牙周膜干细胞与牙周膜细胞甲状旁腺激素受体表达的比较  

Quantitative comparison of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells

作　　者：王烟岚[1] 王萧萧[1] 陈天宇[1] 杨凡巧 戴旭彬 欧乾民 戴霜叶 林雪峰[1] 

机构地区：[1]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室,广东省广州市510006

出　　处：《中国组织工程研究》2015年第10期1562-1569,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81171575)~~

摘　　要：背景:牙周膜干细胞是牙周膜细胞中一类具有自我更新及多向分化潜能的间充质干细胞,经诱导可分化为成骨样细胞或脂肪样细胞。关于牙周膜干细胞是否表达甲状旁腺激素受体以及受体表达量对牙周膜干细胞成骨能力的影响等,目前尚未见相关报道。目的:检测并比较牙周膜干细胞与牙周膜细胞中甲状旁腺激素受体表达的差异,并探讨甲状旁腺激素受体在成骨过程中的作用机制。方法:应用免疫磁珠法筛选分离牙周膜干细胞及牙周膜细胞。通过细胞免疫荧光染色、荧光定量PCR等方法检测并比较牙周膜干细胞与牙周膜细胞中甲状旁腺激素受体的表达差异。应用荧光定量PCR、碱性磷酸酶活性检测及矿化能力分析等方法评价甲状旁腺激素受体对两者成骨能力的影响。结果与结论:应用免疫磁珠法筛选的STRO-1阳性细胞,经细胞免疫荧光染色鉴定STRO-1、CD146、Vimentin等间充质干细胞表面表型均呈阳性,证明为牙周膜干细胞。细胞免疫荧光染色证明,牙周膜干细胞表达甲状旁腺激素受体,其主要分布于细胞膜与胞浆内,与牙周膜细胞及人成骨肉瘤MG63细胞系相似。荧光定量PCR结果表明,牙周膜干细胞甲状旁腺激素受体的表达量约为牙周膜细胞的3.7倍,与MG63细胞相近。经10-7mol/L甲状旁腺激素处理30 min后,牙周膜干细胞的甲状旁腺激素受体表达量明显增加。经成骨诱导后,牙周膜干细胞的甲状旁腺激素受体表达量、成骨相关基因表达量、碱性磷酸酶活性及矿化能力均高于牙周膜细胞。上述实验结果证明,牙周膜干细胞能表达甲状旁腺激素受体,且主要分布于细胞膜和细胞浆内。牙周膜干细胞甲状旁腺激素受体的表达量明显高于牙周膜细胞,而且甲状旁腺激素受体表达量与细胞的成骨能力相关,说明牙周膜干细胞比牙周膜细胞具有更强的成骨能力,是口腔组织工程学中具有�BACKGROUND： Human periodontal ligament stem cells are a kind of mesenchymal stem cells that have self-renewal and multidifferentiation potential. Previous studies have showed that human periodontal ligament stem cells can differentiate into osteoblast-like cells or adipocyte-like cells under appropriate induction. Yet few studies have focused on the expression level of parathyroid hormone 1 receptor which will affect the osteogenicpotential of Human periodontal ligament stem cells.OBJECTIVE： To examine the expression level of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells and to discuss the role of parathyroid hormone 1 receptor in osteogenic differentiation.METHODS： By using magnetic-bead cell sorting, we separated and identified the human periodontal ligament stem cells and human periodontal ligament cells. We examined and compared the m RNA expression level of parathyroid hormone 1receptor in human periodontal ligament stem cells and human periodontal ligament cells by Real-Time PCR. Osteoblastic differentiation was examined through in vitro matrix mineralization by alizarin red staining and alkaline phosphatase assay.RESULTS AND CONCLUSION： Positive immunomagetic sorted cells were positive for STRO-1, CD146, Vimentin,indicating that they were periodontal ligament stem cells. Parathyroid hormone 1 receptor was expressed in human periodontal ligament stem cells and mainly located in cell membrane and cytoplasm which were similar to human periodontal ligament cells and MG63 cells. The expression of parathyroid hormone 1 receptor in human periodontal ligament stem cells was 3.7 times higher than that in human periodontal ligament cells, which was similar to that in MG63 cells. After osteogenic induction, human periodontal ligament stem cells showed a higher expression of parathyroid hormone 1 receptor and osteoblast-related genes as well as the activity of osteoblast alkaline phosphatase and mineralization compared to human periodontal lig

关 键 词：干细胞 牙周膜 受体 甲状旁腺激素 移植 牙周膜干细胞 牙周膜细胞 甲状旁腺激素受体 成骨分化 多向分化 组织工程 牙周炎 再生医学 表面标志物 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]