| 摘要: | 背景：全世界大約有20-50%的人患有牙周病，而牙周病可能使人更易罹患系統性疾病。牙周炎是炎症所導致的牙周組織喪失，當宿主和口腔菌斑生物膜間的共生平衡被打破時就會發生牙周炎。首先，鏈球菌和白色念珠菌會在齦上菌斑中形成玉米芯結構。而後，牙周致病菌附著在結構上並形成小菌落。隨後，牙周致病菌將會繁殖並感染齦下部位。目前，研究口腔菌斑細菌成分的標準方法是透過 16S rRNA 的細菌分析和第三代定序技術。
研究目的：1) 比較健康受試者和牙周炎受試者之間的臨床參數，例如牙菌斑重量、臨床牙周附連喪失和咬合力。2）比較健康和牙周炎受試者的齦上口腔微生物群。
材料和方法：符合納入和排除條件的雙和醫院（台灣新北市）自願患者納入本研究，並由同一位操作者進行口腔檢查、樣本採集和咬合力記錄。口腔檢查部分，記錄DMFT評分、牙菌斑控制記錄、牙菌斑指數（PI）和牙周記錄表（包括牙周囊袋深度、牙齦萎縮、根叉侵犯、牙齒搖動、牙齦探測出血（BOP）和牙齦指數（GI））。另外，以不激活Gracey curettes的方式收集所有牙齒表面的齦上牙菌斑。咬合力紀錄方面，要求受試者以最大咬合力在Dental Prescale II（GC）測試片上咬 3 秒鐘。微生物分析則採用第三代定序技術與分裂式擴增子去噪演算法（DADA2）進行序列錯誤的校正。
結果：對照組有15名受試者，牙周炎組有15名受試者。牙周炎組的平均牙菌斑總重量和平均每顆牙齒的牙菌斑重量均高於對照組（p<0.05）。此外，牙周炎組的平均牙齦探測出血、牙齦指數、牙周囊袋深度和臨床牙周附連喪失均顯著高於對照組（p<0.001）。另一方面，對照組的平均最大咬合力顯示面積和平均咬合力均高於牙周炎組（p<0.05）。就齦上微生物群而言，牙周炎組表現出更高的物種豐富度和均勻度以及更高的Alpha多樣性。健康組和牙周病組的物種組成均以鏈球菌屬為主，而牙周炎組中普雷沃氏菌屬、梭桿菌屬和??單胞菌屬的平均絕對計數顯著高於對照組（p<0.05）。此外，平均臨床牙周附連喪失與普雷沃氏菌屬和梭桿菌屬的絕對計數呈中度正相關，而牙齦探測出血與普雷沃氏菌屬和??單胞菌屬的絕對計數呈中度正相關。
結論：健康和牙周炎受試者齦上牙菌斑的菌群組成存在差異。其中，普雷沃氏菌屬、梭桿菌屬和??單胞菌屬在牙周炎受試者中數量較多，且具有統計顯著性。
BACKGROUND: Around 20-50% of the people worldwide suffers from periodontal disease and it may predispose one to systemic diseases. Periodontitis is the loss of periodontium due to inflammation, which happens when the host and oral plaque biofilm symbiotic balance is broke. Initially, Streptococcus species and Candida albicans form a corncob structure in the supragingival plaque. Then, periodontal pathogens attach to the architecture and develop microcolonies. Later, periodontal pathogens are able to propagate and reinfect the subgingival site. Currently, the standard methodology to investigate the oral plaque bacterial composition is through 16S rRNA-based bacterial profiling with next-generation sequencing.
OBJECTIVE OF THE STUDY: 1) Compare clinical parameter such as plaque weight, clinical attachment loss, and bite force between healthy and periodontitis subjects. 2) Compare the supragingival oral microbiota between healthy and periodontitis subjects.
MATERIALS AND METHODS: Spontaneous patient in Shuang Ho Hospital (New Taipei City, Taiwan) that meet the inclusion and exclusion criteria are enrolled in the study. The same operator performed the oral examination, sample collection, and bite force registration. For oral examination, DMFT score, plaque control record, plaque index (PI), and periodontal charting (including probing depth, gingival recession, furcation involvement, mobility, bleeding on probing (BOP), and gingival index (GI)) are recorded. Additionally, Gracey curettes are used to collect the supragingival plaque of all tooth surfaces without activating it. Besides, the subject is asked to occlude with their maximum bite force for 3 seconds on Dental prescale II (GC) test piece. For microbiota analysis, Third Generation Sequencing with Divisive Amplicon Denoising Algorithm 2 (DADA2) application for sequence error correction are applied.
RESULTS: There were 15 subjects in the control group and 15 in the periodontitis group. Periodontitis group showed higher average total plaque weight and average plaque weight per tooth than that of the control group (p<0.05). Moreover, periodontitis group had significantly higher average bleeding on probing, gingival index, probing depth, and clinical attachment loss than that of the control group (p<0.001). On the other hand, control group showed higher average maximum bite force display area and average bite force than that of the periodontitis group (p<0.05). With regard to supragingival microbiota, periodontitis group demonstrated greater species richness and evenness along with higher Alpha diversity. Streptococcus species were the predominant taxa in both the healthy and periodontitis groups, while the average absolute count of Genus Prevotella, Fusobacterium, and Porphromonas were significantly higher in periodontitis group than in the control group with p-value<0.05. In addition, mean clinical attachment loss was moderately and positively related to Prevotella and Fusobacteriumabsolute count, whereas bleeding on probing was moderately and positively related to Prevotella and Porphyromonas absolute count.
CONCLUSION: There are differences in the supragingival plaque microbiota composition and distribution of healthy and periodontitis subjects, among which, Genus Prevotella, Fusobacterium, and Porphromonas are found to be predominant in the periodontitis subjects with statistical significance. |
