Does the use of Testosterone Enanthate as a Form of Doping in Sports Cause Early Closure of Epiphyseal in Bones?/?El Uso de Enantato de Testosterona como Forma de Dopaje en el Deporte Causa Cierre Epifisiario Temprano de los Huesos?
Anabolic androgenic steroids (AAS) the speed sports in which high performance, power and endurance is needed (Kanayama et al, 2009); by athletes, body builders and weightlifters (Al-Ismail et al, 2002) to increase muscle mass, strength (Kanayama et al.; Attardi et al., 2011; Furstenberger et al., 2012; Khan et al., 2012; Azzazy et al., 2009) and used to be able to achieve high performance (Lood et al., 2012; Hildebrandt et al., 2011; Di Luigi et al, 2012).
AAS damages organs such as liver (Tasgin et al., 2010; Pagonis et al., 2008), heart (Lok et al, 2010; Hassan & Kamal, 2013; Kaushik et al, 2010; Baggish et al, 2010; Tasgin et al, 2011) and kidney (Hassan & Kamal; Neri et al., 2011). Immune system (Khan et al.), endocrine system, urogenital system (Collins & Basaria, 2012) not only cause the problems like that; but also reveal serious side effects on other systems in the body (Bispo et al, 2009) and prompt serious health problems (Di Luigi et al.).
One of the most serious side effects of these substances used for doping by athletes especially, the result of usage at an early age, it causes early epiphyseal closure in long bones and prevent bone from draw out (Al-Ismail et al.; Soma et al., 2008; Lok et al., 2011; Zaman et al., 2012).
The purpose of this research is to investigate among anabolic androgenic steroids short term usage of testosterone enanthate's, used by athletes for doping, morphometric effects on male rat humerus bones.
MATERIAL AND METHOD
The research was carried on 30 male Wistar rats of 35 days (90-120 g). Rats fed ad libitum and were divided into three groups in standard cages. While control group (n= 10) fed without any application, to the peanut oil group (n= 10) the peanut oil (100 [micro]l, Zade peanut oil, Konya-TURKEY), which is used as testosterone enanthate's diluent, executed 5 days intraperitoneally, after 2 days the same application was performed. To the experimental group (n= 10) testosterone enanthate on (Androne[R] 250 Enj, Caspian tamin pharmaceitucal Co, Rasht-IRAN) 10 mg/kg dose (Lok and Yalcin, 2010), was applied intraperitoneally 5 days by dilueting in 100 [micro]l peanut oil and after 2 days break, it was applied again. All processes have been resumed 3 weeks.
Before, at the begining and during the study, one day in a week weight of rats in all groups were measured and weekly weight averages were taken. Weekly dose adjustment was done in the weight measures of testosterone enanthate applied group. All rats were euthanized at the end of 3 weeks with pentobarbital (Nembutal sodium, Abfar). Humerus bones of rats were revealed by the dissection of front extremite bones.
Humerus bones were measured in the right anatomical references (A) length, corpus thickness [B1 (craniocaudal) + B2 (mediolateral) / 2], cortex-cortical bone thickness [C1 (caudal) + C2 (cranial) + C3 (medial) + C4 (lateral) / 4] and medullar diameter-cavum medullare [D1 (craniocaudal) + D2 (mediolateral) / 2] (Lok et al., 2011) each of these points were found and necessary morphometric measures were done (Fig 1)
"Nomina Anatomica Veterinaria" (ICVGAN, 2005) was used in the writing of anatomical terms. It was benefited from SPSS 13.0 package program in data statistical evaluation. Results were presented as Mean [+ or -] SD. In the comparison of data, ANOVA and Duncan test was applied (p<0.05).
The length, thickness of corpus, cortex and medullar diameters of humerus bones of experimental, control and peanut oil groups were compared (Table I).
When the length of humerus bone of rats was examined; in the testosterone enanthate applied group (experiment) growth of the humerus bones stopped and difference found significant (p<0.05). It was found that there is no significant difference at the thickness of corpus and cortex, diameters of medullar of humerus bones between experiment, control and peanut oil groups (p>0.05).
Short term use of testosterone enanthate's morphometric effects on male rats, in puberty period, and length, corpus thickness, cortex thickness and medullar diameter of humerus bone were analyzed.
Rats' length of humerus (testosterone enanthate applied) in experiment group was found shorter than other groups (p<0.05). In the other studies (Lok et al., 2011; Li et al., 2000) of AAS', it was reported that anabolic androgenic steroids causes early epiphyseal closure and avoid bone from growth. Literature findings show parallelism with this study findings.
Among three groups (experiment, control and peanut oil) in this study, no difference was found in medullar diameter, corpus and cortex thickness of humerus bone. In other words; testosterone enanthate had no significant effect on medullar diameter, corpus and cortex thickness of humerus (p>0.05). In the studies done with different research groups, it was reported that AAS' did not have any impact on bone corpus thickness (Lok & Yalcin, 2010), cortex thickness (Windahl et al, 1999) and medullar diameter (Lok et al, 2012; Kim et al, 2003). Literature diagnosis shows parallelism with labour findings.
As a result, short term use of testosterone enanthate's morphometric effects on male rats' humerus bone, in puberty period, causes early epiphyseal closure and shuts down the growth of bone as seen in anabolic androgenic steroids.
Al-Ismail, K.; Torreggiani, W. C.; Munk, P. L. & Nicolaou S. Gluteal mass in a bodybuilder: radiological depiction of a complication of anabolic steroid use. Eur. Radiol., 12(6):1366-9, 2002.
Attardi, B. J.; Marck, B. T.; Matsumoto, A. M.; Koduri, S. & Hild, S. A. Long-term effects of dimethandrolone 17[beta]-undecanoate and 11[beta]-methyl-19-nortestosterone 17[beta]-dodecylcarbonate on body composition, bone mineral density, serum gonadotropins, and androgenic/anabolic activity in castrated male rats. J. Androl., 32(2):183-92, 2011.
Azzazy, H. M.; Mansour, M. M. & Christenson, R. H. Gene doping: of mice and men. Clin. Biochem., 42(6):435-41,2009.
Baggish, A. L.; Weiner, R. B.; Kanayama, G.; Hudson, J. I.; Picard, M. H.; Hutter, A. M. Jr. & Pope, H. G. Jr. Long-term anabolic-androgenic steroid use is associated with left ventricular dysfunction. Circ. Heart Fail., 3(4):472-6, 2010.
Bispo, M.; Valente, A.; Maldonado, R.; Palma, R.; Gloria, H.; Nobrega, J. & Alexandrino, P. Anabolic steroid-induced cardiomyopathy underlying acute liver failure in a young bodybuilder. World J. Gastroenterol., 15(23):2920-2, 2009.
Collins, L. & Basaria, S. Adverse effects of androgen deprivation therapy in men with prostate cancer: a focus on metabolic and cardiovascular complications. Asian J. Androl., 14(2):222-5, 2012.
Di Luigi, L.; Sgro, P.; Aversa, A.; Migliaccio, S.; Bianchini, S.; Botre, F.; Romanelli, F. & Lenzi, A. Concerns about serum androgens monitoring during testosterone replacement treatments in hypogonadal male athletes: a pilot study. J. Sex Med., 9(3):873-86, 2012.
Furstenberger, C.; Vuorinen, A.; Da Cunha, T.; Kratschmar, D. V.; Saugy, M.; Schuster, D. & Odermatt, A. The anabolic androgenic steroid fluoxymesterone inhibits 11[beta]-hydroxysteroid dehydrogenase 2-dependent glucocorticoid inactivation. Toxicol. Sci., 126(2):353-61, 2012.
Hassan, A. F. & Kamal, M. M. Effect of exercise training and anabolic androgenic steroids on hemodynamics, glycogen content, angiogenesis and apoptosis of cardiac muscle in adult male rats. Int. J. Health Sci. (Qassim), 7(1):47-60, 2013.
Hildebrandt, T.; Yehuda, R. & Alfano, L. What can allostasis tell us about anabolic-androgenic steroid addiction? Dev. Psychopathol., 23(3):907-19, 2011.
International Committee on Veterinary Gross Anatomical Nomenclature (ICVGAN). Nomina Anatomica Veterinaria (NAV). 5th ed. Knoxville, ICVGAN & General Assembly of the World Association of Veterinary Anatomists WAVA, 2005.
Kanayama, G.; Brower, K. J.; Wood, R. I.; Hudson, J. I. & Pope, H. G. Jr. Issues for DSM-V: clarifying the diagnostic criteria for anabolic-androgenic steroid dependence. Am. J. Psychiatry, 166(6):642-5, 2009.
Kaushik, M.; Sontineni, S. P. & Hunter, C. Cardiovascular disease and androgens: a review. Int. J. Cardiol., 142(1):8-14, 2010.
Khan, J. I.; Kennedy, T. J. & Christian, D. R. Jr. Basic Principles of Forensic Chemistry. New York, Humana Press/Springer, 2012.
Kim, B. T.; Mosekilde, L.; Duan, Y; Zhang, X. Z.; Tomvig, L.; Thomsen, J. S. & Seeman, E. The structural and hormonal basis of sex differences in peak appendicular bone strength in rats. J. Bone Miner. Res., 18(1):150-5, 2003.
Li, X.; Takahashi, M.; Kushida, K.; Shimizu, S.; Hoshino, H.; Suzuki, M. & Inoue, T. The effects of nandrolone decanoate on bone mass and metabolism in ovariectomized rats with osteopenia. J. Bone Miner. Metab., 18(5):258-63, 2000.
Lok, S. & Yalcin, H. Morphometric effect of nandrolone decanoate used as doping in sport on femur of rats in puberty period. Arch. Budo, 6(4):217-20, 2010.
Lok, S.; Tasgin, E. & Yalcin, H. The morphometrical response of the combined nandrolone and testosterone usage to the femur. Sci. Res. Essays, 6:4867-69, 2011.
Lok, S.; Tasgin, E. & Yalcin, H. Morphometric effects of the combined usage of anabolic androgenic steroids on humerus. Energy Educ. Sci. Technol. Pt. B-Soc. Educ. Stud., 4(4):2625-30, 2012.
Lok, S.; Tasgin, E.; Demir, N. & Ozdemir, M. Long term used testosterone may cause heart and liver damage. J. Anim. Vet. Adv., 9(18):2343-5, 2010.
Lood, Y; Eklund, A.; Garle, M. & Ahlner, J. Anabolic androgenic steroids in police cases in Sweden 1999-2009. Forensic Sci. Int, 219(1-3):199-204, 2012.
Neri, M.; Bello, S.; Bonsignore, A.; Cantatore, S.; Riezzo, I.; Turillazzi, E. & Fineschi, V. Anabolic androgenic steroids abuse and liver toxicity. Mini Rev. Med. Chem., 11(5):430-7, 2011.
Pagonis, T. A.; Koukoulis, G. N.; Hadjichristodoulou, C. S.; Toli, P. N. & Angelopoulos, N. V. Multivitamins and phospholipids complex protects the hepatic cells from androgenic-anabolicsteroids-induced toxicity. Clin. Toxicol. (Phila.), 46(1):57-66, 2008.
Soma, L. R.; Uboh, C. E.; Guan, F. & McDonnell, S. Plasma concentrations of testosterone and 19-nortestosterone (nandrolone) in the nonracing intact male horse by liquid chromatography-mass spectrometry. J. Vet. Pharmacol. Therap., 31(6):587-90, 2008.
Tasgin, E.; Lok, S. & Demir, N. Combined usage of testosterone and nandrolone may cause heart damage. Afr. J. Biotechnol., 10(19)73766-8, 2011.
Tasgin, E.; Lok, S.; Demir, N. & Ozdemir, M. The effect of testosterone used in sportsmen on routine biochemical parameters. J. Anim. Vet. Adv. 9(15):2038-40, 2010.
Windahl, S. H.; Vidal, O.; Andersson, G.; Gustafsson, J. A. & Ohlsson, C. Increased cortical bone mineral content but unchanged trabecular bone mineral density in female ERbeta(-/-) mice. J. Clin. Invest., 104(7):895-901, 1999.
Zaman, F.; Chrysis, D.; Huntjens, K.; Fadeel, B. & Savendahl, L. Ablation of the pro-apoptotic protein Bax protects mice from glucocorticoid-induced bone growth impairment. PLoS One, 7(3): e33168, 2012.
Dr. Sefa Lok
School of Pyhsical Education and Sport
Sefa Lok *
* School of Physical Education and Sport, Selcuk University, Konya, Turkey.
Caption: Fig. 1. Reference points (Right crainal and medial face) of humerus length (A), corpus (B1+B2/2), cortex (C1+C2+C3+C4/4) and medullar calibers (D1+D2/2).
A= Pitch between terminal points of caput humeri and trochlea humeri; B1+B2/ 2= Corpus thickness of humerus;
C1+C2+C3+C4/4= Cortex thickness (cortical bone-substantiate compacta) of humerus level; D1+D2/2= Caliber of cavum medullar of humerus corpus level.
Table I. Comparison of length and thickness of corpus, cortex and medullar diameters of humerus bones (Mean [+ or -] SD). Experimental Control n= 10 n= 10 Length 23.63 [+ or -] 0.52 (a) 24.90 [+ or -] 0.48 (b) Corpus 2.63 [+ or -] 0.09 (a) 2.66 [+ or -] 0.08 (a) Cortex 0.52 [+ or -] 0.02 (a) 0.52 [+ or -] 0.00 (a) Medullar 1.55 [+ or -] 0.09 (a) 1.63 [+ or -] 0.10 (a) Peanut oil n= 10 Length 25.38 [+ or -] 0.81 (b) Corpus 2.66 [+ or -] 0.12 (a) Cortex 0.53 [+ or -] 0.02 (a) Medullar 1.59 [+ or -] 0.10 (a) a, b= Different letters in the same line are statistically significant (Duncan test, p<0.05).