摘要
目的 观察表没食子儿茶素没食子酸酯（epigallocatechin-3-gallate,EGCG）对人牙周膜细胞（human periodontal ligament cell,hPDLC）增殖及成骨分化的影响,探讨其对促进牙周骨组织再生的作用.方法 收集新鲜拔除的前磨牙,分离其牙周韧带并进行hPDLC原代培养.取第4代hPDLC,分别给予0、2、4、6、8、10 μmol/L EGCG处理,处理24、48、72 h时采用细胞增殖与毒性检测试剂盒（cell counting kit,CCK）检测细胞增殖情况,培养第7、14天时检测碱性磷酸酶（alkalinephosphatase,ALP）活性,培养第21天时采用茜素红对矿化结节进行染色及定量分析,处理7d时采用实时荧光定量PCR（real-time quantitative PCR,RT-PCR）检测Runt相关转录因子2（Runt-related transcription factor-2,Runx2）、ALP和Ⅰ型胶原的表达情况.结果 4μmol/L EGCG作用于hPDLC 24 h时促进细胞增殖,8、10 μmol/L EGCG作用于hPDLC 24、72h时抑制细胞增殖.2、4、6、8、10 μmol/L EGCG成骨诱导21 d时均可见红染的矿化物,hPDLC吸光度值分别为0.119±0.001、0.167±0.003、0.173±0.003、0.110±0.001、0.083±0.003,除10 μmol/LEGCG组与无EGCG组（0.077±0.001）间差异无统计学意义外,其余组均显著高于无EGCG组（P值均＜0.05）.各浓度EGCG均可促进细胞Ⅰ型胶原、ALP表达,其中4、6 μmol/L效果最显著;4、6μmol/LEGCG可促进细胞Runx2表达,且4μmol/L组促进效果优于6μmol/L组.结论 4μmol/LEGCG在早期对细胞增殖有促进作用,4～6 μmol/L EGCG能显著增强hPDLC的成骨向分化.
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.
作者
刘洁
逯宜
王珍珍
杜文治
裴丹丹
Liu Jie Lu Yi Wang Zhenzhen Du Wenzhi Pei Dandan(Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stamatology, Xi＇an Jiaotong University, Xi＇an 710004, China （Liu J, Wang ZZ, Pei DD Department of Prosthodontics, College of Stomatology, Xi＇an Jiaotong University, Xi＇an 710004, China （Lu Y, Du W)
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2016年第12期758-764,共7页
Chinese Journal of Stomatology
基金
国家自然科学基金（81400551）
关键词
没食子酸
儿茶索
牙周膜
细胞增殖
成骨分化
Gallic acid
Catechin
Periodontal ligament
Cell proliferation
0steogenic differentiation
作者简介
通信作者：裴丹丹，Email：peidandan1986@126．com，电话：029—87218541
