School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration, Jinan, Shandong, China.
BMC Oral Health. 2024 Jun 26;24(1):733. doi: 10.1186/s12903-024-04497-y.
Human periodontal ligament stem cells (hPDLSCs) are important candidate seed cells for periodontal tissue engineering, but the presence of lipopolysaccharide(LPS) in periodontal tissues inhibits the self-renewal and osteogenic differentiation of hPDLSCs. Our previous studies demonstrated that TAZ is a positive regulator of osteogenic differentiation of hPDLSCs, but whether TAZ can protect hPDLSCs from LPS is still unknown. The present study aimed to explore the regulatory effect of TAZ on the osteogenic differentiation of hPDLSCs in an LPS-induced inflammatory model, and to preliminarily reveal the molecular mechanisms related to the NF-κB signaling pathway.
LPS was added to the culture medium of hPDLSCs. The influence of LPS on hPDLSC proliferation was analyzed by CCK-8 assays. The effects of LPS on hPDLSC osteogenic differentiation were detected by Alizarin Red staining, ALP staining, Western Blot and qRT-PCR analysis of osteogenesis-related genes. The effects of LPS on the osteogenic differentiation of hPDLSCs with TAZ overexpressed or knocked down via lentivirus were analyzed. NF-κB signaling in hPDLSCs was analyzed by Western Blot and immunofluorescence.
LPS inhibited the osteogenic differentiation of hPDLSCs, inhibited TAZ expression, and activated the NF-κB signaling pathway. Overexpressing TAZ in hPDLSCs partly reversed the negative effects of LPS on osteogenic differentiation and inhibited the activation of the NF-κB pathway by LPS. TAZ knockdown enhanced the inhibitory effects of LPS on osteogenesis.
Overexpressing TAZ could partly reverse the inhibitory effects of LPS on the osteogenic differentiation of hPDLSCs, possibly through inhibiting the NF-κB signaling pathway. TAZ is a potential target for improving hPDLSC-based periodontal tissue regeneration in inflammatory environments.
人牙周膜干细胞（hPDLSCs）是牙周组织工程的重要候选种子细胞，但牙周组织中脂多糖（LPS）的存在抑制了 hPDLSCs 的自我更新和成骨分化。我们之前的研究表明，TAZ 是 hPDLSCs 成骨分化的正向调节因子，但 TAZ 是否能保护 hPDLSCs 免受 LPS 的影响尚不清楚。本研究旨在探讨 TAZ 在 LPS 诱导的炎症模型中对 hPDLSCs 成骨分化的调节作用，并初步揭示与 NF-κB 信号通路相关的分子机制。
在 hPDLSCs 的培养基中加入 LPS。通过 CCK-8 检测 LPS 对 hPDLSC 增殖的影响。通过茜素红染色、ALP 染色、Western Blot 和 qRT-PCR 分析成骨相关基因检测 LPS 对 hPDLSC 成骨分化的影响。通过慢病毒过表达或敲低 hPDLSCs 中的 TAZ 分析 LPS 对 hPDLSCs 成骨分化的影响。通过 Western Blot 和免疫荧光分析 hPDLSCs 中的 NF-κB 信号通路。
LPS 抑制 hPDLSCs 的成骨分化，抑制 TAZ 表达，并激活 NF-κB 信号通路。在 hPDLSCs 中过表达 TAZ 部分逆转了 LPS 对成骨分化的负向作用，并抑制了 LPS 激活的 NF-κB 途径。TAZ 敲低增强了 LPS 对成骨的抑制作用。
过表达 TAZ 可部分逆转 LPS 对 hPDLSCs 成骨分化的抑制作用，可能通过抑制 NF-κB 信号通路。TAZ 是改善炎症环境中基于 hPDLSC 的牙周组织再生的潜在靶点。
