–426. https://doi.org/10.1177/1941738118782688

  19. 19.

    Barroso O, Mazzoni I, Rabin O (2008) Hormone abuse in sports: the antidoping perspective. Asian J Androl 10:391–402. https://doi.org/10.1111/j.1745-7262.2008.00402.x

  20. 20.

    Erotokritou-Mulligan I, Eryl Bassett E, Cowan DA et al (2010) The use of growth hormone (GH)-dependent markers in the detection of GH abuse in sport: physiological intra-individual variation of IGF-I, type 3 pro-collagen (P-III-P) and the GH-2000 detection score. Clin Endocrinol 72:520–526. https://doi.org/10.1111/j.1365-2265.2009.03668.x

  21. 21.

    Hoffman JR, Im J, Rundell KW et al (2003) Effect of muscle oxygenation during resistance exercise on anabolic hormone response. Med Sci Sports Exerc 35:1929–1934. https://doi.org/10.1249/01.MSS.0000093613.30362.DF

  22. 22.

    Renno ACM, Silveira Gomes AR, Nascimento RB et al (2007) Effects of a progressive loading exercise program on the bone and skeletal muscle properties of female osteopenic rats. Exp Gerontol 42:517–522. https://doi.org/10.1016/j.exger.2006.11.014

  23. 23.

    Movasaghi Z, Rehman S, Rehman IU (2007) Raman spectroscopy of biological tissues. Appl Spectrosc Rev 42:493–541. https://doi.org/10.1080/05704920701551530

  24. 24.

    Koike TE, Watanabe AY, Kodama FY et al (2018) Physical exercise after immobilization of skeletal muscle of adult and aged rats. Rev Bras Med do Esporte 24:60–63. https://doi.org/10.1590/1517-869220182401172423

  25. 25.

    Aguiar AF, Agati LB, Müller SS et al (2010) Effects of high-impact exercise training on bone mechanical proprieties – an experimental study in female Wistar rats. Acta Ortopédica Bras 18:245–249. https://doi.org/10.1590/S1413-78522010000500002

  26. 26.

    Manchado FDB, Gobatto CA, Contarteze RVL et al (2006) Máxima fase estável de lactato é ergômetro-dependente em modelo experimental utilizando ratos. Rev Bras Med do Esporte 12:259–262. https://doi.org/10.1590/S1517-86922006000500007

  27. 27.

    Longui CA (2008) GH treatment in patients with idiopathic short stature. Arq Bras Endocrinol Metabol 52:750–756. https://doi.org/10.1590/S0004-27302008000500006

  28. 28.

    Tinggaard J, Jensen RB, Sundberg K et al (2014) Ovarian morphology and function during growth hormone therapy of short girls born small for gestational age. Fertil Steril 102:1733–1741. https://doi.org/10.1016/j.fertnstert.2014.09.014

  29. 29.

    Kaminsky P, Walker PM, Deibener J et al (2012) Growth hormone potentiates thyroid hormone effects on post-exercise phosphocreatine recovery in skeletal muscle. Growth Hormon IGF Res 22:240–244. https://doi.org/10.1016/j.ghir.2012.08.001

  30. 30.

    De Mello Malheiro OC, Giacomini CT, Justulin LA et al (2009) Calcaneal tendon regions exhibit different MMP-2 activation after vertical jumping and treadmill running. Anat Rec (Hoboken) 292:1656–1662. https://doi.org/10.1002/ar.20953

  31. 31.

    Castoldi RC, Camargo RCT, Magalhães AJB et al (2013) Concurrent training effect on muscle fibers in Wistar rats. Motriz Rev Educ Fis 19:171–723. https://doi.org/10.1590/S1980-65742013000400008

  32. 32.

    Kendall C, Stone N, Shepherd N et al (2003) Raman spectroscopy, a potential tool for the objective identification and classification of neoplasia in Barrett’s oesophagus. J Pathol 200:602–609. https://doi.org/10.1002/path.1376

  33. 33.

    Souza F de B, Pacheco MTT, VilaVerde AB et al (2003) Avaliação do ácido láctico intramuscular através da espectroscopia Raman: novas perspectivas em medicina do esporte. Rev Bras Med do Esporte 9:388–395. https://doi.org/10.1590/S1517-86922003000600004

  34. 34.

    McManus LL, Bonnier F, Burke G a., et al (2012) Assessment of an osteoblast-like cell line as a model for human primary osteoblasts using Raman spectroscopy. Analyst 137:1559–1569. doi: https://doi.org/10.1039/c2an16209a

  35. 35.

    Cheng W-T, Liu M-T, Liu H-N, Lin S-Y (2005) Micro-Raman spectroscopy used to identify and grade human skin pilomatrixoma. Microsc Res Tech 68:75–79. https://doi.org/10.1002/jemt.20229

  36. 36.

    Gazi E, Dwyer J, Gardner P et al (2003) Applications of Fourier transform infrared microspectroscopy in studies of benign prostate and prostate cancer. A pilot study. J Pathol 201:99–108. https://doi.org/10.1002/path.1421

  37. 37.

    Malini R, Venkatakrishna K, Kurien J et al (2006) Discrimination of normal, inflammatory, premalignant, and malignant oral tissue: a Raman spectroscopy study. Biopolymers 81:179–193. https://doi.org/10.1002/bip.20398

  38. 38.

    Shafer-Peltier KE, Haka AS, Fitzmaurice M et al (2002) Raman microspectroscopic model of human breast tissue: implications for breast cancer diagnosis in vivo. J Raman Spectrosc 33:552–563. https://doi.org/10.1002/jrs.877

  39. 39.

    Castoldi RC, Louzada MJQ, de OBRSM et al (2017) Effects of aerobic, anaerobic, and concurrent training on bone mineral density of rats. Mot rev educ fis 23:71–75. https://doi.org/10.1590/s1980-6574201700010011

  40. 40.

    Val FF de A, Okubo R, Falcai MJ et al (2013) Effects of high-impact exercise training on bone mechanical proprieties - an experimental study in female Wistar rats. Rev Bras Med do Esporte 19:252–255. https://doi.org/10.1590/S1517-86922013000400005

  41. 41.

    Mandair GS, Morris MD (2015) Contributions of Raman spectroscopy to the understanding of bone strength. Bonekey Rep 4:1–8. https://doi.org/10.1038/bonekey.2014.115

  42. 42.

    Torres-del-Pliego E, Vilaplana L, Güerri-Fernández R, Diez-Pérez A (2013) Measuring bone quality. Curr Rheumatol Rep 15:373–378. https://doi.org/10.1007/s11926-013-0373-8

  43. 43.

    Burket J, Gourion-Arsiquaud S, Havill LM et al (2011) Microstructure and nanomechanical properties in osteons relate to tissue and animal age. J Biomech 44:277–284. https://doi.org/10.1016/j.jbiomech.2010.10.018

  44. 44.

    da Silva FF, de Souza RA, Pacheco MTT et al (2011) Effects of different swimming exercise intensities on bone tissue composition in mice: a Raman spectroscopy study. Photomed Laser Surg 29:217–225. https://doi.org/10.1089/pho.2010.2784

  45. 45.

    Inoue-Lima TH, Vasques GA, Scalco RC et al (2019) IGF-1 assessed by pubertal status has the best positive predictive power for GH deficiency diagnosis in peripubertal children. J Pediatr Endocrinol Metab 32:173–179. https://doi.org/10.1515/jpem-2018-0435

  46. 46.

    Davies RD, Parent EC, Steinback CD, Kennedy MD (2018) The effect of different training loads on the lung health of competitive youth swimmers. Int J Exerc Sci 11:999–1018

  47. 47.

    Cruzat VF, Donato Júnior J, Tirapegui J, Schneider CD (2008) Growth hormone and physical exercise: current considerations. Brazilian J Pharm Sci 44:549–562. https://doi.org/10.1590/S1516-93322008000400003

  48. 48.

    Godfrey RJ, Madgwick Z, Whyte GP (2003) The exercise-induced growth hormone response in athletes. Sport Med 33:599–613. https://doi.org/10.2165/00007256-200333080-00005

  49. 49.

    Rozario KS, Lloyd C RF (2015) GH and IGF-1 physiology in childhood. editors. Endotext [Internet]., South Dartmouth (MA)

  50. 50.

    de Rezende Gomes M, Santana de Oliveira Pires I, Alves de Castro I, Tirapegui J (2004) Effect of moderate physical exercise on plasma and tissue levels of insulin-like growth factor–1 in adult rats. Nutr Res 24:555–564. https://doi.org/10.1016/j.nutres.2004.04.003

  51. 51.

    Rawlings JS (2004) The JAK/STAT signaling pathway. J Cell Sci 117:1281–1283. https://doi.org/10.1242/jcs.00963

  52. 52.

    Gent J, van den Eijnden M, van Kerkhof P, Strous GJ (2003) Dimerization and signal transduction of the growth hormone receptor. Mol Endocrinol 17:967–975. https://doi.org/10.1210/me.2002-0261

  53. 53.

    Herrington J (2001) Signaling pathways activated by the growth hormone receptor. Trends Endocrinol Metab 12:252–257. https://doi.org/10.1016/S1043-2760(01)00423-4

  54. 54.

    Gebel A, Lesinski M, Behm DG, Granacher U (2018) Effects and dose–response relationship of balance training on balance performance in youth: a systematic review and meta-analysis. Sport Med 48:2067–2089. https://doi.org/10.1007/s40279-018-0926-0

  55. 55.

    Borde R, Hortobágyi T, Granacher U (2015) Dose–response relationships of resistance training in healthy old adults: a systematic review and meta-analysis. Sport Med 45:1693–1720. https://doi.org/10.1007/s40279-015-0385-9

  56. 56.

    Kraemer WJ, Flanagan SD, Volek JS et al (2013) Resistance exercise induces region-specific adaptations in anterior pituitary gland structure and function in rats. J Appl Physiol 115:1641–1647. https://doi.org/10.1152/japplphysiol.00687.2013

  57. 57.

    Menagh P, Turner R, Jump D et al (2009) Growth hormone regulates the balance between bone formation and bone marrow adiposity. J Bone Miner Res 25:757–768. 091012153414059-44. https://doi.org/10.1359/jbmr.091015

  58. 58.

    Leme JACA, Silveira RF, Gomes RJ et al (2009) Long-term physical training increases liver IGF-I in diabetic rats. Growth Hormon IGF Res 19:262–266. https://doi.org/10.1016/j.ghir.2008.12.004

  59. 59.

    Hojan K, Milecki P, Leszczyński P (2013) The impact of aerobic exercises on bone mineral density in breast cancer women during endocrine therapy. Polish Orthop Traumatol 78:47–51

  60. 60.

    Castoldi RC, Louzada MJQ, De Oliveira BRSM et al (2017) Effects of aerobic, anaerobic, and concurrent training on bone mineral density of rats. Motriz Rev Educ Fis 23:71–75. https://doi.org/10.1590/S1980-6574201700010011

  61. 61.

    Ehrnborg C, Rosén T (2008) Physiological and pharmacological basis for the ergogenic effects of growth hormone in elite sports. Asian J Androl 10:373–383. https://doi.org/10.1111/j.1745-7262.2008.00403.x

  62. 62.

    Ghiasi R, Mohammadi M, Ashrafi Helan J et al (2015) Influence of two various durations of resistance exercise on oxidative stress in the male rat’s hearts. J Cardiovasc Thorac Res 7:149–153. https://doi.org/10.15171/jcvtr.2015.32

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The authors are grateful to the Brazilian Agency of Resources for Higher Education Personnel (CAPES) for supporting the development of this study.


This study was funded by Brazilian Agency of Resources for Higher Education Personnel (CAPES).

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Correspondence to Robson Chacon Castoldi.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. All procedures were approved by the ethics committee for animal use from University of Western São Paulo - UNOESTE (SP, Brazil; protocol no. 2626). This article does not contain any studies with human participants performed by any of the authors.

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Castoldi, R.C., Ozaki, G.A.T., Garcia, T.A. et al. Effects of muscular strength training and growth hormone (GH) supplementation on femoral bone tissue: analysis by Raman spectroscopy, dual-energy X-ray absorptiometry, and mechanical resistance. Lasers Med Sci 35, 345–354 (2020). https://doi.org/10.1007/s10103-019-02821-5

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  • Physical training
  • Growth hormone
  • Bone tissue
  • Muscular strength
  • Anabolics