基质细胞衍生因子1对人牙髓干细胞增殖、迁移和成牙本质能力的影响

发布时间：2018-12-26 08:16

【摘要】：目的:对比基质细胞衍生因子1(stromal cell-derived factor-1,SDF-1)和粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)对人牙髓干细胞(dental pulp stem cell,DPSC)的体外增殖、迁移和成牙本质能力的影响。方法:分别在培养基中加入100μg/L SDF-1或100μg/L G-CSF,采用细胞计数试剂盒(cell counting kit-8,CCK-8)和集落形成实验(colony-forming unit,CFU)检测SDF-1和G-CSF对DPSC增殖的影响;采用划痕实验和Transwell迁移实验检测两者对DPSC迁移能力的影响;对DPSC进行成牙本质诱导,通过碱性磷酸酶(alkaline phosphatase,ALP)染色、测定ALP活性、茜素红染色和real-time RT-PCR检测成牙本质相关基因的表达,以检测两者对DPSC体外成牙本质能力的影响。结果:SDF-1和G-CSF能够轻度提高DPSC的增殖及集落形成能力,但差异无统计学意义。加入SDF-1或G-CSF的实验组划痕汇合速率明显高于对照组(P0.01),但两种因子间差异无统计学意义。Transwell迁移实验中,对照组每视野的迁移细胞数量为(5.0±1.4)个,SDF-1组每视野的迁移细胞数量为(24.3±6.8)个,G-CSF组每视野的迁移细胞数量为(11.8±3.3)个,各组间差异有统计学意义(P0.05)。经成牙本质诱导后,实验组细胞ALP染色加深,ALP活性上升,矿化结节形成数量增加,成牙本质相关基因的表达均显著高于对照组。结论:SDF-1对DPSC的增殖能力影响不显著,但能明显提高DPSC的迁移能力和成牙本质分化能力,效果优于G-CSF。
[Abstract]:Aim: to compare the proliferation of human dental pulp stem cells (dental pulp stem cell,DPSC) by stromal cell derived factor 1 (stromal cell-derived factor-1,SDF-1) and granulocyte colony stimulating factor (granulocyte colony-stimulating factor,G-CSF) in vitro. Effects of migration and dentin formation. Methods: 100 渭 g / L SDF-1 or 100 渭 g / L G-CSF were added to the medium, respectively. The cell count kit (cell counting kit-8,CCK-8) and colony forming assay (colony-forming unit,) were used. CFU) was used to detect the effect of SDF-1 and G-CSF on the proliferation of DPSC. The effects of scratch test and Transwell migration test on the migration ability of DPSC were investigated. DPSC was induced by dentin formation. The activity of ALP was determined by alkaline phosphatase (alkaline phosphatase,ALP) staining. The expression of dentin related genes was detected by alizarin red staining and real-time RT-PCR. To detect the effect of both on dentin formation ability of DPSC in vitro. Results: SDF-1 and G-CSF could slightly improve the proliferation and colony formation of DPSC, but the difference was not statistically significant. The rate of scratch convergence in the experimental group added with SDF-1 or G-CSF was significantly higher than that in the control group (P0.01), but there was no significant difference between the two factors. In the Transwell migration experiment, the number of migration cells per field in the control group was (5.0 卤1.4). The number of migration cells per field was (24.3 卤6.8) in SDF-1 group and (11.8 卤3.3) in G-CSF group (P0.05). After dentin induction, ALP staining deepened, ALP activity increased, the number of mineralized nodules increased, and the expression of dentin related genes in the experimental group was significantly higher than that in the control group. Conclusion: SDF-1 has no significant effect on the proliferation of DPSC, but it can obviously improve the migration ability and dentin differentiation ability of DPSC, and the effect is better than that of G-CSF.
【作者单位】：北京大学口腔医学院·口腔医院儿童口腔科;
【基金】：国家自然科学基金(81170928) 北京大学临床医院合作专项(2013-4-01)资助~~
【分类号】：R781

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