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  • Title: Gene silencing of chordin improves BMP-2 effects on osteogenic differentiation of human adipose tissue-derived stromal cells.
    Author: Schneider H, Sedaghati B, Naumann A, Hacker MC, Schulz-Siegmund M.
    Journal: Tissue Eng Part A; 2014 Jan; 20(1-2):335-45. PubMed ID: 23931154.
    Abstract:
    Although bone morphogenic protein (BMP)-2 is known to potently induce osteogenic differentiation of human mesenchymal stem cells, strong individual differences have been reported. In part, this is due to internal antagonists of BMP-2 for example, noggin and chordin, secreted by differentiating cells. This enabling study was performed to prove the hypothesis that osteogenic effects of BMP-2 can be improved by transient nonviral gene silencing of chordin. We investigated the effect of siRNA against chordin on osteogenic differentiation in human adipose tissue-derived stromal cells (hASC). Cells of two different donors were isolated after liposuction and proliferated for passage 4 or 5. On seeding, hASCs were transfected with siRNA using a commercial liposomal transfection reagent. Subsequently, cells were differentiated in the presence or absence of BMP-2 (100 ng/mL). Noncoding siRNA as well as siRNA against noggin served as a control. Osteogenic differentiation of hASC was determined by alkaline phosphase (ALP) activity and matrix mineralization. ALP activity of hASC treated with siRNA against chordin was increased for cells of both donors. In contrast, silencing of noggin had no effect in any of the donors. In combination with BMP-2, silencing of either chordin or noggin showed strongly improved ALP activity compared with the control group that was also supplemented with BMP-2. Mineralization was observed to start earlier in groups that received siRNA against chordin or noggin and showed increased amounts of incorporated calcium on day 15 compared with the control groups. Silencing chordin in hASCs was successful to increase BMP-2 effects on osteogenic differentiation in both donors, while effects of noggin silencing were reliably observed only in one of the two investigated donors. In contrast to noggin silencing, chordin silencing also increased osteogenic differentiation without supplemented BMP-2.
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