Technetium-99 conjugated with methylene diphosphonate inhibits receptor activator of nuclear factor-κB ligand-induced osteoclastogenesis
Article first published online: 26 SEP 2012
© 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Wiley Publishing Asia Pty Ltd
Clinical and Experimental Pharmacology and Physiology
Volume 39, Issue 10, pages 886–893, October 2012
How to Cite
Gong, W., Dou, H., Liu, X., Sun, L. and Hou, Y. (2012), Technetium-99 conjugated with methylene diphosphonate inhibits receptor activator of nuclear factor-κB ligand-induced osteoclastogenesis. Clinical and Experimental Pharmacology and Physiology, 39: 886–893. doi: 10.1111/j.1440-1681.2012.12006.x
- Issue published online: 26 SEP 2012
- Article first published online: 26 SEP 2012
- Manuscript Revised: 17 AUG 2012
- Manuscript Accepted: 12 AUG 2012
- Manuscript Received: 16 MAY 2012
- National Natural Science Foundation of China. Grant Numbers: 90813036, 81072410
- Scientific Research Foundation of Graduate School of Jiangsu Province. Grant Number: CXZZ11–0036
- Fundamental Research Funds for the Central Universities . Grant Numbers: 1117021406, 1118021406, 1107021460, 1094021410
- Scientific Research Foundation of Graduate School of Nanjing University. Grant Number: 2010 CL04
- receptor activator of nuclear factor-κB ligand (RANKL);
- technetium-99 conjugated with methylene diphosphonate (99Tc-MDP)
- 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.