DNA 형광 염색을 이용한 치은열구상피부착 세균에 관한 연구
Fluorescent detection of bacteria associated with gingival sulcus epithelium
0
조회
0
다운로드
0
인용
0
참고문헌
초록
Purpose: The aim of this study was to compare the number of live and dead bacteria attached to, or within, the stratified squamous epithelium lining the tissue side of the gingival sulcus. Materials and Methods: A total of 50 patients was examined and classified into healthy or diseased sites according to inflammatory status of the gingival tissue. The surface of stratified squamous epithelium was removed by gentle scraping of the gingival sulcus with curettes. The cells were processed in the laboratory by density-gradient centrifugation to separate the epithelial cells from the loose bacteria and debris. The LIVE/DEAD® BacLightTM Bacterial Viability Kit was applied and the specimens were observed by an epifluorescent microscope and the number of bacteria was counted. Results: Live and dead bacteria were stained to green and red, irrespectively. Generally, the number of total bacteria in the diseased sites was significantly higher than in the healthy sites. The mean number of detected bacteria in the diseased sites was 58.6±36.0 (red bacteria 10.4±9.2 / green bacteria 48.2±30.5), while it was 1.5±1.7 in the healthy sites (red bacteria 0.1±0.3 / green bacteria 1.4±1.5). The percentage of red bacteria was 17.5±11.2% in the diseased sites and 2.0±5.8% in the healthy sites. Conclusion: The total number of bacteria in the diseased sites was significantly higher than that of the healthy sites. The ratio and the number of red bacteria were also significantly higher in the diseased sites. (J Korean Acad Periodontol 2008;38:639-644)
Abstract
Purpose: The aim of this study was to compare the number of live and dead bacteria attached to, or within, the stratified squamous epithelium lining the tissue side of the gingival sulcus. Materials and Methods: A total of 50 patients was examined and classified into healthy or diseased sites according to inflammatory status of the gingival tissue. The surface of stratified squamous epithelium was removed by gentle scraping of the gingival sulcus with curettes. The cells were processed in the laboratory by density-gradient centrifugation to separate the epithelial cells from the loose bacteria and debris. The LIVE/DEAD® BacLightTM Bacterial Viability Kit was applied and the specimens were observed by an epifluorescent microscope and the number of bacteria was counted. Results: Live and dead bacteria were stained to green and red, irrespectively. Generally, the number of total bacteria in the diseased sites was significantly higher than in the healthy sites. The mean number of detected bacteria in the diseased sites was 58.6±36.0 (red bacteria 10.4±9.2 / green bacteria 48.2±30.5), while it was 1.5±1.7 in the healthy sites (red bacteria 0.1±0.3 / green bacteria 1.4±1.5). The percentage of red bacteria was 17.5±11.2% in the diseased sites and 2.0±5.8% in the healthy sites. Conclusion: The total number of bacteria in the diseased sites was significantly higher than that of the healthy sites. The ratio and the number of red bacteria were also significantly higher in the diseased sites. (J Korean Acad Periodontol 2008;38:639-644)
참고문헌 (15)
- Michalowicz BS, "Evidence of a substantial genetic basis for risk of adult periodontitis", J Periodontol, 71, pp.1699~1707 (2000)
- Dzink JL, "Socransky SS. The predominant cultivable microbiota of crevicular epithelial cells", Oral Microbiol Immunol, 4, pp.1~5 (1989)
- Hasty DL, "Multiple adhesins of streptococci", Infect Immun, 60, pp.2147~2152 (1992)
- Jenkinson HF, "Cell surface protein receptors in oral streptococci", FEMS Microbiol Lett, 121, pp.133~140 (1994)
- Meyer DH, "Adhesion of oral bacteria to soft tissue", Methods Enzymol, 253, pp.373~385 (1995)
- Moore WE, "The bacteria of periodontal diseases", Periodontol, 5, pp.66~77 (2000)
- Dibart S, "Identification of bacterial species on or in crevicular epithelial cells from healthy and periodontally diseased patients using DNA-DNA hybridization", Oral Microbiol Immunol, 13, pp.30~35 (1998)
- Lamont RJ, "Porphyromonas gingivalis invasion of gingival epithelial cells", Infect Immun, 63, pp.3878~3885 (1995)
- Meyer DH, "Invasion of epithelial cells by Actinobacillus actinomycetemcomitans: a dynamic, multistep process", Infect Immun, 64, pp.2988~2997 (1996)
- Beachey EH, "Bacterial adherence: adhesin-receptor interactions mediating the attachment of bacteria to mucosal surface", J Infect Dis, 143, pp.325~345 (1981)
- Saglie R, "Pattison GL. Bacterial invasion of gingiva in advanced periodontitis in humans", J Periodontol, 53, pp.217~222 (1982)
- Hedges SR, "Epithelial cytokine responses and mucosal cytokine networks", Trends Microbiol, 3, pp.266~270 (1995)
- Childs WC, 3rd, "Use of Percoll density gradients for studying the attachment of bacteria to oral epithelial cells", J Dent Res, 67, pp.826~830 (1988)
- Papaioannou W, "Comparison of fluorescence microscopy and culture assays to quantitate adhesion of Porphyromonas gingivalis to mono- and multi-layered pocket epithelium cultures", J Periodontol, 70, pp.618~625 (1999)
- Vaahtoniemi LH, "Attachment of bacteria to oral epithelial cells in vivo: a possible correlation to gingival health status", J Periodontal Res, 28, pp.308~311 (1993)