Skip to main content
SpringerLink
Log in

Quantitation of estrogen receptors and relaxin binding in human anterior cruciate ligament fibroblasts

  • Articles
  • Cell and Tissue Models
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

The significantly higher incidence of anterior cruciate ligament (ACL) injuries in collegiate women compared with men may result from relative ligament laxity. Differences in estrogen and relaxin activity, similar to that seen in pregnancy, may account for this. We quantified estrogen receptors by flow cytometry and relaxin receptors by radioligand binding assay in human ACL cells and compared the presence of these receptors in males and females. ACL stumps were harvested from seven males and eight females with acute ACL injuries. The tissue was placed in M199 cell culture medium. Outgrowth cultures were obtained, and passage 2 cells were used for all studies. Estrogen receptor determination was performed using flow cytometry. Relaxin binding was performed in ACL cells derived from five female and male patients using I125-labeled relaxin. Estrogen receptors were identified by flow cytometry in 4 to 10% of ACL cells. Mean fluorescence of cells expressing estrogen receptors was approximately twice that of controls, with no significant differences between males and females. Relaxin studies showed low-level binding of I125-relaxin-labeled ACL cells. Relaxin binding was present in four out of five female ACL cells versus one out of five male ACL cells.

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

Similar content being viewed by others

References

  • Arnndt, E.; Dick, R. Knee injury patterns among men and women in collegiate basketball and soccer: NCAA data and review of literature. Am. J. Sports Med. 23:694–701; 1995.

    Google Scholar 

  • Arnold, C.; Van Bell, C.; Rogers, V.; Cooney, T. The relationship between serum relaxin and knee joint laxity in female athletes. Orthopedics 25:669–673; 2002.

    PubMed  Google Scholar 

  • Brotherick, I.; Lennard, T. W.; Wilkinson, S. E.; Cook, S.; Angus, B.; Shenton, B. K. Flow cytometric method for the measurement of epidermal growth factor receptor and comparison with the radio-ligand binding assay. Cytometry 16:262–269; 1994.

    Article  PubMed  CAS  Google Scholar 

  • Calguneri, M.; Bird, H. A.; Wright, V. Changes in joint laxity occurring during pregnancy. Ann. Rheum. Dis. 41:126–128; 1982.

    Article  PubMed  CAS  Google Scholar 

  • Charlton, W. P.; Coslett-Charlton, L. M.; Ciccotti, M. G. Correlation of estradiol in pregnancy and anterior cruciate ligament laxity. Clin. Orthop. 165–170; 2001.

  • Deie, M.; Sakamaki, Y.; Sumen, Y.; Urabe, Y.; Ikuta, Y. Anterior knee laxity in young women varies with their menstrual cycle. Int. Orthop. 26:154–156; 2002.

    Article  PubMed  Google Scholar 

  • Dragoo, J. L.; Lee, R. S.; Benhaim, P.; Finerman, G. A.; Hame, S. L. Relaxin receptors in the human female anterior cruciate ligament. Am. J. Sports. Med. 31:577–584; 2003.

    PubMed  Google Scholar 

  • Galey, S.; Konieczko, E. M.; Arnold, C. A.; Cooney, T. E. Immuohistological detection of relaxin binding to anterior cruciate ligaments. J. Ortho. 26:1201–1204; 2003.

    Google Scholar 

  • Karageanes, S. J.; Blackburn, K.; Vangelos, Z. A. The association of the menstrual cycle with the laxity of the anterior cruciate ligament in adolescent female athletes. Clin. J. Sport Med. 10:162–168; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Liu, S. H.; Al-Shaikh, R.; Panossian, V.; Yang, R. S.; Nelson, S. D.; Soleiman, N.; Finerman, G. A.; Lane, J. M. Primary immunolocalization of estrogen and progesterone target cells in the human anterior cruciate ligament. J. Orthop. Res. 14:526–533; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Liu, S. H.; Al-Shaikh, R. A.; Panossian, V.; Finerman, G. A.; Lane, J. M. Estrogen affects the cellular metabolism of the anterior cruciate ligament: a potential explanation for female athletic, injury. Am. J. Sports Med. 25:704–9; 1997.

    PubMed  CAS  Google Scholar 

  • McShane, J. M.; Balsbaugh, T.; Simpson, Z.; Diamond, J. J.; Bryan, S. T.; Velez, J. Association between the menstrual cycle and anterior cruciate ligament injuries in female athletes. Am. J. Sports Med. 28:131; 2000.

    PubMed  CAS  Google Scholar 

  • Min, G.; Sherwood, O. D. Identification of specific relaxin-binding cells in the cervix, mammary glands, nipples, small intestine, and skin of pregnant pigs. Biol. Reprod. 55:1243–1252; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Moller-Nielsen, J.; Hammar, M. Women's soccer injuries in relation to the menstrual cycle and oral contraceptive use. Med. Sci. Sports Exerc. 21:126–129; 1989.

    PubMed  CAS  Google Scholar 

  • Myklebust, G.; Engebretsen, L.; Braekken, I. H.; Skjolberg, A.; Olsen, O. E.; Bahr, R. Prevention of anterior cruciate ligament injuries in female team handball players: a prospective intervention study over three seasons. Clin. J. Sport Med. 13:71–78; 2003.

    Article  PubMed  Google Scholar 

  • Myklebust, G.; Maehlum, S.; Holm, I.; Bahr, R. A prospective cohort study of anterior cruciate ligament injuries in elite Norwegian team handball. Scand. J. Med. Sci. Sports 8:149–153; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Ohtera, K.; Zobitz, M. E.; Luo, Z. P.; Morrey, B. F.; O'Driscoll, S. W.; Ramin, K. D.; An, K. N. Effect of pregnancy on joint contracture in the rat knee. J. Appl. Physiol. 92:1494–1498; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Palejwala, S.; Stein, D.; Wojtczuk, A.; Weiss, G.; Goldsmith, L. T. Demonstration of a relaxin receptor and relaxin-stimulated tyrosine phosphorylation in human lower uterine segment fibroblasts. Endocrinology 139:1208–1212; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Petersen, L. K.; Vogel, I.; Agger, A. O.; Westergard, J.; Nils, M.; Uldbjerg, N. Variations in serum relaxin (hRLX-2) concentrations during human pregnancy. Acta Obstet. Gynecol. Scand. 74:251–256; 1995.

    PubMed  CAS  Google Scholar 

  • Romani, W.; Patrie, J.; Curl, L. A.; Flaws, J. A. The correlations between estradiol, estrone, estriol, progesterone, and sex hormone-binding globulin and anterior cruciate ligament stiffness in healthy, active females. J. Womens Health (Larchmt) 12:287–298; 2003.

    Article  Google Scholar 

  • Sapadin, A. N.; Fleischmajer, R. Treatment of scleroderma. Arch. Dermatol. 138:99–105; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Schauberger, C. W.; Rooney, B. L.; Goldsmith, L.; Shenton, D.; Silva, P. D.; Schaper A. Peripheral joint laxity increases in pregnancy but does not correlate with serum relaxin levels. Am. J. Obstet. Gynecol. 174:667–671; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Seneviratne, A.; Attia, E.; Williams, R. J.; Rodeo, S. A.; Hannafin, J. A. The effect of estrogen on ovine anterior cruciate ligament fibroblasts: cell proliferation and collagen synthesis. Am. J. Sports Med. 32:1613–1618; 2004.

    Article  PubMed  Google Scholar 

  • Slauterbeck, J.; Clevenger, C.; Lundberg, W.; Burchfield, D. M. Estrogen level alters the failure load of the rabbit anterior cruciate ligament. J. Orthop. Res. 17:405–408; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Slauterbeck, J. R.; Fuzie, S. F.; Smith, M. P.; Clark, R. J.; Xu, K.; Starch, D. W.; Hardy, D. M. The menstrual cycle, sex hormones, and anterior cruciate ligament injury. J. Athl. Train. 37:275–278; 2002.

    PubMed  Google Scholar 

  • Slauterbeck, J. R.; Hardy, D. M. Sex hormones and knee ligament injuries in female athletes. Am. J. Med. Sci. 322:196–199; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Strickland, S. M.; Belknap, T. W.; Turner, S. A.; Wright, T. M.; Hannafin, J. A. Lack of hormonal influences on mechanical properties of sheep knee ligaments. Am. J. Sports Med. 31: 210–215; 2003.

    PubMed  Google Scholar 

  • Triana, S. M.; Bromberg, D. F. ACL injury patterns in women. Orthopedics 20:545–549; quiz 550–551; 1997.

    Google Scholar 

  • Unemori, E. N.; Amento, E. P. Relaxin modulates synthesis and secretion of procollagenase and collagen by human dermal fibroblasts. J. Biol. Chem. 265:10681–10685; 1990.

    PubMed  CAS  Google Scholar 

  • Wojtys, E. M.; Huston, L. J.; Boynton, M. D.; Spindler, K. P.; Lindenfeld, T. N. The effect of the menstrual cycle on anterior cruciate ligament injuries in women as determined by hormone levels. Am. J. Sports Med. 30:182–188; 2002.

    PubMed  Google Scholar 

  • Wojtys, E. M.; Huston, L. J.; Lindenfeld, T. N.; Hewett, T. E.; Greenfield, M. L. Association between the menstrual cycle and anterior cruciate ligament injuries in female athletes. Am J. Sports Med. 26:614–619; 1998.

    PubMed  CAS  Google Scholar 

  • Wolman, R. L. Association between the menstrual cycle and anterior cruciate ligament in female athletes. Am. J. Sports Med. 27:270–271; 1999.

    PubMed  CAS  Google Scholar 

  • Yu, W. D.; Liu, S. H; Hatch, J. D.; Panossian, V.; Finerman, G. A. Effect of estrogen on cellular metabolism of the human anterior cruciate ligament. Clin. Orthop. 366:229–238; 1999.

    Article  PubMed  Google Scholar 

  • Yu, W. D.; Panossian, V.; Hatch, J. D.; Liu, S. H.; Finerman, G. A. Combined effects of estrogen and progesterone on the anterior cruciate ligament. Clin. Orthop. 383:268–81; 2001.

    Article  PubMed  Google Scholar 

  • Zelisko, J. A.; Noble, H. B.; Porter, M. A comparison of men's and women's professional basketbal injuries. Am. J. Sports Med. 10:297–9; 1982.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Orthopedic Surgery and Sports Medicine, The Permanente Medical Group, Inc., Santa Clara, California

    Deborah A. Faryniarz

  2. Sports Medicine and Shoulder Service, Hospital for Special Surgery, New York

    Madhu Bhargava, Erik T. Attia & Jo A. Hannafin

  3. Department of Orthopaedic Surgery, Mayo Clinic, Rochester, Minnesota

    Claudette Lajam

Authors
  1. Deborah A. Faryniarz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Madhu Bhargava
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Claudette Lajam
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Erik T. Attia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jo A. Hannafin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Deborah A. Faryniarz.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Faryniarz, D.A., Bhargava, M., Lajam, C. et al. Quantitation of estrogen receptors and relaxin binding in human anterior cruciate ligament fibroblasts. In Vitro Cell.Dev.Biol.-Animal 42, 176–181 (2006). https://doi.org/10.1290/0512089.1

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1290/0512089.1

Key words

Search

Navigation