南方医科大学学报

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (08): 1020-1022.

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体外培养的人牙囊细胞矿化相关标志物的表达

王浈1, 刘宏伟1, 金岩2, 刘源2, 刘兰宁1   

  1. 1. 南方医科大学南方医院口腔科, 广东, 广州, 510515;
    2. 第四军医大学口腔医学院病理科, 陕西, 西安, 710032
  • 出版日期:2005-08-20 发布日期:2005-08-20
  • 基金资助:
    收稿日期:2004-9-2。
    作者简介:王浈(1973-),女,1995年毕业于江西医学院口腔系,硕士在读,主治医师,工作单位:江西南昌解放军第九四医院,E-mail:wangzhen924@hotmail.com

Expression of mineral-associated markers in human dental follicle cells in vitro

WANG Zhen1, LIU Hong-wei1, JIN Yan2, LIU Yuan2, LIU Lan-ning1   

  1. 1. 南方医科大学南方医院口腔科, 广东, 广州, 510515;
    2. 第四军医大学口腔医学院病理科, 陕西, 西安, 710032
  • Online:2005-08-20 Published:2005-08-20

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摘要： 目的 体外培养人牙囊细胞,探讨牙囊细胞是否具成骨、成牙骨质细胞表型特征,是否可作为牙骨质再生的种子细胞。方法 酶消化联合组织块法获得人牙囊细胞,免疫酶细胞化学染色技术检测其Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白的表达,RT-PCR技术检测细胞的骨涎蛋白、骨钙素、碱性磷酸酶的表达。矿化诱导并连续培养牙囊细胞,行Von kossa染色检测体外矿化形成情况。结果 免疫组化结果:牙囊细胞中Ⅰ型胶原、Ⅲ型胶原、骨桥蛋白、骨粘连蛋白呈不同程度的阳性表达;RT-PCR结果显示骨涎蛋白、骨钙素、碱性磷酸酶表达。经矿化诱导后的牙囊细胞在20d时可形成矿化结节,von Kossa染色阳性。结论 牙囊细胞具有成骨细胞、成牙骨质细胞的部分特性,体外培养的人牙囊细胞具有分泌合成矿化组织的能力,可试将其作为牙周组织再生的种子细胞。

Abstract: Objective To culture human dental follicle cells in vitro and observe their mineralization characteristics. Methods Human dental follicle tissues were digested with bacterial collagenase and cultured to obtain dental follicle cells. Mineralization characteristics of these cells were identified by immunohistochemistry and reverse transcription (RT)-PCR. Dental follicle cells were incubated with mineralization-inducing agents to observe the formation of mineralized nodules. Results Immunohistochemistry staining revealed expressions of type Ⅰ collagen, type Ⅲ collagen, osteopontin and osteonectin in the cytoplasm of dental follicle cells cultured in vitro. Low levels of bone sialoprotein, osteocalcin and alkaline phosphatase mRNA were also detected by RT-PCR. Mineralized nodules were observed after 20 days of incubation with mineralization- inducing agents with positivity for von-Kossa staining. Conclusion Human dental follicle cells cultured in vitro show some features of osteoblasts or cementoblasts and can be used as the seed cells for periodontal tissue engineering.

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