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EGR1 overexpression prevents the downregulation of tendon-associated gene expression in tension released engineered tendons.

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posted on 07.11.2016, 17:30 by Ludovic Gaut, Nicolas Robert, Antony Delalande, Marie-Ange Bonnin, Chantal Pichon, Delphine Duprez

(A) Two week-old fibrin gel constructs made of mouse C3H10T1/2 cells or C3H10T1/2-EGR1 cells were analyzed for tendon gene expression by RT-q-PCR analyses. The mRNA levels of C3H10T1/2 constructs were normalized to 1. Errors bars represent standard errors of the mean of 5 C3HT101/2 constructs and 7 C3H10T1/2-EGR1 constructs. The p values were calculated using the Mann-Whitney test. The mRNA levels of Egr1, Scx, Tnmd and Tgfb2 genes were increased in C3H10T1/2-EGR1 constructs compared to those of C3H10T1/2 constructs. (B) Tension was released in C3H10T1/2 constructs by sectioning one end of the construct. Transcript levels were analyzed by RT-q-PCR analyses in tension-released C3H10T1/2 constructs and compared to C3H10T1/2 constructs. The mRNA levels of C3H10T1/2 constructs were normalized to 1. Errors bars represent standard errors of the mean of 7 C3H10T1/2 constructs and 7 tension-released C3H10T1/2 constructs. The p values were calculated using the Mann-Whitney test. We observed a decrease in the transcript levels of Egr1, Scx, Tnmd, Col1a1, Col1a2 and Tgfb2 genes in tension-released C3H10T1/2 constructs (TR) compared to tensioned C3H10T1/2 constructs (T). (C) The mRNA levels of tendon genes were analyzed in tension-released C3H10T1/2-EGR1 constructs and compared with tensioned C3H10T1/2-EGR1 constructs by RT-q-PCR analyses. The mRNA levels of C3H10T1/2-EGR1 constructs were normalized to 1. Errors bars represent standard errors of the mean of 7 C3HT101/2-EGR1 constructs and 5 tension-released C3H10T1/2-EGR1 constructs. The p values were calculated using the Mann-Whitney test. There was no significant change in tendon gene expression in tension-released C3H10T1/2-EGR1 constructs (TR) compared to tensioned C3H10T1/2-EGR1 constructs (T). T, Tension, TR, Tension release.

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