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- Title
RNA interference for noggin enhances the biological activity of bone morphogenetic proteins in vivo and in vitro.
- Authors
Takayama, Kazushi; Suzuki, Akinobu; Manaka, Tomoya; Taguchi, Susumu; Hashimoto, Yusuke; Imai, Yuuki; Wakitani, Shigeyuki; Takaoka, Kunio
- 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 microg) 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.
- Subjects
OSTEOBLAST metabolism; ALKALINE phosphatase; ANIMAL experimentation; BONE morphogenetic proteins; BONE growth; CARRIER proteins; CELL differentiation; CELL lines; COLLAGEN; COMPARATIVE studies; CYTOLOGICAL techniques; GENE expression; GENES; GENETIC techniques; RESEARCH methodology; MEDICAL cooperation; MICE; RECOMBINANT proteins; RESEARCH; RNA; STEM cells; EVALUATION research; BONE density; SKELETAL muscle; OSTEOBLASTS; OSTEOCALCIN; PHOTON absorptiometry
- Publication
Journal of Bone & Mineral Metabolism, 2009, Vol 27, Issue 4, p402
- ISSN
0914-8779
- Publication type
journal article
- DOI
10.1007/s00774-009-0054-x