血小板衍生生长因子BB对牙周膜干细胞生物学特性的影响

doi:10.19439/j.sjos.2020.04.002

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (4): 343-349.doi: 10.19439/j.sjos.2020.04.002

血小板衍生生长因子BB对牙周膜干细胞生物学特性的影响

邓佳佳^1,2, 潘杰^1,2, 於丽明^1,2, 张维华^1,2, 朱露莹^2,3, 韩欣欣^1,2, 刘月华^1,2

1.复旦大学附属口腔医院,上海市口腔病防治院,上海 200001;
2.复旦大学附属口腔医院口腔生物医学工程实验室,上海 200001;
3.中南大学湘雅口腔医学院·口腔医院, 湖南长沙 410000

收稿日期:2019-01-29 修回日期:2019-04-08 出版日期:2020-08-25 发布日期:2020-09-11
通讯作者: 刘月华,E-mail：liuyuehua@fudan.edu.cn
作者简介:邓佳佳（1992-）,女,在读硕士研究生,E-mail:17211370001@fudan.edu.cn
基金资助:
国家自然科学基金（81470768）; 上海市科委重点基础研究项目（14JC1492800）; 上海市卫生与计划生育委员会重点项目（201640023）

Biological characteristics of periodontal ligament stem cells modified by platelet derived growth factor BB

DENG Jia-jia^1,2, PAN Jie^1,2, YU Li-ming^1,2, ZHANG Wei-hua^1,2, ZHU Lu-ying^2,3, HAN Xin-xin^1,2, LIU Yue-hua^1,2

1. Shanghai Stomatological Hospital, Fudan University. Shanghai 200001;
2. Oral Biomedical Engineering Laboratory, Shanghai Stomatological Hospital, Fudan University. Shanghai 200001;
3. Xiangya School of Stomatology, Xiangya Stomatological Hospital , Central South University. Changsha 410000, Hunan Province, China

Received:2019-01-29 Revised:2019-04-08 Online:2020-08-25 Published:2020-09-11

摘要/Abstract

摘要： 目的：观察血小板衍生生长因子BB（platelet derived growth factor BB, PDGFBB）基因修饰人牙周膜干细胞（human periodontal stem cells, hPDLSCs）的生物学特性,探讨PDGFBB对hPDLSCs细胞增殖、迁移及诱导成骨的影响。方法：分离并扩增hPDLSCs,免疫荧光染色鉴定细胞表面标志和成骨诱导分化能力。通过慢病毒载体介导PDGFBB基因转染hPDLSCs,转染后,采用CCK-8及划痕实验检测其对细胞增殖、迁移的影响。利用实时荧光定量PCR分析PDGFBB基因转染后hPDLSCs细胞内成骨相关基因和成血管相关基因VEGF mRNA的表达水平。采用SPSS 22.0软件包对数据进行统计学分析。结果：采用组织块培养及有限稀释法成功获得hPDLSCs。PDGFBB转染hPDLSCs后,细胞形态良好,与空病毒组和未转染组相比,转染组细胞增殖及迁移能力显著上升,OPN、COL-1和VEGF mRNA表达水平显著上调（P<0.05)。结论：慢病毒载体能在体外将PDGFBB基因转入hPDLSCs,获得持续、稳定表达。PDGFBB可促进hPDLSCs细胞的增殖和迁移,上调成骨和成血管相关基因的表达。

关键词: 骨修复, 血小板衍生生长因子, 牙周膜干细胞, 慢病毒转染

Abstract: PURPOSE: To investigate the biological characteristics of human periodontal stem cells (hPDLSCs) modified with platelet derived growth factor BB(PDGFBB) gene, and to explore its influence on proliferation, migration and osteogenic induction of hPDLSCs. METHODS: hPDLSCs were isolated and amplified, and immunofluorescence staining was performed to identify cell surface markers and osteogenic differentiation ability. hPDLSCs were transfected with PDGFBB gene by lentivirus vector, and the effects on cell proliferation and migration were detected by CCK-8 and scratch test after transfection. Real-time PCR was performed to analyze the mRNA expression levels of osteogenic and angiogenic genes in hPDLSCs cells transfected with PDGFBB gene. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: hPDLSCs were successfully obtained by tissue mass culture and finite dilution method. Compared with the blank virus group and non-transfected group, the proliferation and migration ability of the cells in the transfection group were significantly increased, and the mRNA expression levels of OPN, COL-1 and VEGF were significantly up-regulated(P<0.05). CONCLUSIONS: Lentiviral vector can transfer PDGFBB gene into hPDLSCs in vitro and obtain continuous and stable expression. PDGFBB can promote proliferation and migration of hPDLSCs cells and up-regulate expression of osteogenic and angiogenic genes.

Key words: Bone repair, Platelet derived growth factor, Periodontal ligament stem cells, Lentiviral vector

中图分类号:

R781.4

邓佳佳, 潘杰, 於丽明, 张维华, 朱露莹, 韩欣欣, 刘月华. 血小板衍生生长因子BB对牙周膜干细胞生物学特性的影响[J]. 上海口腔医学, 2020, 29(4): 343-349.

DENG Jia-jia, PAN Jie, YU Li-ming, ZHANG Wei-hua, ZHU Lu-ying, HAN Xin-xin, LIU Yue-hua. Biological characteristics of periodontal ligament stem cells modified by platelet derived growth factor BB[J]. Shanghai Journal of Stomatology, 2020, 29(4): 343-349.

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血小板衍生生长因子BB对牙周膜干细胞生物学特性的影响

Biological characteristics of periodontal ligament stem cells modified by platelet derived growth factor BB

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