氧浓度调节牙髓细胞中HDAC1介导的成骨信号通路调控。

Oxygen concentration modulates HDAC1-Mediated regulation of osteogenic signaling pathways in dental pulp cells.

作者信息

Song Ci, Li Ping, Lin Lin, Cao Ge, Liu Zhao, Liu Fei, Peng Ling, Dai Jingxing, Wu Buling, Chen Ting

机构信息

Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China.

College of Stomatology, Southern Medical University, Guangzhou, China.

出版信息

Front Cell Dev Biol. 2025 Sep 17;13:1627763. doi: 10.3389/fcell.2025.1627763. eCollection 2025.

DOI:10.3389/fcell.2025.1627763

PMID:41041663

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12485506/

Abstract

BACKGROUND

Dental pulp regeneration represents a critical frontier in translational dentistry, with dental pulp stem cells (DPSCs) demonstrating exceptional reparative potential through their multipotent differentiation capacity. While oxygen tension is known to influence cellular physiology, its regulatory mechanisms on DPSC osteo/odontogenic differentiation remain poorly understood.

METHODS

We established physiologically relevant oxygen gradients (3%, 5%, 21% O) to mimic developmental and pathological pulp microenvironments. Cellular proliferation and osteogenic capacity were assessed through flow cytometry, CCK-8 assays, and Live/Dead staining. Differentiation markers (RUNX2, OCN, ALP, DSPP) were quantified via qRT-PCR, immunoblotting, and enzymatic activity assays. Pharmacological inhibition studies using Oltipraz (HIF-1α inhibitor) and Valproic acid (HDAC inhibitor) elucidated pathway interactions. Publicly available transcriptomic datasets were analyzed to identify hypoxia-regulated pathways, and protein interactions were predicted using bioinformatics tools.

RESULTS

Moderate hypoxia (5% O) significantly enhanced DPSC proliferation (p < 0.05 vs. normoxia) and upregulated osteogenic markers at transcriptional (1.8-3.2 fold) and translational levels. Severe hypoxia (3% O) suppressed both proliferation (p < 0.01) and differentiation markers (0.4-0.7 fold). HIF-1α inhibition reversed 5% O-mediated osteogenic enhancement (p < 0.01), while HDAC1 blockade with Valproic acid rescued differentiation capacity under 3% O (1.5-2.1 fold induction). Mechanistically, HDAC1 appeared to influence HIF-1α protein levels in an oxygen-dependent manner, and its inhibition affected pathways consistent with alterations in chromatin remodeling, influencing VEGFA-mediated osteogenic signaling.

CONCLUSION

Our findings establish an oxygen-sensitive HDAC/HIF-1α regulatory axis governing DPSC fate determination. The biphasic response to hypoxia gradients suggests microenvironmental optimization strategies could enhance pulp regenerative outcomes. These insights provide mechanistic foundations for developing HDAC-targeted approaches in endodontic regeneration.

摘要

背景

牙髓再生是转化牙科领域的一个关键前沿领域，牙髓干细胞（DPSC）通过其多能分化能力展现出卓越的修复潜力。虽然已知氧张力会影响细胞生理，但它对DPSC成骨/牙源性分化的调控机制仍知之甚少。

方法

我们建立了生理相关的氧梯度（3%、5%、21% O₂）以模拟发育和病理牙髓微环境。通过流式细胞术、CCK-8 测定和活/死染色评估细胞增殖和成骨能力。通过qRT-PCR、免疫印迹和酶活性测定对分化标志物（RUNX2、OCN、ALP、DSPP）进行定量。使用奥替普拉（HIF-1α抑制剂）和丙戊酸（HDAC抑制剂）的药理抑制研究阐明了信号通路相互作用。分析公开可用的转录组数据集以鉴定缺氧调节的信号通路，并使用生物信息学工具预测蛋白质相互作用。

结果

中度缺氧（5% O₂）显著增强了DPSC增殖（与常氧相比，p < 0.05），并在转录水平（1.8 - 3.2倍）和翻译水平上调了成骨标志物。严重缺氧（3% O₂）抑制了增殖（p < 0.01）和分化标志物（0.4 - 0.7倍）。HIF-1α抑制逆转了5% O₂介导的成骨增强（p < 0.01），而用丙戊酸阻断HDAC1挽救了3% O₂条件下的分化能力（诱导1.5 - 2.1倍）。机制上，HDAC1似乎以氧依赖的方式影响HIF-1α蛋白水平，其抑制作用影响了与染色质重塑改变一致的信号通路，影响了VEGFA介导的成骨信号传导。