FLOW CYTOMETRIC ANALYSES OF CREVICULAR NEUTROPHILS

循环和脑沟中性粒细胞(PMN)功能的研究提示 这些细胞的能力可能会受到全身性和 导致牙周病增加的局部因素。 具体来说，青春期前牙周炎患者，青少年 牙周炎或糖尿病患者的牙周炎与中性粒细胞的相关缺陷 功能。以往对裂隙中性粒细胞功能的研究主要集中在体外 显微镜和细胞培养分析。流式细胞仪快速检测 基于细胞光的悬浮细胞的结构和功能 散射和荧光特性。在传播PMN的时候 用流式细胞仪研究了它们的活性和功能状态 目前尚无应用流式细胞术对裂隙中性粒细胞进行检测的研究。分析 用流式细胞术检测阴沟PMN细胞数量 快速样品分析和更详细的表征。具体的 这项研究的目的是测试流式细胞术方法在 描述缝隙中性粒细胞的功能。最初，我们 将建立流式细胞仪量化活性的能力， 外周血中中性粒细胞的吞噬和呼吸刺激作用 血。这些结果将与公认的光学显微镜进行比较。 技巧。10~(-3)~10~(-6)样品的流式细胞仪分析 还将分析由于规模缩小而产生的影响。最后， 将使用盐水洗涤剂从患者的脑沟收集中性粒细胞 和健康站点提供给牙周炎患者。流式细胞仪 将应用技术来表征其生存能力和功能 这些裂隙中性粒细胞的状态。统计分析将测试 光镜技术与流式细胞术的差异 不同年龄段人群脑沟中性粒细胞功能数据的循环及差异 健康站点与患病站点的对比。流式细胞仪检测技术的应用前景 裂隙中性粒细胞功能状态的研究可能提供一种新的研究工具 在牙周病研究中使用。

Studies of circulating and crevicular neutrophil (PMN's) functions suggest that capabilities of these cells may be depressed by systemic as well as local factors with a resultant increase in periodontal disease. Specifically, patients with prepubertal periodontitis, juvenile periodontitis or periodontitis in diabetics have associated deficits in PMN function. Past studies of crevicular PMN function have focused on in-vitro microscopic and cell culture analyses. The flow cytometer rapidly measures the structure and function of suspended cells based upon cellular light scatter and fluorescence characteristics. While circulating PMN's have had their viability and functional status studied with flow cytometry, there have been no studies of crevicular PMN's using flow cytometry. Analysis of crevicular PMN cell populations using flow cytometry would permit more rapid sample analyses and more detailed characterizations. The specific aim of this study is to test the application of flow cytometric methods to characterize the functional capabilities of crevicular PMN's. Initially we will establish the flow cytometer's ability to quantitate viability, phagocytosis and respiratory stimulation in PMN's isolated from peripheral blood. These results will be compared to accepted light microscopic techniques. Flow cytometric assays of samples containing 10-3 to 10-6 cells also will be analyzed for effects due to reduced scale. Finally, saline washes will be used to collect crevicular PMN's from the diseased and healthy sites to subjects with periodontitis. Flow cytometric techniques will be applied to characterize the viability and functional status of these crevicular PMN's. Statistical analyses will test differences between light microscopic and flow cytometric techniques in circulating as well as differences in crevicular PMN functional data from healthy versus diseased sites. Future application of flow cytometric tests of crevicular PMN functional status could provide a new investigative tool for use during studies of periodontal diseases.