Villanueva-Castellote Álvaro, Llena Puy Carmen, Cerda-Diéguez Miguel, Mira Álex, Ferrer María D
Department of Stomatology, Universitat de València, Valencia, Spain.
Genomics&Health Department, Foundation for the Promotion of Health and Biomedical Research of Valencia Region (FISABIO), Valencia, Spain.
J Oral Microbiol. 2022 Dec 22;15(1):2160536. doi: 10.1080/20002297.2022.2160536. eCollection 2023.
To develop an in vitro model for real-time monitoring of endodontic biofilm growth and evaluate the ex vivo effect of antibiotics on biofilm growth.
Root canal samples were taken from 40 patients and inoculated into 96-well plates in a system that measures biofilm growth through electrical impedance. Biofilm bacterial composition at the genus and species level was analyzed by Illumina sequencing. ANCOM-BC corrected data were used to compare bacterial composition after antibiotic treatment through compositional analysis, and to compare microbiological with clinical data.
The stationary phase was reached at 8 hours. The biofilm formed had a similar bacterial composition to the inoculum, and Enterococcus faecalis was virtually absent from the samples. The bacterial composition and the effect of antibiotics were sample-dependent. Metronidazole was the antibiotic that most inhibited biofilm formation and azithromycin the one that inhibited it in the highest percentage of cases. The antibiotic effect could not be related to the biofilm original bacterial composition.
The impedance system allowed real-time monitoring of endodontic biofilm formation, and we propose it as a model for ex vivo evaluation of the whole biofilm susceptibility to antimicrobials, as opposed to evaluating antibiotic sensitivity of specific bacterial isolates.
建立一种用于实时监测牙髓生物膜生长的体外模型,并评估抗生素对生物膜生长的体外效应。
从40名患者身上采集根管样本,并接种到96孔板中,该系统通过电阻抗测量生物膜生长。通过Illumina测序分析生物膜在属和种水平上的细菌组成。使用ANCOM-BC校正数据,通过成分分析比较抗生素治疗后的细菌组成,并将微生物学数据与临床数据进行比较。
8小时时达到稳定期。形成的生物膜的细菌组成与接种物相似,样本中几乎没有粪肠球菌。细菌组成和抗生素的效果因样本而异。甲硝唑是最能抑制生物膜形成的抗生素,阿奇霉素在最高比例的病例中能抑制生物膜形成。抗生素的效果与生物膜原来的细菌组成无关。
阻抗系统能够实时监测牙髓生物膜的形成,我们建议将其作为一种体外评估整个生物膜对抗微生物药物敏感性的模型,而不是评估特定细菌分离株的抗生素敏感性。