Alpay Ozenci, Tevfik Aslan, Zeliha Sahin, Ozlem Ozbey, Nuray Acar, Ismail Ustunel

Sıçan eklem kıkırdağında kortikosteroide bağlı gelişen kondrosit apoptozu üzerinde zoledronik asidin koruyucu etkisi

Amaç: Bu çalışmamızda sistemik kortikosteroid uygulamasının in vivo eklem kıkırdağı kondrositleri üzerindeki apoptotik etkilerini ve zoledronik asidin kortikosteroide bağlı apoptoz üzerindeki olası etkileri olup olmadığını değerlendirmeyi amaçladık. Çalışma planı: Yirmi dört adet Wistar sıçanı rastgele 3 gruba ayrıldı. Kontrol grubunda, intramusküler izotonik tuz çözeltisi haftalık olarak enjekte edildi. İkinci grupta ise, kas içine 10 mg/kg dozunda kortikosteroid (metilprednizolon) enjeksiyonu 8 hafta boyunca haftada bir uygulandı. Üçüncü grupta, 10 mg/kg intramusküler kortikosteroid (metilprednizolon) enjeksiyonu 8 hafta boyunca haftada bir doz uygulanırken, ek olarak 0.1 mg/kg zoledronik asit 0, 21 ve 42. günde deri altına enjekte edildi. Tedavinin sonunda her gruptan femur başı örnekleri alındı ve apoptotik kondrositleri saptamak için TUNEL yöntemi kullanıldı. Karşılaştırmalı analizler için ANOVA yöntemi ve Tukey testinden yararlanıldı. Bulgular: Kortikosteroid grupla diğer iki grup (kontrol grubu için p=0.005 ve kortikosteroid + zoledronik asit grubu için p=0.047) arasında anlamlı bir fark vardı. Zoledronik asit tedavisi eklem kıkırdağında kortikosteroide bağlı gelişen apoptotik kondrosit sayısını anlamlı olarak azalttı (p<0.05). Çıkarımlar: Zoledronik asit, kondrositlerde kortikosteroide bağlı gelişen apoptoz nedeniyle oluşabilecek eklem kıkırdağı bozulmasını önleyici bir potansiyele sahip olabilir.

Anahtar Kelimeler:

Apoptoz; kıkırdak; kondrosit; kortikosteroid; zoledronik asit.

Protective effect of zoledronic acid on corticosteroid-induced chondrocyte apoptosis

Objective: The aim of this study was to assess the apoptotic effects of systemic corticosteroid application on the articular cartilage chondrocytes in vivo and to investigate the potential effects of zoledronic acid on corticosteroid-induced apoptosis.Methods: Twenty-four Wistar rats were randomly divided into 3 groups. In the control group, intramuscular isotonic salt solution was injected weekly. In the second group, a dose of 10 mg/kg intramuscular corticosteroid (methylprednisolone) injection was applied weekly for 8 weeks. In the third group, a dose of 10 mg/kg intramuscular corticosteroid (methylprednisolone) injection was applied weekly for 8 weeks and 0.1 mg/kg zoledronic acid was injected subcutaneously on days 0, 21 and 42. Femoral head specimens from each group were obtained at the end of the treatment and the TUNEL method was applied to detect apoptotic chondrocytes. Comparison analyses were performed using the ANOVA method and Tukey’s test.Results: There was a significant difference between the corticosteroid group and two other groups (control group: p=0.005; corticosteroid + zoledronic acid group: p=0.047). Zoledronic acid treatment significantly decreased the number of corticosteroid-induced apoptotic chondrocytes in the joint cartilage (p<0.05).Conclusion: Zoledronic acid may have the potential to prevent joint cartilage deterioration due to the corticosteroid-induced apoptosis of the chondrocytes.

Keywords:

Apoptosis, cartilage, chondrocyte, corticosteroid zoledronic acid,

PDF

___

Annefeld M. The dose dependent effect of glycosaminoglycan peptide complex on corticosteroid-induced disordered metab- olism in cartilage tissue of rats. [Article in German] Z Rheumatol 1989;48:188-93.
Chechik O, Dekel S. Bilateral chondrolysis of the hip follow- ing liver transplantation. Skeletal Radiol 2009;38:297-300.
Higuchi M, Masuda T, Susuda K, Ishii S, Abe K. Ultrastructure of the articular cartilage after systemic admin- istration of hydrocortisone in the rabbit: an electron micro- scopic study. Clin Orthop Relat Res 1980;(152):296-302.
Itani T, Kanai K, Watanabe J, Ogawa R, Kanamura S. Quantitative analysis of rough endoplasmic reticulum in chondrocytes of articular and tracheal cartilage of rabbits fol- lowing the systemic administration of hydrocortisone. J Anat 1992;181:357-63.
Mankin HJ, Zarins A, Jaffe WL. The effect of systemic corti- costeroids on rabbit articular cartilage. Arthritis Rheum 1972; 15:593-9.
Podbielski A, Raiss R. Dose related effects of dexamethasone treatment on the ultrastructure of articular cartilage in rats. Agents Actions 1986;17:322-4.
Shaw NE, Lacey E. The influence of corticosteroids on nor- mal and papain-treated articular cartilage in the rabbit. J Bone Joint Surg Br 1973;55:197-205.
Silbermann M, Finkelbrand S. Reversibility of systemic corti- costeroid-induced mandibular osteoarthritis: an experimental study in A/J mice. J Oral Surg 1980;38:660-3.
Weiss A, Raz E, Silbermann M. Effects of systemic glucocor- ticoids on the degradation of glycosaminoglycans in the mandibular condylar cartilage of newborn mice. Bone Miner 1986;1:335-46.
Kyrylkova K, Kyryachenko S, Leid M, Kioussi C. Detection of apoptosis by TUNEL assay. Methods Mol Biol 2012;887:41-7.
Loo DT. In situ detection of apoptosis by the TUNEL assay: an overview of techniques. Methods Mol Biol 2011;682:3-13.
Farkas B, Kvell K, Czömpöly T, Illés T, Bárdos T. Increased chondrocyte death after steroid and local anesthetic combina- tion. Clin Orthop Relat Res 2010;468:3112-20.
Chrysis D, Zaman F, Chagin AS, Takigawa M, Sävendahl L. Dexamethasone induces apoptosis in proliferative chondrocytes through activation of caspases and suppression of the Akt-phos- phatidylinositol 3’-kinase signaling pathway. Endocrinology 2005;146:1391-7.
Hossain MA, Park J, Choi SH, Kim G. Dexamethasone induces apoptosis in proliferative canine tendon cells and chondrocytes. Vet Comp Orthop Traumatol 2008;21:337-42.
Mocetti P, Silvestrini G, Ballanti P, Patacchioli FR, Di Grezia R, Angelucci L, et al. Bcl-2 and Bax expression in cartilage and bone cells after high-dose corticosterone treatment in rats. Tissue Cell 2001;33:1-7.
Nakazawa F, Matsuno H, Yudoh K, Watanabe Y, Katayama R, Kimura T. Corticosteroid treatment induces chondrocyte apoptosis in an experimental arthritis model and in chondro- cyte cultures. Clin Exp Rheumatol 2002;20:773-81.
Silvestrini G, Ballanti P, Patacchioli FR, Mocetti P, Di Grezia R, Wedard BM, et al. Evaluation of apoptosis and the gluco- corticoid receptor in the cartilage growth plate and metaphy- seal bone cells of rats after high-dose treatment with corticos- terone. Bone 2000;26:33-42.
Sanchez CP, He YZ. Alterations in the growth plate cartilage of rats with renal failure receiving corticosteroid therapy. Bone 2002;30:692-8.
Dombrecht EJ, De Tollenaere CB, Aerts K, Cos P, Schuerwegh AJ, Bridts CH, et al. Antioxidant effect of bispho- sphonates and simvastatin on chondrocyte lipid peroxidation. Biochem Biophys Res Commun 2006;348:459-64.
Van Offel JF, Schuerwegh AJ, Bridts CH, Stevens WJ, De Clerck LS. Effect of bisphosphonates on viability, prolifera- tion, and dexamethasone-induced apoptosis of articular chon- drocytes. Ann Rheum Dis 2002;61:925-8.
Voulgaris A, Liapi C, Papadopoulos J. Effect of low-dose methylprednisolone on calcium balance and bone composi- tion in rats. J Endocrinol Invest 1997;20:659-63.
Little DG, Peat RA, Mcevoy A, Williams PR, Smith EJ, Baldock PA. Zoledronic acid treatment results in retention of femoral head structure after traumatic osteonecrosis in young Wistar rats. J Bone Miner Res 2003;18:2016-22.
Zometa.com [Internet]. [Cited 2013 Mar 28]. Available from: http://www.us.zometa.com
Taillandier J, Alemanni M, Samuel D, Bismuth H, Lioté F. Rapid multifocal chondrolysis after liver transplantation in four patients. Ann Rheum Dis 2006;65:118-20.
Barnhart BC, Alappat EC, Peter ME. The CD95 type I/type II model. Semin Immunol 2003;15:185-93.
Singleton JR, Baker BL, Thorburn A. Dexamethasone inhibits insulin-like growth factor signaling and potentiates myoblast apoptosis. Endocrinology 2000;141:2945-50.
Shirai T, Kobayashi M, Nishitani K, Satake T, Kuroki H, Nakagawa Y, et al. Chondroprotective effect of alendronate in a rabbit model of osteoarthritis. J Orthop Res 2011;29:1572-7.
Kobayashi M, Squires GR, Mousa A, Tanzer M, Zukor DJ, Antoniou J, et al. Role of interleukin-1 and tumor necrosis fac- tor a in matrix degradation of human osteoarthritic cartilage. Arthritis Rheum 2005;52:128-35.
Komuro H, Olee T, Kühn K, Quach J, Brinson DC, Shikhman A, et al. The osteoprotegerin/receptor activator of nuclear factor kappaB/receptor activator of nuclear factor kappaB ligand system in cartilage. Arthritis Rheum 2001;44: 2768-76.
van Lent PL, Grevers L, Lubberts E, de Vries TJ, Nabbe KC, Verbeek S, et al. Fcgamma receptors directly mediate carti- lage, but not bone, destruction in murine antigen-induced arthritis: uncoupling of cartilage damage from bone erosion and joint inflammation. Arthritis Rheum 2006;54:3868-77.
Boos N, Nerlich AG, Wiest I, von der Mark K, Ganz R, Aebi M, et al. Immunohistochemical analysis of type X-collagen expression in osteoarthritis of the hip joint. J Orthop Res 1999;17:495-502.
Pufe T, Petersen W, Tillmann B, Mentlein R. The splice vari- ants VEGF121 and VEGF189 of the angiogenic peptide vas- cular endothelial growth factor are expressed in osteoarthritic cartilage. Arthritis Rheum 2001;44:1082-8.
Plotkin LI, Weinstein RS, Parfitt AM, Roberson PK, Manolagas SC, Bellido T. Prevention of osteocyte and osteoblast apoptosis by bisphosphonates and calcitonin. J Clin Invest 1999;104:1363-74.
Plotkin LI, Aguirre JI, Kousteni S, Manolagas SC, Bellido T. Bisphosphonates and estrogens inhibit osteocyte apoptosis via distinct molecular mechanisms downstream of extracellular signal-regulated kinase activation. J Biol Chem 2005;280: 7317-25.
Plotkin LI, Bivi N, Bellido T. A bisphosphonate that does not affect osteoclasts prevents osteoblast and osteocyte apoptosis and the loss of bone strength induced by glucocorticoids in mice. Bone 2011;49:122-7.
Vermes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death. J Immunol Methods 2000;243:167-90.
D’Lima DD, Kuhn K, Lotz MK. Detection of apoptosis in cartilage in situ and in isolated chondrocytes. Methods Mol Med 2004;100:275-90.
Ozbey O, Sahin Z, Ozenci AM, Acar N, Ustunel I. The effect of systemic corticosteroid treatment on the immunolocalisa- tion of Notch-1, Delta, CD105 and CD166 in rat articular cartilage. Acta Histochem 2010;112:424-31.