MTA、iRoot SP和AH Plus 对人牙周膜干细胞增殖及分化的影响

doi:10.19439/j.sjos.2020.05.001

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (5): 449-455.doi: 10.19439/j.sjos.2020.05.001

MTA、iRoot SP和AH Plus 对人牙周膜干细胞增殖及分化的影响

郑晶晶^1,*, 康馨允^2,*, 李少明³, 刘嘉成³, 孙凯³, 高岭³, 侯铁舟²

1.青岛大学附属医院牙体牙髓病科,山东青岛 266555;
2.西安交通大学附属口腔医院牙体牙髓病科,陕西西安 710000;
3.青岛大学附属医院口腔颌面外科,山东青岛 266555

收稿日期:2018-12-24 修回日期:2019-04-23 出版日期:2020-10-25 发布日期:2020-11-02
通讯作者: 侯铁舟,E-mail：tiezhou@mail.xjtu.edu.cn
作者简介:郑晶晶（1984-）,女,博士,主治医师,E-mail：zhengjingjing.1984@163.com;康馨允（1991-）,女,硕士,住院医师,E-mail：1286324702@qq.com。^*并列第一作者
基金资助:
国家自然科学基金 (81600880); 青岛市源头创新计划(应用研究专项-青年专项)(17-1-1-45-jch)

Effects of MTA, iRoot SP and AH Plus on proliferation and differentiation of human periodontal ligament stem cells

ZHENG Jing-jing¹, KANG Xin-yun², LI Shao-ming³, LIU Jia-cheng³, SUN Kai³, GAO Ling³, HOU Tie-zhou²

1. Department of Endodontics, Affiliated Hospital of Qingdao University. Qingdao 266555, Shandong Province;
2. Department of Endodontics, Stomatological Hospital, College of Medicine, Xi'an Jiaotong University. Xi'an 710000, Shaanxi Province;
3. Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Qingdao University. Qingdao 266555, Shandong Province, China

Received:2018-12-24 Revised:2019-04-23 Online:2020-10-25 Published:2020-11-02

摘要/Abstract

摘要： 目的：利用分子生物学方法检测MTA、iRoot SP和AH Plus对人牙周膜干细增殖及成骨的影响。方法：利用单克隆方法培养牙周膜干细胞,MTT检测和Annexin V-FITC/PI双染分别检测MTA、iRootSP和AH Plus 对人牙周膜干细胞增殖和凋亡的影响,茜素红染色、qRT-PCR检测MTA、iRoot SP和AH Plus对牙周膜干细胞成骨能力的影响。采用SPSS 21.0 软件包对数据进行统计学分析。结果：MTA在24 h和48 h呈轻度毒性,72 h无毒性; AH Plus在24 h呈轻度毒性,48 h和72 h无毒性;3 d内iRoot SP相对于MTA和AH Plus对细胞的增殖活性影响最小,几乎无毒性;MTA组和iRoot SP组矿化结节显著多于AH Plus组和对照组。2组在7、14、21 d时OC、RUNX2、COL1A、ALP基因表达均显著高于对照组,且iRoot SP组矿化基因表达最高（P<0.05）。结论：在体外,硬固后的iRoot SP对人牙周膜干细胞几乎无毒性。硬固后的iRoot SP和MTA均有一定的骨诱导能力,但在相同时间内,iRoot SP的作用更明显。

关键词: 牙周膜干细胞, iRoot SP, 细胞毒性, 成骨, 矿化

Abstract: PURPOSE: To explore the effects of MTA, iRoot SP and AH Plus on periodontal ligament stem cells. METHODS: The periodontal ligament stem cells were cloned by limiting dilution culture method. The effects of MTA, iRoot SP and AH Plus on proliferation and apoptosis of periodontal ligament stem cells were detected by MTT and Annexin-V-FITC/PI double staining. Alizarin and qRT-PCR were used to evaluate the effect of MTA, iRoot SP and AH plus on osteogenesis of periodontal ligament stem cells. SPSS 21.0 software package was used for statistical analysis. RESULTS: MTA showed mild toxicity at 24 and 48 hours, AH Plus showed mild toxicity at 24 h. iRoot SP was the least (P<0.05) compared to MTA and AH Plus. The effect of three kinds of materials on apoptosis of periodontal ligament stem cells gradually decreased with the prolongation of time. Compared with the control group, the three kinds of materials were toxic at 3 d, the toxicity of MTA was the strongest and the toxicity of iRoot SP was the lowest(P<0.05). Mineralization nodules in MTA and iRoot SP group were significantly higher than those in AH Plus and control group. The expression of OC, RUNX2, COL1A and ALP gene was higher at 7, 14, 21 d than in the control group and the expression of iRoot SP mineralization was the greatest(P<0.05). CONCLUSIONS: The hardened iRoot SP is non-toxic to human periodontal ligament stem cells. Osteogenic ability and mineralization capacity of hardened iRoot SP on human periodontal ligament stem cells are better than MTA.

Key words: Periodontal ligament stem cells, iRoot SP, Cytotoxicity, Osteogenesis, Mineralization

中图分类号:

R781.4

郑晶晶, 康馨允, 李少明, 刘嘉成, 孙凯, 高岭, 侯铁舟. MTA、iRoot SP和AH Plus 对人牙周膜干细胞增殖及分化的影响[J]. 上海口腔医学, 2020, 29(5): 449-455.

ZHENG Jing-jing, KANG Xin-yun, LI Shao-ming, LIU Jia-cheng, SUN Kai, GAO Ling, HOU Tie-zhou. Effects of MTA, iRoot SP and AH Plus on proliferation and differentiation of human periodontal ligament stem cells[J]. Shanghai Journal of Stomatology, 2020, 29(5): 449-455.

参考文献

[1] Lee SJ, Monsef M, Torabinejad M.Sealing ability of a mineral trioxide aggregate for repair of lateral root perforations[J]. J Endod, 1993, 19(11): 541-544.
[2] 李羽弘, 韦曦. iRoot BP和iRoot BP Plus应用于牙髓治疗的研究现状[J]. 中华口腔医学研究杂志, 2016, 10(3): 208-211.
[3] Seo BM, Miura M, Gronthos S, et al.Investigation of multipotent postnatal stem cells from human periodontal ligament[J]. Lancet, 2004, 364(9429): 149-155.
[4] Lv FJ, Tuan RS, Cheung KM, et al.Concise review: the surface markers and identity of human mesenchymal stem cells[J]. Stem Cells, 2014, 32(6): 1408-1419.
[5] Shi S, Bartold PM, Miura M, et al.The efficacy of mesenchymal stem cells to regenerate and repair dental structures[J]. Orthod Craniofac Res, 2005, 8(3): 191-199.
[6] Lin LM, Rosenberg PA.Repair and regeneration in endodontics[J]. Int Endod J, 2011, 44(10): 889-906.
[7] Morsczeck C, Gotz W, Schierholz J, et al.Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth[J]. Matrix Biol, 2005, 24(2): 155-165.
[8] Gandolfi MG, Shah SN, Feng R, et al.Biomimetic calcium-silicate cements support differentiation of human orofacial mesenchymal stem cells[J]. J Endod, 2011, 37(8): 1102-1108.
[9] Giacomino CM, Wealleans JA, Kuhn N, et al.Comparative biocompatibility and osteogenic potential of two bioceramic sealers[J]. J Endod. 2019, 45(1): 51-56.
[10] 赵志鹏. 一种新型环氧基树脂根管糊剂的体外细胞毒性研究[J]. 国外医学·口腔医学分册, 2002, 29(4): 227.
[11] 张伟. iRoot SP生物相容性及根尖封闭能力的研究[D]. 武汉: 武汉大学, 2011.
[12] Zhang W, Walboomers XF, Wolke JG, et al.Differentiation ability of rat postnatal dental pulp cells in vitro[J]. Tissue Eng, 2005, 11(3-4): 357-368.
[13] Seo MS, Hwang KG, Lee J, et al.The effect of mineral trioxide aggregate on odontogenic differentiation in dental pulp stem cells[J]. J Endod, 2013, 39(2): 242-248.
[14] Yokose S, Kadokura H, Tajima Y, et al.Establishment and characterization of a culture system for enzymatically released rat dental pulp cells[J]. Calcif Tissue Int, 2000, 66(2): 139-144.
[15] Adachi M, Characterization of osteoblast precursor cells isolated from colchicine treated rats[J]. Bull Tokyo Med Dent Univ, 1987, 34(2): 41-51.
[16] Iwata T, Mino C, Kawata T.In vitro proliferation of periodontal ligament-like tissue on extracted teeth[J]. Arch Oral Biol, 2017, 75(1): 31-36.
[17] Roholl PJ, Blauw E, Zurcher C.Evidence for a diminished maturation of preosteoblasts into osteoblasts during aging in rats: an ultrastructural analysis[J]. Bone Miner Res, 1994, 9(3): 355-366.
[18] Zhu Z, Xie Q, Huang Y, et al.Aucubin suppresses titanium particles-mediated apoptosis of MC3T3-E1 cells and facilitates osteogenesis by affecting the BMP2/Smads/RunX2 signaling pathway[J]. Mol Med Rep, 2018, 18(3): 2561-2570.
[19] Kawai A.A newly established human osteosarcoma cell line with osteoblastic properties[J]. Clin Orthop Relat Res, 1990, (259): 256-267.
[20] Marelli B, Ghezzi CE, Barralet JE, et al.Three-dimensional mineralization of dense nanofibrillar collagen-bioglass hybrid scaffolds[J]. Biomacromolecules, 2010, 11(6): 1470-1479.

MTA、iRoot SP和AH Plus 对人牙周膜干细胞增殖及分化的影响

Effects of MTA, iRoot SP and AH Plus on proliferation and differentiation of human periodontal ligament stem cells

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

本文评价

[1]	王冕, 姚莉莉, 韩爽, 笪海芹, 侯颖. 双波长激光联合生物陶瓷材料iRoot SP辅助磨牙根管治疗效果分析[J]. 上海口腔医学, 2024, 33(4): 421-425.
[2]	王静, 徐娜, 任慧迪. TNF-α通过ERK1/2-Runx2信号通路调控SHED成骨分化能力的实验研究[J]. 上海口腔医学, 2024, 33(2): 135-140.
[3]	赵姗, 张凌, 李胜男, 康楠, 孟箭, 李晓东. 掺镁纳米多孔钛涂层的生物特性及促成骨分化作用[J]. 上海口腔医学, 2024, 33(1): 6-12.
[4]	沈忆芬, 刘超, 汤颖, 杨涛, 顾永春. ROS/JNK/caspase 3信号轴在薄荷味电子烟烟液诱导牙周膜干细胞凋亡中的作用[J]. 上海口腔医学, 2024, 33(1): 40-48.
[5]	苏娟娟, 陈洪婷, 王旭, 靳强, 王琳. PGE2、COX-2表达与牙槽骨成骨活性、内氧水平的关系[J]. 上海口腔医学, 2024, 33(1): 85-89.
[6]	李莉, 连文伟. 炎症微环境作用下BMP9通过ERK5/KLF4信号通路促进牙周膜干细胞成骨分化[J]. 上海口腔医学, 2023, 32(6): 583-589.
[7]	虞美琳, 吴海苗, 李贵飞, 胡孟飏, 陈栋. 海藻酸钠-去铁胺/壳聚糖微球对大鼠骨髓间充质干细胞成骨分化的影响[J]. 上海口腔医学, 2023, 32(4): 356-362.
[8]	杨习良, 郑天霞, 李文, 王武良, 尹紫涵, 白宇宏. 硅酸盐根管封闭剂的体外细胞毒性、成骨潜力和抗菌活性评价[J]. 上海口腔医学, 2023, 32(3): 246-250.
[9]	胡熹, 罗俊. miR-497-5p在前成骨细胞MC3T3-E1分化和矿化中的作用及机制探讨[J]. 上海口腔医学, 2023, 32(1): 17-22.
[10]	迪丽达尔·塔西甫拉提, 牛雅琪, 热依沙·阿布都克依木, 王玲. 长链非编码RNA AWPPH通过调控Notch信号通路对人牙周膜细胞增殖和成骨向分化的影响[J]. 上海口腔医学, 2023, 32(1): 23-27.
[11]	潘巧婷, 臧晓龙, 孙照薇, 潘梦琪, 朱鑫美, 李志勇. 信号素4D在大鼠双膦酸盐相关颌骨坏死发生中的作用探讨[J]. 上海口腔医学, 2022, 31(6): 625-631.
[12]	胡逸鹏, 欧晓艳, 钟鸿梅. miR-124对牙髓间充质干细胞成骨分化的影响及机制探讨[J]. 上海口腔医学, 2022, 31(5): 466-470.
[13]	段昕, 刘菲, 刘珂珂, 王松松, 张云涛. 嵌入BMP-2蛋白微球双重缓释纳米纤维支架对MC3T3-E1细胞增殖、分化的影响[J]. 上海口腔医学, 2022, 31(5): 476-482.
[14]	付洪海, 孙乐刚, 丁昌成, 马向瑞, 黄玉梅. miR-31-5p对牙髓干细胞HIF-1α/BNIP3信号通路及成骨相关因子表达的影响[J]. 上海口腔医学, 2022, 31(3): 237-242.
[15]	董家辰, 束蓉. 炎症微环境对人牙周膜成纤维细胞增殖与成骨分化的影响[J]. 上海口腔医学, 2022, 31(3): 243-247.