Effect of Tensile Frequency on the Osteogenic Differentiation of Periodontal Ligament Stem Cells

PURPOSE: The role of periodontal ligament stem cells (PDLSCs) in mediating osteogenesis involved in orthodontic tooth movement (OTM) is well established. However, various relevant in vitro studies vary in the frequency of tension. The effect of tensile frequency on the mechanotransduction of PDLSCs is not clear. The current study aimed to determine the effect of different tensile frequencies on the osteogenic differentiation of PDLSCs and to identify important mechano-sensitivity genes. METHODS: Human PDLSCs were isolated, identified, and subjected to cyclic equibiaxial tensile strain of 12% at different frequencies of 0.1 Hz, 0.5 Hz, 0.7 Hz, or static cultures. Osteogenic differentiation of PDLSCs was assessed by using Western blotting. High-throughput sequencing was used to identify differential mRNA expression. Short time-series expression miner (STEM) was utilized to describe the frequency patterns of the mRNAs. The functions and enriched pathways were identified, and the hub genes were identified and validated. RESULTS: We found that the osteoblastic differentiation capacity of PDLSCs increased with tensile frequency in the range of 0.1–0.7 Hz. Eight frequency-tendency gene expression profiles were identified to be statistically significant. Tensile frequency-specific expressed genes, such as SALL1 and EYA1, which decreased with the increase in tensile frequency, were found. CONCLUSION: The osteoblastic differentiation of PDLSCs under mechanical tensile force is frequency dependent. EYA1 and SALL1 were identified as potential important tensile frequency-sensitive genes, which may contribute to the cyclic tension-induced osteogenic differentiation of PDLSCs in a frequency-dependent manner.