Sequential application of bFGF and BMP-2 facilitates osteogenic differentiation of human periodontal ligament stem cells

Wenyan Kang; Qianyu Liang; Lingqian Du; Lingling Shang; Ting Wang; Shaohua Ge

doi:10.1111/jre.12644

Sequential application of bFGF and BMP-2 facilitates osteogenic differentiation of human periodontal ligament stem cells

J Periodontal Res. 2019 Aug;54(4):424-434. doi: 10.1111/jre.12644. Epub 2019 Mar 9.

Authors

Wenyan Kang^{1

2}, Qianyu Liang^{1

2}, Lingqian Du³, Lingling Shang^{1

2}, Ting Wang^{1

2}, Shaohua Ge^{1

2}

Affiliations

¹ Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, China.
² Department of Periodontology, School of Stomatology, Shandong University, Jinan, China.
³ Department of Stomatology, The Second Hospital of Shandong University, Jinan, China.

PMID: 30851068
DOI: 10.1111/jre.12644

Abstract

Background and objectives: Basic fibroblast growth factor (bFGF) promotes cells proliferation and chemotaxis and maintains stemness while inhibits mineralized nodule formation. Bone morphogenetic protein 2 (BMP-2) shows great potential in promoting bone formation. However, sequential application of these two growth factors on periodontal ligament stem cells (PDLSCs) has not been explored. In this study, we aimed to identify the optimal concentration and time of bFGF on PDLSCs proliferation, migration and then investigate the sequential delivery of bFGF and BMP-2 on osteogenic differentiation of PDLSCs in vitro.

Materials and methods: Periodontal ligament stem cells were isolated by limiting dilution method. Dose-dependent additive effects of bFGF and BMP-2 on PDLSCs were detected. Cell counting assay, cell migration assay, alkaline phosphatase (ALP) activity assay, Alizarin red staining, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot analysis were used to determine different application modalities of bFGF and BMP-2 on proliferation, migration, and osteogenic differentiation of PDLSCs.

Results: 50 ng/mL bFGF significantly promoted PDLSCs proliferation and chemotaxis while time-dependently inhibited BMP-2 induced ALP activity. Sequential application of 25 ng/mL bFGF for first 3 days and followed with 50 ng/mL BMP-2 for another 9, 18, and 25 days significantly promoted PDLSCs osteogenic differentiation. Compared with bFGF and BMP-2 simultaneous group, sequential application of bFGF and BMP-2 group significantly enhanced ALP activity, osteogenesis-related genes and proteins expression and mineral deposition.

Conclusion: Sequential application of bFGF and BMP-2 synergistically promoted osteogenic differentiation of PDLSCs, and this sequential application modality of growth factors would provide a new strategy for periodontal regeneration.

Keywords: basic fibroblast growth factor; bone morphogenetic protein; osteogenic differentiation; periodontal ligament stem cells.

MeSH terms

Bone Morphogenetic Protein 2 / pharmacology*
Cell Differentiation*
Cell Proliferation
Cells, Cultured
Dose-Response Relationship, Drug
Fibroblast Growth Factor 2 / pharmacology*
Humans
Osteogenesis*
Periodontal Ligament / chemistry*
Stem Cells / cytology*
Stem Cells / drug effects

Substances

Bone Morphogenetic Protein 2
Fibroblast Growth Factor 2

Abstract

MeSH terms

Substances

Grants and funding