THE EFFECT OF DEXAMETHASONE ON CALCIUM HOMEOSTASIS AND THE EXPRESSION OF TGF-B1 OF WISTAR RAT’S FEMUR BONE POST TRANSPLANTATION WITH DEMINERALIZED FREEZE-DRIED BOVINE BONE XENOGRAFT
Abstract
This research aims to evaluate the safety of dexamethasone use as an anti-inflammatory and analgesic on the handling of femur fracture of rats by bone graft method. The sample used was 32 male Wistar rats aged 2 months were divided into two groups, 16 rats for each. Group I was fractured rats and fixed with intramedullary pin without bone graft, whereas Group II was fractured and fixed with intramedullary pin and demineralized freeze-dried bovine bone xenograft (DFDBBX). Group I and II were then divided into two groups for each, Group IA, IB, IIA, and IIB, eight rats for each. Group IA and IIA were not injected with dexamethasone, whereas Group IB and IIB were injected with dexamethasone 0.5 mg/kg BW/ day intramuscularly. At the end of the first week, blood was collected from eight rats (two rats for each) of Group IA, IIA, IB, and IIB for total leucocytes and blood Calcium analysis. Urine was also collected for urine Calcium analysis. Rats were then killed to collect the femur for histopathologic analysis using HE staining and immunohistochemistry to analyze the expression of TGF?-1 using Streptavidin-biotin method. Subsequent sampling was conducted at the end of 2nd, 4th, and 8th weeks of treatment, respectively. The total data of leucocytes, blood, and urine Calcium were statistically analyzed using a 2x2x4 factorial method, while the histopathologic and immunohistochemistry specimens were analyzed descriptively under the microscope based on the alteration of the bones. The statistical analysis showed that dexamethasone increased the concentration of urine Calcium (p<0,05), the concentration of blood Calcium insignificantly (p>0,05) affected by dexamethasone but cause homeostatic calcium disorder and the total leucocytes significantly (p<0,05) affected by grafted materials. Histopathologic analysis of the femur showed that there was an increased infiltration of inflammatory cells in the fracture area from the group untreated with dexamethasone. Whereas the immunohistochemistry analysis showed positive results from all groups. Based on the whole analysis, it can be concluded that although dexamethasone depressed inflammation process, it can also disturb the fracture healing process.
Downloads
References
Becker W, Clokie C, Sannerby L, Urist MR, Becker BB. 1998. Histologic finding after implantation and evaluation of different grafting materials and titanium miro screws into extraction socket: case reports. J. Periodontal. 69: 414-21.
Bloom W, Fawcett D. 1994. A Textbook of Histology. 12th Ed. Chapman and Hall. New York. Pp. 194-233.
Bolander ME. 1992. Regulation of fracture repair by growth factors. Pro. Soc. Exp. Biol. Med. 200: 165-170.
Bullock BL, Rosendahl PP. 1984. Pathophysiology Adaptations and Alterations in Function. Little, Brown and Company. Boston/Toronto. Pp. 547-577.
Campbell WB. 1991. Lipid-Derived Autacoids: Eicosanoids and Platelet-Activating Factor. Dalam: Goodman and Gilman's The Pharmacological Basis of Therapeutics. Ed 8. Editor: Gilman, A.G. et al. New York: Pergamon Press. I, Pp. 600-611.
Chen G, Deng C, Li YP. 2012. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int. J. Biol. Sci. 8(2):272-288.
Cunningham, J.G. 1992. Teksbook of Veterinary Physiology. W.B. Saunders Company. Pp. 416-423
Djojosoebagio S. 1990. Fisologi Kelenjar Endokrin. Departemen pendidikan dan Kebudayaan. Direktorat jenderal Pendidikan Tinggi. Pusat Antar Universitas. Ilmu Hayati IPB. Pp. 147-227.
Faler BJ, Mascata RA, Plummer D, Mishra L, Sidawy AN. 2006. Transforming growth factor B and wound healing. Perspect. Vasc. Surg. Endovasc. Ther. 18: 55.
Finkemeier CG. 2002. Bone grafting and bone graft substitutes. J. Bone. Joint. Surg. Am. 84: 454-464.
Hamish RD, Steven JB. 2000. A Guide to Canine and Feline Orthopaedic Surgery. 4th Ed. Blackwell Science ltd. Blackwell Publishing Company.
Haynes RC. 1991. Adrenocorticotropic Hormone. Dalam: Goodman and Gilman's The Pharmacological Basis of Therapeutics. Ed 8. Editor: Gilman, A.G. et al. New York: Pergamon Press. II, Pp. 1443.
Janssens K, Dijke PT, Janssens S, Hul WV. 2005. Transforming growth factor-β1 to the bone. Endoc. Rev. 26(6):743-774.
Kaewloet LN. 2008. The Correlation Between Affecting Factors and Wound Healing Rate in Diabetic Foot Ulcer. Faculty of Graduate Studies. Mahidot University. India.
Lorenz HP, Longaker MT. 2005. Wounds: Biology, pathology and management. department of surgery. Ann. Surg. 217(4): 391-396.
Maiorama C, Simon M. 2003. Advanced Techniques for Bone Regeneration with Bio-Oss and Bio-Gide. R.C. Libri, Srl, Milano-Italy. Pp.73-88.
Masulili SLC, Maulani C, Sukardi I. 2008. Evaluasi radiografis cangkok tulang alograft dan membran periosteum pada terapi regenaratif untuk periodontitis agresif. Majalah Kedokteran Gigi. 15(2): 169-174.
Pilitsis JG, Lucas DR, Rengachary SR. 2002. Bone healing and spinal fusion. Neurosurg. Focus. 13(6): article 1.
Plata DV, Scheyer ET, Mellonig JT. 2002. Clinical comparison of an enamel matrix derivative used alone or in combination with a bovine-derived xenograft for treatment of periodontal osseus defect in humans. J. Periodontal. 73: 433-440.
Poetker DM, Reh DD. 2010. A comprehensive review of the adverse effects of systemic corticosteroid. Otolaryngol. Clin. N. Am. 43: 753-768.
Puricelli E, Corsetti A, Ponzoni D, Martins GL, Leite GM, Santos LA. 2010. Characterization of bone repair in rat femur after treatment with calcium phosphate cement and autogenous bone graft. J. Head Face Med. 6: 10.
Ratna S. 2011. Kortikosteroid dalam tata laksana uveitis: mekanisme kerja, aplikasi klinis dan efek samping. J. Indon. Med. Assoc. 61(6): 265-269.
Rozman P, Bolta Z. 2007. Use of platelet growth factor in treating wounds and soft tissue injuries. Acta. Dermatovenerol. Alp. Pannonica. Adriat. 16(4):156-165.
Smith, E.L., Hill, R.L., Lehman, I.R., Lefkowitz, R.J., Handler, P. And White, A. 1983. Priciples of Bichemistry : Mamalian Biochemistry. Ed. 7. MacGraw Hill Book Co. New York. Pp. 441-467.
Stephan EB, Jang D, Lynch S, Bush P, Dziak R. 1999. Anorganic bovine bone supports osteoblastic cell attachment and proliferation. J. Periodontal. 70: 364-369.
Tzakas P, Wong BYL, Logan AG, Rubin LA, Cole DEC. 2005. Transforming growth factor beta-1 (TGFB-1) and peak bone mass: Association between intragenic polymorphisms and quantitative ultrasound of the heel. BMC. 6: 29.
Virolainen P, Elima K, Metsaranta M, Aro HT, Vuorio E. 1998. Incorporation of cortical bone allografts an autografts in rats. Expression patterns of mRNAs for the TGF-βs. Acta. Orthop. Sacnd. 69(5): 537-544.
Werner, S and Grose, R. 2003. regulation of wound healing by growth factors and cytokines. London research institute. London. United Kingdom. Physiol. Rev 83: 835-870
Wildmann B, Kadow RA, Haas NP, Schmidmaier G. 2007. Quantification of various growth factors in different demineralized bone matrix preparation. J. Biomed Mater. Res. A. 81(2): 437-442.
Wu CH, Yang MY, Chan KC, Chung PJ, Ou TT, Wang CJ. 2010. Improvment in high fat diet induced obesity and body fat accumulation by a nelumbo nucifera leaf flavonoid-rich extract in mice. J. Agris. Food Cem. 58(11): 7075-7081.
Yasear AY, Hamouda SA. 2009. Effect of Dexamethasone on Osteoclast Formation in The Alveolar Bone of Rabbits. Iraqi J. Vet. Sci. 23(1): 13-16.
Yunanthi RAE, Soesilowati ASK, Murdiastuti K. 2010. Perbedaan efektivitas antara demineralized freeze-dried bone allograft dan demineralized freeze-dried bovine bone xenograft pada perawatan kerusakan intraboni. J. Kedokteran Gigi. (1): 45-54.
Yuniarti, W.M., Yudaniayanti, I.S. dan Triakoso, N. 2008. Pengaruh Pemberian Suplemen Kalsium Karbonat Dosis Tinggi pada Tikus Putih Ovariohisterektomi terhadap Mineralisasi Ginjal. J. Vet. Vol 9, No.2: 73-78.
Yuniwarti EYW, Saraswati T. 2002. Aktifitas kalsifikasi tulang tibia broiler jantan dan betina setelah pemberian 1,25-dehidroxycholecalcifeol. Laporan penelitian dosen muda, Universitas Diponegoro, Semarang.
