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Abstract Objective To explore the mechanism of continuous stretch stress inducing differentiation of tendon stem cells (TSCs) into osteoblasts.Methods TSCs were treated with different degrees of Strain for 48 h to observe the cell morphology,and the cell relative viability rate was detected by CCK-8 method.RT-PCR and Western-blot were used to detect the differentiation,stem cell maintenance-related transcription factors SOX2,OCT4 gene and protein expression,osteoblast RUNX2(Runt related transcription factor 2),DLX5 and COL1a relative expression and RUNX2 protein expression.Results The cell relative viability by CCK-8 method showed that the cell relative viability rate increased in each Strain treatment group,but the increase was only statistically significant when the Strain was 6% (P<0.05).Compared with the control group,when the Strain reached 4%,the expression of SOX2 gene and protein decreased significantly (P<0.05).For OCT4 gene,expression was significantly decreased after 6% Strain treatment (P<0.05).The osteogenic gene RUNX2 increased only in the 2% Strain group and the 6% Strain group (P<0.05).However,COL1a only increased in the 6% Strain group (P<0.05).RUNX2 protein was detected only in the 6% Strain group,and the difference was statistically significant (P<0.05).After further testing by alkaline phosphatase,it was found that 4% Strain and 6% Strain can effectively improve the process of osteogenic differentiation.Conclusion Moderately mild tension helps to improve the survival rate of TSCs cells,and moderate tension can also accelerate the differentiation of TSCs and accelerate its ability to differentiate into osteoblasts by regulating the expression level SOX2、OCT4 genes and their proteins,the relative expression of osteoblast RUNX2、DLX5 and COL1a,and the protein expression of RUNX2.
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