Skip to main content
SpringerLink
Log in

Effect of dihydrotestosterone on cultured human tenocytes from intact supraspinatus tendon

  • Experimental Study
  • Published:
Knee Surgery, Sports Traumatology, Arthroscopy Aims and scope

Abstract

The role of hormones in the pathogenesis of tendinopathy is not well recognised, even though the use of anabolic steroids is correlated with a higher incidence of spontaneous tendon ruptures. The aim of this study was to investigate the effects of dihydrotestosterone (DHT) on human tenocyte cultures from the intact supraspinatus tendon of male subjects. Cultured human tenocytes were seeded into culture plates at a density of 5 × 104 cells per well and incubated for 24 h. Then, 10−9 M–10−7 M DHT or Dulbecco’s modified Eagle’s medium (DMEM) only (control) was added to the culture plate wells. Cell morphology assessment and cell proliferation tests were performed 48, 72 and 96 h after DHT treatment. DHT-treated tenocytes showed an increased proliferation rate at DHT concentration higher than 10−8 M. Differences in cell numbers between control and DHT-treated cells were statistically significant (P < 0.05) after 48 and 72 h of treatment with DHT concentrations of 10−8 and 10−7 M. The tenocytes treated with DHT (10−8 and 10−7 M) became more flattened and polygonal compared to control cells that maintained their fibroblast-like appearance during the experiment at each observation time. In conclusion, in vitro, progressive increasing concentration of DHT at doses greater than 10−8 M had direct effects on male human tenocytes, increasing cell number after 48 and 72 h of treatment, and leading to a dedifferentiated phenotype after 48 h of treatment. This effect can be important during tendon-healing and repair, when active proliferation is required. Our results represent preliminary evidence for a possible correlation between testosterone abuse and shoulder tendinopathy.

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

Similar content being viewed by others

References

  1. Ames PR, Longo UG, Denaro V, Maffulli N (2008) Achilles tendon problems: not just an orthopaedic issue. Disabil Rehabil 30:1646–1650

    Article  PubMed  Google Scholar 

  2. Andres BM, Murrell GA (2008) Treatment of tendinopathy: what works, what does not, and what is on the horizon. Clin Orthop Relat Res 466:1539–1554

    Article  PubMed  Google Scholar 

  3. Arnoczky SP, Lavagnino M, Egerbacher M, Caballero O, Gardner K (2007) Matrix metalloproteinase inhibitors prevent a decrease in the mechanical properties of stress-deprived tendons: an in vitro experimental study. Am J Sports Med 35:763–769

    Article  PubMed  Google Scholar 

  4. Arnoczky SP, Lavagnino M, Egerbacher M, Caballero O, Gardner K, Shender MA (2008) Loss of homeostatic strain alters mechanostat “set point” of tendon cells in vitro. Clin Orthop Relat Res 466:1583–1591

    Article  PubMed  Google Scholar 

  5. Benz DJ, Haussler MR, Thomas MA, Speelman B, Komm BS (1991) High-affinity androgen binding and androgenic regulation of alpha 1(I)-procollagen and transforming growth factor-beta steady state messenger ribonucleic acid levels in human osteoblast-like osteosarcoma cells. Endocrinology 128:2723–2730

    Article  CAS  PubMed  Google Scholar 

  6. Bhasin S, Storer TW, Berman N, Callegari C, Clevenger B, Phillips J, Bunnell TJ, Tricker R, Shirazi A, Casaburi R (1996) The effects of supraphysiologic doses of testosterone on muscle size and strength in normal men. N Engl J Med 335:1–7

    Article  CAS  PubMed  Google Scholar 

  7. De Mos M, van El B, DeGroot J, Jahr H, van Schie HT, van Arkel ER, Tol H, Heijboer R, van Osch GJ, Verhaar JA (2007) Achilles tendinosis: changes in biochemical composition and collagen turnover rate. Am J Sports Med 35:1549–1556

    Article  PubMed  Google Scholar 

  8. Egerbacher M, Arnoczky SP, Caballero O, Lavagnino M, Gardner KL (2008) Loss of homeostatic tension induces apoptosis in tendon cells: an in vitro study. Clin Orthop Relat Res 466:1562–1568

    Article  PubMed  Google Scholar 

  9. Er F, Gassanov N, Brandt MC, Madershahian N, Hoppe UC (2009) Impact of dihydrotestosterone on L-type calcium channels in human ventricular cardiomyocytes. Endocr Res 34:59–67

    Article  CAS  PubMed  Google Scholar 

  10. Evans NA, Bowrey DJ, Newman GR (1998) Ultrastructural analysis of ruptured tendon from anabolic steroid users. Injury 29:769–773

    Article  CAS  PubMed  Google Scholar 

  11. Franceschi F, Longo UG, Ruzzini L, Morini S, Battistoni F, Dicuonzo G, Maffulli N, Denaro V (2008) Circulating substance P levels and shoulder joint contracture after arthroscopic repair of the rotator cuff. Br J Sports Med 42:742–745

    Article  CAS  PubMed  Google Scholar 

  12. Franceschi F, Longo UG, Ruzzini L, Papalia R, Rizzello G, Denaro V (2007) To detach the long head of the biceps tendon after tenodesis or not: outcome analysis at the 4-year follow-up of two different techniques. Int Orthop 31:537–545

    Article  PubMed  Google Scholar 

  13. Franceschi F, Longo UG, Ruzzini L, Rizzello G, Denaro V (2007) Arthroscopic management of calcific tendinitis of the subscapularis tendon. Knee Surg Sports Traumatol Arthrosc 15:1482–1485

    Article  PubMed  Google Scholar 

  14. Franceschi F, Longo UG, Ruzzini L, Rizzello G, Maffulli N, Denaro V (2008) No advantages in repairing a type II superior labrum anterior and posterior (SLAP) lesion when associated with rotator cuff repair in patients over age 50: a randomized controlled trial. Am J Sports Med 36:247–253

    Article  PubMed  Google Scholar 

  15. Franceschi F, Longo UG, Ruzzini L, Rizzello G, Maffulli N, Denaro V (2007) The Roman Bridge: a “double pulley—suture bridges” technique for rotator cuff repair. BMC Musculoskelet Disord 8:123

    Article  PubMed  Google Scholar 

  16. Franceschi F, Longo UG, Ruzzini L, Rizzello G, Maffulli N, Denaro V (2008) Soft tissue tenodesis of the long head of the biceps tendon associated to the Roman Bridge repair. BMC Musculoskelet Disord 9:78

    Article  PubMed  Google Scholar 

  17. Franceschi F, Ruzzini L, Longo UG, Martina FM, Zobel BB, Maffulli N, Denaro V (2007) Equivalent clinical results of arthroscopic single-row and double-row suture anchor repair for rotator cuff tears: a randomized controlled trial. Am J Sports Med 35:1254–1260

    Article  PubMed  Google Scholar 

  18. Gardner K, Arnoczky SP, Caballero O, Lavagnino M (2008) The effect of stress-deprivation and cyclic loading on the TIMP/MMP ratio in tendon cells: an in vitro experimental study. Disabil Rehabil 30:1523–1529

    Article  PubMed  Google Scholar 

  19. Hartgens F, Kuipers H (2004) Effects of androgenic–anabolic steroids in athletes. Sports Med 34:513–554

    Article  PubMed  Google Scholar 

  20. Inhofe PD, Grana WA, Egle D, Min KW, Tomasek J (1995) The effects of anabolic steroids on rat tendon. An ultrastructural, biomechanical, and biochemical analysis. Am J Sports Med 23:227–232

    Article  CAS  PubMed  Google Scholar 

  21. Jenkins C, Milsted A, Doane K, Meszaros G, Toot J, Ely D (2007) A cell culture model using rat coronary artery adventitial fibroblasts to measure collagen production. BMC Cardiovasc Disord 7:13

    Article  PubMed  Google Scholar 

  22. Kane TP, Ismail M, Calder JD (2008) Topical glyceryl trinitrate and noninsertional Achilles tendinopathy: a clinical and cellular investigation. Am J Sports Med 36:1160–1163

    Article  PubMed  Google Scholar 

  23. Karousou E, Ronga M, Vigetti D, Passi A, Maffulli N (2008) Collagens, proteoglycans, MMP-2, MMP-9 and TIMPs in human achilles tendon rupture. Clin Orthop Relat Res 466:1577–1582

    Article  PubMed  Google Scholar 

  24. Knobloch K (2008) The use of a neovascularization score to predict clinical severity in Achilles tendinopathy. Am J Sports Med 36:395–397

    Article  PubMed  Google Scholar 

  25. Knobloch K, Grasemann R, Spies M, Vogt PM (2008) Midportion achilles tendon microcirculation after intermittent combined cryotherapy and compression compared with cryotherapy alone: a randomized trial. Am J Sports Med 36:2128–2138

    Article  PubMed  Google Scholar 

  26. Knobloch K, Schreibmueller L, Longo UG, Vogt PM (2008) Eccentric exercises for the management of tendinopathy of the main body of the Achilles tendon with or without an AirHeeltrade mark Brace. A randomized controlled trial. B: effects of compliance. Disabil Rehabil 30:1692–1696

    Article  PubMed  Google Scholar 

  27. Knobloch K, Schreibmueller L, Longo UG, Vogt PM (2008) Eccentric exercises for the management of tendinopathy of the main body of the Achilles tendon with or without the AirHeeltrade mark Brace. A randomized controlled trial. A: effects on pain and microcirculation. Disabil Rehabil 30:1685–1691

    Article  PubMed  Google Scholar 

  28. Lavagnino M, Arnoczky SP, Elvin N, Dodds J (2008) Patellar tendon strain is increased at the site of the jumper’s knee lesion during knee flexion and tendon loading: results and cadaveric testing of a computational model. Am J Sports Med 36:2110–2118

    Article  PubMed  Google Scholar 

  29. Longo UG, Franceschi F, Ruzzini L, Rabitti C, Morini S, Maffulli N, Denaro V (2009) Characteristics at Haematoxylin and Eosin staining of ruptures of the long head of the biceps tendon. Br J Sports Med 43:603–607

    Article  CAS  PubMed  Google Scholar 

  30. Longo UG, Franceschi F, Ruzzini L, Rabitti C, Morini S, Maffulli N, Denaro V (2008) Histopathology of the supraspinatus tendon in rotator cuff tears. Am J Sports Med 36:533–538

    Article  PubMed  Google Scholar 

  31. Longo UG, Franceschi F, Ruzzini L, Rabitti C, Morini S, Maffulli N, Forriol F, Denaro V (2007) Light microscopic histology of supraspinatus tendon ruptures. Knee Surg Sports Traumatol Arthrosc 15:1390–1394

    Article  PubMed  Google Scholar 

  32. Longo UG, Garau G, Denaro V, Maffulli N (2008) Surgical management of tendinopathy of biceps femoris tendon in athletes. Disabil Rehabil 30:1602–1607

    Article  PubMed  Google Scholar 

  33. Longo UG, Olivia F, Denaro V, Maffulli N (2008) Oxygen species and overuse tendinopathy in athletes. Disabil Rehabil 30:1563–1571

    Article  PubMed  Google Scholar 

  34. Longo UG, Ramamurthy C, Denaro V, Maffulli N (2008) Minimally invasive stripping for chronic Achilles tendinopathy. Disabil Rehabil 30:1709–1713

    Article  PubMed  Google Scholar 

  35. Maffulli N, Ajis A, Longo UG, Denaro V (2007) Chronic rupture of tendo Achillis. Foot Ankle Clin 12:583–596

    Article  PubMed  Google Scholar 

  36. Maffulli N, Longo UG (2008) Conservative management for tendinopathy: is there enough scientific evidence? Rheumatology (Oxford) 47:390–391

    Article  CAS  Google Scholar 

  37. Maffulli N, Longo UG (2008) How do eccentric exercises work in tendinopathy? Rheumatology (Oxford) 47:1444–1445

    Article  CAS  Google Scholar 

  38. Maffulli N, Testa V, Capasso G, Oliva F, Panni AS, Longo UG, King JB (2008) Surgery for chronic Achilles tendinopathy produces worse results in women. Disabil Rehabil 30:1714–1720

    Article  PubMed  Google Scholar 

  39. Magra M, Maffulli N (2005) Matrix metalloproteases: a role in overuse tendinopathies. Br J Sports Med 39:789–791

    Article  CAS  PubMed  Google Scholar 

  40. Magra M, Maffulli N (2005) Molecular events in tendinopathy: a role for metalloproteases. Foot Ankle Clin 10:267–277

    Article  PubMed  Google Scholar 

  41. Malliaras P, Richards PJ, Garau G, Maffulli N (2008) Achilles tendon Doppler flow may be associated with mechanical loading among active athletes. Am J Sports Med 36:2210–2215

    Article  PubMed  Google Scholar 

  42. Markova MS, Zeskand J, McEntee B, Rothstein J, Jimenez SA, Siracusa LD (2004) A role for the androgen receptor in collagen content of the skin. J Invest Dermatol 123:1052–1056

    Article  CAS  PubMed  Google Scholar 

  43. Marqueti RC, Parizotto NA, Chriguer RS, Perez SE, Selistre-de-Araujo HS (2006) Androgenic–anabolic steroids associated with mechanical loading inhibit matrix metallopeptidase activity and affect the remodeling of the achilles tendon in rats. Am J Sports Med 34:1274–1280

    Article  PubMed  Google Scholar 

  44. Miles JW, Grana WA, Egle D, Min KW, Chitwood J (1992) The effect of anabolic steroids on the biomechanical and histological properties of rat tendon. J Bone Joint Surg Am 74:411–422

    CAS  PubMed  Google Scholar 

  45. Millar NL, Wei AQ, Molloy TJ, Bonar F, Murrell GA (2008) Heat shock protein and apoptosis in supraspinatus tendinopathy. Clin Orthop Relat Res 466:1569–1576

    Article  PubMed  Google Scholar 

  46. Raivio T, Tapanainen JS, Kunelius P, Jänne OA (2002) Serum androgen bioactivity during 5alpha-dihydrotestosterone treatment in elderly men. J Androl 23:919–921

    CAS  PubMed  Google Scholar 

  47. Ray R, Herring CM, Markel TA, Crisostomo PR, Wang M, Weil B, Lahm T, Meldrum DR (2008) Deleterious effects of endogenous and exogenous testosterone on mesenchymal stem cell VEGF production. Am J Physiol Regul Integr Comp Physiol 294:R1498–R1503

    CAS  PubMed  Google Scholar 

  48. Schwarz R, Colarusso L, Doty P (1976) Maintenance of differentiation in primary cultures of avian tendon cells. Exp Cell Res 102:63–71

    Article  CAS  PubMed  Google Scholar 

  49. Scott A, Lian O, Bahr R, Hart DA, Duronio V, Khan KM (2008) Increased mast cell numbers in human patellar tendinosis: correlation with symptom duration and vascular hyperplasia. Br J Sports Med 42:753–757

    Article  CAS  PubMed  Google Scholar 

  50. Sharma P, Maffulli N (2006) Biology of tendon injury: healing, modeling and remodeling. J Musculoskelet Neuronal Interact 6:181–190

    CAS  PubMed  Google Scholar 

  51. Sharma P, Maffulli N (2008) Tendinopathy and tendon injury: the future. Disabil Rehabil 30:1733–1745

    Article  PubMed  Google Scholar 

  52. Singh R, Artaza JN, Taylor WE, Gonzalez-Cadavid NF, Bhasin S (2003) Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway. Endocrinology 144:5081–5088

    Article  CAS  PubMed  Google Scholar 

  53. Xu Y, Murrell GA (2008) The basic science of tendinopathy. Clin Orthop Relat Res 466:1528–1538

    Article  PubMed  Google Scholar 

  54. Yao L, Bestwick CS, Bestwick LA, Maffulli N, Aspden RM (2006) Phenotypic drift in human tenocyte culture. Tissue Eng 12:1843–1849

    Article  CAS  PubMed  Google Scholar 

  55. Yoo HG, Kim JS, Lee SR, Pyo HK, Moon HI, Lee JH, Kwon OS, Chung JH, Kim KH, Eun HC, Cho KH (2006) Perifollicular fibrosis: pathogenetic role in androgenetic alopecia. Biol Pharm Bull 29:1246–1250

    Article  CAS  PubMed  Google Scholar 

Download references

Conflict of interest statement

The authors declare no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Orthopaedic and Trauma Surgery, Campus Biomedico University, Via A. Del Portillo, 200, Rome, Italy

    Vincenzo Denaro, Laura Ruzzini, Umile Giuseppe Longo & Francesco Franceschi

  2. “Giovanni XXIII” Cancer Research Center-Institute of General Pathology, Catholic University, Rome, Italy

    Barbara De Paola, Achille Cittadini & Alessandro Sgambato

  3. Centre for Sports and Exercise Medicine, Queen Mary University of London, Mile End Hospital, 275 Bancroft Road, London, E1 4DG, UK

    Nicola Maffulli

Authors
  1. Vincenzo Denaro
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laura Ruzzini
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Umile Giuseppe Longo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Francesco Franceschi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Barbara De Paola
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Achille Cittadini
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nicola Maffulli
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Alessandro Sgambato
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nicola Maffulli.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Denaro, V., Ruzzini, L., Longo, U.G. et al. Effect of dihydrotestosterone on cultured human tenocytes from intact supraspinatus tendon. Knee Surg Sports Traumatol Arthrosc 18, 971–976 (2010). https://doi.org/10.1007/s00167-009-0953-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00167-009-0953-3

Keywords

Search

Navigation