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

doi:10.3969/j.issn.1671-167X.2016.01.005

北京大学学报(医学版) ›› 2016, Vol. 48 ›› Issue (1): 23-29. doi: 10.3969/j.issn.1671-167X.2016.01.005

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

温泉，赵玉鸣，王媛媛，王旭，凌龙，葛立宏△

（北京大学口腔医学院·口腔医院儿童口腔科，北京100081）

出版日期:2016-02-18 发布日期:2016-02-18
通讯作者: 葛立宏△ E-mail:gelh0919@126.com
基金资助:
国家自然科学基金（81170928）和北京大学临床医院合作专项（2013-4-01）资助

Effects of stromal cell-derived factor-1 on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cells

WEN Quan, ZHAO Yu-ming, WANG Yuan-yuan, WANG Xu, LING Long, GE Li-hong△

(Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing 100081, China)

Online:2016-02-18 Published:2016-02-18
Contact: GE Li-hong△ E-mail:gelh0919@126.com
Supported by:
Supported by the National Natural Science Foundation of China (81170928) and the Cooperation Projects of Clinical Hospital of Peking University (2013-4-01)

摘要/Abstract

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100 μg/L SDF-1或100 μg/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（P<0.01），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（P<0.05）。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。

关键词: 趋化因子CXCL12, 粒细胞集落刺激因子, 牙髓, 成体干细胞, 牙髓再生

Abstract:

Objective： To compare the effects of stromal cell-derived factor-1 (SDF-1) and granulocyte colony-stimulating factor (G-CSF) on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cell (DPSC) in vitro. Methods: DPSCs were cultured in vitro and treated with either 100 μg/L SDF-1 or 100 μg/L G-CSF. Cell counting kit-8 (CCK-8) and colony-forming unit (CFU) were used to detect the effect of SDF-1 and G-CSF on the proliferation ability of DPSC. Cell migration of DPSC was determined by wound healing assay and Transwell migration assay. The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP) staining, ALP activity and alizarin red S staining. The expression of odontoblastic-related genes such as dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) were quantified by real-time RT-PCR. Results: SDF-1 and G-CSF promoted the proliferation of DPSC slightly, but the difference was not statistically significant. Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01), but there was no significant difference between the two factors. In Transwell migration assay, the number of migrated cells of the control group was 5.0±1.4 per sight, while the SDF-1 group was 24.3±6.8 per sight and the G-CSF group was 11.8±3.3 per sight, suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF, and SDF-1 was more effective than G-CSF (P<0.05). Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining. Higher ALP activity, more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment. Conclusion: SDF-1 had no significant effect on the proliferation of DPSC, but could significantly promote cell migration and odontoblastic differentiation of DPSC. Its effect on DPSC was better than G-CSF.

Key words: Chemokine CXCL12, Granulocyte colony-stimulating factor, Dental pulp, Adult stem cells, Regeneration

中图分类号:

R329.28

温泉，赵玉鸣，王媛媛，王旭，凌龙，葛立宏. 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[J]. 北京大学学报(医学版), 2016, 48(1): 23-29.

WEN Quan, ZHAO Yu-ming, WANG Yuan-yuan, WANG Xu, LING Long, GE Li-hong. Effects of stromal cell-derived factor-1 on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cells[J]. Journal of Peking University(Health Sciences), 2016, 48(1): 23-29.

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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

Effects of stromal cell-derived factor-1 on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cells

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