大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响

doi:10.13701/j.cnki.kqyxyj.2023.06.013

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (6): 534-540.DOI: 10.13701/j.cnki.kqyxyj.2023.06.013

大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响

姜震, 颜燕宏, 韩雪, 蒋备战^*

同济大学口腔医学院·同济大学附属口腔医院儿童口腔科,上海牙组织修复与再生工程技术研究中心上海 200072

收稿日期:2022-12-01 出版日期:2023-06-28 发布日期:2023-06-21
通讯作者: *蒋备战,E-mail:jiangbeizhan@tongji.edu.cn
作者简介:姜震(1996~ ),男,浙江宁波人,硕士在读,主要从事儿童口腔医学研究。
基金资助:
国家自然科学基金(编号:82000995);上海市卫生健康委员会科研项目(编号:202040095)上海申康医院发展中心新兴前沿技术联合攻关项目(编号:SHDC12023115)

Effects of Escherichia Coli Lipopolysaccharide on Biological Behaviors of Rat Dental Follicle Stem Cells

JIANG Zhen, YAN Yanhong, HAN Xue, JIANG Beizhan^*

Department of Pediatrics, School and Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China

Received:2022-12-01 Online:2023-06-28 Published:2023-06-21

摘要/Abstract

摘要： 目的:探究大肠杆菌脂多糖(E.coli LPS)对大鼠牙囊干细胞(rDFSCs)生物学行为的影响。方法:体外培养鉴定rDFSCs,使用不同浓度(0、1、10 μg/mL) E.coli LPS处理细胞,通过qRT-PCR检测炎症因子的表达;CCK-8法检测细胞增殖情况;划痕实验和Transwell实验检测细胞迁移能力;碱性磷酸酶(ALP)染色和茜素红染色分别评估rDFSCs的矿化情况,并利用qRT-PCR检测其成骨相关基因表达水平。结果:相比于对照组,LPS组IL-6、IL-1β及TNF-α的mRNA表达水平升高(P＜0.05);1 μg/mL LPS促进rDFSCs增殖及迁移能力(P＜0.05),而10 μg/mL LPS对其增殖、迁移影响不明显;LPS处理后ALP染色降低、矿化结节数量减少,且成骨标志物RUNX2、COL1及OPN表达下降(P＜0.05)。结论:E.coli LPS模拟的炎症微环境下,rDFSCs成骨分化能力受到明显抑制,低浓度(1 μg/mL) LPS刺激可促进rDFSCs的增殖和迁移。

关键词: 脂多糖, 牙囊干细胞, 细胞增殖, 细胞迁移, 成骨分化

Abstract: Objective: To investigate the effects of Escherichia coli lipopolysaccharide (E. coli LPS) on biological behaviors of rat dental follicle stem cells (rDFSCs). Methods: rDFSCs were cultured and identified, and then were treated with different concentrations of E. coli LPS (0, 1, 10 μg/mL). The mRNA levels of pro-inflammatory genes were measured by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR). The effects of LPS on proliferation and migration ability of rDFSCs were assessed by CCK-8 and scratch test plus transwell assay, respectively. The osteogenic capability of rDFSCs was detected by alkaline phosphatase (ALP) staining and alizarin red staining, and their osteogenic-related gene mRNA levels were measured by qRT-PCR. Results: The gene expressions of IL-6, IL-1 β, and TNF-α were significantly upregulated under 1 or 10 μg/mL LPS treatment (P＜0.05). LPS at 1 μg/mL significantly promoted the proliferation and migration of rDFSCs (P＜0.05), while a higher concentration (10 μg/mL) did not show any improvement. ALP staining intensity and the number of mineralized nodules decreased, and the expressions of osteogenic markers RUNX2, COL1, and OPN were inhibited in LPS-treated group (P＜0.05).Conclusion: The osteogenic ability of rDFSCs was significantly inhibited under inflammatory microenvironment simulated by E.coli LPS, and LPS promoted the proliferation and migration of rDFSCs at low concentration.

Key words: lipopolysaccharide, dental follicle stem cells, cell proliferation, cell migration, osteogenic differentiation

姜震, 颜燕宏, 韩雪, 蒋备战. 大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响[J]. 口腔医学研究, 2023, 39(6): 534-540.

JIANG Zhen, YAN Yanhong, HAN Xue, JIANG Beizhan. Effects of Escherichia Coli Lipopolysaccharide on Biological Behaviors of Rat Dental Follicle Stem Cells[J]. Journal of Oral Science Research, 2023, 39(6): 534-540.

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大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响

Effects of Escherichia Coli Lipopolysaccharide on Biological Behaviors of Rat Dental Follicle Stem Cells

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