Abstract
As a biocompatible and low cytotoxic nanomaterial, graphene oxide (GO) has captured tremendous interests in tissue engineering. However, little is known about the behavior of dental stem cells on GO. This study was to evaluate the bioactivity of human periodontal ligament stem cells (PDLSCs) on GO coated titanium (GO-Ti) substrate in vitro as compared to sodium titanate (Na-Ti) substrate. By scanning electron microscope (SEM), confocal laser scanning microscope (CLSM), methylthiazol tetrazolium (MTT) assay, alkaline phosphatase (ALP) activity, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis, we investigated the attachment, morphology, proliferation and osteogenic differentiation of PDLSCs on these two substrates. When seeded on GO-Ti substrate, PDLSCs exhibited significantly higher proliferation rate, ALP activity and up-regulated gene expression level of osteogenesis-related markers of collagen type I (COL-I), ALP, bone sialoprotein (BSP), runt related transcription factor 2 (Runx2) and osteocalcin (OCN) compared with those on Na-Ti substrate. Moreover, GO promoted the protein expression of BSP, Runx2 and OCN. These findings suggest that the combination of GO and PDLSCs provides a promising construct for regenerative dentistry.
Highlights
Titanium surfaces, which is demonstrated by the expression of bone forming related proteins[28]
We explored the use of graphene oxide (GO) sheets as a stem cell culture substratum to provide suitable environments for the proliferation and differentiation of human PDLSCs
The surface structure of GO-coated titanium (GO-Ti) was characterized by Scanning electron microscope (SEM) and different sizes of GO deposited on the titanium substrate
Summary
Titanium surfaces, which is demonstrated by the expression of bone forming related proteins[28]. Surface roughness and hydrophilicity of titanium can affect differential Wnt pathways and signaling molecules, targeting the osteogenic differentiation of PDLSCs29. A recent study demonstrated that a composite film of GO could enhance cell adhesion, proliferation and mesenchymal phenotype expression of PDLSCs30. In spite of these studies, the behavior of dental stem cells, especially the response of PDLSCs to GO-coated titanium (GO-Ti) substrate is not fully understood. We fabricated GO-Ti substrate as a cell culture platform. We analyzed the effects of GO-Ti substrate on cell morphology, proliferation and osteogenic differentiation potentials of PDLSCs
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