Histopathological study of osteoporotic changes after experimental creation of osteoporosis by ovariectomy in the rat

Subject Areas : Veterinary Clinical Pathology

داریوش Mohajeri ¹ , مهران Mesgari ² , Ali Rezaie ³ , Amin Balila ⁴

1 - Department of Pathobiology, Faculty of Veterinary Medicine, Islamic Azad University of Tabriz, Tabriz, Iran
2 - Drug Applied Research Center, Tabriz University of Medical Science, Tabriz, Iran
3 - Department of Clinical Sciences, Faculty of Veterinary Medicine, Islamic Azad University of Tabriz, Tabriz, Iran
4 - Graduated of Veterinary Medicine, Islamic Azad University of Tabriz, Tabriz, Iran

Received: 2007-04-24 Accepted : 2007-11-11 Published : 2007-08-23

Keywords: Rat, Histopathology, Ovariectomy, Osteoporosis,

Abstract :

In order to study the histopathology of osteoporosis during menopause, 80 female Sprague-Dawley rats with approximate age of 10 weeks were allocated to 10 groups of 8 rats each (C1, C2 and C3 as control, S1, S2 and S3 as sham, T1, T2 and T3 as treatment groups and C0 as one pure control group). The nutritional and maintenance conditions were similar in all groups. Treatment groups (T1, T2 and T3) were bilaterally ovariectomized using right and left flank approach. Sham groups (S1, S2 and S3) underwent operation in which ovaries were exposed from the same approach but left intact. Treatment (T1, T2 and T3) and their related control groups (C1, C2 and C3) were euthanized at 5, 12 and 21 weeks respectively after ovariectomy. After necropsy, both tibia and femur bones were totally removed and fixed in 10%  buffered formalin, decalcified and tissue specimens stained with H&E for histopathological study. Microscopically, osteoporosis was prominent in treatment groups T2 and T3 although it was more severe in treatment group T3. Statistically, differences between ovariectomized groups (T1, T2 and T3) regarding the severity of osteoporosis with 99% confidence were significant (P<0.01). Significant differences between treatment group T1 and control groups were not observed (P>0.05).

References:

1. Adami, S., Bufalino, L. and Cervetti, R. (1997): Ipriflavone prevents radial bone loss in postmenopausal women with low bone mass over 2 years. Osteoporos Int. 7:119–25.


2. Agnusdei, D. and Bufalino, L. (1997): Efficacy of ipriflavone in established osteoporosis and long-term safety. Calcif. Tissue Int. 61:S23–7.


3. Avioli, L.V. (1997): The future of ipriflavone in the management of osteoporotic syndromes. Calcif. Tissue Int. 61:S33–5.


4. Brubaker, K.D. and Gray, C.V. (1997): Evidence for plasma membrane estrogen receptors and rapid signaling events in osteoclasts. J. Bone. Miner. Res., 12, pp: 134.


5. Calomme, M., Geusens, P., Demeester, N., Behets, G. J.,  D’Haese, P.,  Sindambiwe, J. B., et al. (2006): Partial prevention of long-term femoral bone loss in aged ovariectomized rats supplemented with choline-stabilized orthosilicic acid. Journal of Calcified Tissue International. 78(4): 227-232.


6. Carlton, W.W. and McGavin, M.D. (2002): Thomson’s Special Veterinary Pathology, 4th edn. Mosby ,Londn ., pp: 436-437.


7. Dempster, D.W., Birchman, R., Lindsay, R. and Shen, U. (1995): Temporal changes in cancelous bone structure immediately after ovariectomy. Bone, 16, pp: 157-163.


8. Gallagher, J.C., Kable, W.T. and Goldgar, D. (1991): Effect of progestin therapy on cortical and trabecular bone: Comparison with estrogen. Am. J. Med., 90, pp: 171-178.


9. Gallagher, J.C., Riggs, B.L. and DeLuca, H.F.(1980): Effect of estrogen on calcium absorption and serum vitamin D metabolites in postmenopausal osteoporosis. Journal of Clinical Endocrinology & Metabolism. 51:1359-1364.


10. Eriksen, E.F., Mosekilde, L. and Melsen, F. (1985): Trabecular bone resorption depth deceases with age: Differences between normal males and females. Bone, 6, pp: 141-146.


11. Gambacciani, M., Ciaponi, M. and Cappagli, B. (1997): Effects of combined low dose of the isoflavone derivative ipriflavone and estrogen replacement on bone mineral density and metabolism in postmenopausal women. Maturitas. 28:75–81.


12. Gennari, C., Adami, S. and Agnusdei, D. (1667): Effect of chronic treatment with ipriflavone in postmenopausal women with low bone bass. Calcif Tissue Int. 61:S19–22.


13. Gennari, C., Agnusdei, D. and Crepaldi, G. (1998): Effect of ipriflavone—a synthetic derivative of natural isoflavones—on bone mass loss in the early years after menopause. Menopause. 5(1):9–15.


14. Hanabayashi, T., Imai, A. and Tamaya, T. (1995): Effects of ipriflavone and estriol on postmenopausal osteoporotic changes. Int. J. Gynaecol. Obstet. 51:63–4.


15. Hau, J. ( 2003): Hand book of laboratory animal science. 2nd ed. CRC press, pp: 112-120.


16. Head, K.A. (1999): Ipriflavone: an important bone-building isoflavone. Altern .Med. Rev. 4(1):10–22.


17. Junqueira, L.C., Carino, J. and Kelley, R.O. (1992): Basi Hitology. 7^th ed. Appleton & Lange Company, pp: 189-213.


18. Kim. S.K. and Rhee, M.H. (2003): Studies on the effects of biomedicinal agents on serum concentration of Ca++,P andALP activity in osteoprosis-induced rats. J. Am. Coll. Nutr. 4(2):151-4.


19. Kobayashi, M., Hara, K. and Akiyama, Y. (2002): Effects of Vitamin K2 (Menatetrenone) on Calcium Balance,in Ovariectomized Rats. Jpn. J. Pharmacol. 88, 55 – 61.


20. Lindsay, R. and Cosman, F. (2005): Osteoprosis. In Kasper, Principles of internal medicine. Mc Graw, London. 226-245.


21. Lotz, J.C., Kroeber, M.W., Heilmann, M., Pericherla, K., Kimmel, D., et al. (2000): Tibial plateau fracture as a measure of early estrogen-dependent bone fragility in rats.18, pp: 326-332.


22. Melis, G.B., Paoletti, A.M. and Bartolini, R. (1992): Ipriflavone and low doses of estrogens in the prevention of bone mineral loss in climacterium. Bone Miner. 19:S49–56.


23. Nieves, J.W., Komar, L., Cosman, F. and Lindsay, R. (1998): Calcium potentiates the effect of estrogen and calcitonin on bone mass: review and analysis. Am. J. Clin. Nutr. 67:18–24.


24. Nozaki, M., Hashimoto, K. and Inoue, Y. (1998): Treatment of bone loss in oophorectomized women with a combination of ipriflavone and conjugated equine estrogen. Int. J. Gynaecol. Obstet. 62(1):69–75.


25. Ohta, H., Komukai, S. and Makita, K. (1999): Effects of 1-year ipriflavone treatment on lumbar bone mineral density and bone metabolic markers in postmenopausal women with low bone mass. Horm. Res. 51:178–83.


26. Sahota, O. (2000): Osteoporosis and the role of vitamin D and calcium-vitamin D deficiency, vitamin D insufficiency and vitamin D sufficiency. Ageing. 29:301–4.


27. Saka Kura, C.E., Giro, G., Goncalves, D., Pereira, R.M., Orrico, S.R., et al. (2006): Radiographic assessment of bone density around integrated titanium implant after ovariectomy in rats. 17, pp: 134-146.


28. Sastry, G.A. and Rao, P.R. (2002): Veterinary Pathology. 7th edn. CBS, pp: 468-469.


29. Stephan, C., Blum, S.N., Heaton, B.M., Bowman, M.H. and Scott, C.M. (2003): Dietary soy protein maintains some indices of bone mineraldensity and bone formation in aged ovariectomized rats.American nutritional. 1244-1249.


30. Takehiko, U., Toshiya, T., Kuniro, T. and Hitoshi, I. (2001): Comprative study on reduction of  bone loss and lipid metabolism abnormality in ovariectomized rats by soyisoflavones,daidzin,genestin and glystin boil. Pharm. 368-379.


31. Toombs, J.P. (1985): Evaluation of keys hypothesis in the feline tibia: An experimental model for augmented bone healing studies. Am. J. Res., 46, pp: 512.


32. Watkins, B.A. Reinwald, S. and Seifert, M.F. (2005): Protective actions of soy isoflavones and n-3PUF ason bone mass in ovariectomized rats. Nutr. Biochem. 479-488.


33. Zhang, Y., Lai, W., Leung, P., Wu, C., Yao, X., et al.  (2006): Effects of fructus ligustri lucidi extract on bone turnover and calcium balance in ovariectomized Rats. Biol. Pharm. Bull. 29(2): 291-296.