超氧化物歧化酶2清除ROS,通过调节牙槽骨缺损大鼠的Smad3促进人牙周膜干细胞的成骨分化。

IF 4.2 2区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE
Wei Qiu, Qian Sun, Na Li, Zehao Chen, Hongle Wu, Zhao Chen, Xiaolan Guo, Fuchun Fang
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引用次数: 0

摘要

背景:人牙周膜干细胞(hPDLSCs)的成骨分化是牙槽骨再生中的一个重要事件。氧化应激可能是hPDLSC成骨的主要抑制因子。超氧化物歧化酶2(SOD2)是一种关键的抗氧化酶,但其对hPDLSC成骨分化的影响尚不清楚。方法:用流式细胞仪检测几种表面标志物,并用碱性磷酸酶(ALP)、茜素红S和油红O染色验证hPDLSCs的分化潜力。通过实时定量聚合酶链反应(RT-qPCR)、蛋白质印迹和ALP染色检测hPDLSCs的成骨指标。此外,通过显微CT、苏木精和伊红以及Masson染色对大鼠牙槽骨缺损模型进行分析。通过ROS测定试剂盒评估细胞内活性氧(ROS)水平。最后,通过RT-qPCR和蛋白质印迹(WB)检测hPDLSCs中SOD2、Smad3和p-Smad3的表达。结果:SOD2对ALP、BMP6、BMP5的基因和蛋白表达具有正调控作用,和hPDLSCs中的RUNX2(p2 O2下调hPDLSCs中的成骨指标(ALP、BMP6、RUNX2)(p结论:SOD2在体内外均对hPDLSC的成骨分化具有正向调控作用。从机制上讲,SOD2通过调节Smad3的磷酸化清除ROS来促进hPDLSC的成骨细胞分化。本研究为治疗牙槽骨再生提供了理论依据。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Superoxide dismutase 2 scavenges ROS to promote osteogenic differentiation of human periodontal ligament stem cells by regulating Smad3 in alveolar bone-defective rats

Background

Osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) is an essential event in alveolar bone regeneration. Oxidative stress may be the main inhibiting factor of hPDLSC osteogenesis. Superoxide dismutase 2 (SOD2) is a key antioxidant enzyme, but its effect on hPDLSC osteogenic differentiation is unclear.

Methods

Several surface markers were detected by flow cytometry, and the differentiation potential of hPDLSCs was validated by alkaline phosphatase (ALP), Alizarin Red S, and Oil Red O staining. Osteogenic indicators of hPDLSCs were detected by real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and ALP staining. Furthermore, alveolar bone defect rat models were analyzed through micro-CT, hematoxylin and eosin, and Masson staining. The intracellular reactive oxygen species (ROS) level was evaluated by a ROS assay kit. Finally, the expression of SOD2, Smad3, and p-Smad3 in hPDLSCs was detected by RT-qPCR and Western blotting (WB).

Results

SOD2 positively regulated the gene and protein expressions of ALP, BMP6, and RUNX2 in hPDLSCs (p < 0.05). Ideal bone formation and continuous cortical bone were obtained by transplanting LV-SOD2 hPDLSCs (lentivirus vector for overexpressing SOD2 in hPDLSCs) in vivo. Exogenous H2O2 downregulated osteogenic indicators (ALP, BMP6, RUNX2) in hPDLSCs (p < 0.05); this was reversed by overexpression of SOD2. WB results showed that the Smad3 and p-Smad3 signaling pathways participated in the osteogenic process of SOD2 in hPDLSCs.

Conclusion

SOD2 positively regulated hPDLSC osteogenic differentiation in vitro and in vivo. Mechanistically, SOD2 promotes hPDLSC osteogenic differentiation by regulating the phosphorylation of Smad3 to scavenge ROS. This work provides a theoretical basis for the treatment of alveolar bone regeneration.

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来源期刊
Journal of periodontology
Journal of periodontology 医学-牙科与口腔外科
CiteScore
9.10
自引率
7.00%
发文量
290
审稿时长
3-8 weeks
期刊介绍: The Journal of Periodontology publishes articles relevant to the science and practice of periodontics and related areas.
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