Data_Sheet_2_Oral and Fecal Microbiome in Molar-Incisor Pattern Periodontitis.docx

In order to improve our understanding on the microbial complexity associated with Grade C/molar-incisor pattern periodontitis (GC/MIP), we surveyed the oral and fecal microbiomes of GC/MIP and compared to non-affected individuals (Control). Seven Afro-descendants with GC/MIP and seven age/race/gender-matched controls were evaluated. Biofilms from supra/subgingival sites (OB) and feces were collected and submitted to 16S rRNA sequencing. Aggregatibacter actinomycetemcomitans (Aa) JP2 clone genotyping and salivary nitrite levels were determined. Supragingival biofilm of GC/MIP presented greater abundance of opportunistic bacteria. Selenomonas was increased in subgingival healthy sites of GC/MIP compared to Control. Synergistetes and Spirochaetae were more abundant whereas Actinobacteria was reduced in OB of GC/MIP compared to controls. Aa abundance was 50 times higher in periodontal sites with PD≥ 4 mm of GC/MIP than in controls. GC/MIP oral microbiome was characterized by a reduction in commensals such as Kingella, Granulicatella, Haemophilus, Bergeyella, and Streptococcus and enrichment in periodontopathogens, especially Aa and sulfate reducing Deltaproteobacteria. The oral microbiome of the Aa JP2-like+ patient was phylogenetically distant from other GC/MIP individuals. GC/MIP presented a higher abundance of sulfidogenic bacteria in the feces, such as Desulfovibrio fairfieldensis, Erysipelothrix tonsillarum, and Peptostreptococcus anaerobius than controls. These preliminary data show that the dysbiosis of the microbiome in Afro-descendants with GC/MIP was not restricted to affected sites, but was also observed in supragingival and subgingival healthy sites, as well as in the feces. The understanding on differences of the microbiome between healthy and GC/MIP patients will help in developing strategies to improve and monitor periodontal treatment.

为加深我们对C级磨牙-切牙型牙周炎（Grade C/molar-incisor pattern periodontitis, GC/MIP）相关的微生物群落复杂性的认识，本研究对GC/MIP患者的口腔与粪便微生物组进行了调查，并与未患病个体（对照组）开展对比。本研究共纳入7名患有GC/MIP的非洲裔后裔患者，以及7名年龄、种族、性别匹配的健康对照者。研究人员采集了龈上/龈下位点生物膜（以下简称OB）与粪便样本，提交进行16S rRNA测序；同时完成了伴放线放线杆菌（Aggregatibacter actinomycetemcomitans, Aa）JP2克隆基因分型检测，并测定了唾液亚硝酸盐水平。
实验结果显示，GC/MIP患者的龈上生物膜中机会致病菌丰度更高。与对照组相比，GC/MIP患者的龈下健康位点内月形单胞菌属（Selenomonas）丰度升高。相较于对照组，GC/MIP患者的OB样本中互养菌门（Synergistetes）与螺旋体门（Spirochaetae）丰度更高，而放线菌门（Actinobacteria）丰度降低。在GC/MIP患者牙周探诊深度（PD）≥4mm的牙周位点中，Aa的丰度较对照组高出50倍。
GC/MIP患者的口腔微生物组以共生菌丰度降低为特征，涉及的共生菌包括金氏菌属（Kingella）、颗粒链菌属（Granulicatella）、嗜血杆菌属（Haemophilus）、伯杰氏菌属（Bergeyella）与链球菌属（Streptococcus）；同时患者体内牙周致病菌丰度显著升高，尤其是Aa与硫酸盐还原变形菌门（sulfate reducing Deltaproteobacteria）。携带Aa JP2样克隆的患者，其口腔微生物组的系统发育特征与其他GC/MIP患者存在显著差异。GC/MIP患者粪便中产硫化物细菌丰度更高，包括费氏脱硫弧菌（Desulfovibrio fairfieldensis）、扁桃体丹毒丝菌（Erysipelothrix tonsillarum）与厌氧消化链球菌（Peptostreptococcus anaerobius）。
本初步研究数据表明，非洲裔后裔GC/MIP患者的微生物群失调并非仅局限于患病牙周位点，同样可见于龈上、龈下健康位点以及粪便样本中。深入解析健康个体与GC/MIP患者的微生物组差异，将有助于开发改善牙周治疗效果并监测治疗进程的相关策略。