Journal of Prevention and Treatment for Stomatological Diseases ›› 2021, Vol. 29 ›› Issue (10): 656-662.DOI: 10.12016/j.issn.2096-1456.2021.10.002

• Basic Study • Previous Articles     Next Articles

Effects of graphene on the proliferation, migration and morphology of dental pulp stem cells

SUN Jingxuan(),LI Yanping,PAN Shuang,HE Lina,SUN Xiangyu,ZHANG Shuang,NIU Yumei()   

  1. The First Affiliated Hospital of Harbin Medical University, School of Stomatology, Harbin Medical University, Harbin 150001, China
  • Received:2021-02-10 Revised:2021-04-06 Online:2021-10-20 Published:2021-06-23
  • Contact: Yumei NIU
  • Supported by:
    National Natural Science Foundation of China(81970924);Postdoc-toral Fund Project of China(2018M641871)

石墨烯对牙髓干细胞增殖、迁移能力及细胞形态的影响

孙靖宣(),李艳萍,潘爽,何丽娜,孙翔宇,张爽,牛玉梅()   

  1. 哈尔滨医科大学附属第一医院,哈尔滨医科大学口腔医学院,黑龙江 哈尔滨(150001)
  • 通讯作者: 牛玉梅
  • 作者简介:孙靖宣,硕士研究生,Email: 2018021032@hrbmu.edu.cn
  • 基金资助:
    国家自然科学基金项目(81970924);中国博士后基金项目(2018M641871)

Abstract:

Objective To investigate the effects of graphene on the proliferation, migration and cell morphology of dental pulp stem cells (DPSCs). Methods Graphene powder was prepared by the oxidation-reduction method, and a 0.5 mg/mL graphene dispersion was prepared. Raman spectroscopy and atomic force microscopy were used to characterize the structure and surface morphology of graphene. DPSCs were isolated and cultured in vitro. MTT assay was used to detect the effects of different concentrations of graphene dispersions (0, 1, 5, 10, 20, 50, 100 μg/mL) on the proliferation and wound healing assay was used to detected the migration abilities of DPSCs. The effects of graphene on the morphology of DPSCs were observed by immunofluorescence staining. Results In the present study, compared with the control group (0 μg/mL), the proliferation of DPSCs in the 100 μg/mL group was inhibited at 72 h (P < 0.05), and the proliferation of DPSCs in the other groups was not significantly affected (P > 0.05). Graphene dispersions at 10 and 20 μg/mL promoted the migration of DPSCs (P < 0.05). After being cultured in 20 μg/mL graphene dispersions for 3 days, the DPSCs showed a large and orderly cytoskeletal structure, and the spread area of cells was not significantly different from that of the control group (0 μg/mL) (P > 0.05), while some cells had the morphological characteristics of nerve cells. Conclusion Graphene has good biocompatibility and is expected to be a suitable material for tissue engineering within fitting concentration.

Key words: graphene, graphene oxide, structure characterization, Raman spectrum, atomic force microscopy, dental pulp stem cells, cell proliferation, cell migration, tissue engineering

摘要:

目的 探讨石墨烯对牙髓干细胞(dental pulp stem cells,DPSCs)增殖、迁移及细胞形态等生物学行为的影响。方法 采用氧化还原法制备石墨烯粉末,配制成0.5 mg/mL的石墨烯分散液。拉曼光谱及原子力显微镜对石墨烯进行结构表征及表面形态的观察。体外分离并培养DPSCs,MTT实验检测不同浓度(0、1、5、10、20、50、100 μg/mL)的石墨烯分散液对DPSCs增殖能力的影响,划痕实验检测石墨烯对于DPSCs的迁移能力的影响,通过免疫荧光染色观察石墨烯对DPSCs细胞形态的影响。结果 与对照组(0 μg/mL)相比,72 h内仅100 μg/mL石墨烯分散液组在72 h对DPSCs的增殖有抑制作用(P < 0.05),其余各组对DPSCs的增殖均没有显著的影响(P>0.05);10 μg/mL及20 μg/mL的石墨烯分散液可促进DPSCs的迁移(P<0.05);20 μg/mL的石墨烯分散液培养3 d后,DPSCs可见粗大且有序排列的细胞骨架结构,细胞铺展面积与对照组(0 μg/mL)相比没有显著性差异(P>0.05),部分细胞具有神经细胞的形态特点。结论 适宜浓度的石墨烯具有良好的生物相容性,有望成为组织工程学理想的材料。

关键词: 石墨烯, 氧化石墨烯, 结构表征, 拉曼光谱, 原子力显微镜, 牙髓干细胞, 细胞增殖, 细胞迁移, 组织工程

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