可溶性环氧化物水解酶抑制剂对暴露于HEMA的人牙髓干细胞细胞迁移及成牙分化的影响

doi:10.13701/j.cnki.kqyxyj.2024.02.014

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (2): 166-171.DOI: 10.13701/j.cnki.kqyxyj.2024.02.014

可溶性环氧化物水解酶抑制剂对暴露于HEMA的人牙髓干细胞细胞迁移及成牙分化的影响

罗加欣, 黄旌, 王渝灵, 冯燕, 党海霞^*

西南医科大学附属口腔医院口颌面修复重建和再生泸州市重点实验室四川泸州 646000

收稿日期:2023-09-27 出版日期:2024-02-28 发布日期:2024-02-26
通讯作者: *党海霞,E-mail:dentist198771@163.com
作者简介:罗加欣(1991~ ),女,四川乐山人,硕士在读,研究方向:牙髓再生。
基金资助:
国家自然科学基金(编号:81771032) 西南医科大学青年基金(编号:00031520) 西南医科大学口腔医院一般项目(编号:201906)

Effect of Soluble Epoxide Hydrolase Inhibitor on Migration and Odontogenic Differentiation of Human Dental Pulp Stem Cells Exposed to HEMA

LUO Jiaxin, HUANG Jing, WANG Yuling, FENG Yan, DANG Haixia^*

Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, The Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, China

Received:2023-09-27 Online:2024-02-28 Published:2024-02-26

摘要/Abstract

摘要： 目的:探讨在树脂单体甲基丙烯酸羟乙酯(2-hydroxyethyl methacrylate,HEMA)暴露环境下,可溶性环氧化物水解酶抑制剂(TPPU)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)迁移及成牙分化的影响。方法:分离提取hDPSCs,并利用茜素红染色和流式细胞仪鉴定其干性。以无添加药物的诱导培养基为对照组,以HEMA、HEMA+TPPU为实验组,利用CCK-8检测细胞活性;利用细胞划痕实验检测细胞迁移;利用碱性磷酸酶染色法进行染色,并检测碱性磷酸酶(ALP)活性;利用实时荧光定量聚合酶链反应(RT-qPCR)检测成牙分化相关基因Runt相关转录因子2(Runx2)、牙本质基质蛋白-1(DMP-1)和牙本质涎磷蛋白(DSPP)的相对表达;利用茜素红染色检测矿化结节形成情况。结果:HEMA组细胞活性较对照组显著降低(P<0.05),HEMA+TPPU组较HEMA组增加(P<0.05)。HEMA组细胞迁移率较对照组显著降低(P<0.05),HEMA+TPPU组较HEMA组增加(P<0.05)。在成牙诱导分化过程中,用2 mmol/L的HEMA处理细胞后,ALP活性较对照组降低(P<0.05),相关成牙分化基因表达也显著降低(P<0.05)。而HEMA+TPPU组ALP活性较HEMA组显著升高(P<0.05),且基因Runx2、DMP-1、DSPP表达也显著增加(P<0.05)。此外茜素红染色结果也显示HEMA+TPPU组和对照组的相对钙含量均较HEMA 组显著升高(P<0.05)。结论:HEMA可在一定程度上抑制hDPSCs的细胞迁移和成牙分化能力,而TPPU可部分逆转HEMA对细胞迁移和成牙分化能力的抑制作用。

关键词: 可溶性环氧化物水解酶抑制剂, HEMA, 牙髓干细胞, 细胞迁移, 成牙分化

Abstract: Objective: To investigate the effect of soluble epoxide hydrolase inhibitor (TPPU) on the migration and odontogenic differentiation of human dental pulp stem cells (hDPSCs) exposed to 2-hydroxyethyl methacrylate (HEMA). Methods: hDPSCs were isolated and extracted, and then identified by alizarin red staining and flow cytometry. hDPSCs without drug culture were used as the control group, and HEMA and HEMA+TPPU were used as the experimental group. hDPSCs activity was detected by CCK-8. hDPSCs migration was detected by Scratch test. ALP activity was detected and be quantified by alkaline phosphatase staining and quantitative test kit. Real-time quantitative fluorescent PCR (RT-qPCR) was used to detect the relative expression of Runx2, DMP-1, and DSPP related to odontogenic differentiation. Alizarin red staining was used to detect the formation of mineralized nodules. Results: The cell activity of HEMA group was lower than that of control group (P<0.05), and the cell activity of HEMA+TPPU group was higher than that of HEMA group (P<0.05). The cell mobility of HEMA group was significantly decreased compared with that of control group (P<0.05), and the cell mobility of HEMA+TPPU group was significantly increased compared with that of HEMA group (P<0.05). After the cells were treated with 2 mmol/L HEMA in the process of odontogenic differentiation, alkaline phosphatase activity was decreased when compared with control group (P<0.05), and the expression of odontogenic differentiation related genes was significantly decreased (P<0.05). The activity of alkaline phosphatase in the HEMA+TPPU group was significantly higher than that in HEMA group (P<0.05), and the expressions of Runx2, DMP-1, and DSPP were also significantly increased (P<0.05). In addition, alizarin red staining showed that the relative calcium contents of HEMA+TPPU group and control group were significantly higher than that of HEMA group (P<0.05). Conclusion: HEMA can inhibit the cell migration and odontogenic differentiation of hDPSCs to some extent, and TPPU can partially reverse the inhibitory effect of HEMA on the cell migration and odontogenic differentiation of hDPSCs.

Key words: soluble epoxide hydrolase inhibitor, HEMA, dental pulp stem cells, cell migration, dontogenic differentiation

罗加欣, 黄旌, 王渝灵, 冯燕, 党海霞. 可溶性环氧化物水解酶抑制剂对暴露于HEMA的人牙髓干细胞细胞迁移及成牙分化的影响[J]. 口腔医学研究, 2024, 40(2): 166-171.

LUO Jiaxin, HUANG Jing, WANG Yuling, FENG Yan, DANG Haixia. Effect of Soluble Epoxide Hydrolase Inhibitor on Migration and Odontogenic Differentiation of Human Dental Pulp Stem Cells Exposed to HEMA[J]. Journal of Oral Science Research, 2024, 40(2): 166-171.

参考文献

[1] Van Landuyt KL, Nawrot T, Geebelen B, et al. How much do resin-based dental materials release? A meta-analytical approach [J]. Dent Mater, 2011, 27(8):723-747.
[2] Chen RS, Lee MS, Hu YJ, et al. The effects of low-dose 2-hydroxyethyl methacrylate on apoptosis and survival in human dental pulp cells [J]. J Formos Med Assoc, 2021, 120(6):1332-1339.
[3] Baldion PA, Velandia-Romero ML, Castellanos JE. Dental resin monomers induce early and potent oxidative damage on human odontoblast-like cells [J]. Chem Biol Interact, 2021, 333:109336.
[4] Pagano S, Coniglio M, Valenti C, et al. Biological effects of resin monomers on oral cell populations: descriptive analysis of literature [J]. Eur J Paediatr Dent, 2019, 20(3):224-232.
[5] Wuersching SN, Högg C, Kohl L, et al. Leaching components and initial biocompatibility of novel bioactive restorative materials [J]. Dent Mater, 2023, 39(3):293-304.
[6] Williams DW, Wu H, Oh JE, et al. 2-Hydroxyethyl methacrylate inhibits migration of dental pulp stem cells [J]. J Endod, 2013, 39(9):1156-1160.
[7] Kwon JH, Park HC, Zhu T, et al. Inhibition of odontogenic differentiation of human dental pulp cells by dental resin monomers [J]. Biomater Res, 2015, 19:8.
[8] Diomede F, Tripodi D, Trubiani O, et al. HEMA effects on autophagy mechanism in human dental pulp stem cells [J]. Materials (Basel), 2019, 12(14):2285.
[9] Rand AA, Barnych B, Morisseau C, et al. Cyclooxygenase-derived proangiogenic metabolites of epoxyeicosatrienoic acids [J]. Proc Natl Acad Sci U S A, 2017, 114(17):4370-4375.
[10] Napimoga MH, Rocha EP, Trindade-da-Silva CA, et al. Soluble epoxide hydrolase inhibitor promotes immunomodulation to inhibit bone resorption [J]. J Periodontal Res, 2018, 53(5): 743-749.
[11] Alizadehgharib S, Östberg AK, Dahlgren U. Effects of the methacrylate/acrylate monomers HEMA, TEGDMA, DEGDA, and EMA on the immune system [J]. Clin Exp Dent Res, 2017, 3(6):227-234.
[12] Diomede F, Marconi GD, Guarnieri S, et al. A novel role of ascorbic acid in anti-inflammatory pathway and ROS generation in HEMA treated dental pulp stem cells [J]. Materials (Basel), 2019, 13(1):130.
[13] Liu T, Li T, Chen X, et al. EETs/sEHi alleviates nociception by blocking the crosslink between endoplasmic reticulum stress and neuroinflammation in a central poststroke pain model [J]. J Neuroinflammation, 2021, 18(1):211.
[14] Lee SM, Kim SY, Kim JH, et al. Depth-dependent cellular response from dental bulk-fill resins in human dental pulp stem cells [J]. Stem Cells Int, 2019, 2019: 1251536.
[15] Alvarez ML, Lorenzetti F. Role of eicosanoids in liver repair, regeneration and cancer [J]. Biochem Pharmacol, 2021, 192:114732.
[16] 党海霞,霍欣茹,王福,等.14,15-环氧二十碳三烯酸促进颅骨缺损骨再生[J].口腔医学研究,2016,32(7):692-695.
[17] Dang H, Chen W, Chen L, et al. TPPU inhibits infammation-induced excessive autophagy to restore the osteogenic diferentiation potential of stem cells and improves alveolar ridge preservation [J]. Sci Rep, 2023, 13(1):1574.
[18] Williams DW, Wu H, Oh JE, et al. HEMA inhibits migration of dental pulp stem cells [J]. J Endod, 2013, 39(9):1156-1160.
[19] 孙琴,吕迪,陈美玲.HEMA 对人乳牙牙髓干细胞分化潜能的影响[J].临床口腔医学杂志,2023,39(5):279-282.

可溶性环氧化物水解酶抑制剂对暴露于HEMA的人牙髓干细胞细胞迁移及成牙分化的影响

Effect of Soluble Epoxide Hydrolase Inhibitor on Migration and Odontogenic Differentiation of Human Dental Pulp Stem Cells Exposed to HEMA

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	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.
[2]	贺桢茹, 郑雪晴, 杨春晖, 汪鑫, 李阳, 宋亚玲. Fam83h突变对小鼠成骨细胞增殖和迁移的影响[J]. 口腔医学研究, 2023, 39(5): 435-439.
[3]	谢元栋, 李泽华, 刘晓莉, 胡玲, 詹骆宁, 李毅. 牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究[J]. 口腔医学研究, 2022, 38(8): 736-741.
[4]	潘露, 梁燕. 乳牙牙髓干细胞在口腔颌面疾病治疗中的应用[J]. 口腔医学研究, 2022, 38(7): 605-608.
[5]	曹威, 刘梦佳, 张清彬. SIRT 1通过下调PPAR-γ受体提升人乳牙牙髓干细胞骨向分化效能[J]. 口腔医学研究, 2022, 38(2): 169-175.
[6]	黄梦妤, 刘生波. 牙髓再生技术应用于成熟恒牙的挑战及应对策略[J]. 口腔医学研究, 2022, 38(10): 922-925.
[7]	王凯丽, 张巍巍, 李艳萍, 何丽娜, 张爽, 潘爽, 牛玉梅. 沉默整合素α6通过激活FOXO1信号通路促进人牙髓干细胞的成牙本质向分化[J]. 口腔医学研究, 2022, 38(10): 930-935.
[8]	牛玉梅, 孙翔宇. 牙髓干细胞对面神经损伤修复研究进展[J]. 口腔医学研究, 2021, 37(3): 185-190.
[9]	谢惠兰, 方芳, 林毅. Chir99021调控Wnt/β-catenin信号通路干预干细胞成骨/成牙分化的研究[J]. 口腔医学研究, 2021, 37(12): 1150-1155.
[10]	关孟莹, 何丽娜, 潘爽, 李艳萍, 刘会梅, 牛玉梅. 经典Wnt信号通路对hDPSCs增殖、迁移和成牙本质向分化的影响[J]. 口腔医学研究, 2020, 36(9): 861-865.
[11]	马兰, 禹怡君, 刘含笑, 刘超, 苗雷英. 黄连素促人牙髓干细胞成骨/成牙本质细胞向分化[J]. 口腔医学研究, 2020, 36(7): 639-643.
[12]	王薛藤, 聂敏海, 左东川, 邓浩, 曾锦. TRPM4通道对人口腔颊黏膜成纤维细胞增殖和迁移的影响[J]. 口腔医学研究, 2020, 36(5): 490-495.
[13]	殷晓薇, 张爽, 邓皓天, 何丽娜, 李艳萍, 张巍巍, 潘爽, 牛玉梅. 整合素α6对hDPSCs增殖和成牙本质向分化的影响[J]. 口腔医学研究, 2020, 36(12): 1132-1136.
[14]	张维甲, 潘爽, 李艳萍, 何丽娜, 吴作栋, 牛玉梅. 兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究[J]. 口腔医学研究, 2020, 36(10): 968-972.
[15]	闫娜, 黄涛, 张中月, 张淑华, 王丽华, 韩国良. 富血小板纤维蛋白对牙髓干细胞增殖和成骨分化的功能研究[J]. 口腔医学研究, 2020, 36(10): 973-977.