人牙周膜干细胞与牙周膜细胞生物学特性的比较

Differences between biological characteristics of human periodontal ligament stem cells and human periodontal ligament cells

背景:牙周膜干细胞生物作用是目前牙周病治疗研究的热点,牙周膜成纤维细胞是其分化的终末功能细胞之一,也是其主要的支持细胞,两者生物学特性的差异研究鲜有报道。目的:比较牙周膜干细胞与牙周膜细胞生物学特性的差异。方法:用组织块法体外对牙周膜细胞以及单细胞克隆分离纯化后的人牙周膜干细胞两种细胞分别进行显微镜下形态观察,CCK8法检测并绘制2种细胞的生长曲线。流式细胞分析比较2种细胞的细胞周期以及细胞表面标记物的表达、实时PCR对2种细胞碱性磷酸酶、增殖细胞核抗原和Scleraxis基因进行检测。结果与结论:牙周膜干细胞与牙周膜成纤维细胞外观差别明显,人牙周膜干细胞的生长曲线培养前5 d要低于牙周膜细胞,但在5 d后明显高于牙周膜细胞。人牙周膜干细胞与牙周膜细胞的细胞周期分别为41.1%和23.9%。表面标记物检测结果显示2种细胞虽有相似的表达,但在表达率差异有有显著性意义。实时荧光定量PCR结果显示,人牙周膜干细胞在碱性磷酸酶、增殖细胞核抗原以及Scleraxis基因的表达检测均高于牙周膜细胞。表明牙周膜干细胞在成骨增殖等生物学功能上比牙周膜细胞具有更强的潜能。

BACKGROUND：The biological function of human periodontal ligament stem cells is a hot area of research in the treatment of periodontal disease. Human periodontal ligament cells are one of the end cells derived from human periodontal ligament stem cells;meanwhile, it can also provide supports to the development of human periodontal ligament stem cells. However, few studies are reported about the difference of biological characteristics between human periodontal ligament stem cells and human periodontal ligament cells. OBJECTIVE：To compare the differences of biological characteristics between human periodontal ligament stem cells and human periodontal ligament cells. METHODS：The human periodontal ligament stem cells and human periodontal ligament cells were isolated and purified using tissue explant method and cellclone method, respectively, and then were observed under light microscope to compare the differences of morphology. cellproliferation curves of human periodontal ligament stem cells and human periodontal ligament cells were drawn respectively with cellcounting kit 8 assay. Flow cytometry analysis was used to detect their cellcircles and their surface markers expressions. The alkaline phosphatase gene, proliferating cellnuclear antigen gene and Scleraxis gene of human periodontal ligament stem cells and human periodontal ligament cells were detected by Real-time PCR assay.RESULTS AND CONCLUSION：The human periodontal ligament stem cells and human periodontal ligament cells showed a notable difference in morphology under the light microscope observation. During the first 5 days, the cellproliferation curve of human periodontal ligament stem cells was lower than that of human periodontal ligament cells, but 5 days later, the curve of human periodontal ligament stem cells was significantly higher than that of human periodontal ligament cells. The cellcircles of human periodontal ligament stem cells and human periodontal ligament cells were 41.1%and 23.9%, respectively. The surface markers of human periodontal ligament stem cells and human periodontal ligament cells were similar, but their expression rates had significant difference. The expressions of alkaline phosphatase gene, proliferating cellnuclear antigen gene and Scleraxis gene of human periodontal ligament stem cells were significantly higher than those of human periodontal ligament cells. The above results suggest that human periodontal ligament stem cells have much stronger potential ability than human periodontal ligament cells in osteogensis and cellproliferation.