Department of Periodontics, Stomatological Hospital, General Hospital of Ningxia Medical University, Yinchuan 750003, Ningxia, China.
Medical Laboratory, General Hospital of Ningxia Medical University, Yinchuan 750003, Ningxia, China.
J Steroid Biochem Mol Biol. 2023 Sep;232:106347. doi: 10.1016/j.jsbmb.2023.106347. Epub 2023 Jun 17.
Periodontitis is a chronic inflammatory disease caused by Porphyromonas gingivalis and other bacteria, and human periodontal ligament stem cells (hPDLSCs) are a promising candidate for the treatment of periodontal supporting tissue defects. This study aimed to investigate the effect of 1α,25-dihydroxyvitamin D [1,25(OH)VitD] on osteogenic differentiation of hPDLSCs in an in vitro periodontitis model and whether it can improve inflammatory status. hPDLSCs were in vitro isolated and identified. After treatment with 1,25(OH)VitD and ultrapure pure Porphyromonas gingivalis lipopolysaccharide (LPS-G), the viability of hPDLSCs was detected using Cell Counting Kit-8, the expressions of osteogenic markers and inflammatory genes using Western blotting and quantitative reverse transcription PCR (qRT-PCR), the levels of inflammatory factors in cells using enzyme linked immunosorbent assay (ELISA), and the fluorescence signal intensity of osteoblastic markers and inflammatory genes in cells using immunofluorescence assay. It was found that 1,25(OH)VitD reversed the inhibition of hPDLSCs proliferation by LPS-G; LPS-G exhibited inhibitory effect on ALP, Runx2, and OPN expressions, and such inhibitory effect was significantly weakened when co-acting with 1,25(OH)VitD. Meanwhile, LPS-G upregulated the expressions of inflammatory genes IL-1β and Casp1, whereas 1,25(OH)VitD antagonized such an effect and improved the inflammatory status. In conclusion, 1,25(OH)VitD can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation and suppress LPS-G-induced upregulation of inflammatory gene expressions.
牙周炎是由牙龈卟啉单胞菌和其他细菌引起的慢性炎症性疾病，人牙周膜干细胞（hPDLSCs）是治疗牙周支持组织缺损的有前途的候选物。本研究旨在探讨 1α,25-二羟维生素 D [1,25(OH)VitD] 在体外牙周炎模型中对 hPDLSCs 成骨分化的影响，以及是否能改善炎症状态。体外分离和鉴定 hPDLSCs。用 1,25(OH)VitD 和超纯纯牙龈卟啉单胞菌脂多糖（LPS-G）处理 hPDLSCs 后，用细胞计数试剂盒-8 检测 hPDLSCs 的活力，用 Western blot 和定量逆转录 PCR（qRT-PCR）检测成骨标志物和炎症基因的表达，用酶联免疫吸附测定法（ELISA）检测细胞内炎症因子的水平，并用免疫荧光法检测细胞内成骨标志物和炎症基因的荧光信号强度。结果发现，1,25(OH)VitD 逆转了 LPS-G 对 hPDLSCs 增殖的抑制作用；LPS-G 对 ALP、Runx2 和 OPN 的表达有抑制作用，而与 1,25(OH)VitD 共同作用时，这种抑制作用明显减弱。同时，LPS-G 上调了炎症基因 IL-1β 和 Casp1 的表达，而 1,25(OH)VitD 拮抗了这种作用并改善了炎症状态。综上所述，1,25(OH)VitD 可逆转 LPS-G 对 hPDLSCs 增殖和成骨分化的抑制作用，并抑制 LPS-G 诱导的炎症基因表达上调。
