Study on the calcification by periodental ligament fibroblasts in the periodonta regeneration

在牙周再生方面，我们重点研究了成纤维细胞生长因子(成纤维细胞生长因子)对人牙周膜成纤维细胞(HPLF)增殖和趋化作用的调节以及1α，25-二羟基维生素D3(Vit.D3)对成骨功能的调节。为了了解细胞如何在相同的生长因子刺激下迁移或增殖的机制，我们感兴趣的是检测经历迁移的人成纤维细胞的不同时相。向成纤维细胞生长因子迁移的细胞未掺入溴脱氧尿苷，提示处于S期的细胞未被诱导迁移。迁移的细胞群中未见分裂细胞。但是，迁移的细胞表达c-myc和c-fos mRNA，提示这些细胞退出了细胞周期的G0期。根据这一结果，我们推测正在进行迁移的细胞是…更可能是在细胞周期的G1期。已有研究表明维生素T。D3可提高具有成骨细胞样细胞功能的HPLF的碱性磷酸酶(ALP)活性和骨钙素(OC)产量。评估VIT的作用。在HPLF的碱性磷酸酶(ALP)活性和OC产量方面，我们检测了不同培养阶段1α，25-二羟基维生素D3受体(VDR)基因表达、ALP活性和OC产量的动态变化。随着培养细胞密度的增加，VDR基因表达水平、ALP活性和OC产量均增加。维特。D3显著提高了各培养细胞密度下的ALP活性和OC产量。然而，VIT。D3不增加VDR基因的表达水平。当细胞密度稀疏时，多层红细胞培养上清可促进VDR基因在细胞内的表达。这些稀疏的培养相细胞不表达VDR基因和Vit。D3对这些细胞的碱性磷酸酶(ALP)活性和OC产量几乎没有影响。因此，如果稀疏细胞事先用培养上清液刺激，维生素T。D3极强地提高了OC产量。这些结果表明维生素T。D3可提高OC产量，并依赖于HPLF培养细胞中VDR基因的表达水平，以及多层红细胞培养上清中含有增加VDR基因表达的因子。较少

In the aspect of periodontal regeneration, we focused on the regulation of the proliferation and the chemotaxis by fibroblast growth factor (FGF) and that of osteogenic functions by 1alpha, 25-dihydroxyvitamin D3 (vit.D3) in human periodontal ligament fibroblast (HPLF).Numbers of growth factors have been shown to induce cell migraiton as well as cell proliferation. To understand the mechanisms how the cells were driven to migration or proliferation upon same growth factor stimuli, we are interested in detecting the different phases of the cell of human fibroblasts undergoing migration. Cells migrated against FGF did not incorporate bromodeoxyuridine, suggesting that the cells in S phase were not induced to migrate. No dividing cells were found in the migrated cell population. However, migrated cells were found to express c-myc and c-fos mRNA,suggesting that these cells were exit from G0 phase of the cell cycle. Based upon the results, we conclude that the cells undergoing migration wer … More e most likely in G1 phase of the cell cycle.It has been shown that vit. D3 enhances alkaline phosphatase (ALP) activity and osteocalcin (OC) productivity in HPLF which have osteoblast-like cell functions. To evaluate the role of vit. D3 in ALP activity and OC productivity in HPLF,we examined the kinetics of gene expression of 1alpha, 25-dihydroxyvitamin D3 receptor (VDR), ALP activity and OC productivity at different phases of culture. The level of VDR gene expression, ALP activity and OC productivity were enhanced as culture cell density increased. Vit. D3 significantly enhanced ALP activity and OC productivity in each culture cell density. However, vit. D3 did not increase the level of VDR gene expression. The supernatant obtained from the culture of multilayred cells enhance the expression of the VDR genes in cells when the culture cell density was sparse. These sparse culture phase cells did not express the VDR gene and vit. D3 hardly affect ALP activity and OC productivity in these cells. Thus, if the sparse cells were previously stimulated with the culture supernatant, vit. D3 strongly enhanced OC productivity. These results suggest that vit. D3 enhances OC productivity and depends upon the level of VDR gene expression in HPLF culture cells and that the culture supernatant from multilayred cells contains the factors which increase VDR gene expression. Less