Technetium-99 Conjugated with Methylene Diphosphonate Inhibits RANKL-induced Osteoclastogenesis

Clinical and Experimental Pharmacology and PhysiologyVolume 39, Issue 10 p. 886-893 Original Article Technetium-99 conjugated with methylene diphosphonate inhibits receptor activator of nuclear factor-κB ligand-induced osteoclastogenesis Wei Gong, Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, ChinaSearch for more papers by this authorHuan Dou, Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, ChinaSearch for more papers by this authorXianqin Liu, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, ChinaSearch for more papers by this authorLingyun Sun, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, ChinaSearch for more papers by this authorYayi Hou, Corresponding Author Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China Jiangsu Key Laboratory of Molecular Medicine, Nanjing, ChinaCorrespondence: Dr Yayi Hou, Immunology and Reproductive Biology Laboratory, Medical School & State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China. Email: yayihou@nju.edu.cnSearch for more papers by this author Wei Gong, Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, ChinaSearch for more papers by this authorHuan Dou, Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, ChinaSearch for more papers by this authorXianqin Liu, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, ChinaSearch for more papers by this authorLingyun Sun, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, ChinaSearch for more papers by this authorYayi Hou, Corresponding Author Immunology and Reproductive Biology Laboratory, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China Jiangsu Key Laboratory of Molecular Medicine, Nanjing, ChinaCorrespondence: Dr Yayi Hou, Immunology and Reproductive Biology Laboratory, Medical School & State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China. Email: yayihou@nju.edu.cnSearch for more papers by this author First published: 26 September 2012 https://doi.org/10.1111/j.1440-1681.2012.12006.xCitations: 4Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Summary In the present study, we investigated the effects of technetium-99 conjugated with methylene diphosphonate (99Tc-MDP), an agent used in radionuclide therapy, on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis and explored the underlying mechanisms. The murine macrophage cell line RAW264.7 and bone marrow-derived-macrophages from C57BL/6 mice (BMM) were used as models for osteoclastogenesis in vitro. The expression of some key factors in RANKL (50 ng/mL)-induced osteoclastogenesis in RAW264.7 cells was investigated by flow cytometry and real-time reverse transcription–polymerase chain reaction (RT-PCR). To detect multinucleated osteoclast formation, RAW264.7 cells were induced with RANKL for 4 days, whereas BMM were induced by 50 ng/mL RANKL and 20 ng/mL macrophage colony-stimulating factor for 7 days, before being stained with tartrate-resistant acid phosphatase. Osteoclastogenesis was evaluated using the osteoclast markers CD51, matrix metalloproteinase (MMP)-9 and cathepsin K. At 0.01 μg/mL, 99Tc-MDP significantly inhibited RANKL-induced osteoclastogenesis without any cytotoxicity. In addition, 99Tc-MDP abolished the appearance of multinucleated osteoclasts. Real-time RT-PCR analysis of transcription factor expression revealed that 99Tc-MDP inhibited the expression of c-Fos and nuclear factor of activated T cells. In addition, 99Tc-MDP inhibited the expression of the inflammatory factors interleukin (IL)-6, tumour necrosis factor-α and IL-1β. Finally, 99Tc-MDP inhibited the activation of mitogen-activated protein kinases in RAW264.7 cells following RANKL stimulation. In conclusion, 99Tc-MDP possesses anti-osteoclastogenic activity against RANKL-induced osteoclast formation. Citing Literature Supporting Information Filename Description cep12006-sup-0001-FigS1-S2.docxWord document, 5 MB Figure S1. Expression of CD51 in receptor activator of nuclear factor- κB ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells.Figure S2. Anti-osteoclastogenic activity and toxicity of technetium-99 conjugated with methylene diphosphonate (99Tc-MDP) on bone marrow-derived-macrophages (BMM). Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article. Volume39, Issue10October 2012Pages 886-893 RelatedInformation