Abstract：
BACKGROUND: Subgingival dental plaque is an ecosystem playing a key role in supporting both oral health and systemic health. Menopause-related changes have the potential to disrupt its balance, which is crucial to postmenopausal well-being. Our study explored how circulating estradiol levels correlate with subgingival microbial composition using checkerboard DNA-DNA hybridization in premenopausal and postmenopausal women. We also demonstrated that combining this method with 16S ribosomal RNA (rRNA) sequencing insights remains valuable for examining subgingival ecology.
METHODS: We assessed 40 bacterial species in 77 premenopausal and 81 postmenopausal women using checkerboard DNA-DNA hybridization and measured serum estradiol with enzyme-linked immunosorbent assay (ELISA). Women were categorized by subgingival dysbiosis severity using a modified Subgingival Microbial Dysbiosis Index (mSMDI). Six women from each normobiotic and dysbiotic subgroup across premenopausal and postmenopausal women underwent 16S rRNA sequencing analysis.
RESULTS: DNA checkerboard analysis revealed that most observed variability in individual bacterial proportions is associated with periodontitis. Two species, Leptotrichia buccalis and Streptococcus constellatus, exhibited differences related to estradiol levels within the premenopausal group (p = 0.055 and p = 0.009, respectively). 16S rRNA sequencing confirmed the mSMDI\'s validity in categorizing normobiotic and dysbiotic states. Menopausal status was not associated with a dysbiotic shift in the subgingival microbiome despite significantly more attachment loss in postmenopausal compared to premenopausal women.
CONCLUSIONS: Our results indicate that decreased estradiol levels or increased attachment loss during menopause are not associated with changes in species abundance or dysbiotic shifts in women. The mSMDI may be a useful tool for classifying subgingival ecology based on its normobiotic or dysbiotic inclination.
METHODS: We assessed 40 bacterial species in 77 premenopausal and 81 postmenopausal women using checkerboard DNA-DNA hybridization and measured serum estradiol with enzyme-linked immunosorbent assay (ELISA). Women were categorized by subgingival dysbiosis severity using a modified Subgingival Microbial Dysbiosis Index (mSMDI). Six women from each normobiotic and dysbiotic subgroup across premenopausal and postmenopausal women underwent 16S rRNA sequencing analysis.
RESULTS: DNA checkerboard analysis revealed that most observed variability in individual bacterial proportions is associated with periodontitis. Two species, Leptotrichia buccalis and Streptococcus constellatus, exhibited differences related to estradiol levels within the premenopausal group (p = 0.055 and p = 0.009, respectively). 16S rRNA sequencing confirmed the mSMDI\'s validity in categorizing normobiotic and dysbiotic states. Menopausal status was not associated with a dysbiotic shift in the subgingival microbiome despite significantly more attachment loss in postmenopausal compared to premenopausal women.
CONCLUSIONS: Our results indicate that decreased estradiol levels or increased attachment loss during menopause are not associated with changes in species abundance or dysbiotic shifts in women. The mSMDI may be a useful tool for classifying subgingival ecology based on its normobiotic or dysbiotic inclination.
摘要：
背景：龈下牙菌斑是一个生态系统，在支持口腔健康和全身健康方面发挥着关键作用。更年期相关的变化有可能破坏其平衡，这对绝经后的健康至关重要。我们的研究探索了绝经前和绝经后妇女使用棋盘DNA-DNA杂交的循环雌二醇水平如何与龈下微生物组成相关。我们还证明，将此方法与16S核糖体RNA（rRNA）测序见解相结合，对于检查龈下生态学仍然很有价值。
方法：我们使用棋盘DNA-DNA杂交方法评估了77名绝经前和81名绝经后妇女的40种细菌，并使用酶联免疫吸附测定（ELISA）测量了血清雌二醇。使用改良的龈下微生物菌群失调指数（mSMDI），按牙龈下菌群失调的严重程度对妇女进行分类。来自绝经前和绝经后妇女的每个正常生物和生态失调亚组的六名妇女进行了16SrRNA测序分析。
结果：DNA棋盘分析显示，大多数观察到的个体细菌比例的变异性与牙周炎有关。两个物种,布氏鳞毛和星座链球菌，在绝经前组中表现出与雌二醇水平相关的差异（分别为p=0.055和p=0.009）。16SrRNA测序证实了mSMDI在对正常生物和异常生物状态进行分类方面的有效性。尽管与绝经前妇女相比，绝经后的附着丧失明显更多，但绝经后的状态与龈下微生物组的生态失调变化无关。
结论：我们的结果表明，绝经期间雌二醇水平降低或附着丧失增加与女性物种丰度变化或生态失调变化无关。mSMDI可能是根据其正常生物或生态失调倾向对龈下生态进行分类的有用工具。
方法：我们使用棋盘DNA-DNA杂交方法评估了77名绝经前和81名绝经后妇女的40种细菌，并使用酶联免疫吸附测定（ELISA）测量了血清雌二醇。使用改良的龈下微生物菌群失调指数（mSMDI），按牙龈下菌群失调的严重程度对妇女进行分类。来自绝经前和绝经后妇女的每个正常生物和生态失调亚组的六名妇女进行了16SrRNA测序分析。
结果：DNA棋盘分析显示，大多数观察到的个体细菌比例的变异性与牙周炎有关。两个物种,布氏鳞毛和星座链球菌，在绝经前组中表现出与雌二醇水平相关的差异（分别为p=0.055和p=0.009）。16SrRNA测序证实了mSMDI在对正常生物和异常生物状态进行分类方面的有效性。尽管与绝经前妇女相比，绝经后的附着丧失明显更多，但绝经后的状态与龈下微生物组的生态失调变化无关。
结论：我们的结果表明，绝经期间雌二醇水平降低或附着丧失增加与女性物种丰度变化或生态失调变化无关。mSMDI可能是根据其正常生物或生态失调倾向对龈下生态进行分类的有用工具。
