浓缩生长因子(CGF)对人根尖牙乳头干细胞生物学特性的影响

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

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的临床应用提供实验依据。方法:体外分离培养、纯化根尖牙乳头干细胞,通过流式细胞仪、茜素红染色及油红O染色鉴定hSCAPs细胞表面抗原分子及多分化潜能;取静脉血差速离心制备异体CGF,CCK8法检测CGF培养下hSCAPs细胞增殖率,实时定量聚合酶链反应(qPCR)检测成骨基因表达变化,碱性磷酸酶活性(ALP)实验检测CGF培养下hSCAPs ALP活性,Transwell法检测CGF培养下hSCAPs迁移能力。结果:原代培养的hSCAPs阳性表达间充质干细胞表面分子,并具有多向分化潜能;相较于对照组,异体CGF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。结论:异体CGF在hSCAPs增殖能力、迁移能力及成骨向分化能力方面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 homogenous concentrate growth factor(CGF)on the proliferation,differentiation and migration 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 homogenous CGF,then hSCAPs were cultured with CGF.CCK8 was used to detect the proliferation of hSCAPs,the alkaline phosphatase activity was used to detect the ALP activity,qPCR was used to detect the mRNA expression of osteogenetic/odontogenic related genes,and cell migration was detected by Transwell assay after treated with homogenous CGF.Results:HSCAPs were positively expressed mesenchymal stem cell markers,and hold multi-directional differentiation ability.Compared with the control group,homogenous CGF significantly enhance the proliferation and migration ability of hSCAPs,and significantly increase the ALP activity and the expression of osteogenic/odontogenic related genes.Conclusion:Homogenous CGF significantly promote the proliferation,migration and osteogenesis/odontogenesis of hSCAPs,suggesting that homogenous CGF plays an important role in the application of hSCAPs tissue repair.