Adenovirus-mediated transfer of siRNA against Runx2/Cbfa1 inhibits the formation of heterotopic ossification in animal model.
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.
Lin
Lin
L
Institute of Sports Medicine, Peking University Third Hospital, Haidian District, Beijing 100083, PR China.
Chen
Lianxu
L
Wang
Haijun
H
Wei
Xuelei
X
Fu
Xin
X
Zhang
Jiying
J
Ma
Kangtao
K
Zhou
Chunyan
C
Yu
Changlong
C
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*
D050659:Core Binding Factor Alpha 1 Subunit; D017931:DNA Primers; D034741:RNA, Small Interfering