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RhoA controls Wnt upregulation on microstructured titanium surfaces

Lookup NU author(s): Dr Silvia Mazzotta

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Abstract

Rough topography enhances the activation of Wnt canonical signaling in vitro, and this mediates its effects on cell differentiation. However, the molecular mechanisms underlying topography-dependent control of Wnt signaling are still poorly understood. As the small GTPase RhoA controls cytoskeletal reorganization and actomyosin-induced tensional forces, we hypothesized that RhoA could affect the activation of Wnt signaling in cells on micropatterned titanium surfaces. G-LISA assay revealed that RhoA activation was higher in C2C12 cells on rough (SLA) surfaces under basal conditions than on smooth (Polished) titanium. Transfection with dominant negative RhoA decreased Wnt activation by normalized TCF-Luc activity on SLA, whilst transfection with constitutively active RhoA increased TCF-Luc activation on Polished titanium. One mM Myosin II inhibitor Blebbistatin increased RhoA activation but decreased Wnt activation on SLA surfaces, indicating that tension-generating structures are required for canonical Wnt modulation on titanium surfaces. Actin inhibitor Cytochalasin markedly enhanced RhoA and TCF-Luc activation on both surfaces and increased the expression of differentiation markers in murine osteoblastic MC3T3 cells. Taken together, these data show that RhoA is upregulated in cells on rough surfaces and it affects the activation of Wnt canonical signaling through Myosin II modulation.


Publication metadata

Author(s): Lumetti S, Mazzotta S, Ferrillo S, Piergianni M, Piemontese M, Passeri G, Macaluso GM, Galli C

Publication type: Article

Publication status: Published

Journal: BioMed Research International

Year: 2014

Volume: 2014

Online publication date: 14/05/2014

Acceptance date: 20/03/2014

ISSN (print): 2314-6133

ISSN (electronic): 2314-6141

Publisher: Hindawi

URL: http://dx.doi.org/10.1155/2014/401859

DOI: 10.1155/2014/401859

PubMed id: 401859


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