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RNA interference for noggin enhances the biological activity of bone morphogenetic proteins in vivo and in vitro

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Abstract

Noggin is a major extracellular antagonist to bone morphogenetic proteins (BMPs) which binds to BMPs and blocks binding of them to BMP-specific receptors and negatively regulates BMP-induced osteoblastic differentiation. In this study, we investigated the effect of noggin silencing by transfection of small interfering RNA (siRNA) on BMP-induced osteoblastic differentiation in vitro and ectopic bone formation in vivo induced by recombinant human BMP-2 (rhBMP-2). Noggin mRNA expression was up-regulated in response to rhBMP-2 in C2C12 cells, a myoblastic cell line, in dose- and time-dependent fashion as determined by real-time RT-PCR assay. Silencing of noggin expression by transfection of noggin siRNA suppressed BMP-stimulated noggin expression, resulting in acceleration of BMP-induced osteoblastic differentiation. For in vivo noggin silencing, siRNA was injected locally into back muscles and transfected into local cells by electroporation, where rhBMP-2-retaining (5 μg) collagen disks had been surgically placed. The implants were harvested at 2 weeks after surgery from experimental and control group mice and analyzed by radiological and histological methods. As a result, bone mineral content of ossicles ectopically induced by rhBMP-2 was significantly increased by silencing of noggin. Our findings suggest that silencing of noggin enhances the osteoblastic differentiation of BMP-responding cells in vitro and new bone formation induced by rhBMP-2 in vivo by eliminating negative regulation of the effects of BMP. RNA interference might be useful for intensifying the effects of BMP in promoting new bone (callus) formation in repair of damaged bone.

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Acknowledgments

This work was supported in part by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (Project Grants 16109009 and 1679085 to KT, and 19791018 to YI). Authors would like to thank Ms A. Inagaki, K. Hata, K. Kamei and Y. Hanamoto for their technical assistances.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abenoku, Osaka, 545-8585, Japan

    Kazushi Takayama, Akinobu Suzuki, Tomoya Manaka, Susumu Taguchi, Yusuke Hashimoto, Yuuki Imai, Shigeyuki Wakitani & Kunio Takaoka

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  1. Kazushi Takayama
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  2. Akinobu Suzuki
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  3. Tomoya Manaka
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  4. Susumu Taguchi
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  8. Kunio Takaoka
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Correspondence to Yuuki Imai.

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Takayama, K., Suzuki, A., Manaka, T. et al. RNA interference for noggin enhances the biological activity of bone morphogenetic proteins in vivo and in vitro. J Bone Miner Metab 27, 402–411 (2009). https://doi.org/10.1007/s00774-009-0054-x

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  • DOI: https://doi.org/10.1007/s00774-009-0054-x

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