MEHMET ERDEM, Deniz GULABİ, Murat AŞÇI, Bora BOSTAN, Taner GÜNEŞ, REŞİT DOĞAN KÖSEOĞLU

The effects of melatonin and caffeic acid phenethyl ester (CAPE) on fracture healing under ischemic conditions

Amaç:Çalışmada melatonin ve kafeik asit fenetil ester (CAPE) antioksidan moleküllerinin iskemik ortamda kırık iyileşmesi üzerine etkisinin incelenmesi amaçlandı. Çalışma planı: Kırk dört erkek Wistar-albino sıçan tibiasında kırık oluşturulup intramedüller pinleme ile tespit uygulandı. Daha sonra sıçanlar rastgele kırık, kırık-iskemi, kırık-iskemi-melatonin ve kırık-iskemi-CAPE gruplarına ayrıldı. İskemi gruplarındaki sıçanların femoral arterleri 4.5 saat süreyle klemplendi. Cerrahi girişimden 6 hafta sonra sıçanlar sakrifiye edildi ve radyografik, histolojik ve biyomekanik değerlendirmeler yapıldı. Bulgular: Radyografik ve histolojik olarak, 6. hafta sonunda kırık-iskemi-CAPE grubu kırık-iskemi grubundan anlamlı olarak iyi sonuçlar verdi. Tüm gruplarda, tüm kırıkların radyolojik ve histolojik olarak tam iyileştiği saptandı. Maksimum kırılma kuvvetlerinin (N) karşılaştırılmasında, gruplar arasında (kırık-iskemi

Melatonin ve kafeik asit fenetil ester’in iskemik şartlarda kırık iyileşmesi üzerine etkisi: Deneysel çalışma

Objective:The aim of this study was to investigate the effects of antioxidant molecules Melatonin and Caffeic Acid Phenethyl Ester (CAPE) on fracture healing under ischemic conditions. Methods: A right tibia fracture was created and fixed with an intramedullary pin in forty four male Wistar-albino rats. The rats were then randomly allocated to fracture, fracture-ischemia, fracture- ischemia-melatonin, and fracture-ischemia-CAPE groups. Ischemia was created by clamping femoral arteries four and a half hours. Animals were killed and radiographic, histological and biomechanical evaluation was performed sixth week after surgery. Results: The radiological and histological scores of the fracture-ischemia-CAPE group were significantly better than the fracture- ischemia group at 6th week follow-up. Complete radiographical and histological healing of all fractures was detected in all groups. There was a significant difference between the maximum fracture force between the groups (fracture-ischemia<fracture-ischemia-melatonin<fracture<fracture-ischemia-CAPE) (p<0.005). Although difference was not statistically significant between fracture and fracture-ischemia-CAPE groups, all other groups revealed statistically significant difference with respect to toughness (N/mm). Fracture-ischemia group revealed the lowest toughness. Conclusion: Ischemia adversely affects the fracture healing of rat tibias. Melatonin and CAPE eradicate adverse effects of ischemia. Possible adverse effects of ischemia on fracture healing can be eradicated with melatonin and CAPE in patients with tibia fractures associated with vascular injury or compartment syndrome.

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1. Cheeseman KH. Mechanisms and effects of lipid peroxi- dation. Mol Aspects Med 1993;14:191-7. CrossRef
2. Clanton TL, Zuo L, Klawitter P. Oxidants and skeletal muscle function: physiologic and pathophysiologic impli- cations. Proc Soc Exp Biol Med 1999;222:253-62. CrossRef
3. Turgut A, Göktürk E, Köse N, Kaçmaz M, Oztürk HS, Seber S, et al. Oxidant status increased during fracture healing in rats. Acta Orthop Scand 1999;70:487-90. CrossRef
4. Yeler H, Tahtabas F, Candan F. Investigation of oxidative stress during fracture healing in the rats. Cell Biochem Funct 2005;23:137-9. CrossRef
5. Göktürk E, Turgut A, Bayçu C, Günal I, Seber S, Gülbas Z. Oxygen-free radicals impair fracture healing in rats. Acta Orthop Scand 1995;66:473-5. CrossRef
6. Erdem M, Bostan B, Güneş T, Özkan F, Sen C, Özyurt H, et al. Protective effects of melatonin on ischemia-reperfu- sion injury of skeletal muscle. Eklem Hastalik Cerrahisi 2010;21:166-71.
7. Rodriguez C, Mayo JC, Sainz RM, Antolín I, Herrera F, Martín V, et al. Regulation of antioxidant enzymes: a sig- nificant role for melatonin. J Pineal Res 2004;36:1-9. CrossRef
8. Ozyurt H, Irmak MK, Akyol O, Söğüt S. Caffeic acid phenethyl ester changes the indices of oxidative stress in serum of rats with renal ischaemia-reperfusion injury. Cell Biochem Funct 2001;19:259-63. CrossRef
9. Allegra M, Reiter RJ, Tan DX, Gentile C, Tesoriere L, Livrea MA. The chemistry of melatonin’s interaction with reactive species. J Pineal Res 2003;34:1-10. CrossRef
10. Okatani Y, Wakatsuki A, Reiter RJ, Enzan H, Miyahara Y. Protective effect of melatonin against mitochondrial in- jury induced by ischemia and reperfusion of rat liver. Eur J Pharmacol 2003;469:145-52. CrossRef
11. Sener G, Sehirli AO, Keyer-Uysal M, Arbak S, Ersoy Y, Yeğen BC. The protective effect of melatonin on re- nal ischemia-reperfusion injury in the rat. J Pineal Res 2002;32:120-6. CrossRef
12. Sud’ina GF, Mirzoeva OK, Pushkareva MA, Korshu- nova GA, Sumbatyan NV, Varfolomeev SD. Caffeic acid phenethyl ester as a lipoxygenase inhibitor with antioxi- dant properties. FEBS Lett 1993;329:21-4. CrossRef
13. An Y, Friedman RJ, Parent T, Draughn RA. Production of a standard closed fracture in the rat tibia. J Orthop Trau- ma 1994;8:111-5. CrossRef
14. Skjeldal S, Grøgaard B, Reikerås O, Müller C, Torvik A, Svindland A. Model for skeletal muscle ischemia in rat hindlimb: evaluation of reperfusion and necrosis. Eur Surg Res 1991;23:355-65. CrossRef
15. Histing T, Anton C, Scheuer C, Garcia P, Holstein JH, Klein M, et al. Melatonin impairs fracture healing by sup- pressing RANKL-mediated bone remodeling. J Surg Res 2012;173:83-90. CrossRef
16. Lane JM, Sandhu HS. Current approaches to experimen- tal bone grafting. Orthop Clin North Am 1987;18:213- 25.
17. Huddleston PM, Steckelberg JM, Hanssen AD, Rouse MS, Bolander ME, Patel R. Ciprofloxacin inhibition of experimental fracture healing. J Bone Joint Surg Am 2000;82:161-73.
18. Wang WZ, Fang XH, Stephenson LL, Baynosa RC, Khiabani KT, Zamboni WA. Microcirculatory effects of melatonin in rat skeletal muscle after prolonged ischemia. J Pineal Res 2005;39:57-65. CrossRef
19. Toledo-Pereyra LH, Lopez-Neblina F, Toledo AH. Re- active oxygen species and molecular biology of ischemia/ reperfusion. Ann Transplant 2004;9:81-3.
20. Ozyurt H, Ozyurt B, Koca K, Ozgocmen S. Caffeic acid phenethyl ester (CAPE) protects rat skeletal muscle against ischemia-reperfusion-induced oxidative stress. Vascul Pharmacol 2007;47:108-12. CrossRef
21. Histing T, Marciniak K, Scheuer C, Garcia P, Holstein JH, Klein M, et al. Sildenafil accelerates fracture healing in mice. J Orthop Res 2011;29:867-73. CrossRef
22. Spiro AS, Khadem S, Jeschke A, Marshall RP, Pogoda P, Ignatius A, et al. The SERM raloxifene improves di- aphyseal fracture healing in mice. J Bone Miner Metab 2013;31:629-36. CrossRef
23. Bukata SV. Systemic administration of pharmacologi- cal agents and bone repair: what can we expect. Injury 2011;42:605-8. CrossRef
24. Cicek E, Gokalp O, Varol R, Cesur G. Influence of elec- tromagnetic fields on bone fracture in rats: role of CAPE. Biomed Environ Sci 2009;22:157-60. CrossRef
25. Collin-Osdoby P, Li L, Rothe L, Anderson F, Kirsch D, Oursler MJ, Osdoby P. Inhibition of avian osteoclast bone resorption by monoclonal antibody 121F: a mechanism involving the osteoclast free radical system. J Bone Miner Res 1998;13:67-78. CrossRef
26. Cetinus E, Kilinç M, Uzel M, Inanç F, Kurutaş EB, Bilgic E, et al. Does long-term ischemia affect the oxidant sta- tus during fracture healing? Arch Orthop Trauma Surg 2005;125:376-80. CrossRef
27. Roth JA, Kim BG, Lin WL, Cho MI. Melatonin promotes osteoblast differentiation and bone formation. J Biol Chem 1999;274:22041-7. CrossRef
28. Reiter RJ. Pineal melatonin: cell biology of its synthe- sis and of its physiological interactions. Endocr Rev 1991;12:151-80. CrossRef
29. Halıcı M, Öner M, Güney A, Canöz Ö, Narin F, Halıcı C. Melatonin promotes fracture healing in the rat model. Eklem Hastalik Cerrahisi 2010;21:172-7.
30. Elmali N, Ayan I, Türköz Y, Mizrak B, Germen B, Bora A. Effect of caffeic acid phenethyl ester on cartilage in experi- mental osteoarthritis. Rheumatol Int 2002;22:222-6. CrossRef
31. Ilhan A, Koltuksuz U, Ozen S, Uz E, Ciralik H, Akyol O. The effects of caffeic acid phenethyl ester (CAPE) on spinal cord ischemia/reperfusion injury in rabbits. Eur J Cardiothorac Surg 1999;16:458-63. CrossRef
32. Gurel A, Armutcu F, Sahin S, Sogut S, Ozyurt H, Gulec M, et al. Protective role of alpha-tocopherol and caffeic acid phenethyl ester on ischemia-reperfusion injury via ni- tric oxide and myeloperoxidase in rat kidneys. Clin Chim Acta 2004;339:33-41. CrossRef
33. Ang ES, Pavlos NJ, Chai LY, Qi M, Cheng TS, Steer JH, et al. Caffeic acid phenethyl ester, an active component of honeybee propolis attenuates osteoclastogenesis and bone resorption via the suppression of RANKL-induced NF- kappaB and NFAT activity. J Cell Physiol 2009;221:642-9.