中国实用口腔科杂志

中国实用口腔科杂志

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根尖牙乳头干细胞促进人脐静脉内皮细胞血管生成的研究

庄雪莹，马铭辰，余思，刘双伟，刘雪梅，刘尧   

  1. 中国医科大学口腔医学院儿童口腔科，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:2020-01-15 发布日期:2020-03-02
  • 基金资助:
    国家自然科学基金面上项目（81771059）；辽宁省科学技术计划项目（2018225061）；中国医科大学创新创业训练计划项目（201910159129）

  • Online:2020-01-15 Published:2020-03-02

摘要： 目的    血管早期生成是组织修复与再生的关键。本研究探究根尖牙乳头干细胞（stem cells from apical papilla，SCAP）对人脐静脉内皮细胞（human umbilical vein endothelical cells，HUVECs）血管生成能力的影响，为促进组织再生提供新的策略。方法　SCAP原代提取，流式细胞术鉴定干细胞表面标记物，茜素红S和油红O染色，鉴定SCAP的多向分化潜能。应用Transwell培养体系，将SCAP和HUVECs共培养，Western blot和管腔形成实验检测HUVECs血管生成相关蛋白表达和成管能力。采用SPSS 20.0软件对数据进行统计学分析。结果    　SCAP表达间充质干细胞表面标记物CD29、CD44、CD105、CD146，不表达造血干细胞表面标记物CD34、CD45，体外诱导培养后可见大量矿化结节和脂滴形成。SCAP和HUVECs共培养后，HUVECs中血管内皮生长因子VEGFA的蛋白表达水平显著提高（P < 0.001），同时HUVECs的管腔形成能力明显增强（P < 0.01）。结论    SCAP能够促进HUVECs血管生成，且与其旁分泌作用相关。

关键词: 根尖牙乳头干细胞, 人脐静脉血管内皮细胞, 血管生成, 旁分泌

Abstract: Objective    Angiogenesis is the key to tissue repair and regeneration. This study is to investigate the effects of stem cells from apical papilla（SCAP）on the angiogenesis of human umbilical vein endothelial cells（HUVECs），and to provide new strategies for promoting angiogenesis in the process of tissue regeneration. Methods    The primary SCAP were isolated and the surface markers of stem cells were identified by flow cytometry analysis. The multipotent differentiation potentials of SCAP were detected by Alizarin red S and Oil red O staining. SCAP and HUVECs were co-cultured through Transwell，and the angiogenesis ability of HUVECs was tested by Western blot and tube formation assay. The data were analyzed by SPSS 20.0 software. Results    SCAP expressed the mesenchymal stem cell surface markers，including CD29，CD44，CD105 and CD146，but failed to express the hematopoietic stem cells surface markers CD34 and CD45. Mineralized nodules and lipid droplets could be observed after osteogenic and adipogenic induction of SCAP in vitro. After co-culture of SCAP and HUVECs，the protein expression of VEGFA was significantly enhanced（P < 0.001）. Meanwhile，the tube formation ability of HUVECs was enhanced（P < 0.01）. Conclusion    SCAP can promote HUVECs angiogenesis，which is related to its paracrine effects.

Key words: stem cells from apical papilla, human umbilical vein endothelial cells, angiogenesis, paracrine