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  • Title: Wrinkled topography regulates osteogenesis via autophagy-mediated Wnt/β-catenin signaling pathway in MC3T3-E1 cells.
    Author: Zhang Q, Zhang Q, Yan X, Wang L, Yuan X.
    Journal: Arch Oral Biol; 2023 Jul; 151():105700. PubMed ID: 37094411.
    Abstract:
    OBJECTIVE: In this study, we aimed to evaluate the effects of different dimensional wrinkled in topography on the osteogenic differentiation of MC3T3-E1 cells and explored the underlying mechanisms. DESIGN: Polydimethylsiloxane (PDMS) with a wrinkled topography was synthesized using an elastomer base and crosslinking while observing by atomic force microscopy. MC3T3-E1 proliferation was detected by Cell Counting Kit-8(CCK-8) assays and the cell morphology was determined by phalloidin staining. Osteogenetic genes expression levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. In addition, Autophagy-related genes expression levels were evaluated by immunostaining and western blotting in MC3T3-E1 in order to assess the induction of autophagy. RESULTS: In this experiment, the 0.7 µm amplitude and 3 µm wavelength (W3) group increased the expression of osteogenic markers, whereas the 4.3 µm amplitude and 27 µm wavelength (W27) group showed inhibition. Both the cytoplasm and the nucleus of β-catenin, compared with those of the Flat, W3 increased, whereas W27 decreased. At the same time, the autophagy was consistent with the influence of the topography on osteogenic differentiation. Moreover, using CQ or RAPA significantly inhibited or promoted autophagy, as well as partially decreasing or increasing osteogenesis, respectively. Infecting siRNA-β-catenin decreased the expression of RUNX2 and OSX in MC3T3-E1 cells both treated with CQ and RAPA. CONCLUSIONS: Wrinkled topographies activated the autophagy-mediated Wnt/β-catenin signaling pathway and affected the osteogenic differentiation of MC3T3-E1 cells. The introduction of aligned topographies on biomaterial scaffolds could provide physical cues with which modulate MC3T3-E1 responses for bone engineering constructs.
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