BACTERIAL SPECIES ON OR IN GINGIVAL EPITHELIAL CELLS

作为理解相互作用的重要第一步 龈下菌种和牙龈上皮细胞，是要确定 从人类身上回收的上皮细胞上或其中存在多种物种 牙周缝隙或牙周袋。这个项目的具体目标是 主要工作如下：1)分离鉴定可培养细菌 附着或侵入来源于 牙周间隙或牙周袋，2)寻找不可培养的 侵入或附着于上皮细胞的物种，3)计数 优势细菌种类，使用DNA探针，这些DNA探针位于On或In 来自不同年龄、不同种族/民族受试者的上皮细胞 群体和不同的健康或疾病状况以及4)使用光线和 电子显微镜以确定物种、数量和位置(On 和/或进入)与角膜缘上皮细胞相关的细菌。 将从牙龈样本中分离出上皮细胞 不同年龄、性别和性别受试者牙周间隙或牙周袋 种族/民族群体。最初的培养微生物学研究将是 对32名受试者的样本进行了研究。可培养的细菌从 这样的样本将被用来定义一组DNA探针，用于后续的 明确的目标。聚合酶链式反应技术将被用于寻找不可培养的物种 从采集的绒毛上皮细胞和DNA探针中提取 添加到文化研究定义的电池中。全套的 探测器将被用来检查从60名黑人，60名 西班牙裔，60名亚裔和60名白人。每个种族/民族都将 由同等数量的男性和女性，年轻的和年长的 牙周健康和牙周病的受试者。4至8个 将从每个受试者身上提取上皮细胞样本，并对其进行检测 他们使用DNA探针检测了多达50种物种的含量。最后，上皮性 细胞样本将通过光学和电子显微镜进行检查 不同形态类型细菌的数量和位置。细菌细胞 将在这些样本中通过免疫细胞化学和/或原位鉴定 杂交技术。

As an essential first step in understanding the interaction between subgingival species and gingival epithelial cells, is to determine which species are present on or in epithelial cells recovered from the human gingival crevice or periodontal pocket. The Specific Aims of this project will be as follows: 1) to isolate and identify cultivable bacterial species which are attached to or invade into epithelial cells derived from the gingival crevice or periodontal pocket, 2) to seek uncultivable species which invade into or adhere to epithelial cells, 3) to enumerate predominant bacterial species, using DNA probes, which are on or in epithelial cells derived from subjects of different age and racial/ethnic groups and different states of health or disease and 4) to use light and electron microscopy to determine the species, numbers and location (on and/or in) of bacteria associated with crevicular epithelial cells. Epithelial cells will be isolated from samples taken from the gingival crevice or periodontal pocket of subjects of different ages, gender and racial/ethnic group. Initial cultural microbiology studies will be performed on samples from 32 subjects. Cultivable bacteria recovered from such samples will be used to define a set of DNA probes for subsequent Specific Aims. PCR techniques will be used to seek uncultivable species from harvested crevicular epithelial cells and DNA probes to such species added to the battery defined by cultural studies. The complete set of probes will be used to examine samples from a larger group of 60 Black, 60 Hispanic, 60 Asiatic and 60 White subjects. Each racial/ethnic group will consist of equal numbers of male and female, young and old and periodontally healthy and periodontally diseased subjects. 4 to 8 epithelial call samples will be taken from each subject and assayed for their content of up 50 species using the DNA probes. Finally, epithelial cell samples will be examined by light and electron microscopy for the number and location of bacteria of different morphotypes. Bacterial cells will be identified in such samples by immunocytochemical and/or in situ hybridization techniques.