| The purpose of the study reported here was to analyze the detection Porphyromonas gingivalis in buccal epithelial cells and subgingival plaque from perio-dontally healthy subjects and chronic periodontitis. We tested that Porphyromonas gingivalis can exist within mucosal cells at sites remote from the gingival crevice.METHODS1. Study populationSubjects were recruited from those seeking treating at the stomatology of China Medical University. There were 40 subjects who were included in the healthy group and 39 subjects who were included in the diseased group in the study. The subjects were excluded if they smoke and received professional teeth cleaning or systemic medication, including any drugs, during the 3 months prior to the study.2. Measurement of clinical periodontal statusClinical parameters were measured by a single examiner and include plaque index (PLI) , sulcus bleeding index (SBI) , tooth mobility, radiographic bone loss and probing depth ( PD ).3. Sample collectionCells were obtained from mucosa of both cheeks with sterile cytological brushes. Subgingival plaque samples were taken from the first molar with sterile teeth point. Samples were placed into sterile eppendorf tubes containing 1ml PBS (PH7.4) and frozen for later analysis.4. DNA extractionGenomic DNA was extracted with phenol and chloroform from samples. The concentration and purity of DNA were calculated on the basis of absorbance at 260nm and 280nm with ultraviolet spectrophotometer.5. Statistical analysisRegressing analysis showed the change among different clinical periodontal parameters and t - test showed the significance in DNA concentration between mucosa and subgingival plaque.6. Polymerase chain reactionThe extracted DNA was amplified with universal primers and Pg species -specific primer.7. Samples were scored as positive or negative for Pg based on a clear band of the expected size. Integrated density value was measured by computer based on band of agarose, and calculated the relative quantity of Pg.8. Prevalence of Pg in buccal epithelial cells and subgingival plaque from different periodontal status and the correlation between relative quantity of Pg and clinical parameters was analyzed.RESULTS1. There was linear among clinical parameters. PLI, SBI, teeth mobility were moderately related toPD (P<0.01, r=0.528, 0. 587, 0.424 respectively) . Bone loss was highly related to teeth mobility (P<0.01, r=0.723).2. The concentration of DNA of healthy group was 36.58g /ml in subgingival plaque samples and 24. 5g /ml in buccal mucosa. The concentration of DNA of diseased group was 41. 26g /ml in subgingival plaque samples and 26. 39g /ml in buccal mucosa. There was not significant between different groups.3. Pg was detected in 37. 5% (15 of 40) of subgingival plaque samples, 32. 5% (13 of 40) of buccal mucosa samples in the healthy subjects. But it was found in 69. 23% (27of 39) of subgingival plaque samples, 46. 15% (18 of 39 ) of buccal mucosa samples in the periodontitis group. Highly statistically significant differences were observed between detection of Pg of subgingival plaque samples in the healthy and periodontitis groups (P < 0.05).4. In the healthy subjects, the relative quantity of Pg was 23.93% in sub-gingival plaque samples and 22.3% in the buccal mucosa samples. In the perio-dontal group, the relative quantity of Pg was 32. 16% in subgingival plaque samples and 25. 57% in the buccal mucosa samples. Significant differences were observed between buccal mucosa and subgingival plaque in the periodontitis (P <0.05). The same relations were observed between healthy subjects and periodontitis in the subgingival plaque (P <0.05).5. Correlation analysis showed that buccal mucosa was significantly and positively related to subgingival plaque in the relative quantity of Pg in the periodontitis. PD was highly related to the quantity of Pg.CONCLUSION1. There were a lot of bacteria in the buccal mucosa and subgingival plaque, diseased sites and non - dis...
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