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Abstract：
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：人牙周膜干细胞（hPDLSCs）是牙周组织工程的重要候选种子细胞，但是牙周组织中脂多糖（LPS）的存在抑制了hPDLSCs的自我更新和成骨分化。我们以前的研究表明，TAZ是hPDLSCs成骨分化的正调节因子，但TAZ是否能保护hPDLSCs免受LPS的侵害尚不清楚。本研究旨在探讨TAZ对LPS诱导的炎症模型中hPDLSCs成骨分化的调控作用。初步揭示与NF-κB信号通路相关的分子机制。
方法:将LPS添加到hPDLSCs的培养基中。通过CCK-8测定分析LPS对hPDLSC增殖的影响。用茜素红染色检测LPS对hPDLSC成骨分化的影响,ALP染色,成骨相关基因的WesternBlot和qRT-PCR分析。分析了LPS对TAZ过表达或通过慢病毒敲低的hPDLSCs成骨分化的影响。通过蛋白质印迹和免疫荧光分析hPDLSCs中的NF-κB信号传导。
结果：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对成骨分化的负面影响，并抑制了LPS对NF-κB通路的激活。TAZ敲除增强了LPS对成骨的抑制作用。
结论：过表达TAZ可以部分逆转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标。
方法:将LPS添加到hPDLSCs的培养基中。通过CCK-8测定分析LPS对hPDLSC增殖的影响。用茜素红染色检测LPS对hPDLSC成骨分化的影响,ALP染色,成骨相关基因的WesternBlot和qRT-PCR分析。分析了LPS对TAZ过表达或通过慢病毒敲低的hPDLSCs成骨分化的影响。通过蛋白质印迹和免疫荧光分析hPDLSCs中的NF-κB信号传导。
结果：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对成骨分化的负面影响，并抑制了LPS对NF-κB通路的激活。TAZ敲除增强了LPS对成骨的抑制作用。
结论：过表达TAZ可以部分逆转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标。
