没食子酸表没食子儿茶素酯促进人牙周膜细胞增殖及成骨分化的影响。
Influence of epigallocatechin-3-gallate in promoting proliferation and osteogenic differentiation of human periodontal ligament cells.
机构信息
Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.
Department of Prosthodontics, College of Stomatology, Xi'an Jiaotong University, 98 Xiwu Road, Xi'an, 710004, Shaanxi, China.
出版信息
BMC Oral Health. 2019 May 2;19(1):73. doi: 10.1186/s12903-019-0768-7.
BACKGROUND
Epigallocatechin-3-gallate (EGCG) was recently proposed to have the potential to regulate bone metabolism, however, its influence on osteogenesis remains controversial. The present study aimed to investigate the effects of EGCG on the proliferation and osteogenesis of human periodontal ligament cells (hPDLCs).
METHODS
Cells were cultured in osteogenic medium and treated with EGCG at various concentrations. Cell proliferation was analyzed using a CCK-8 assay and acridine orange (AO)/ethidium bromide (EB) staining. Flow cytometry was used to measure the intracellular reactive oxygen species (ROS) potential of hPDLCs. The expression levels of osteogenic marker genes and proteins in hPDLCs, including type I collagen (COL1), runt-related transcription factor 2 (RUNX2), osteopontin (OPN), and osterix (OSX), were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. In addition, alkaline phosphatase (ALP) activity was monitored both quantitatively and qualitatively. Extracellular matrix mineralization was further analyzed by alizarin red S staining.
RESULTS
The results showed that EGCG concentrations from 6 to 10 μM increased the ROS level and inhibited the cell proliferation of hPDLCs. EGCG concentrations from 2 to 8 μM effectively increased extracellular matrix mineralization, in which 4 and 6 μM EGCG generated the most mineralizing nodules. The ALP activity and the mRNA and protein expression levels of the tested osteogenic markers were most strongly up-regulated by treatment with 4 and 6 μM EGCG.
CONCLUSIONS
The present study demonstrated that EGCG might promote the osteogenesis of hPDLCs in a dose-dependent manner, with concentrations of 4 and 6 μM EGCG showing the strongest osteogenic enhancement without cytotoxicity, indicating a promising role for EGCG in periodontal regeneration in patients with deficient alveolar bone in the future.
背景
表没食子儿茶素没食子酸酯（EGCG）最近被提出具有调节骨代谢的潜力，但它对成骨的影响仍存在争议。本研究旨在探讨 EGCG 对人牙周膜细胞（hPDLCs）增殖和成骨的影响。
方法
细胞在成骨培养基中培养，并以不同浓度的 EGCG 处理。使用 CCK-8 测定法和吖啶橙（AO）/溴化乙锭（EB）染色分析细胞增殖。使用流式细胞术测量 hPDLCs 的细胞内活性氧（ROS）水平。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹分析测定 hPDLCs 中成骨标志物基因和蛋白的表达水平，包括 I 型胶原（COL1）、 runt 相关转录因子 2（RUNX2）、骨桥蛋白（OPN）和骨形成蛋白（OSX）。此外，还定量和定性监测碱性磷酸酶（ALP）活性。通过茜素红 S 染色进一步分析细胞外基质矿化。
结果
结果表明，6-10μM 的 EGCG 浓度增加了 ROS 水平并抑制了 hPDLCs 的细胞增殖。2-8μM 的 EGCG 浓度有效地增加了细胞外基质矿化，其中 4 和 6μM 的 EGCG 产生了最多的矿化结节。ALP 活性以及测试的成骨标志物的 mRNA 和蛋白表达水平均被 4 和 6μM 的 EGCG 强烈上调。
结论
本研究表明，EGCG 可能以剂量依赖的方式促进 hPDLCs 的成骨作用，4 和 6μM 的 EGCG 浓度在没有细胞毒性的情况下表现出最强的成骨增强作用，表明 EGCG 在未来牙槽骨不足的牙周再生患者中有很好的应用前景。
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