Lattar Santiago M, Schneider Rene Peter, Eugenio Vidal Jorge, Padilla Gabriel
Cell Biology of Microorganism Laboratory, Department of Microbiology, Institute of Biological Science, Federal University of Minas Gerais, Belo Horizonte, BH, Brazil.
Polytechnic School, University of São Paulo, São Paulo, SP, Brazil.
Braz J Microbiol. 2024 Dec;55(4):3921-3932. doi: 10.1007/s42770-024-01550-4. Epub 2024 Oct 31.
This study aimed to determine the protective role of the high release of C. albicans extracellular DNA (eDNA) in a polymicrobial biofilm formed by S. aureus and S. mutans in the course of DNase I treatment. A tube-flow biofilm bioreactor was developed to mimic biofilm formation in the oral cavity. eDNA release was quantified by real-time PCR (qPCR) and confocal microscopy analysis were used to determine the concentration and distribution of eDNA and intracellular DNA (iDNA). The mean amount of eDNA released by each species in the polymicrobial was higher than that in monospecies biofilms (S. aureus: 3.1 × 10 ng/μl polymicrobial versus 5.1 × 10 ng/μl monospecies; S. mutans: 3 × 10 ng/μl polymicrobial versus 2.97 × 10 ng/μl monospecies; C. albicans: 8.35 ng/μl polymicrobial versus 4.85 ng/μl monospecies). The large amounts of eDNA released by C. albicans (96%) in polymicrobial biofilms protects the S. aureus and S. mutans cells against the degradation by DNase I and dampens the effect of clindamycin.
本研究旨在确定白色念珠菌细胞外DNA(eDNA)的高释放量在金黄色葡萄球菌和变形链球菌形成的多微生物生物膜中,于脱氧核糖核酸酶I(DNase I)处理过程中的保护作用。开发了一种管流生物膜生物反应器以模拟口腔中的生物膜形成。通过实时聚合酶链反应(qPCR)对eDNA释放进行定量,并使用共聚焦显微镜分析来确定eDNA和细胞内DNA(iDNA)的浓度及分布。多微生物生物膜中各物种释放的eDNA平均量高于单物种生物膜(金黄色葡萄球菌:多微生物生物膜中为3.1×10 ng/μl,单物种生物膜中为5.1×10 ng/μl;变形链球菌:多微生物生物膜中为3×10 ng/μl,单物种生物膜中为2.97×10 ng/μl;白色念珠菌:多微生物生物膜中为8.35 ng/μl,单物种生物膜中为4.85 ng/μl)。多微生物生物膜中白色念珠菌释放的大量eDNA(96%)可保护金黄色葡萄球菌和变形链球菌细胞免受DNase I的降解,并减弱克林霉素的作用。