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Bone morphogenetic protein-2 (BMP-2) gene delivery is expected to be useful as a way to overcome protein delivery problems, such as the need for a scaffold and high dose of BMP-2 protein, and using non-viral vectors is known to be a safer gene delivery system than that using viral vectors for clinical application. However, the efficiency of the BMP-2 expression is very low when non-viral vectors are used. In the present study, we clarified the feasibility of RANKL-binding peptide, which has recently been recognized as a bone anabolic reagent, in bone formation induced by BMP-2 gene transfer using non-viral vectors. We induced ectopic bone in the gastrocnemius muscle of C57BL/6 mice by a single injection of GFP/BMP-2-expressing vector. Subcutaneous administration of RANKL-binding peptide increased the ectopic bone mass on days 14 and 21 after plasmid injection, and histological analyses showed the stimulation of bone formation activity of the ectopic bone compared to the vehicle-treated control. Micro computed tomography images showed that RANKL-binding peptide improved the microstructure of ectopic bone. Our results suggest that RANKL-binding peptide may be effective in promoting bone formation induced by BMP-2 gene transfer using non-viral vectors. *Correspondence to: Kazuhiro Aoki, Professor, Department of Basic Oral Health Engineering, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan, Tel: +813-5803-4641, E-mail: kazu.hpha@tmd.ac.jp

RANKL-binding peptide promotes ectopic bone formation induced by BMP-2 gene transfer in murine gastrocnemius muscle