Frizzled 6 disruption suppresses osteoblast differentiation induced by nanotopography through the canonical Wnt signaling pathway

• Published in: Journal of cellular physiology Vol. 235; no. 11; pp. 8293 - 8303
• Main Authors: Abuna, Rodrigo Paolo Flores, Oliveira, Fabiola Singaretti, Adolpho, Leticia Faustino, Fernandes, Roger Rodrigo, Rosa, Adalberto Luiz, Beloti, Marcio Mateus
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.11.2020
• Subjects: Animals; Biomedical materials; Calcium ions; Calcium signalling; Calcium-binding protein; Calmodulin; Catenin; Cell differentiation; Cell Differentiation - physiology; Cell Line; CRISPR; Differentiation (biology); Disruption; frizzled; Frizzled protein; Frizzled Receptors - metabolism; Kinases; Metal Nanoparticles; Mice; nanotopography; osteoblast; Osteoblastogenesis; Osteoblasts - metabolism; Osteogenesis - physiology; Protein kinase; Receptors; Signal transduction; Signaling; Surface Properties; Titanium; Titanium - chemistry; Titanium - pharmacology; Transcription; Transfection; Wnt; Wnt protein; Wnt Signaling Pathway - physiology; nanotopography; osteoblast; Wnt; frizzled; titanium
• ISSN: 0021-9541; 1097-4652; 1097-4652
• DOI: 10.1002/jcp.29674

This study aimed to investigate if wingless‐related integration site (Wnt) signaling participates in the high osteogenic potential of titanium with nanotopography (Ti‐Nano). We showed that among the several components of the Wnt signaling pathway, Frizzled 6 (Fzd6) was the transcript most intensely modulated by nanotopography compared with the untreated Ti surface (Ti‐Machined). Then, we investigated whether and how Fzd6 participates in the regulation of osteoblast differentiation caused by nanotopography. The Fzd6 silencing with CRISPR–Cas9 transfection in MC3T3‐E1 cells induced a more pronounced inhibition of osteoblast differentiation of cells cultured on nanotopography than those cultured on Ti‐Machined. The analysis of the expression of calcium‐calmodulin‐dependent protein kinase II and β‐catenin demonstrated that Fzd6 disruption inhibited the osteoblast differentiation induced by Ti‐Nano by preventing the activation of Wnt/β‐catenin but not that of Wnt/Ca2+ signaling, which is usually triggered by the receptor Fzd6. These findings elucidate the biological function of Fzd6 as a receptor that triggers Wnt/β‐catenin signaling and the cellular mechanisms modulated by nanotopography during osteoblast differentiation. Here, we investigated the participation of wingless‐related integration site (Wnt) signaling in the high osteogenic potential of titanium with nanotopography (Ti‐Nano). Among several components of the Wnt signaling pathway, the receptor Frizzled 6 (Fzd6) was hugely modulated by nanotopography compared with the nontreated titanium surface. The Fzd6 disruption reduced the osteogenicity of nanotopography by impairing Wnt/β‐catenin but not Wnt/Ca2+ signaling in cells cutured on such a surface.