Investigation for establishment of periodontal regenerative therapy using stem cells in cultured periodontal ligament cells

2008年：存在于侧群（SP）中的各种类型的干细胞强烈渗出Hoechst 33342染料。骨髓中的SP细胞表达人干细胞（HSC）的细胞表面标记物，并且高度富集HSC活性。本研究旨在从人牙周膜细胞中分离SP细胞，并对其特性进行研究。不幸的是，目前的数据得出结论，HPDL细胞含有SP细胞，而SP细胞不富集间充质干细胞（MSC）-like properties.In 2009，Twist和骨形态发生蛋白（BMP）拮抗剂可能在PDL的恒定性的维持中发挥重要作用，通过其负调控PDL中存在的细胞的成骨细胞分化。在这项研究中，这些分子表达的变化，诱导成骨细胞分化的HPDL细胞进行了评估。我们进一步研究了BMP拮抗剂对成骨细胞分化的调节作用。目前的结果表明，在生理条件下， ...更多信息 条件下，扭转和BMP拮抗剂; gremlin和卵泡抑素，这是组成性表达的HPL细胞，负调控成骨细胞分化的细胞。此外，HPL细胞中成骨细胞分化的诱导同时增强扭曲表达，而其抑制gremlin和卵泡抑素的表达，结果，成骨细胞分化可能被调节。PDL含有成纤维细胞，其可能参与PDL纤维的形成和维持，以及具有生成牙槽骨和牙骨质的潜力的成骨细胞，然而，用于识别和分离每种细胞类型的特异性标志物尚未阐明。本研究的目的是使用流式细胞术从HPDL细胞中分离碱性磷酸酶（ALP）阳性和阴性亚群，并鉴定2个亚群之间在促有丝分裂活性、成骨潜能和核心蛋白聚糖以及双糖蛋白聚糖表达方面的功能差异。本研究结果提示，ALP阳性的HPDL细胞具有成骨细胞样表型，而ALP阴性的HPDL细胞是成纤维细胞群，成骨能力较弱。此外，双糖蛋白聚糖可能正调控ALP阳性HPDL细胞中的成骨细胞分化，而核心蛋白聚糖，其表达通过成骨诱导而增加，可能负调控两个亚群中的矿化。2010年，为了研究PDL中存在的MSC的特征，通过FACS从HPDL分离的CD 271 +/CD 90+细胞进行单细胞培养，并收集具有MSC样性质的克隆。因此，CD 271 +/CD 90+可作为从HPDL中采集MSC的标志物。少

In 2008 : The various types of stem cells exist in side population(SP) effused strongly Hoechst 33342 dye. The SP cells in bone marrow express cell surface markers of human stem cells(HSCs) and are highly enriched for HSC activity. In this study, we clarified the characteristics of SP cells isolated from human periodontal ligament(HPDL) cells. Unfortunately, the present data concluded that HPDL cells contain SP cells, whereas the SP cells are not enriched for mesenchymal stem cell(MSC)-like properties.In 2009, Twist and bone morphogenetic protein(BMP) antagonists may play an important role in maintenance of constancy of PDL, by which negatively regulate osteoblastic differentiation of cells existing in PDL. In this study, changes of these molecules expression by inducing osteoblastic differentiation were assessed in HPDL cells. We further investigated regulatory effects of BMP antagonists on osteoblastic differentiation in the cells. The present results suggest that under physiological … More condition, twist and BMP antagonists ; gremlin and follistatin, which are constitutively expressed in HPL cells, negatively regulate osteoblastic differentiation of the cells. Further, the induction of osteoblastic differentiation in HPL cells simultaneously enhances twist expression, while it suppresses expression of gremlin and follistatin, as the result, the osteoblastic differentiation may be modulated.PDL contains fibroblastic cells, which may participate in formation and maintenance of PDL fibers, and osteoblastic cells with a potential to generate alveolar bone and cementum, however, the specific markers for recognition and isolation of each cell type have not been clarified. The purpose of this study was to isolate alkaline phosphatase(ALP)-positive and-negative subpopulations from HPDL cells using flowcytometry and to identify functional difference between the 2 subsets in terms of mitogenic activity, osteogenic potential and expression of decorin as well as biglycan. The present results suggest that ALP-positive HPDL cells have osteoblast-like phenotype, while ALP-negative HPDL cells are fibroblastic cell population with weak osteogenic potential. Further, biglycan may positively regulate osteoblastic differentiation in ALP-positive HPDL cells, while decorin, whose expression is increased by osteogenic induction, may negatively regulate mineralization in both subpopulations.In 2010, to investigate characteristics of MSC existed in PDL, CD271+/CD90+cells, which were isolated from HPDL by FACS, were single cell cultured, and the clones with MSC-like properties were collected. Therefore, CD271+/CD90+may be a marker to collect MSC from HPDL. Less