Skip to main content

Advertisement

SpringerLink
Log in

Longitudinal Examination of Bone Loss in Male Rats After Moderate–Severe Contusion Spinal Cord Injury

  • Original Research
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

To elucidate mechanisms of bone loss after spinal cord injury (SCI), we evaluated the time-course of cancellous and cortical bone microarchitectural deterioration via microcomputed tomography, measured histomorphometric and circulating bone turnover indices, and characterized the development of whole bone mechanical deficits in a clinically relevant experimental SCI model. 16-weeks-old male Sprague–Dawley rats received T9 laminectomy (SHAM, n = 50) or moderate–severe contusion SCI (n = 52). Outcomes were assessed at 2-weeks, 1-month, 2-months, and 3-months post-surgery. SCI produced immediate sublesional paralysis and persistent hindlimb locomotor impairment. Higher circulating tartrate-resistant acid phosphatase 5b (bone resorption marker) and lower osteoblast bone surface and histomorphometric cancellous bone formation indices were present in SCI animals at 2-weeks post-surgery, suggesting uncoupled cancellous bone turnover. Distal femoral and proximal tibial cancellous bone volume, trabecular thickness, and trabecular number were markedly lower after SCI, with the residual cancellous network exhibiting less trabecular connectivity. Periosteal bone formation indices were lower at 2-weeks and 1-month post-SCI, preceding femoral cortical bone loss and the development of bone mechanical deficits at the distal femur and femoral diaphysis. SCI animals also exhibited lower serum testosterone than SHAM, until 2-months post-surgery, and lower serum leptin throughout. Our moderate–severe contusion SCI model displayed rapid cancellous bone deterioration and more gradual cortical bone loss and development of whole bone mechanical deficits, which likely resulted from a temporal uncoupling of bone turnover, similar to the sequalae observed in the motor-complete SCI population. Low testosterone and/or leptin may contribute to the molecular mechanisms underlying bone deterioration after SCI.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Bauman WA, Cardozo CP (2015) Osteoporosis in individuals with spinal cord injury. PMR 7(2):188–201

    Article  Google Scholar 

  2. Dauty M, Perrouin Verbe B, Maugars Y, Dubois C, Mathe JF (2000) Supralesional and sublesional bone mineral density in spinal cord-injured patients. Bone 27(2):305–309

    Article  CAS  PubMed  Google Scholar 

  3. Eser P, Frotzler A, Zehnder Y, Wick L, Knecht H, Denoth J, Schiessl H (2004) Relationship between the duration of paralysis and bone structure: a pQCT study of spinal cord injured individuals. Bone 34(5):869–880

    Article  CAS  PubMed  Google Scholar 

  4. Frotzler A, Berger M, Knecht H, Eser P (2008) Bone steady-state is established at reduced bone strength after spinal cord injury: a longitudinal study using peripheral quantitative computed tomography (pQCT). Bone 43(3):549–555

    Article  PubMed  Google Scholar 

  5. Frisbie JH (1997) Fractures after myelopathy: the risk quantified. J Spinal Cord Med 20(1):66–69

    Article  CAS  PubMed  Google Scholar 

  6. Morse LR, Battaglino RA, Stolzmann KL, Hallett LD, Waddimba A, Gagnon D, Lazzari AA, Garshick E (2009) Osteoporotic fractures and hospitalization risk in chronic spinal cord injury. Osteoporos Int 20(3):385–392

    Article  CAS  PubMed  Google Scholar 

  7. Grassner L, Klein B, Maier D, Buhren V, Vogel M (2017) Lower extremity fractures in patients with spinal cord injury characteristics, outcome and risk factors for non-unions. J Spinal Cord Med. https://doi.org/10.1080/10790268.2017.1329915

    Article  PubMed  Google Scholar 

  8. Jiang SD, Jiang LS, Dai LY (2007) Changes in bone mass, bone structure, bone biomechanical properties, and bone metabolism after spinal cord injury: a 6-month longitudinal study in growing rats. Calcif Tissue Int 80(3):167–175

    Article  CAS  PubMed  Google Scholar 

  9. Sharif-Alhoseini M, Khormali M, Rezaei M et al (2017) Animal models of spinal cord injury: a systematic review. Spinal Cord 55(8):714–721

    Article  CAS  PubMed  Google Scholar 

  10. Morse L, Teng YD, Pham L et al (2008) Spinal cord injury causes rapid osteoclastic resorption and growth plate abnormalities in growing rats (SCI-induced bone loss in growing rats). Osteoporos Int 19(5):645–652

    Article  CAS  PubMed  Google Scholar 

  11. Morse LR, Xu Y, Solomon B, Boyle L, Yoganathan S, Stashenko P, Battaglino RA (2011) Severe spinal cord injury causes immediate multi-cellular dysfunction at the chondro-osseous junction. Transl Stroke Res 2(4):643–650

    Article  PubMed  PubMed Central  Google Scholar 

  12. Yarrow JF, Ye F, Balaez A et al (2014) Bone loss in a new rodent model combining spinal cord injury and cast immobilization. J Musculoskelet Neuronal Interact 14(3):255–266

    CAS  PubMed  Google Scholar 

  13. Voor MJ, Brown EH, Xu Q, Waddell SW, Burden RL Jr, Burke DA, Magnuson DS (2012) Bone loss following spinal cord injury in a rat model. J Neurotrauma 29(8):1676–1682

    Article  PubMed  PubMed Central  Google Scholar 

  14. Lin T, Tong W, Chandra A et al (2015) A comprehensive study of long-term skeletal changes after spinal cord injury in adult rats. Bone Res 3:15028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Lin CY, Androjna C, Rozic R, Nguyen BT, Parsons B, Midura RJ, Lee YS (2018) Differential adaptations of the musculoskeletal system following spinal cord contusion and transection in rats. J Neurotrauma 35(15):1737–1744

    Article  PubMed  PubMed Central  Google Scholar 

  16. Jiang SD, Jiang LS, Dai LY (2006) Spinal cord injury causes more damage to bone mass, bone structure, biomechanical properties and bone metabolism than sciatic neurectomy in young rats. Osteoporos Int 17(10):1552–1561

    Article  PubMed  Google Scholar 

  17. Liu D, Zhao CQ, Li H, Jiang SD, Jiang LS, Dai LY (2008) Effects of spinal cord injury and hindlimb immobilization on sublesional and supralesional bones in young growing rats. Bone 43(1):119–125

    Article  CAS  PubMed  Google Scholar 

  18. Jiang SD, Shen C, Jiang LS, Dai LY (2007) Differences of bone mass and bone structure in osteopenic rat models caused by spinal cord injury and ovariectomy. Osteoporos Int 18(6):743–750

    Article  PubMed  Google Scholar 

  19. Devivo MJ (2012) Epidemiology of traumatic spinal cord injury: trends and future implications. Spinal Cord 50(5):365–372

    Article  CAS  PubMed  Google Scholar 

  20. Basso DM, Beattie MS, Bresnahan JC (1996) Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection. Exp Neurol 139(2):244–256

    Article  CAS  PubMed  Google Scholar 

  21. Yarrow JF, Conover CF, Beggs LA et al (2014) Testosterone dose dependently prevents bone and muscle loss in rodents after spinal cord injury. J Neurotrauma 31(9):834–845

    Article  PubMed  PubMed Central  Google Scholar 

  22. Yarrow JF, Phillips EG, Conover CF et al (2017) Testosterone plus finasteride prevents bone loss without prostate growth in a rodent spinal cord injury model. J Neurotrauma 34(21):2972–2981

    Article  PubMed  Google Scholar 

  23. Beggs LA, Ye F, Ghosh P et al (2015) Sclerostin inhibition prevents spinal cord injury-induced cancellous bone loss. J Bone Miner Res 30(4):681–689

    Article  CAS  PubMed  Google Scholar 

  24. Yarrow JF, Conover CF, Purandare AV, Bhakta AM, Zheng N, Conrad B, Altman MK, Franz SE, Wronski TJ, Borst SE (2008) Supraphysiological testosterone enanthate administration prevents bone loss and augments bone strength in gonadectomized male and female rats. Am J Physiol Endocrinol Metab 295(5):E1213–E1222

    Article  CAS  PubMed  Google Scholar 

  25. McCoy SC, Yarrow JF, Conover CF et al (2012) 17beta-Hydroxyestra-4,9,11-trien-3-one (Trenbolone) preserves bone mineral density in skeletally mature orchiectomized rats without prostate enlargement. Bone 51(4):667–673

    Article  CAS  PubMed  Google Scholar 

  26. Carbone LD, Chin AS, Burns SP, Svircev JN, Hoenig H, Heggeness M, Bailey L, Weaver F (2014) Mortality after lower extremity fractures in men with spinal cord injury. J Bone Miner Res 29(2):432–439

    Article  PubMed  Google Scholar 

  27. Reiter AL, Volk A, Vollmar J, Fromm B, Gerner HJ (2007) Changes of basic bone turnover parameters in short-term and long-term patients with spinal cord injury. Eur Spine J 16(6):771–776

    Article  PubMed  Google Scholar 

  28. Jiang SD, Yan J, Jiang LS, Dai LY (2011) Down-regulation of the Wnt, estrogen receptor, insulin-like growth factor-I, and bone morphogenetic protein pathways in osteoblasts from rats with chronic spinal cord injury. Joint Bone Spine 78(5):488–492

    Article  CAS  PubMed  Google Scholar 

  29. Zhao W, Li X, Peng Y et al. (2018) Sclerostin antibody reverses the severe sublesional bone loss in rats after chronic spinal cord injury. Calcif Tissue Int. https://doi.org/10.1007/s00223-018-0439-8

    Article  PubMed  PubMed Central  Google Scholar 

  30. Modrowski D, del Pozo E, Miravet L (1992) Dynamics of circulating osteocalcin in rats during growth and under experimental conditions. Horm Metab Res 24(10):474–477

    Article  CAS  PubMed  Google Scholar 

  31. Han B, Copeland M, Geiser AG, Hale LV, Harvey A, Ma YL, Powers CS, Sato M, You J, Hale JE (2007) Development of a highly sensitive, high-throughput, mass spectrometry-based assay for rat procollagen type-I N-terminal propeptide (PINP) to measure bone formation activity. J Proteome Res 6(11):4218–4229

    Article  CAS  PubMed  Google Scholar 

  32. Ito M, Nishida A, Koga A, Ikeda S, Shiraishi A, Uetani M, Hayashi K, Nakamura T (2002) Contribution of trabecular and cortical components to the mechanical properties of bone and their regulating parameters. Bone 31(3):351–358

    Article  CAS  PubMed  Google Scholar 

  33. Bauman WA, La Fountaine MF, Cirnigliaro CM, Kirshblum SC, Spungen AM (2017) Testicular responses to hCG stimulation at varying doses in men with spinal cord injury. Spinal Cord 55(7):659–663

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Sullivan SD, Nash MS, Tefera E, Tinsley E, Blackman MR, Groah S (2017) Prevalence and etiology of hypogonadism in young men with chronic spinal cord injury: a cross-sectional analysis from two university-based rehabilitation centers. PMR 9(8):751–760

    Article  Google Scholar 

  35. Maimoun L, Lumbroso S, Paris F, Couret I, Peruchon E, Rouays-Mabit E, Rossi M, Leroux JL, Sultan C (2006) The role of androgens or growth factors in the bone resorption process in recent spinal cord injured patients: a cross-sectional study. Spinal Cord 44(12):791–797

    Article  CAS  PubMed  Google Scholar 

  36. Park AJ, Battaglino RA, Nguyen NMH, Morse LR (2018) Associations between lean mass and leptin in men with chronic spinal cord injury: results from the FRASCI-muscle study. PLoS ONE 13(6):e0198969

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Phillips EG, Beggs LA, Ye F, Conover CF, Beck DT, Otzel DM, Ghosh P, Bassit ACF, Borst SE, Yarrow JF (2018) Effects of pharmacologic sclerostin inhibition or testosterone administration on soleus muscle atrophy in rodents after spinal cord injury. PLoS ONE 13(3):e0194440

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Powell D, Affuso O, Chen Y (2017) Weight change after spinal cord injury. J Spinal Cord Med 40(2):130–137

    Article  PubMed  Google Scholar 

  39. Doubelt I, Totosy de Zepetnek J, MacDonald MJ, Atkinson SA (2015) Influences of nutrition and adiposity on bone mineral density in individuals with chronic spinal cord injury: A cross-sectional, observational study. Bone Rep 2:26–31

    Article  PubMed  PubMed Central  Google Scholar 

  40. Sabour H, Norouzi Javidan A, Latifi S, Shidfar F, Vafa MR, Emami Razavi SH, Larijani B, Heshmat R (2015) Relationship between leptin and adiponectin concentrations in plasma and femoral and spinal bone mineral density in spinal cord-injured individuals. Spine J 15(1):1–9

    Article  PubMed  Google Scholar 

  41. Turner RT, Kalra SP, Wong CP, Philbrick KA, Lindenmaier LB, Boghossian S, Iwaniec UT (2013) Peripheral leptin regulates bone formation. J Bone Miner Res 28(1):22–34

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Baek K, Bloomfield SA (2009) Beta-adrenergic blockade and leptin replacement effectively mitigate disuse bone loss. J Bone Miner Res 24(5):792–799

    Article  CAS  PubMed  Google Scholar 

  43. Qin W, Li X, Peng Y et al (2016) Sclerostin antibody preserves the morphology and structure of osteocytes and blocks the severe skeletal deterioration after motor-complete spinal cord injury in rats. J Bone Miner Res 31(7):1482

    Article  PubMed  Google Scholar 

  44. Gifre L, Vidal J, Carrasco JL, Filella X, Ruiz-Gaspa S, Muxi A, Portell E, Monegal A, Guanabens N, Peris P (2015) Effect of recent spinal cord injury on wnt signaling antagonists (sclerostin and dkk-1) and their relationship with bone loss. A 12-month prospective study. J Bone Miner Res 30(6):1014–1021

    Article  CAS  PubMed  Google Scholar 

  45. Qin W, Li X, Peng Y et al (2015) Sclerostin antibody preserves the morphology and structure of osteocytes and blocks the severe skeletal deterioration after motor-complete spinal cord injury in rats. J Bone Miner Res 30(11):1994–2004

    Article  CAS  PubMed  Google Scholar 

  46. Pruss H, Tedeschi A, Thiriot A et al (2017) Spinal cord injury-induced immunodeficiency is mediated by a sympathetic-neuroendocrine adrenal reflex. Nat Neurosci 20(11):1549–1559

    Article  CAS  PubMed  Google Scholar 

  47. Zhang Y, Guan Z, Reader B et al (2013) Autonomic dysreflexia causes chronic immune suppression after spinal cord injury. J Neurosci 33(32):12970–12981

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Samantaray S, Das A, Matzelle DC, Yu SP, Wei L, Varma A, Ray SK, Banik NL (2016) Administration of low dose estrogen attenuates persistent inflammation, promotes angiogenesis, and improves locomotor function following chronic spinal cord injury in rats. J Neurochem 137(4):604–617

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Yarrow JF, McCoy SC, Ferreira JA, Pingel JE, Conrad BP, Wronski TJ, Williams AA, Borst SE, Brown M (2012) A rehabilitation exercise program induces severe bone mineral deficits in estrogen-deficient rats after extended disuse. Menopause 19(11):1267–1276

    Article  PubMed  Google Scholar 

  50. Yarrow JF, Wronski TJ, Borst SE (2015) Testosterone and adult male bone: actions independent of 5 alpha-reductase and aromatase. Exerc Sport Sci Rev 43(4):222–230

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Office of Research and Development, Rehabilitation Research and Development (RR&D) Service, Department of Veterans Affairs (SPiRE 1I21RX001273-01 and PECASE #B9280-O) to JFY, and by resources provided by the North Florida/South Georgia Veterans Health System. The work reported herein does not represent the views of the US Department of Veterans Affairs or the US Government.

Author information

Authors and Affiliations

  1. Brain Rehabilitation Research Center, Malcom Randall Department of Veterans Affairs Medical Center, North Florida/South Georgia Veterans Health System, 1601 SW Archer Road, Research 151, Gainesville, FL, 32608, USA

    Dana M. Otzel, Yi Zhang & Prodip K. Bose

  2. Research Service, Malcom Randall Department of Veterans Affairs Medical Center, North Florida/South Georgia Veterans Health System, 1601 SW Archer Road, Research 151, Gainesville, FL, 32608, USA

    Christine F. Conover, Fan Ye, Ean G. Phillips, Taylor Bassett, Russell D. Wnek, Micah Flores, Andrea Catter, Payal Ghosh, Alexander Balaez, Jason Petusevsky & Joshua F. Yarrow

  3. Department of Orthopedics and Rehabilitation, University of Florida, PO Box 112727, Gainesville, FL, 32611, USA

    Cong Chen

  4. University of Florida College of Medicine, Jacksonville, FL, 32209, USA

    Yongxin Gao

  5. Department of Physiological Sciences, University of Florida, PO Box 100144, Gainesville, FL, 32610, USA

    Jessica M. Jiron, Prodip K. Bose, Thomas J. Wronski & J. Ignacio Aguirre

  6. Department of Neurology, University of Florida, HSC PO Box 100236, Gainesville, FL, 32610, USA

    Prodip K. Bose

  7. Department of Applied Physiology and Kinesiology, University of Florida, PO Box 118205, Gainesville, FL, 32611, USA

    Stephen E. Borst

  8. Division of Endocrinology, Diabetes, and Metabolism, University of Florida College of Medicine, 1600 SW Archer Road, Gainesville, FL, 32610, USA

    Joshua F. Yarrow

Authors
  1. Dana M. Otzel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine F. Conover
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fan Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ean G. Phillips
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Taylor Bassett
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Russell D. Wnek
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Micah Flores
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Andrea Catter
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Payal Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Alexander Balaez
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jason Petusevsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Cong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Yongxin Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Yi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Jessica M. Jiron
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Prodip K. Bose
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Stephen E. Borst
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Thomas J. Wronski
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. J. Ignacio Aguirre
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Joshua F. Yarrow
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Joshua F. Yarrow.

Ethics declarations

Conflict of interest

Dana M. Otzel, Christine F. Conover, Fan Ye, Ean G. Phillips, Taylor Bassett, Russell D. Wnek, Micah Flores, Andrea Catter, Payal Ghosh, Alexander Balaez, Jason Petusevsky, Cong Chen, Yongxin Gao, Yi Zhang, Jessica M. Jiron, Prodip K. Bose, Stephen E. Borst, Thomas J. Wronski, J. Ignacio Aguirre and Joshua F. Yarrow declares that they have no conflict of interest.

Human and Animal Rights and Informed Consent

All experimental procedures conformed to the ILAR Guide to the Care and Use of Experimental Animals and were approved by the Institutional Animal Care and Use Committee at the Malcom Randall VA Medical Center.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Otzel, D.M., Conover, C.F., Ye, F. et al. Longitudinal Examination of Bone Loss in Male Rats After Moderate–Severe Contusion Spinal Cord Injury. Calcif Tissue Int 104, 79–91 (2019). https://doi.org/10.1007/s00223-018-0471-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-018-0471-8

Keywords

Search

Navigation