Effect of enamel matrix proteins on odontogenic differentiation of human dental pulp cells in vitro
10.3760/cma.j.issn.1002-0098.2012.s1.013
- VernacularTitle:釉基质蛋白对人牙髓细胞成牙本质分化的影响
- Author:
Hui-Ru ZOU
1
;
Jian-Guo WANG
;
Lan-Cheng ZHANG
;
Zong-Chang QIN
;
Jun SHEN
;
Brookes STEVEN
Author Information
1. 天津市口腔医院牙体牙髓病二科
- Keywords:
Cell differentiation;
Alkaline phosphatase;
Tooth remineralization;
Enamel matrix proteins;
Human dental pulp cells
- From:
Chinese Journal of Stomatology
2012;47(z1):57-62
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of enamel matrix proteins (EMP) on odontogenic differentiation of human dental pulp cells (HDPC) in vitro.Methods HDPC were isolated by explant culture methods and expanded to passage 4 for experiments.The cells were seeded into 6-well plates (2 × 105 cells/well) and divided into 5 groups,treated with 1,10 or 100 mg/L EMP,10-8 mol/L dexamethasone and 100 mg/L ascorbic acid (Dex-AA) or basal media respectively.Alkaline phosphatase (ALP) activity was measureded after up to 10 days in culture.The mRNA expression of dentin matrix protein-1 (DMP-1)and dentin sialophosphoprotein (DSPP) were determined by quantitative real-time reverse transcriptase polymerase chain reaction.Calcified nodule formation was evaluated by alizarin red staining and spectroscopic method.Results The ALP activity levels of all 5 groups were increased in a time-dependent manner.There was no significant difference among each group after 1 day.After 5 days,the ALP activity was increased significantly especially for the 10 mg/L EMP,100 mg/L EMP and Dex-AA groups,values were 7.573 ± 0.267,6.119 ± 0.502 and 5.846 ± 0.096,respectively (P ＜ 0.05).After 10 days,the increase of ALP activity in the 10 mg/L EMP(21.035 ± 0.149) and Dex-AA groups(13.223 ± 0.797) was remarkable,compared with the negative control group (5.825 ± 0.404) (P ＜ 0.01).Statistical differences were detected for DMP-1 and DSPP mRNA expressions in 10mg/L EMP group(14.791 ± 0.164,12.238 ± 0.421),compared with negative control group (4.959 ± 0.184,2.645 ± 0.570) (P ＜ 0.01).Phase contrast microscopy revealed orange-red stained nodules in 1 mg/L and 10 mg/L EMP groups.Such staining was not that much obvoius in the negative control group.After quantifying the staining,it can be clearly seen that 10 mg/L EMP enhanced calcium deposition [(191.8 ± 2.0) μmol/L] significantly,compared to the negative control[(81.1 ± 8.1) μmol/L] (P ＜ 0.01).Conclusions This study showed that optimized concentration of EMP can stimulate odontoblastic differentiation of HDPC,which suggests that EMP may be useful for directing HDPC down the odontoblastic lineage in dentine tissue engineering.
