title: Enhanced osteogenic differentiation of MC3T3-E1 cells on grid-topographic surface and evidence for involvement of YAP mediator.
abstract: Numerous studies have shown that surface topography can promote cell-substrate associations and deeply influence cell fate. The intracellular mechanism or how micro- or nano-patterned extracellular signal is ultimately linked to activity of nuclear transcription factors remains unknown. It has been reported that Yes-associated protein (YAP) can respond to extracellular matrix microenvironment signals, thus regulates stem cell differentiation process. We propose that YAP may play a role in mediating the topography induced cell differentiation. To this end, we fabricated polydimethylsiloxane (PDMS) micropatterns with grid topology (GT) (3 μm pattern width, 2 μm pattern interval length, 7 μm pattern height); nonpatterned PDMS substrates were used as the planar controls. The MC3T3-E1 cells were then cultured on these surfaces, respectively, in osteogenic inducing medium. Cell differentiation in terms of osteogenesis related gene expression, protein levels, alkaline phosphatase activity and extracellular matrix mineralization was assessed. It was shown that the cells on GT surfaces had stronger osteogenesis capacity. In addition, expression level of YAP was increased when MC3T3-E1 cells grew on GT substrates, which was similar to the levels of osteogenic differentiation markers. It was also shown that YAP knockdown attenuated GT substrates-induced MC3T3-E1 differentiation, which reduced the osteogenic differentiation effect of the GT substrates. Collectively, our findings indicate that GT substrates-induced MC3T3-E1 differentiation may be associated with YAP. This paper provides new target points for transcriptional mechanism research of microenvironment induced cell differentiation and a useful approach to obtain more biofunctionalization scaffolds for tissue engineering.
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China.
authors.forename: He
authors.initials: H
authors.lastname: Gong
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China.
authors.forename: Yubo
authors.initials: Y
authors.lastname: Fan
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002113:Calcification, Physiologic; D018797:Cell Cycle Proteins; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D013499:Surface Properties; D054457:Tissue Scaffolds / Q000737:chemistry; D000091102:YAP-Signaling Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D001672:Biocompatible Materials; D018797:Cell Cycle Proteins; D004129:Dimethylpolysiloxanes; D010750:Phosphoproteins; D000091102:YAP-Signaling Proteins; C486223:Yap1 protein, mouse; C013830:baysilon; D000469:Alkaline Phosphatase
keywords: YAP; osteogenesis precursor cells; osteogenic differentiation; substrate; topography