Exogenous heparin binds and inhibits bone morphogenetic protein 6 biological activity

Brkljačić, Jelena and Pauk, Martina and Erjavec, Igor and Cipčić, Antonio and Grgurević, Lovorka and Zadro, Renata and Inman, Gareth J. and Vukičević, Slobodan (2013) Exogenous heparin binds and inhibits bone morphogenetic protein 6 biological activity. International Orthopaedics, 37 (3). pp. 529-41. ISSN 0341-2695

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PURPOSE: The purpose of this study was to explore the effect of heparin on bone morphogenetic protein 6 (BMP6) osteogenic activity. ----- METHODS: Western blot analysis was used to confirm the binding of BMP6 to heparin and to observe its effect on BMP6 signaling in C2C12-BRE-Luc myoblasts. Real-time RT-PCR was performed for the expression analysis of alkaline phosphatase (ALP) and osteocalcin (OC) in C2C12 myoblasts treated with BMP6 and heparin for 72 hours. Rat ectopic bone formation assay was performed to explore the effect of heparin on BMP6 osteogenic activity. Two weeks following implantation the implants were analysed morphologically and histologically. A mouse osteoporotic model was used to test the ability of BMP6 to improve the bone quality in vivo in the presence of heparin, followed by DEXA and μCT analyses. Blood coagulation was tested in rats previously treated with BMP6. ----- RESULTS: BMP6 specifically bound to heparin and induced Smad1/5/8 phosphorylation which was inhibited by heparin. After 48 and 72 hours of treatment, heparin inhibited BMP6-induced ALP and OC expression in C2C12 cells. Heparin dose dependently inhibited BMP6-induced new bone and cartilage formation in the rat ectopic bone formation assay, while in osteoporotic mice heparin inhibited the BMP6 potential to improve the bone quality as evidenced by decreased bone mineral density and trabecular bone parameters. Interestingly, BMP6 prevented the effect of heparin on the blood coagulation parameters. ----- CONCLUSION: The interaction of BMP6 with heparin might contribute to the heparin-induced osteoporosis and blood coagulation.

Item Type: Article
MeSH: Animals ; Anticoagulants/pharmacology ; Bone Morphogenetic Protein 6/antagonists & inhibitors ; Bone Morphogenetic Protein 6/metabolism ; Cell Line ; Disease Models, Animal ; Heparin/pharmacology ; Mice ; Osteogenesis/drug effects ; Osteoporosis/physiopathology ; Rats ; Rats, Sprague-Dawley
Departments: Katedra za anatomiju i kliničku anatomiju
Depositing User: Marijan Šember
Status: Published
Brkljačić, JelenaUNSPECIFIED
Cipčić, AntonioUNSPECIFIED
Grgurević, LovorkaUNSPECIFIED
Vukičević, SlobodanUNSPECIFIED
Date: March 2013
Date Deposited: 18 Jan 2016 12:16
Last Modified: 18 Jan 2016 12:16
Subjects: /
Related URLs:
URI: http://medlib.mef.hr/id/eprint/2376

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