Biochemical and Biophysical Research Communications
S100A4 inhibition by RNAi up-regulates osteoblast related genes in periodontal ligament cells
Section snippets
Materials and methods
Cell culture. Human PDL (hPDL) cells were obtained from healthy human teeth indicated for extraction for orthodontic treatment according to the report described by Somerman et al. [16]. Periodontal tissue was removed from the middle third of the root using a sterile scalpel. The tissue was rinsed five times with growth media (α-modified minimal essential medium, α-MEM; Gibco) and transferred to culture dishes.
Cultures were maintained in α-MEM supplemented with 10% fetal bovine serum (FBS;
Expression of S100A4 after siRNA transfection in hPDL cells
mRNA expression of S100A4 in cultured hPDL cells 72 h after siRNA transfection was detected by semi-quantitative RT-PCR methods. Non-sense siRNA was also transfected as a control. The results of the semi-quantitative RT-PCR analysis of S100A4 are shown in Fig. 1. S100A4 mRNA was expressed in cultured hPDL cells. The expression was down-regulated after siRNA transfection, while no change was observed after non-sense siRNA transfection.
Immunostaining of S100A4 in cultured hPDL cells with or
Discussion
PDL cells are known to be a heterogeneous cell population and are indispensable for the regeneration of periodontal tissues including the unmineralized PDL and two mineralized tissues, i.e., cementum and bone [18].
S100A4 was detected in cultured hPDL cells at the protein and mRNA levels. Duarte et al. [14] have suggested that a decrease in S100A4 expression may be associated with a terminal osteoblastic differentiation and/or the initiation of mineralized matrix formation. Other reports have
Acknowledgments
This study was supported by Japan Society for the Promotion of Science (JSPS-B-4656) and Grant-in-Aid for Scientific Research of the Ministry of Education, Science, Sports and Culture of Japan (13470459).
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