天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (2): 137-141.doi: 10.11958/20150183

• 细胞与分子生物学 • 上一篇    下一篇

人牙龈成纤维细胞和牙周膜韧带细胞多向分化潜能的比较

苏瑞， 宋立婷， 董允允， 邓嘉胤， 蒋少云△   

  1. 天津医科大学口腔医院牙周科 （邮编 300070）
  • 收稿日期:2015-09-22 修回日期:2015-10-21 出版日期:2016-02-15 发布日期:2016-02-15
  • 通讯作者: △通讯作者 E-mail: sjiang@tmu.edu.cn E-mail:midekexiner@126.com
  • 基金资助:
    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）

The comparison on pluripotent differentiation between human gingival fibroblasts and human periodontal ligament cells in vitro

SU Rui, SONG Liting, DONG Yunyun, DENG Jiayin, JIANG Shaoyun △   

  1. Department of Periodontology, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • Received:2015-09-22 Revised:2015-10-21 Published:2016-02-15 Online:2016-02-15
  • Contact: △Corresponding Author E-mail: sjiang@tmu.edu.cn E-mail:midekexiner@126.com

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摘要： 目的 体外培养人牙周膜韧带细胞 （HPDLCs） 和人牙龈成纤维细胞 （HGFs）， 对比两者成骨、 成软骨以及成脂的多向分化潜能的差异。方法 运用酶消化结合组织块法体外培养 HPDLCs 和 HGFs， 选择生长状态良好的 3~4 代 HPDLCs 和 HGFs， 进行成骨、 成软骨、 成脂诱导， 未分化诱导的细胞作为对照组。分别用茜素红染色、 油红 O 染色以及阿利新蓝染色分别检测两种细胞的成骨、 成软骨及成脂分化能力。半定量逆转录聚合酶链反应 （RT-PCR） 检测 HPDLCs 和 HGFs 中相关标志基因骨钙素(OCN)、 Ⅰ型胶原蛋白 （Col 1）、 runt 相关转录因子 2 （RUNX2）、 过氧化物酶体增殖物激活受体γ2 （PPARγ2）、 X 型胶原蛋白 （Col 10） mRNA 的表达。结果 成骨诱导两种细胞培养至 28 d 时， 细胞周围均有红染钙结节形成， HPDLCs 形成的钙结节明显多于 HGFs； 成软骨诱导两种细胞 14 d 时均可见胞质蓝染的细胞， HGFs 较 HPDLCs 明显； 成脂诱导两种细胞 21 d 时， 可见红色脂肪滴形成， HPDLCs 形成的脂肪颗粒明显少于 HGFs。HGFs 和 HPDLCs 分化培养 7 d 和 14 d 后， 均有 OCN、 Col 1、 RUNX2、 PPARγ2 和 Col 10 的表达， 在 14 d 时的表达均高于 7 d； HPDLCs 中 OCN、 Col 1、 RUNX2 的表达高于 HGFs， PPARγ2、 Col 10 的表达低于 HGFs（均 P＜ 0.05）。结论 HPDLCs 的成骨能力较 HGFs 强， 成软骨和成脂能力较 HGFs 弱。

关键词: 人牙周膜韧带细胞, 人牙龈成纤维细胞, 细胞分化, 成骨分化, 成脂分化, 成软骨分化

Abstract: Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic medium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP⁃ DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly higher in HPDLCs than those in HGFs (P＜0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P＜0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was significantly higher in HGFs than that of HPDLCs (P＜0.01). Conclusion HPDLCs has the better potency of osteogenic differetiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.

Key words: human periodontal ligament cells, human gingival fibroblasts, cell differentiation, osteogenic differentiation, adipogenic differentiation, chondrogenic differentiation