Identification of the unknown periodontal ligament-related molecules

未知牙周膜相关分子的鉴定

• 批准号: 18592269
• 负责人: KOBAYASHI Makoto
• 金额: $ 2.49万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18592269/
• 关键词: periodontal ligament cells; tendon; ligament markers; periodontal ligament markers; side population cells; mesenchymal stem cells; potential of multi-lineage differentiation; 軟骨分化誘導; TGF-β3; 腱・靭帯マーカー分子; 幹細胞; 中胚葉系前駆細胞

To identify the unknown periodontal ligament-related molecules, we performed the following studies.1. The changes of the tendon/ligament markers expression by induction of chondrogenic differentiation and TGF-β3 treatment in human periodontal ligament cells (HPLC)Many tendon/ligament marker molecules may play a role in healing and maintenance of constancy of periodontal ligament as well as tendon/ligament. Further, transforming growth factor-β(TGF-β) stimulates tendon healing. In the present study, we assessed the changes of the tendon/ligament markers expression by induction of chondrogenic differentiation and TGF-β3 treatment in HPLC. When HPLC were aggregated into micropellets and incubated with chondrogenic differentiatin medium with TGF-β3, chondrocyte-like lacunae formation was not detected in HPLC pellets. However, the expression levels of periostin, periodontal ligament associated protein-1 (PLAP-1), biglycan and decorin genes were higher, but growth and differentiation factor- … More 5 (GDF-5) and cartilage oligomeric protein (COMP) gene expression were lower levels, as compared with these pellets cultured with control medium. Further, scleraxis gene expression did not change throughout the culture periods. In monolayer cultures of HPLC, TGF-β3 stimulated expression of periostin, PLAP-1, biglycan, scleraxis and COMP, but suppressed GDF-5 expression, while, this cytokine had no effect on decorin expression. These results demonstrate that the expression of these tendon/ligament marker molecules in HPLC is modulated under the conditions. which induce chondrogenesis. as well as by the TGF-β3 stimulation.2. Characteristics of side population cells existing in HPLC.The various types of stem cells exist in side population (SP) effused strongly Hoechst 33342 dye. The SP cells in bone marrow express various mesenchymal stem cell (MSC) markers and are highly enriched for MSC activity. In the present study, we determined the characteristics of SP cells isolated from HPLC. SP cells and non-SP (main population : MP) cells were isolated from HPLC stained with Hoechst 33342 using flowcytometry. The frequency of SP cells existed in HPLC was mean 0.07% of total cells. Then, the gene expression profile, PDL-marker gene expression, activity of cell proliferation and multi-lineage differentiation were determined in SP cells and MP cells. Expression of CD73, a MSC marker gene, in SP cells was higher levels than that in MP cells, while MP cells expressed higher levels of extracellular molecule genes characterized in HPL (type I/III collagen, PLAP-1, periostin, decorin). Further, SP cells had higher cell proliferation activity than MP cells. However, activities of differentiation into osteoblasts and adipocytes were not different between the 2 cell types. Therefore, MSC-like cells may not be enriched in the SP fraction of HPLC. Less

为了鉴定未知的牙周韧带相关分子，我们进行了以下研究.人牙周膜细胞诱导成软骨分化和TGF-β3处理后肌腱/韧带标志物表达的变化（HPLC）许多肌腱/韧带标志物分子可能在牙周膜和肌腱/韧带的愈合和稳定性维持中发挥作用。此外，转化生长因子-β（TGF-β）刺激肌腱愈合。在本研究中，我们通过诱导软骨分化和TGF-β3处理，在HPLC中评估了肌腱/韧带标记物表达的变化。当HPLC聚集成微球并与含有TGF-β3的软骨分化培养基孵育时，在HPLC微球中未检测到软骨细胞样陷窝形成。而骨膜蛋白、牙周膜相关蛋白-1（PLAP-1）、双糖链蛋白聚糖（biglycan）和核心蛋白聚糖（decorin）基因的表达水平较高，而生长分化因子- ...更多信息 5（GDF-5）和软骨寡聚蛋白（COMP）基因表达水平较低。此外，巩膜轴基因表达在整个培养期间没有变化。在HPLC单层培养中，TGF-β3刺激periostin、PLAP-1、biglycan、scleraxis和COMP的表达，但抑制GDF-5的表达，而该细胞因子对decorin的表达无影响。这些结果表明，这些肌腱/韧带标记分子在HPLC中的表达在所述条件下被调节。其诱导软骨形成。以及TGF-β3刺激。HPLC中存在的侧群细胞的特征：在Hoechst 33342染料强烈渗出的侧群（SP）中存在各种类型的干细胞。骨髓中的SP细胞表达各种间充质干细胞（MSC）标志物，并且高度富集MSC活性。在本研究中，我们确定了SP细胞从HPLC分离的特性。使用流式细胞术从用Hoechst 33342染色的HPLC分离SP细胞和非SP（主要群体：MP）细胞。SP细胞在HPLC中的出现率平均为0.07%。检测SP细胞和MP细胞的基因表达谱、PDL标记基因表达、细胞增殖活性和多向分化能力。SP细胞中MSC标记基因CD 73的表达水平高于MP细胞，而MP细胞表达以HPL为特征的细胞外分子基因（I/III型胶原、PLAP-1、骨膜蛋白、核心蛋白聚糖）的水平更高。此外，SP细胞比MP细胞具有更高的细胞增殖活性。然而，向成骨细胞和脂肪细胞的分化活性在两种细胞类型之间没有差异。因此，MSC样细胞可能不会在HPLC的SP级分中富集。少

项目成果

培養ヒト歯根膜細胞における各種腱・靭帯マーカー遺伝子の発現制御.

培养的人牙周膜细胞中各种肌腱和韧带标记基因表达的调节。

• 发表时间: 2008
• 期刊: Dental Med Res 28
• 作者: 三井将;小林誠;ら
• 通讯作者: ら

培養ヒト歯根膜細胞における各種腱・靱帯マーカー遺伝子の発現制御.

培养的人牙周膜细胞中各种肌腱和韧带标记基因表达的调节。

• 发表时间: 2007
• 作者: 三井将;小林誠;ら
• 通讯作者: ら

培養ヒト歯根膜細胞中に存在するSide Population細胞の特徴.

培养的人牙周膜细胞中存在的侧群细胞的特征。

• 发表时间: 2008
• 作者: 小出容子;小林誠;ら
• 通讯作者: ら

The changes of the tendon/ligament markers expression by induction of chondrogenic differentiation and TGF-β3 treatment in human periodontal ligament cells

诱导软骨分化和TGF-β3处理人牙周膜细胞肌腱/韧带标志物表达的变化

• 发表时间: 2008
• 期刊: Dental Med Res 28(3)
• 作者: Mitsui M;Kobayashi M;et al.
• 通讯作者: et al.

Characteristics of side population cells existing in periodontal ligament cells.

牙周膜细胞中存在的侧群细胞的特征。

• 发表时间: 2008
• 作者: Koide Y;Kobayashi M;et al.
• 通讯作者: et al.