CGF对人根尖牙乳头干细胞增殖分化及迁移能力的影响  

Effects of CGF on proliferation, differentiation and migration of human stem cells from apical papilla

作　　者：张伟祥 白雪 冯保静[1] ZHANG Wei-xiang;BAI Xue;FENG Bao-jing(Department of Prosthodontie,the Six-People's Hospital of Anyang City,Anyang 455000 China)

机构地区：[1]安阳市第六人民医院修复科,安阳455000 [2]安阳市第六人民医院儿童牙病科,安阳455000

出　　处：《中华老年口腔医学杂志》2021年第3期134-139,共6页Chinese Journal of Geriatric Dentistry

基　　金：安阳市科技计划攻关项目(项目编号:2020-168)。

摘　　要：目的:体外研究异体浓缩生长因子(Concentrate Growth Factors,CGF)对人根尖牙乳头干细胞(human stem cells from apical papilla,hSCAPs)增殖、迁移及成骨分化能力的影响,为异体CGF临床应用提供实验依据。方法:体外分离、纯化根尖牙乳头干细胞,流式细胞仪检测根尖牙乳头干细胞表面抗原分子,成骨及成脂分化诱导后茜素红染色及油红O染色检测细胞多向分化潜能;静脉取血变速离心制作自体及异体CGF,并使用CGF培养hSCAPs,CCK8法检测1、3、5、7天hSCAPs细胞增殖能力,碱性磷酸酶活性(ALP)实验检测细胞ALP活性,实时定量聚合酶链反应(qPCR)检测细胞ALP、OCN、BSP、DSPP mRNA水平的表达,Transwell小室检测细胞迁移能力。结果:原代培养的hSCAPs阴性表达CD34、CD45,阳性表达CD90、CD29和CD146,并具有成骨向及成脂向分化潜能;相较于对照组,自体及异体CGF均可明显增强hSCAPs增殖及迁移能力,并能显著增加细胞ALP活性及成骨/成牙相关基因的表达,自体CGF对细胞迁移能力的促进作用略强于异体组,而增殖及分化能力在自体和异体组间无显著性差异。结论:自体及异体CGF在hSCAPs增殖能力、迁移能力及成骨向分化能力方面均具有显著的促进作用,提示异体CGF在牙髓损伤修复及组织工程应用中具有重要的潜在作用。Objective:To investigate the effects of autogenous and homogenous concentrate growth factor(CGF)on the proliferation migration and differentiation of stem cells from apical papilla(hSCAPs)in vitro.which provided experimental basis for clinical application of allogeneic CGF.Methods:hSCAPs were isolated and cultured in vitro.Cell surface antigen molecules of HSCAPs were detected by flow cytometry,and their multidirectional differentiation ability was detected by alizarin red staining and oil red O staining Venous blood was centrifuged at variable speed to make autologous and allogeneic CGF,then hSCAPs were cultured with CGF.CCK8 was used to detect the proliferation of hSCAPs cells at 1,3,5 and 7 days,the alkaline phosphatase activity test was used to detect the ALP activity,qPCR was used to detect the mRNA expression of osteogenetic/odontogenic related genes(ALP,OCN,BSP,DSPP),and cell migration was detect by Transwell assay after treated with autogenous and homogenous CGF.Results:Flow cytometry showed that HSCAPs were negative for CD34 and CD45,and positive for CD90,CD29 and CD146,and had osteogenic and adipogenic differentiation potential.Compared with the control group,both autologous and allogeneic CGF could significantly enhance the proliferation and migration ability of HSCAPs,and significantly increase the ALP activity and the expression of osteogenic/odontogenic related genes.The promotion effect of autologous CGF on the migration ability of HSCAPs was slightly stronger than that of the allogeneic group,but there was no significant difference in the proliferation and differentiation ability between the autologous and allogeneic groups.Conclusion:Both autologous and allogeneic CGF can significantly promote the proliferation,migration and osteogenesis/odontogenesis of hSCAPs,suggesting that allogeneic CGF plays an important role in the application of hSCAPs tissue repair.

关 键 词：浓缩生长因子(CGF) 人根尖牙乳头干细胞(hSCAPs) 分化 增殖 迁移 

分 类 号：R783[医药卫生—口腔医学]