Department of Orthodontics, School of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Department of Periodontics, School of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Arch Oral Biol. 2017 Jun;78:100-108. doi: 10.1016/j.archoralbio.2017.01.019. Epub 2017 Feb 7.
Periodontium regeneration is one of the most important processes for periodontitis therapy. Human periodontal ligament cells (hPDLCs) play a vital role in the repair and regeneration of periodontal tissues. Our study aimed to investigated the mechanisms underlying the promotion of hPLDCs osteogenic differentiation by baicalein.
hPDLCs were obtained from periodontal ligament (PDL) tissues by primary culture. The MTT assay was used to determine the growth curves of hPDLCs treated with different concentrations of baicalein (1.25, 2.5, 5, or 10μM). Alkaline phosphatase (ALP) staining and Alizarin red S staining were performed to assess osteogenic differentiation of hPDLCs administered baicalein. Osteogenic differentiation-related gene and protein expression levels and Wnt/β-catenin pathway signal changes were assessed by qRT-PCR and Western blotting analysis.
The results showed that baicalein decreased the growth of hPDLCs slightly and increased ALP activity and calcium deposition in a dose-dependent manner. The expression of runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), Osterix (OSX) and osteocalcin (OCN) were elevated after baicalein administration. Moreover, baicalein strongly activated the Wnt/β-catenin pathway and up-regulated the expression of β-catenin, lymphoid enhancer factor 1 (LEF1) and Cyclin D1. Dickkopf-related protein 1 (DKK-1) significantly reversed the effects of baicalein on hPDLCs.
Our findings indicated that baicalein enhanced the osteogenic differentiation of hPDLCs via the activation of the Wnt/β-catenin signaling pathway, which may represent a potential candidate for periodontitis therapy.
牙周组织再生是牙周炎治疗的最重要过程之一。人牙周韧带细胞(hPDLCs)在牙周组织的修复和再生中起着至关重要的作用。我们的研究旨在探讨白杨素促进 hPDLCs 成骨分化的机制。
通过原代培养从牙周韧带(PDL)组织中获得 hPDLCs。MTT 法测定不同浓度白杨素(1.25、2.5、5 或 10μM)处理的 hPDLCs 生长曲线。碱性磷酸酶(ALP)染色和茜素红 S 染色评估白杨素处理的 hPDLCs 的成骨分化。通过 qRT-PCR 和 Western blot 分析评估成骨分化相关基因和蛋白表达水平以及 Wnt/β-catenin 通路信号变化。
结果表明,白杨素轻微抑制 hPDLCs 的生长,并且呈剂量依赖性增加 ALP 活性和钙沉积。白杨素处理后,RUNX2、骨形态发生蛋白 2(BMP2)、成骨特异性转录因子 2(OSX)和骨钙素(OCN)的表达增加。此外,白杨素强烈激活 Wnt/β-catenin 通路,上调β-catenin、淋巴增强因子 1(LEF1)和细胞周期蛋白 D1 的表达。Dickkopf 相关蛋白 1(DKK-1)显著逆转了白杨素对 hPDLCs 的作用。
我们的研究结果表明,白杨素通过激活 Wnt/β-catenin 信号通路增强 hPDLCs 的成骨分化,这可能是牙周炎治疗的潜在候选药物。