天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (7): 713-716.doi: 10.11958/j.issn.0253-9896.2015.07.003

• 细胞与分子生物学 • 上一篇    下一篇

体外诱导人牙龈成纤维细胞多向分化潜能的实验研究

蒋少云，陶玉飞，李阳，宋立婷，朱东望，邓嘉胤   

  1. 天津医科大学口腔医院牙周科 （邮编 300070）
  • 收稿日期:2015-02-27 修回日期:2015-04-02 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 邓嘉胤 E-mail: yazhou2991@126.com E-mail:jiayin_d@yahoo.com.cn
  • 作者简介:蒋少云 （1972）， 女， 副教授， 博士， 主要从事牙周病发病机制及牙周组织工程学的研究
  • 基金资助:
    诱导性多潜能干细胞修复牙周骨组织缺损的实验研究

Study on differentiation pluripotency of human gingival fibroblasts induced in vitro

JIANG Shaoyun, TAO Yufei, LI Yang, SONG Liting, ZHU Dongwang, DENG Jiayin   

  1. Department of Periodontics, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • Received:2015-02-27 Revised:2015-04-02 Published:2015-07-15 Online:2015-07-15
  • Contact: DENG Jiayin E-mail: yazhou2991@126.com E-mail:jiayin_d@yahoo.com.cn
  • Supported by:
    天津市应用基础及前沿技术研究计划-自然科学基金重点项目 （12JCZDJC22700）

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摘要： 目的 探索人牙龈成纤维细胞（hGFs）是否具有多向分化潜能， 为组织工程学提供新的细胞来源。方法 经志愿者知情同意后收集健康牙龈组织， 组织块法培养 hGFs。取第 3 代 hGFs 进行成骨、 成软骨和成脂诱导， 未分化诱导的细胞为对照组。分别用碱性磷酸酶 （ALP） 染色和茜素红染色、 阿利新蓝染色、 油红 O 染色检测细胞成骨、 成软骨和成脂能力。实时定量聚合酶链反应 （PCR） 检测细胞中成骨分化标志基因 ALP、 runt 相关转录因子 2 （Runx2）、成软骨分化标志基因聚集蛋白聚糖（AGR）和成脂分化标志基因过氧化物酶体增殖物激活受体γ2（PPARγ2）的表达。结果 成骨诱导组培养至 7 d 时 ALP 染色可见大量的蓝紫色沉淀， 28 d 时细胞周围有大量红染的钙结节沉积，而对照组无钙结节, 培养至 14 d 细胞中 ALP 和 Runx2 表达明显高于对照组（P < 0.01）。14 d 时成软骨诱导组阿利新蓝阳性， 成脂诱导组可见红色的脂肪滴， 而对照组 2 种染色为阴性， AGR、 PPARγ2 表达均明显高于对照组（P <0.01）。结论 hGFs 具有成骨、 成软骨和成脂分化的多向分化潜能。

关键词: 牙龈, 成纤维细胞, 成脂分化, 细胞分化, 人牙龈成纤维细胞, 成骨分化, 成软骨分化

Abstract: Objective To investigate the pluripotency of human gingival fibroblasts (hGFs), and provide a novel cell source for tissue engineering. Methods With informed consent from volunteers, fresh and healthy gingiva were collected. The hGFs were obtained from the gingiva by tissue culture. The third passage of hGFs was cultured in osteogenic medium, chondrogenic medium and adipogenic medium. Cells without differentiation were taken as control. Cells were examined by alkaline phosphatase (ALP) staining, Alizarin red staining, Alcian blue staining and oil red O staining for detecting of the ability of differentiation pluripotency. Real-time polymerase chain reaction was applied to examine the expression of osteogenic marker genes ALP, runt-related transcript factor 2 (Runx2), chondrogenic marker aggrecan (AGR) and adipogenic marker peroxisome proliferator-activated receptor gamma 2 (PPARγ2).Results The hGFs cultured in osteogenic medium showed massive violet deposit at day 7 and calcium nodulus at day 28, meanwhile, the expressions of ALP and Runx2 were higher than those of control (P < 0.01). In chondrogenic group cells were found blue deposit at day 14. In adipogenic group lipidfilled droplets stained with oil red O were found in cells at day 14. However, hGFs in control group had no any positive staining. Furthermore, expressions of AGR and PPARγ2 were significantly higher than those of control (P < 0.01). Conclusion Human gingival fibroblasts have the pluripotency of osteogenic, adipogenic and chondrogenic differentiation.

Key words: gingiva, fibroblasts, adipogenesis, cell differentiation, human gingival fibroblasts, osteogenic differentiation, chondrogenic differentiation