Eluted zinc ions stimulate osteoblast differentiation and mineralization in human dental pulp stem cells for bone tissue engineering

Kazuyuki Yusa; Osamu Yamamoto; Mitsuyoshi Iino; Hiroshi Takano; Masayuki Fukuda; Zhiwei Qiao; Toshihiro Sugiyama

doi:10.1016/j.archoralbio.2016.07.010

Eluted zinc ions stimulate osteoblast differentiation and mineralization in human dental pulp stem cells for bone tissue engineering

Arch Oral Biol. 2016 Nov:71:162-169. doi: 10.1016/j.archoralbio.2016.07.010. Epub 2016 Aug 5.

Authors

Kazuyuki Yusa¹, Osamu Yamamoto², Mitsuyoshi Iino³, Hiroshi Takano⁴, Masayuki Fukuda⁴, Zhiwei Qiao⁵, Toshihiro Sugiyama⁵

Affiliations

¹ Department of Biochemistry, Akita University Graduate School of Medicine, 1-1-1 Hondo, Akita, Akita, 010-8543, Japan; Department of Dentistry, Oral and Maxillofacial-Plastic and Reconstructive Surgery, School of Medicine, Yamagata University, 2-2-2 Iida-Nishi, Yamagata, Yamagata, 990-9585, Japan. Electronic address: k-yusa@med.id.yamagata-u.ac.jp.
² Department of Bio-System Engineering, Graduate School of Science and Engineering, Yamagata University, 4-3-16 Johnan, Yonezawa, Yamagata, 992-8510, Japan.
³ Department of Dentistry, Oral and Maxillofacial-Plastic and Reconstructive Surgery, School of Medicine, Yamagata University, 2-2-2 Iida-Nishi, Yamagata, Yamagata, 990-9585, Japan.
⁴ Division of Dentistry and Oral Surgery, Akita University School of Medicine, 1-1-1 Hondo, Akita, Akita, 010-8543, Japan.
⁵ Department of Biochemistry, Akita University Graduate School of Medicine, 1-1-1 Hondo, Akita, Akita, 010-8543, Japan.

PMID: 27521529
DOI: 10.1016/j.archoralbio.2016.07.010

Abstract

Objective: Zinc is an essential element for proliferation, differentiation and survival in various cell types. In a previous study, we found that zinc ions released from zinc-modified titanium surfaces (eluted zinc ions; EZ) stimulate cell viability, osteoblast marker gene expression and calcium deposition in human bone marrow-derived mesenchymal cells (hBMCs). The aim of the present study was to investigate the effects of EZ on osteoblast differentiation among dental pulp stem cells (DPSCs) in vitro.

Method: In this study, we evaluated the effects of EZ on osteogenesis in DPSCs. Osteoblast and osteoclast marker gene expression was evaluated by real-time PCR. We also evaluated alkaline phosphatase (ALP) staining and calcium deposition.

Results: We found that EZ stimulated osteoblast marker gene (type I collagen, alkaline phosphatase (ALP), osteocalcin (OCN) and Runx2) expression, vascular endothelial growth factor A (VEGF-A), and TGF-beta signaling pathway-related gene expression after 7days of incubation. Osteoclastogenesis occurs in a receptor for activated nuclear-factor kappa B ligand (RANKL)/osteoprotegerin (OPG)-independent manner. Real-time PCR analysis revealed that EZ did not affect RANKL or OPG mRNA expression. It was also revealed that EZ induced alkaline phosphatase (ALP) staining and calcium deposition in DPSCs. Collectively, these results demonstrate the potential for clinical application to prospective treatment of bone diseases.

Keywords: Bone tissue engineering; Dental pulp stem cells; Osteoblast differentiation; Zinc ion; Zinc-modified titanium.

MeSH terms

Alkaline Phosphatase / biosynthesis
Bone Regeneration / drug effects*
Cell Differentiation / drug effects*
Collagen Type I / biosynthesis
Core Binding Factor Alpha 1 Subunit / biosynthesis
Dental Pulp / cytology*
Gene Expression
Humans
In Vitro Techniques
Ions / pharmacology
Mesenchymal Stem Cells / drug effects*
Osteoblasts / drug effects*
Osteocalcin / biosynthesis
Osteogenesis / drug effects*
Real-Time Polymerase Chain Reaction
Tissue Engineering / methods*
Titanium / pharmacology
Transforming Growth Factor beta / biosynthesis
Vascular Endothelial Growth Factor A / biosynthesis
Zinc / pharmacology*

Substances

Collagen Type I
Core Binding Factor Alpha 1 Subunit
Ions
Transforming Growth Factor beta
Vascular Endothelial Growth Factor A
Osteocalcin
Titanium
Alkaline Phosphatase
Zinc