State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Clinical Research Center for Oral Diseases, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.
Biomed Res Int. 2019 Jun 23;2019:8735952. doi: 10.1155/2019/8735952. eCollection 2019.
HPDLSCs derived from periodontal ligament tissues contribute to tooth development and tissue regeneration. Exploring the effects of long noncoding RNAs (lncRNAs) in the process of osteogenic differentiation of periodontal ligament stem cells would provide novel therapeutic strategies for tissue regeneration. The expression levels of lncRNA, which significantly changed during osteogenic differentiation, were observed by real-time quantitative PCR (q-PCR). Then, we screened for osteogenic-related lncRNA, which was initially named lncRNA-TWIST1. Moreover, we detected the mRNA expression levels of TWIST1 and osteogenesis-related genes after upregulating and downregulating lncRNA-TWIST1 in PPDLSCs (periodontal mesenchymal stem cells from periodontitis patients) and HPDLSCs (periodontal mesenchymal stem cells from healthy microenvironment), respectively. The osteogenic degree was verified by detecting ALP activity and alizarin red staining. LncRNA-TWIST1 decreased the mRNA levels of TWIST1 and promoted osteogenic differentiation in PPDLSCs, which was confirmed by the increase in osteogenesis-related gene levels (Runx2, ALP, and OCN), the increase in ALP activity, and the formation of more osteogenic nodules. In contrast, downregulating lncRNA-TWIST1 decreased the expression of osteogenesis-related genes, ALP activity, and osteogenic nodules both in PPDLSCs and in HPDLSCs. LncRNA-TWIST1 promoted osteogenic differentiation both in PPDLSCs and in HPDLSCs by inhibiting the TWIST1 expression. LncRNA-TWIST1 may be a novel therapeutic strategy to regenerate dental tissues.
牙周膜干细胞来源于牙周组织，有助于牙齿发育和组织再生。探索长链非编码 RNA（lncRNA）在牙周膜干细胞成骨分化过程中的作用，可为组织再生提供新的治疗策略。通过实时定量 PCR（q-PCR）观察到成骨分化过程中 lncRNA 表达水平显著变化。然后，我们筛选出与成骨相关的 lncRNA，最初命名为 lncRNA-TWIST1。此外，我们分别在 PPDLSCs（来自牙周炎患者的牙周膜间充质干细胞）和 HPDLSCs（来自健康微环境的牙周膜间充质干细胞）中上调和下调 lncRNA-TWIST1 后，检测 TWIST1 和骨形成相关基因的 mRNA 表达水平。通过检测碱性磷酸酶（ALP）活性和茜素红染色来验证成骨程度。lncRNA-TWIST1 降低了 PPDLSCs 中 TWIST1 的 mRNA 水平，并促进了成骨分化，这通过增加成骨相关基因（Runx2、ALP 和 OCN）水平、增加 ALP 活性以及形成更多的成骨结节得到证实。相反，下调 lncRNA-TWIST1 降低了 PPDLSCs 和 HPDLSCs 中成骨相关基因、ALP 活性和成骨结节的表达。lncRNA-TWIST1 通过抑制 TWIST1 的表达促进 PPDLSCs 和 HPDLSCs 的成骨分化。lncRNA-TWIST1 可能是一种再生牙齿组织的新治疗策略。
