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.
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.
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.
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.
Calguneri, M.; Bird, H. A.; Wright, V. Changes in joint laxity occurring during pregnancy. Ann. Rheum. Dis. 41:126–128; 1982.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Sapadin, A. N.; Fleischmajer, R. Treatment of scleroderma. Arch. Dermatol. 138:99–105; 2002.
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.
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.
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.
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.
Slauterbeck, J. R.; Hardy, D. M. Sex hormones and knee ligament injuries in female athletes. Am. J. Med. Sci. 322:196–199; 2001.
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.
Triana, S. M.; Bromberg, D. F. ACL injury patterns in women. Orthopedics 20:545–549; quiz 550–551; 1997.
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.
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.
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.
Wolman, R. L. Association between the menstrual cycle and anterior cruciate ligament in female athletes. Am. J. Sports Med. 27:270–271; 1999.
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.
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.
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.
Author information
Authors and Affiliations
Corresponding author
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1290/0512089.1