摘要
背景:人脱落乳牙牙髓干细胞广泛应用于组织修复再生领域,但组织再生效果在实际应用中存在局限性。利用mi RNA干预人脱落乳牙牙髓干细胞的定向分化是未来组织修复再生的重要发展方向。目的:探讨mi R-26b对乳牙牙髓干细胞成神经及成血管分化能力的影响。方法:从人脱落乳牙中分离提取乳牙牙髓干细胞,并将其向神经及血管方向诱导分化,分别检测成神经标志物Nestin、NSE、βⅢ-Tubulin和成血管标志物CD31、VEGFR2、ANG-1及mi R-26b的表达。然后将乳牙牙髓干细胞分为空白对照组、mi R-26过表达组和阴性对照组,RT-q PCR、细胞免疫荧光染色及Western blot检测各组乳牙牙髓干细胞成神经及成血管诱导后相关标志物的表达变化。结果与结论:(1)乳牙牙髓干细胞具有多向分化潜能,能向成骨、成脂、成神经及成血管方向分化;(2)在乳牙牙髓干细胞成神经及成血管分化过程中mi R-26b表达水平升高;(3)相较于空白对照组和阴性对照组,mi R-26b过表达组乳牙牙髓干细胞成神经相关基因βⅢ-Tubulin、Nestin、NSE及成血管相关基因CD31、VEGFR2、ANG-1的m RNA表达显著升高(P<0.01),βⅢ-Tubulin、Nestin及CD31、VEGFR2蛋白表达显著升高(P <0.01)。以上结果表明,miR-26b过表达能促进人脱落乳牙牙髓干细胞成神经和成血管诱导分化。
BACKGROUND:Stem cells from human exfoliated deciduous teeth are widely used in the field of tissue repair and regeneration,but the tissue regeneration effect has limitations in practical applications.Using miRNA to intervene in the directional differentiation of stem cells from human exfoliated deciduous teeth is an important development direction for tissue repair and regeneration in the future.OBJECTIVE:To investigate the effect of miR-26b on the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth.METHODS:Dental pulp stem cells were isolated and extracted from human exfoliated deciduous teeth,and induced to differentiate into nerves and blood vessels.The expressions of neurogenic markers Nestin,NSE,βIII-Tubulin,and angiogenic markers CD31,VEGFR2,ANG-1 and miR-26b were detected.Dental pulp stem cells were divided into blank control group,miR-26 overexpression group,and negative control group.RT-qPCR,cell immunofluorescence staining,and western blot assay were used to detect the expression changes of related markers of neurogenic and angiogenic induction of stem cells from human exfoliated deciduous teeth in each group.RESULTS AND CONCLUSION:(1)Stem cells from human exfoliated deciduous teeth had multi-directional differentiation potential and could differentiate into osteogenesis,adipogenesis,neurogenesis,and vasculogenesis.(2)The expression level of miR-26b increased during the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth.(3)Compared with the blank control group and negative control group,the mRNA expression of neuroblast-related genesβIII-Tubulin,Nestin,NSE and angiogenesis-related genes CD31,VEGFR2,and ANG-1 in stem cells from human exfoliated deciduous teeth in the miR-26b overexpression group was significantly increased(P<0.01),βIII-Tubulin,Nestin,CD31,and VEGFR2 protein expression was significantly increased(P<0.01).The above results show that overexpression of miR-26b can promote the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth.
作者
周绍兰
袁媛园
潘露
徐文
瞿姝熳
Zhou Shaolan;Yuan Yuanyuan;Pan Lu;Xu Wen;Qu Shuman(School of Stomatology,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)
出处
《中国组织工程研究》
北大核心
2025年第36期7769-7775,共7页
Chinese Journal of Tissue Engineering Research
基金
贵州省卫生健康委科学技术基金项目(gzwjkj2021-1-353),项目负责人:袁媛园
贵州医科大学附属口腔医院学科项目(KQYY-2021-2),项目负责人:袁媛园。
关键词
乳牙牙髓干细胞
miR-26b
成神经分化
成血管分化
牙髓再生
工程化干细胞
stem cells from deciduous teeth
miR-26b
neurogenic differentiation
angiogenic differentiation
pulp regeneration
engineered stem cells
