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Bioactivity of periodontal ligament stem cells on sodium titanate coated with graphene oxide.

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.

SUBMITTER: Zhou Q

PROVIDER: S-EPMC4725920 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature

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Bioactivity of periodontal ligament stem cells on sodium titanate coated with graphene oxide.

Zhou Qi Q Yang Pishan P Li Xianlei X Liu Hong H Ge Shaohua S

Scientific reports 20160114

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 ...[more]

PMID: 26763307

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Project description:The objective of the present study was to evaluate the capacity of a tissue-engineered complex of human osteoprotegerin (hOPG)-transfected periodontal ligament stem cells (PDLSCs) seeding on beta-tricalcium phosphate (?-TCP) to regenerate alveolar bone defects in New Zealand rabbits.PDLSCs were isolated from rabbit periodontal ligament tissues and expanded in vitro to enrich PDLSC numbers, and their proliferative activities and differentiation capability were evaluated under specific induction conditions. Lentiviral vector containing hOPG and enhanced green fluorescent protein (EGFP) was constructed by using Gateway technology and transfected into rabbit PDLSCs. The expression of hOPG was determined with quantitative real-time reverse transcription-polymerase chain reaction and Western blot. The PDLSCs with or without engineered hOPG were seeded on ?-TCP scaffolds prior to transplantation. Morphological characterization of cells and materials was done by scanning electron microscope. Twenty rabbits with alveolar bone defects were randomly allocated into four groups and transplanted with ?-TCP, PDLSCs/?-TCP, and hOPG-transfected PDLSCs/?-TCP or were left untreated as a control. Animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis.PDLSCs expressed STRO-1 and vementin and favored osteogenesis and adipogenesis in conditioned media. Expressions of hOPG were significantly upregulated after transfection of the lentiviral vector into PDLSCs. PDLSCs attached and spread well on ?-TCP, and there was no significant difference in growth of PDLSCs on ?-TCP between the hOPG transfection group and the non-transfection group. The histological observation and histomorphometric analysis showed that the hOPG-transfected PDLSCs/?-TCP complex exhibited an earlier mineralization and more bone formation inside the scaffold than control, ?-TCP, and PDLSCs/?-TCP complexes. Implantation of hOPG-transfected PDLSCs contributed to new bone formation as determined by EGFP gene expression under circularly polarized light microscopy.The present study demonstrated the feasibility of ?-TCP scaffolds for primary PDLSC culture and expression of hOPG gene in vitro and in vivo, and hOPG-transfected PDLSCs could serve as a potential cell source for periodontal bone regeneration, which may shed light on the potential of systemic hOPG gene therapy in combination with PDLSC tissue engineering as a good candidate in periodontal tissue engineering for alveolar bone regeneration.

Dataset Information

Bioactivity of periodontal ligament stem cells on sodium titanate coated with graphene oxide.

Publications

Bioactivity of periodontal ligament stem cells on sodium titanate coated with graphene oxide.

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