缺氧诱导因子1α基因修饰牙髓干细胞的体外成骨及成血管分化  

Osteogenic and angiogenic differentiation of dental pulp stem cells modified by hypoxia-inducible factor-1 alpha gene in vitro

作　　者：唐娟 于栋林 刘郭琦 宋娇娇 左金华[2] 付洪海[2] Tang Juan;Yu Donglin;Liu Guoqi;Song Jiaojiao;Zuo Jinhua;Fu Honghai(Binzhou Medical College,Binzhou 256603,Shandong Province,China;Department of Oral and Maxillofacial Surgery,Affiliated Hospital of Binzhou Medical College,Binzhou 256603,Shandong Province,China)

机构地区：[1]滨州医学院,山东省滨州市256603 [2]滨州医学院附属医院口腔颌面外科,山东省滨州市256603

出　　处：《中国组织工程研究》2023年第24期3865-3870,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金(ZR2018PH023),项目负责人:付洪海;滨州医学院徐荣祥再生医学课题(BY2020XRX011),项目负责人:付洪海。

摘　　要：背景:如何有效地修复和重建临界骨缺损是临床医学的难题。牙髓干细胞作为转基因靶细胞修复受损组织或器官具有一定优势。目的:探究缺氧诱导因子1α对牙髓干细胞体外成骨及成血管的作用。方法:选取临床上12-18岁患者因正畸治疗需要拔除的健康、完整双尖牙的牙髓组织分离、培养牙髓干细胞,实验组将缺氧诱导因子1α基因转染至牙髓干细胞,对照组不做任何处理,采用qPCR检测缺氧诱导因子1α的基因表达以及Western blot检测成骨及成血管因子的蛋白表达。结果与结论:(1)牙髓干细胞为短梭形的成纤维样细胞,细胞呈集落式生长。(2)缺氧诱导因子1α基因稳定转染至牙髓干细胞内,随着时间延长缺氧诱导因子1α基因表达逐渐增高,至14 d达到高峰,21 d明显降低。(3)与对照组相比,实验组的成血管相关因子表达水平更高。转染后1 d,血管内皮生成因子、基质细胞衍生因子1、促血管生成素2、胎盘生长因子蛋白表达明显增高,4 d达到高峰,随后实验组成血管相关蛋白表达虽略有降低,但与对照组相比明显增高。(4)与对照组相比,实验组的成骨相关因子表达水平更高。转染后1 d,人骨形态发生蛋白2、骨钙蛋白、Runt相关转录因子2蛋白表达明显增高,4 d达到高峰。(5)结果表明,缺氧诱导因子1α能够促进牙髓干细胞成骨及成血管因子的表达。BACKGROUND:How to effectively repair and reconstruct critical bone defects has become a difficult problem in clinical medicine.Dental pulp stem cells have certain advantages as transgenic target cells to repair damaged tissues or organs.OBJECTIVE:To investigate the effect of hypoxia-inducible factor-1αon osteogenesis and angiogenesis of dental pulp stem cells in vitro.METHODS:The pulp tissues of healthy and intact bicuspid teeth extracted by orthodontic treatment in patients aged 12 to 18 years were isolated to culture dental pulp stem cells.The hypoxia-inducible factor-1αgene was transfected into dental pulp stem cells in the experimental group.The control group did not do any treatment.The expression of the hypoxia-inducible factor 1αgene was detected by qPCR and the expression of osteogenic and angiogenic factors was detected by western blot assay.RESULTS AND CONCLUSION:(1)Dental pulp stem cells were short spindle-shaped fibroblast-like cells,and the cells grew in colonies.(2)Hypoxia-inducible factor-1αgene was stably transfected into dental pulp stem cells.The expression of the hypoxia-inducible factor-1αgene increased gradually with the extension of time,reached a peak at 14 days,and significantly decreased at 21 days.(3)Compared with the control group,the expression level of angiogenesis-related factors in the experimental group was higher.The protein expression levels of vascular endothelial growth factor,stromal cell-derived factor 1,angiopoietin 2,and placental growth factor were significantly increased at 1 day after gene transfection and reached a peak at 4 days.After that,the expression of angiogenesis-associated protein in the experimental group decreased slightly,but was significantly higher than that in the control group.(4)Compared with the control group,the expression level of osteogenic-related factors in the experimental group was higher.The protein expression levels of human bone morphogenetic protein 2,osteocalcin,and Runt-related transcription factor 2 were significantly increased at 1

关 键 词：牙髓干细胞 缺氧诱导因子1Α 血管内皮生长因子 骨形态发生蛋白2 生长因子 基质细胞衍生因子1 血管生成素2 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R782.12