Ma Lan, Li Min, Xuan Guihong, Dai Ying
Department of Stomatology, Shaoxing People's Hospital, Zhongxing North Road NO.568, Shaoxing, 312000, China.
Odontology. 2025 Apr 18. doi: 10.1007/s10266-025-01097-2.
Periodontitis is a chronic inflammatory disease that leads to the loss of periodontal supporting tissue. Furthermore, human periodontal ligament stem cells (hPDLSCs) are identified as candidate cells for the regeneration of periodontal and alveolar bone tissues. N6-Methyladenosine (m6A) performs a vital role in osteoporosis and bone metabolism. However, the role and mechanism of Methyltransferase-like 14 (METTL14) in the osteogenic differentiation of PDLSCs from periodontitis sufferers (pPDLSCs) is unclear. In this research, GSE223924 database analyzed the expression of METTL14 and Wnt Family Member 3A (WNT3A) in gingival tissue samples of 10 healthy subjects, 10 patients with periodontitis and peri-implantitis. RT-qPCR and western blot detected METTL14, COL1A1, Runx2, ALP, and WNT3A mRNA level and protein level. Osteogenic differentiation was evaluated by Alizarin Red S staining and ALP activity. MeRIP and dual-luciferase reporter assays verified interaction between METTL14 and WNT3A. GSE223924 database showed METTL14 was differentially expressed in patients with periodontitis and peri-implantitis. Furthermore, our data verified that METTL14 and WNT3A expression were decreased in pPDLSCs and were upregulated by osteogenic induction. METTL14 promoted osteogenic differentiation of pPDLSCs. METTL14 regulated WNT3A mRNA expression via m6A methylation. METTL14 facilitates osteogenic differentiation of pPDLSCs via modulating WNT3A, providing a possible target for improving alveolar bone regeneration outcomes.Highlights 1. METTL14 expression was decreased in pPDLSCs 2. METTL14 knockdown negatively regulated the osteogenic differentiation of pPDLSCs 3. WNT3A mRNA was a m6A-methylated target by METTL14.
牙周炎是一种导致牙周支持组织丧失的慢性炎症性疾病。此外，人牙周膜干细胞（hPDLSCs）被确定为牙周和牙槽骨组织再生的候选细胞。N6-甲基腺苷（m6A）在骨质疏松症和骨代谢中发挥着至关重要的作用。然而，甲基转移酶样14（METTL14）在牙周炎患者（pPDLSCs）的牙周膜干细胞成骨分化中的作用和机制尚不清楚。在本研究中，GSE223924数据库分析了10名健康受试者、10名牙周炎患者和种植体周围炎患者的牙龈组织样本中METTL14和Wnt家族成员3A（WNT3A）的表达。RT-qPCR和蛋白质印迹法检测了METTL14、COL1A1、Runx2、ALP和WNT3A的mRNA水平和蛋白质水平。通过茜素红S染色和碱性磷酸酶（ALP）活性评估成骨分化。甲基化RNA免疫沉淀（MeRIP）和双荧光素酶报告基因检测验证了METTL14与WNT3A之间的相互作用。GSE223924数据库显示，METTL14在牙周炎和种植体周围炎患者中差异表达。此外，我们的数据证实，METTL14和WNT3A在pPDLSCs中的表达降低，并在成骨诱导后上调。METTL14促进pPDLSCs的成骨分化。METTL14通过m6A甲基化调节WNT3A mRNA表达。METTL14通过调节WNT3A促进pPDLSCs的成骨分化，为改善牙槽骨再生结果提供了一个可能的靶点。研究亮点：1. METTL14在pPDLSCs中的表达降低；2. METTL14基因敲低负向调节pPDLSCs的成骨分化；3. WNT3A mRNA是METTL14的m6A甲基化靶点。
