title: Adenovirus-mediated transfer of siRNA against Runx2/Cbfa1 inhibits the formation of heterotopic ossification in animal model.
abstract: The heterotopic ossification of muscles, tendons, and ligaments is a common problem faced by orthopaedic surgeons. Runx2/Cbfa1 plays an essential role during the osteoblast differentiation and is considered as a molecular switch in osteoblast biology. RNA interference technology is a powerful tool for silencing endogenous or exogenous genes in mammalian cells. In this study, we investigated the effect of Runx2/Cbfa1-specific siRNA on osteoblast differentiation and mineralization in osteoblastic cells, and then constructed adenovirus containing siRNA against Runx2/Cbfa1 (Ad-Runx2-siRNA) to inhibit the formation of heterotopic ossification induced by BMP4, demineralized bone matrix, and trauma in animal model. Our results showed that the Runx2/Cbfa1-specific siRNA could inhibit the expression of Runx2/Cbfa1 at the level of mRNA and protein. Analysis of the expression of osteoblast maturation genes including type I collagen, osteopontin, bone sialoprotein, and osteocalcin, alkaline phosphatase activity, and matrix mineralization (von kossa) revealed that osteoblast differentiation was inhibited in cultured primary mouse osteoblasts transduced with Ad-Runx2-siRNA. Furthermore, adenovirus-mediated transfer of siRNA against Runx2/Cbfa1 could inhibit the formation of heterotopic ossification induced by BMP4, demineralized bone matrix, and trauma in animal model. It is likely that the inhibition of Runx2/Cbfa1 by RNAi could be developed as a powerful approach to prevent or treat heterotopic ossification.
authors.affiliation: Institute of Sports Medicine, Peking University Third Hospital, Haidian District, Beijing 100083, PR China.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Lianxu
authors.initials: L
authors.lastname: Chen
authors.forename: Haijun
authors.initials: H
authors.lastname: Wang
authors.forename: Xuelei
authors.initials: X
authors.lastname: Wei
authors.forename: Xin
authors.initials: X
authors.lastname: Fu
authors.forename: Jiying
authors.initials: J
authors.lastname: Zhang
authors.forename: Kangtao
authors.initials: K
authors.lastname: Ma
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhou
authors.forename: Changlong
authors.initials: C
authors.lastname: Yu
mesh_terms: D000256:Adenoviridae / Q000378:metabolism*; D000818:Animals; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004195:Disease Models, Animal; D018507:Gene Expression Regulation, Developmental; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D051379:Mice; D009999:Ossification, Heterotopic / Q000517:prevention & control; D010006:Osteoblasts / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D017931:DNA Primers; D034741:RNA, Small Interfering