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

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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