Zhang Xu, Hu Wen, Hua Hua
Department of Emergency and General Dentistry, Changsha Stomatological Hospital, No. 389 Youyi Road, Tianxin District, Changsha, 41000, Hunan Province, People's Republic of China.
Hum Cell. 2025 Jul 15;38(5):130. doi: 10.1007/s13577-025-01262-3.
Periodontal ligament stem cells (PDLSCs) constitute a promising source for successful periodontal regeneration. This study aims to explore roles of metformin, krüppel-like factor 2 (KLF2), and miR-181a-5p in mediating osteogenic differentiation of human PDLSCs (hPDLSCs) following lipopolysaccharide (LPS) stimulation. The osteogenic differentiation potential of hPDLSCs isolated from human premolar root samples were examined by alkaline phosphatase (ALP) staining, ALP activity assay, Alizarin red S staining, and Western blotting of osteogenic markers. Metformin pretreatment at dose of 100 μM significantly resulted in increased ALP activity, elevated protein expressions of osteogenic markers, and more generated mineralized matrix in hPDLSCs with LPS stimulation. KLF2 and miR-181a-5p were found to be increased by metformin pretreatment at dose of 100 μM in hPDLSCs with stimulation but not in hPDLSCs without LPS stimulation. The interaction between the KLF2 and the promoter of miR-181a-5p was noted by the dual-luciferase reporter assay. KLF2 knockdown or miR-181a-5p inhibition notably abrogated the improvements of osteogenic differentiation by metformin pretreatment in LPS-stimulated hPDLSCs. The findings of the study indicate metformin protects hPDLSCs against impaired osteogenic differentiation of hPDLSCs after LPS stimulation by KLF2-mediated activation of miR-181a-5p under inflammation conditions.
牙周膜干细胞（PDLSCs）是牙周成功再生的一个有前景的细胞来源。本研究旨在探讨二甲双胍、Krüppel样因子2（KLF2）和miR-181a-5p在介导脂多糖（LPS）刺激后人牙周膜干细胞（hPDLSCs）成骨分化中的作用。通过碱性磷酸酶（ALP）染色、ALP活性测定、茜素红S染色和成骨标志物的蛋白质印迹法检测从人前磨牙牙根样本中分离的hPDLSCs的成骨分化潜能。100μM剂量的二甲双胍预处理显著提高了LPS刺激的hPDLSCs的ALP活性，增加了成骨标志物的蛋白表达，并产生了更多矿化基质。发现100μM剂量的二甲双胍预处理可使受刺激的hPDLSCs中KLF2和miR-181a-5p增加，但未刺激的hPDLSCs中则无此现象。双荧光素酶报告基因检测显示KLF2与miR-181a-5p启动子之间存在相互作用。在LPS刺激的hPDLSCs中，敲低KLF2或抑制miR-181a-5p可显著消除二甲双胍预处理对成骨分化的改善作用。该研究结果表明，在炎症条件下，二甲双胍通过KLF2介导的miR-181a-5p激活保护hPDLSCs免受LPS刺激后hPDLSCs成骨分化受损的影响。
