Gu Ke, Feng Xiao-Mei, Sun Shao-Qing, Hao Xing-Yao, Wen Yong
Department of Implantology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, No. 44-1 Wenhua Road West, Jinan 250012, Shandong Province, China.
Stomatological Hospital, School of Medicine, Nankai University, Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction, Tianjin 300041, China.
World J Stem Cells. 2024 Nov 26;16(11):926-943. doi: 10.4252/wjsc.v16.i11.926.
Human periodontal ligament stem cells (PDLSCs) regenerate oral tissue. expansion causes replicative senescence in stem cells. This causes intracellular reactive oxygen species (ROS) accumulation, which can impair stem cell function. Tissue engineering efficiency is reduced by exogenous ROS stimulation, which causes premature senescence under oxidative stress. Melatonin (MT), a powerful free radical scavenger, can delay PDLSCs senescence but may not maintain stemness under oxidative stress. This experiment examined the effects of hydrogen peroxide-induced oxidative stress on PDLSCs' apoptosis, senescence, and stemness.
To determine if MT can reverse the above effects along with the underlying molecular mechanisms involved.
PDLSCs were isolated from human premolars and cultured in different conditions. Flow cytometry was used to characterize the cell surface markers of PDLSCs. Hydrogen peroxide was used to induce oxidative stress in PDLSCs. Cell cycle, proliferation, apoptosis, differentiation, ROS, and senescence-associated β-galactosidase activity were assessed by various assays. Reverse transcription-polymerase chain reaction and western blot were used to measure the expression of genes and proteins related to stemness and senescence.
MT increases Yes-associated protein expression and maintains cell stemness in an induced inflammatory microenvironment, which may explain its therapeutic effects. We examined how MT affects PDLSCs aging and stemness and its biological mechanisms.
Our study reveals MT's role in regulating oxidative stress in PDLSCs and Yes-associated protein-mediated activity, providing insights into cellular functions and new therapeutic targets for tissue regeneration.
人牙周膜干细胞(PDLSCs)可使口腔组织再生。细胞扩增会导致干细胞发生复制性衰老。这会引起细胞内活性氧(ROS)积累,进而损害干细胞功能。外源性ROS刺激会降低组织工程效率,导致在氧化应激下过早衰老。褪黑素(MT)是一种强大的自由基清除剂,可延缓PDLSCs衰老,但在氧化应激下可能无法维持干细胞特性。本实验研究了过氧化氢诱导的氧化应激对PDLSCs凋亡、衰老和干细胞特性的影响。
确定MT是否能逆转上述影响以及相关的潜在分子机制。
从人前磨牙中分离出PDLSCs,并在不同条件下培养。采用流式细胞术对PDLSCs的细胞表面标志物进行表征。用过氧化氢诱导PDLSCs发生氧化应激。通过各种检测方法评估细胞周期、增殖、凋亡、分化、ROS和衰老相关β-半乳糖苷酶活性。采用逆转录-聚合酶链反应和蛋白质免疫印迹法检测与干细胞特性和衰老相关的基因和蛋白质表达。
MT可增加Yes相关蛋白表达,并在诱导的炎症微环境中维持细胞干细胞特性,这可能解释了其治疗作用。我们研究了MT如何影响PDLSCs衰老和干细胞特性及其生物学机制。
我们的研究揭示了MT在调节PDLSCs氧化应激和Yes相关蛋白介导的活性中的作用,为细胞功能和组织再生的新治疗靶点提供了见解。