中国实用口腔科杂志

中国实用口腔科杂志

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干扰素-γ对脱落乳牙干细胞增殖和凋亡影响研究

戴　璐，刘雪梅，杨宁，陈旭，刘尧   

  1. 中国医科大学口腔医学院·附属口腔医院儿童口腔科，辽宁省口腔疾病重点实验室，辽宁省口腔疾病临床医学研究中心，辽宁  沈阳  110002
  • 出版日期:2019-02-15 发布日期:2019-03-27
  • 基金资助:

    国家自然科学基金青年科学基金项目（81600825）；军事口腔医学国家重点实验室开放课题（2017KA04）；沈阳市科技计划项目（17-231-1-50）

  • Online:2019-02-15 Published:2019-03-27

摘要：

目的　研究促炎症细胞因子干扰素-γ（interferon-gamma，IFN-γ）对脱落乳牙干细胞（stem cells from exfoliated deciduous teeth，SHED）增殖和凋亡的影响，为揭示免疫微环境在组织再生中的作用及潜在机制提供理论基础和实验依据。方法　原代分离培养人SHED，用不同浓度（10、50、100、200 ng/mL）的IFN-γ处理SHED记为IFN-γ处理组，未加入IFN-γ的记为对照组。应用CCK-8实验检测细胞增殖情况，流式细胞术和甲苯胺蓝染色检测细胞凋亡情况。采用SPSS 20.0软件进行统计学分析。结果　原代培养SHED表达间充质干细胞表面标记物CD73、CD90、CD105，体外诱导培养具有成骨、成神经和成脂的多向分化潜能。不同浓度IFN-γ均显著抑制SHED的增殖（F = 169.7，P < 0.0001），且可能具有浓度依赖性；随IFN-γ处理时间的延长，其对SHED增殖的抑制作用越强（F = 4.81，P = 0.0007）；高浓度IFN-γ处理组（100、200 ng/mL）SHED凋亡率和存活率与对照组相比，差异有统计学意义（均P < 0.05）。结论　IFN-γ能够抑制SHED增殖以及诱导其凋亡，从而损伤SHED的生物学性能，降低其骨再生能力。

关键词: 脱落乳牙干细胞, 干扰素-&gamma, 细胞增殖, 细胞凋亡

Abstract:

Objective　To study the effects of pro-inflammatory cytokine interferon-gamma（IFN-γ）on proliferation and apoptosis of stem cells from exfoliated deciduous teeth（SHED），and provide theoretical basis and experimental basis for revealing the role and potential mechanism of immune microenvironment in tissue regeneration. Methods　The primary SHED was isolated and cultured，and the treatment group called IFN-γ treatment group was treated with IFN-γ at different concentrations（10，50，100，200 ng/mL），and the control group was not treated with IFN-γ. Cell proliferation was detected by CCK-8 assay，and apoptosis was detected by flow cytometry and toluidine blue staining. SPSS20.0 software was used for statistical analysis. Results　Primary cultured SHED expressed mesenchymal stem cell surface markers CD73，CD90，CD105，and induced multi-directional differentiation potential of osteoblasts，neuroblasts and adipogenesis in vitro. Different concentrations of IFN-γ significantly inhibited SHED proliferation（F = 169.7，P < 0.0001），and may be concentration-dependent. With the prolongation of IFN-γ treatment time，IFN-γ inhibited the proliferation of SHED（F = 4.81，P = 0.0007） more strongly；the apoptotic rate and survival rate of the high-concentration IFN-γ treatment group（100，200 ng/mL）were significantly different from those of the control group（all P < 0.05）. Conclusion　IFN-γ can inhibit the proliferation of SHED and induce apoptosis, thereby impairing the biological properties of SHED and reducing its bone regeneration ability.

Key words: stem cells from exfoliated deciduous teeth, SHED；interferon-gamma, IFN-γ；cell proliferation；cell apoptosis