Muchova Maria, Balacco Dario L, Grant Melissa M, Chapple Iain L C, Kuehne Sarah A, Hirschfeld Josefine
Periodontal Research Group, School of Dentistry, Institute of Clinical Sciences, University of Birmingham, Birmingham, United Kingdom.
Front Oral Health. 2022 Mar 15;3:853618. doi: 10.3389/froh.2022.853618. eCollection 2022.
Development of dysbiosis in complex multispecies bacterial biofilms forming on teeth, known as dental plaque, is one of the factors causing periodontitis. () is recognised as a key microorganism in subgingival dental plaque, and is linked to periodontitis as well as colorectal cancer and systemic diseases. Five subspecies of have been identified: , and . Differential integration of subspecies into multispecies biofilm models has been reported, however, biofilm forming ability of individual subspecies is largely unknown. The aim of this study was to determine the single-subspecies biofilm forming abilities of ATCC type strains. Static single subspecies biofilms were grown anaerobically for 3 days on untreated or surface-modified (sandblasting, artificial saliva, fibronectin, gelatin, or poly-L-lysine coating) plastic and glass coverslips. Biofilm mass was quantified using crystal violet (CV) staining. Biofilm architecture and thickness were analysed by scanning electron microscopy and confocal laser scanning microscopy. Bioinformatic analysis was performed to identify orthologues of known adhesion proteins in subspecies. Surface type and treatment significantly influenced single-subspecies biofilm formation. Biofilm formation was overall highest on poly-L-lysine coated surfaces and sandblasted glass surfaces. Biofilm thickness and stability, as well as architecture, varied amongst the subspecies. Interestingly, ssp. did not form a detectable, continuous layer of biofilm on any of the tested substrates. Consistent with limited biofilm forming ability ssp. showed the least conservation of the adhesion proteins CmpA and Fap2 . Here, we show that biofilm formation by is subspecies- and substrate-specific. Additionally, ssp. does not appear to form stable single-subspecies continuous layers of biofilm . Understanding the differences in single-subspecies biofilm formation may shed light on multi-species biofilm formation mechanisms and may reveal new virulence factors as novel therapeutic targets for prevention and treatment of -mediated infections and diseases.
在牙齿上形成的复杂多物种细菌生物膜（即牙菌斑）中出现的微生物群落失调是导致牙周炎的因素之一。（某微生物名称）被认为是龈下牙菌斑中的关键微生物，与牙周炎以及结直肠癌和全身性疾病有关。已鉴定出该微生物的五个亚种：（具体亚种名称1）、（具体亚种名称2）、（具体亚种名称3）、（具体亚种名称4）和（具体亚种名称5）。已有报道不同亚种在多物种生物膜模型中的差异整合情况，然而，单个亚种的生物膜形成能力在很大程度上尚不清楚。本研究的目的是确定该微生物ATCC型菌株的单亚种生物膜形成能力。静态单亚种生物膜在未处理或经表面改性（喷砂、人工唾液、纤连蛋白、明胶或聚-L-赖氨酸包被）的塑料和玻璃盖玻片上厌氧培养3天。使用结晶紫（CV）染色对生物膜质量进行定量。通过扫描电子显微镜和共聚焦激光扫描显微镜分析生物膜结构和厚度。进行生物信息学分析以鉴定该微生物亚种中已知粘附蛋白的直系同源物。表面类型和处理显著影响单亚种生物膜形成。生物膜形成总体上在聚-L-赖氨酸包被表面和喷砂玻璃表面上最高。生物膜厚度和稳定性以及结构在各亚种之间有所不同。有趣的是，（具体亚种名称）在任何测试底物上均未形成可检测到的连续生物膜层。与有限的生物膜形成能力一致，（该亚种）显示粘附蛋白CmpA和Fap2的保守性最低。在此，我们表明该微生物的生物膜形成具有亚种特异性和底物特异性。此外，（具体亚种名称）似乎不会形成稳定的单亚种连续生物膜层。了解该微生物单亚种生物膜形成的差异可能有助于阐明多物种生物膜形成机制，并可能揭示新的毒力因子作为预防和治疗该微生物介导的感染和疾病的新型治疗靶点。
