Development of periodontal disease-diagnosis kit by nanotechnology and the application for information on health network

我们利用纳米技术开发了牙周病诊断试剂盒，并发现它将是研究牙周病与全身功能障碍之间关系的有用工具。我们还通过微通道芯片的开发，建立了流体系统中巨噬细胞体外斑块形成的评估方法。在微通道中，内毒素刺激后RAW264.7细胞的斑块形成显著增加。本研究试图阐明黏附分子在斑块形成中的作用。最初，证实了脂多糖刺激的RAW264.7细胞中斑块形成的时间依赖性增加。用流式细胞仪和Western blotting检测RAW264.7细胞表面黏附分子的表达。在内毒素刺激的0h，ICAM-1的表达水平很低。然而，在内毒素刺激的细胞中，这些水平在刺激后6小时和12小时显著升高。LFA-1和…的表达水平刺激后0h，未刺激和内毒素刺激的细胞中d-L-选择素的含量均较高。内毒素刺激12h后，LFA-1水平显著升高。然而，在内毒素刺激的细胞中，L-选择素的水平并没有增加。Western印迹分析检测到脂多糖刺激RAW264.7细胞2小时后ICAM-1的表达。这些结果表明，脂多糖可促进RAW264.7细胞斑块的形成，上调ICAM-1和LFA-1的表达，但不上调L-选择素的表达，这与前人关于单核细胞黏附分子LFA-1、Mac-1和ICAM-1表达增加的结果一致。综上所述，这项研究证实了在我们的微通道芯片上，巨噬细胞在流动的液体流中形成了斑块。目前的研究表明，ICAM-1和LFA-1在内毒素刺激的巨噬细胞聚集过程中起重要作用。我们的微通道芯片是一种适合于体外评估包括牙周炎在内的动脉粥样硬化病因的工具。较少

We developed the periodontal disease-diagnosis kit by nanotechnology and found that it would be useful tool to examine the relationship between the periodontal diseases and systemic dysfunctions. We also established the method involving the evaluation of in vitro plaque formation by macrophage cells in a fluid system via development of the micro-channel chip. In the micro-channel, plaque formation by RAW264.7 cells increased significantly following LPS stimulation. The current study attempted to elucidate the role of adhesion molecules in plaque formation. Initially, the time-dependent increase in plaque formation in LPS-stimulated RAW264.7 cells was confirmed. Expression of adhesion molecules on RAW264.7 cells was examined with flow cytometry and Western blotting analysis. Expression levels of ICAM-1 were very low in LPS-stimulated cells at 0h of stimulation. However, these levels were elevated markedly in LPS-stimulated cells at 6 and 12h of stimulation. Expression levels of LFA-1 an … More d L-selectin were medium in un-stimulated and LPS-stimulated cells at 0h of stimulation. Levels of LFA-1 were elevated significantly in LPS-stimulated cells at 12h of stimulation. However, L-selectin levels did not increase in LPS-stimulated cells. Western blot analysis detected ICAM-1 in RAW264.7 cells stimulated with LPS for 2h. These finding indicated that LPS increases plaque formation and up-regulates expression of ICAM-1 and LFA-1, but not that of L-selectin, in RAW264.7 cells, which are accord with the results of the previous report showing the increased expression of monocyte adhesion molecules, such as LFA-1, Mac-1 and ICAM-1. Taken together, this study confirmed plaque formation by macrophages in a flowing liquid stream on our micro-channel chip. The current investigation indicated that ICAM-1 and LFA-1 play an important role in cell aggregation of LPS-stimulated macrophages. Our micro-channel chip is a suitable tool for the in vitro evaluation of etiological factors of atherosclerosis including periodontitis. Less