Abstract：
Endodontic infections involve a multispecies biofilm, making it difficult to choose an antimicrobial treatment. Characteristics such as the pathogens involved and number of microorganisms, nutrients, material surface to develop the biofilm, flow and oxygenation conditions are important for biofilm development using in vitro models.
OBJECTIVE: To develop a standardized biofilm model, which replicates the main features (chemical, microbiological, and topographical) of an infected root canal tooth to detect components as treatment target.
METHODS: Clinical strains of Enterococcus faecalis, Candida albicans, and Actinomyces israelii were isolated, and a multispecies biofilm was developed using continuous laminar flow reactors under anaerobic conditions in human dental roots. The microbiological composition was determined by counting colony-forming units and scanning electron microscope micrographs. In addition, the chemical composition of the exopolymeric matrix was determined by vibrational Raman spectroscopy and liquid chromatography of biofilm supernatant treated with enzyme.
RESULTS: E. faecalis turned out to be the main microorganism in mature biofilm, this was related to the presence of β-galactosidase detected by vibrational Raman spectroscopy. After the enzymatic treatment of the extracellular polymeric substance, the presence of mannose and glucose was established.
CONCLUSIONS: The present work contributes to better understanding of standard conditions to develop a multispecies biofilm in human dental roots, which could have an impact on the generation of new root canal disinfection techniques in endodontic pathologies.
OBJECTIVE: To develop a standardized biofilm model, which replicates the main features (chemical, microbiological, and topographical) of an infected root canal tooth to detect components as treatment target.
METHODS: Clinical strains of Enterococcus faecalis, Candida albicans, and Actinomyces israelii were isolated, and a multispecies biofilm was developed using continuous laminar flow reactors under anaerobic conditions in human dental roots. The microbiological composition was determined by counting colony-forming units and scanning electron microscope micrographs. In addition, the chemical composition of the exopolymeric matrix was determined by vibrational Raman spectroscopy and liquid chromatography of biofilm supernatant treated with enzyme.
RESULTS: E. faecalis turned out to be the main microorganism in mature biofilm, this was related to the presence of β-galactosidase detected by vibrational Raman spectroscopy. After the enzymatic treatment of the extracellular polymeric substance, the presence of mannose and glucose was established.
CONCLUSIONS: The present work contributes to better understanding of standard conditions to develop a multispecies biofilm in human dental roots, which could have an impact on the generation of new root canal disinfection techniques in endodontic pathologies.
摘要：
牙髓感染涉及多物种生物膜，这使得选择抗菌治疗变得困难。所涉及的病原体和微生物数量等特征，营养素,材料表面形成生物膜，流动和氧合条件对于使用体外模型的生物膜发育很重要。
目的：为了建立标准化的生物膜模型，它复制了主要特征(化学，微生物,和地形）感染的根管牙齿，以检测成分作为治疗目标。
方法：粪肠球菌临床菌株，白色念珠菌,以色列放线菌被隔离，并在人牙根的厌氧条件下使用连续层流反应器开发了多物种生物膜。通过计数菌落形成单位和扫描电子显微镜显微照片确定微生物组成。此外,通过振动拉曼光谱和用酶处理的生物膜上清液的液相色谱法确定外聚合物基质的化学组成。
结果：E.粪便被证明是成熟生物膜中的主要微生物，这与振动拉曼光谱检测到的β-半乳糖苷酶的存在有关。胞外聚合物的酶处理后，建立了甘露糖和葡萄糖的存在。
结论：本工作有助于更好地理解在人类牙根中形成多物种生物膜的标准条件，这可能会对根管病理学中新的根管消毒技术的产生产生影响。
目的：为了建立标准化的生物膜模型，它复制了主要特征(化学，微生物,和地形）感染的根管牙齿，以检测成分作为治疗目标。
方法：粪肠球菌临床菌株，白色念珠菌,以色列放线菌被隔离，并在人牙根的厌氧条件下使用连续层流反应器开发了多物种生物膜。通过计数菌落形成单位和扫描电子显微镜显微照片确定微生物组成。此外,通过振动拉曼光谱和用酶处理的生物膜上清液的液相色谱法确定外聚合物基质的化学组成。
结果：E.粪便被证明是成熟生物膜中的主要微生物，这与振动拉曼光谱检测到的β-半乳糖苷酶的存在有关。胞外聚合物的酶处理后，建立了甘露糖和葡萄糖的存在。
结论：本工作有助于更好地理解在人类牙根中形成多物种生物膜的标准条件，这可能会对根管病理学中新的根管消毒技术的产生产生影响。
