BACTERIAL SPECIES ON OR IN GINGIVAL EPITHELIAL CELLS

作为理解这两个因素之间相互作用的重要的第一步， 龈下物种和牙龈上皮细胞，是为了确定 种存在于从人回收的上皮细胞上或中 牙龈缝隙或牙周袋。本项目的具体目标 具体如下：1）分离鉴定可培养细菌 附着或侵入上皮细胞的物种， 龈沟或牙周袋，2）寻求不可培养的 侵入或粘附于上皮细胞的物种，3）计数 优势细菌物种，使用DNA探针，这是对或在 上皮细胞来源于不同年龄和种族/民族的受试者 组和不同的健康或疾病状态和4）使用光， 电子显微镜，以确定物种，数量和位置（上 和/或在）与龈沟上皮细胞相关的细菌中。 将从牙龈样本中分离上皮细胞 不同年龄、性别和 种族/民族团体。初步的微生物培养研究将 对来自32名受试者的样本进行了检测。可培养细菌从 这些样本将用于确定一组DNA探针， 具体目标。PCR技术将用于寻找不可培养的物种 从收获的沟上皮细胞和DNA探针到这些物种 添加到文化研究定义的电池中。的完整集合 探针将被用来检查样本从一个更大的组60黑色，60 西班牙裔、60名亚裔和60名白色受试者。 每个种族/族裔群体将 由同等数量的男性和女性，年轻人和老年人组成， 牙周健康和牙周疾病受试者。4至8 将从每个受试者中采集上皮细胞样品 他们的内容多达50种使用的DNA探针。 最后，上皮细胞 将通过光学和电子显微镜检查细胞样品， 不同形态类型细菌的数量和位置。细菌细胞 将通过免疫细胞化学和/或原位 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.