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[Effects of bioactive glass and extracted dentin proteins on human dental pulp cells]
1Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081,China.
Objective:
To investigate the proliferation, odontogenic differentiation and mineralization of human dental pulp cells (HDPCs) on bioactive glass(BG) and extracted dentin proteins(EDP).
Methods:
Primary HDPCs were isolated from third molars by enzyme digestion and were cultured in Dulbecco's minimum essential medium (DMEM). Then the 4th generation of HDPCs was cultured with DMEM, which contained BG-EDP, BG, and EDP, respectively. Meanwhile HDPCs were cultured in DMEM as control group. Proliferation of HDPCs was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) colorimetric assay. Odontogenic differentiation was determined by alkaline phosphatase (ALP) activity assay and real-time PCR. Mineralization was investigated by Alizarin red staining and cetylpyridinium chloride (CPC) assay.
Results:
The proliferation of HDPCs was increased significantly in BG-EDP group on 3,7,and 9 d (optical density value: 1.36±0.06, 2.52±0.20, 2.72±0.29) compared with BG (optical density value: 1.20±0.26, 2.33±0.26, 2.50±0.30), EDP(optical density value: 1.13±0.15, 2.10±0.13, 2.38±0.22) and control group (optical density value: 0.84±0.17, 1.84±0.18, 1.95±0.19), P<0.05. After 7 days, ALP activity of BG-EDP group had no statistical difference compared with EDP group and control group; the expression of odontogenic differentiation genes (DSPP, DMP-1) showed no difference among all the groups(P>0.05). After 14 days, ALP activity of BG-EDP group (56.67±1.83) was significantly upregulated compared with EDP group (41.98±9.71) and control group (30.82±6.70), P<0.05, but had no statistical difference compared with BG group (56.29±6.20), P>0.05; DSPP gene expression was upregulated significantly in BG-EDP group (5.79±1.94) compared with the other groups (P<0.05); DMP-1 gene expression of BG-EDP group (3.87±1.87) increased but had no statistical difference compared with the other groups (P>0.05). The alizarin red staining showed more mineral nodules in BG-EDP group, the cetylpyridinium chloride semi-quantification presented higher calcification in BG-EDP group (0.27±0.01) compared with the other groups (P<0.05).
Conclusion:
Compared with either BG or EDP, BG-EDP significantly promotes the proliferation, odontogenic differentiation and mineralization of HDPCs.