qRT-PCR results of cell differentiation showed that miR-221 overexpression significantly increased the expression of
Runx2 and Ocn on days 7 and 14, whereas miR-221 suppression showed opposite results (P<0.05, Figure 4A and B).
In vitro studies have shown that IL-1 and TNF-a enhance the expression of Wnt signaling and BMP2 in osteoblasts.[21] IL-1 can also stimulate ALP activity and mineralization by inducing a mechanism that is independent of
Runx2 in VSMCs.[22] IL-6 can promote TNF-a expression and increase
Runx2 expression associated with sodium-dependent phosphate transporter 1 (PiT1) and aid calcium deposition in mice.[23] Persistent activation of the inflammatory response leads to activation of inflammation-related signaling pathways, macrophages, and T-lymphocytes, thereby leading to the osteoblast-like differentiation of VSMCs.[24] Studies have shown that levels of pro-inflammatory factors in the peripheral blood such as CRP and TNF-a are increased significantly in CAC patients.
Similarly, the expression of ALP, COL1 and
RUNX2, markers of fibroblast differentiation and bone formation, was also not significantly different between the groups, although the expression of
RUNX2 in the cells exposed to PC 15% was notably higher.
The role of DMP-1 signaling has been well studied; after endocytosis by preodontoblasts, DMP-1 induces calcium release from the endoplasmic reticulum, activating the p38 pathway and
RUNX2 nuclear translocation, which are responsible for initiating the transcription of odontogenesis-related genes [24].
Remarkably, together with enhanced level of
RUNX2, ALP, and OSX, there was an overexpression of CEMP1, which is a novel cementum component exclusively expressing for cementoblasts and their progenitors.
Xu, "Downregulated LncRNA-ANCR promotes osteoblast differentiation by targeting EZH2 and regulating
Runx2 expression," Biochemical and Biophysical Research Communications, vol.
The slices were incubated in a humid atmosphere with diluted primary antibodies (Dkk-1, GSK-3[beta], [beta]-catenin, PPAR[gamma]2, and
Runx2) at 37[degrees]C for 45 min, washed 3 times with PBS, and then incubated with biotin-conjugated secondary antibody at 37[degrees]C for 45 min.
The mRNA expression of the osteogenic differentiation factors
RUNX2, OPN, and OCN increased gradually as the culture time increased.
OC, ON, Col I, and
Runx2 are major phenotypic markers for preosteoblast differentiation during bone formation.