Effect of epigallocatechin-3-gailate on the proliferation and osteogenesis of human periodontal ligament cells
10.3760/cma.j.issn.1002-0098.2016.12.011
- VernacularTitle:表没食子儿茶素没食子酸酯对人牙周膜细胞增殖及成骨分化的影响
- Author:
Jie LIU
1
;
Yi LU
;
Zhenzhen WANG
;
Wenzhi DU
;
Dandan PEI
Author Information
1. 710004,西安交通大学口腔医学院陕西省颅颌面精准医学研究重点实验室
- Keywords:
Gallic acid;
Catechin;
Periodontal ligament;
Cell proliferation;
0steogenic differentiation
- From:
Chinese Journal of Stomatology
2016;51(12):758-764
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the effect of epigallocatechin-3-gallate (EGCG) treatment on the proliferation and osteogenic differentiation of human periodontal ligament cell (hPDLC) and to explore the potential role of EGCG in promoting periodontal hard tissue regeneration.Methods The hPDLC was isolated from periodontal ligament tissue obtained from freshly extracted human teeth.The effect of treatments with various concentrations of EGCG (0 μmol/L,2 μmol/L,4 μmo]/L,6 μmol/L,8 μmol/L and 10 μmol/L) on cell proliferations were determined by cell counting kits (CCK) after 24-,48-and 72-hour-incubations,respectively.Osteogenic differentiation abilities of hPDLCs were assessed by using alkaline phosphatase (ALP) activity tests after 7-and 14-day-incubations,respectively.The mineralized nodules were quantitatively examined and analyzed by using alizarin red staining after 21-day-incubation.The real-time PCR (RT-PCR) assays were conducted fordetecting the expressions of Runt related transcription factor-2 (Runx2),ALP and collagen type Ⅰ (COL Ⅰ) after 7-day-incubation.Results Treatment with 4 μ mol/L EGCG increased hDPLC proliferation at 24 h,while 8 μmol/L or 10 μ mol/L EGCG treatment groups showed inhibiting effects at 24 h and 72 h,respectively.Findings of alizarin redstaining showed orange to red colored extracellular mineralized nodules in all groups.The the A values of 2,4,6,8,10 μ mol/L EGCG groups were 0.119±0.001,0.167±0.003,0.173±0.003,0.110±0.001 and 0.083±0.003,respectively.A values of 2-8 μmol/L EGCG groups were significantly higher than that of the control group,however there was no significant difference of the A values betweenl0 μmol/L EGCG group and the control group (0.077±.0.001).Treatments with 2-10 μmol/L EGCG could significantly increase the mRNA expressions of COL Ⅰ and ALP with the highest values in 4-6 μ mol/L EGCG treatment groups.Although treatments with 4 and 6 μmol/L EGCG both could increase the mRNA expressions of Runx2,the result in 4 μmol/L group was much better than that of 6 μmol/L group.Conclusions Treatment of 4 μmol/L EGCG could promote hPDLC proliferation at early stageand treatments with 4-6 μ mol/L EGCG could significantly promote the osteogenesis of hPDLCs which might play a promising role in periodontal hard tissue regeneration.
