Int J Stomatol

Int J Stomatol ›› 2020, Vol. 47 ›› Issue (1): 24-31.doi: 10.7518/gjkq.2020006

• Original Articles • Previous Articles     Next Articles

Effects of enamel matrix derivative on proliferation and osteogenic differentiation of human periodontal ligament stem cells

Yu Xiaohong1,Liu Yu2(),Zeng Lian1,Yang Yanling1,Wang Zhou1,Li Wei1   

  1. 1. Dept. of Oral Medicine, The Second People’s Hospital of Yunnan Province, Kunming 650012, China;
    2. Dept. of Oral and Maxillofacial Surgery, The Affiliated Stomatology Hospital, Kunming Medical University, Kunming 650031, China
  • Received:2019-05-19 Revised:2019-10-10 Online:2020-01-01 Published:2020-01-01
  • Contact: Yu Liu E-mail:383321966@qq.com
  • Supported by:
    This study was supported by Health Science and Technology Project in Yunnan Province(2018NS0006)

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Abstract:

Objective To investigate the effects of enamel matrix derivative (EMD) on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and the potential mechanism. Methods The primary cultured hPDLSCs were identified by flow cytometry, and the third generation cells were selected for experiments. The cell counting kit-8 assay was performed to detect the effect of different EMD concentrations (0, 20, 50 and 100 mg·L-1) on the proliferation of hPDLSCs. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the effect of EMD on the osteogenic differentiation of hPDLSCs in different concentrations (0, 20, 50 and 100 mg·L-1). The effect of different EMD concentrations (0, 20, 50 and 100 mg·L-1) on collagen synjournal and mineralised nodule formation of hPDLSCs were detected through trichrome and Von Kosa’s staining. Western blotting and qRT-PCR were performed to detect β-catenin, RunX2, CaMKⅡ and NLK expressions after different concentrations of EMD and DDK1 were applied to hPDLSCs.Results EMD significantly promoted the proliferation of hPDLSCs, which increased with the increase in concentration and stimulation time of EMD. After EMD treatment with hPDLSCs, the formation of mineralised nodules and collagen synjournal increased significantly. Furthermore, the expressions of osteocalcin, collagenⅠand RunX2 were elevated. Moreover, the expressions of β-catenin, RunX2, CaMKⅡ and NLK were significantly increased with EMD treatment and inhibited by DDK1.Conclusion EMD can promote the proliferation and osteogenic differentiation of hPDLSCs in vitro through the Wnt/β-catenin signalling pathway.

Key words: enamel matrix derivative, human periodontal ligament stem cell, osteogenic differentiation

CLC Number: 

  • Q254

TrendMD: 

Tab 1

Primer sequence for qRT-PCR"

名称 正向引物 反向引物 产物长度/bp
Ⅰ型胶原 TGACGAGACCAAGAACTGCC GCACCATCATTTCCACGAGC 785
RunX2 GCGGTGCAAACTTTCTCCAG GACTCTGTTGGTCTCGGTGG 97
骨钙素 CATGAGAGCCCTCACACTCC CTCCTGAAAGCCGATGTGGT 256
β-连环蛋白 TAGGTAAGGTGGGTGAGGGT AGGCCCGATTCGGAAGTCT 87
CaMKⅡ ACCTGTGGATATCTGGGCCT TCTGCCTGCCAACTGAGAAG 380
NLK ATCTCCTTTGCAGGATGTTGGT TCATCTAGGTAGGGGTGGGC 82
β-肌动蛋白 TATGGAATCCTGTGGCAT GTGTTGGCATAGAGGTCTT 87

Fig 1

Morphological characteristics of human periodontal ligament stem cells"

Fig 1

Flow cytometry detection of surface antigens of human perio-donatal ligament stem cells"

Fig 3

Effects of enamel matrix derivatives on proliferation of periodontal ligament stem cell"

Fig 4

Expression of osteogensis-related genes of periodontal ligament stem cells detected by qRT-PCR"

Fig 5

Collagen synjournal and mineralization nodule formation of periodontal ligament stem cells detected by Trichrome staining and alizarin red staining"

Fig 6

Expression of β-catenin, RunX2, CaMKⅡ and NLK in periodontal ligament stem cells detected by qRT-PCR and Western blot"

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